CN1261801A - Allelic polygene diagnosis of reward deficiency syndrome and treatment - Google Patents

Abstract

Enhancement of attentional processing is attained by administration of an endorphinase inhibitor or enkephalinase inhibitor and optionally, a dopamine precursor, or a serotonin precursor, a GABA precursor, or an endorphin or enkephalinase releaser, or certain herbal compounds including Rhodiola rosea extract (Pharmaline) and/or Huperzine. These components promote restoration of normal neurotransmitter function and the components combined enhance the release of dopamine at the nucleus accumbens and are non-addictive. Use of the dopamine precursors L-phenylalanine, or L-Tyrosine, the enkephalinase inhibitor D-phenylalanine, and/or the serotonin precursor -hydroxytryptophan and a natural acetylcholenesterase inhibitor and chromium salts (i.e. picolinate, nicotinate, etc.) is especially preferred, but not limited to assist in relieving symptoms associated with brain phenylalanine deficiency.

Description

The allele polygene diagnosis and the treatment of recompense deficit syndrome

Background of invention

Have right among the present invention according to fund 1-ROI-DA08417 number of National Institute of Drug Abuse and Nicotiana tabacum L. correlational study disease plan fund 4RT-0110 government.1, invention field

The present invention partly relates to the combination of some withdrawal composition and some all participate in the specific gene typing of the gene of recompense behavior neurotransmitter function.One aspect of the present invention be understand in brain center edge system (meso-limbic system) some neurotransmitter of having set up how collaborative work activation nervous pathway, cause good feel, and exploitation influences the material of these nervous pathways.The present invention partly relates to and utilizes precursor aminoacid and some medical herbs chemical compound to improve attention processing procedure and memory and increase attention in healthy individual, and promotes to lose weight and control surfeit.The invention discloses utilize the neurotransmitter Gene Polymorphisms to various neurological unusual and the various diagnostic methods of behavior and the Therapeutic Method that utilizes patient that combination treatment of the present invention is identified.Diagnostic method to polygenic character is also disclosed.2, the description of association area

In the past few decades, some brain regions and the neurotransmitter of recompense can have been determined to participate in for the research of chemicals dependency Basic of Biology.Particularly, the dependence of ethanol, opioid and cocaine be it seems that depending on one overlaps common biochemical mechanism (Cloninger, 1983; Blum, 1978; Blum, 1989).The brain depths participate in a kind of neuron paths of limbic system and be called volt nuclear and pallidal two zones taking medicine (drugs) people's recompense express in (Wise and Bozarth, 1984) of key seemingly.The chronic use that has confirmed cocaine, morphine and ethanol causes several biochemistrys in the edge dopamine system to adapt to (Ortiz etc., 1996).

The center edge dopamine system connect in the brain the special frame cortex of frontal lobe of high-level structure (in the prefrontal area behind forehead) with the amygdaloid body of brain central authorities and with volt nuclear, it has been proved to be a main activity site in the addiction in zooscopy.Participate in multiple addicted various brain paths and come together in some dopaminergic receptor (D1, D2, D3, D4, D5), as if wherein the D2 site is the most outstanding.As if though the material of every kind of abuse acts on the different piece of path, last result is identical: dopamine is released (Koob and Bloom, 1988) in volt nuclear and Hippocampus.Seemingly these strengthen the main neurotransmitter of the recompense in site to dopamine.

Metabolic several behaviors, i.e. sexual abnormality (Gessa and Tagiamonte, 1975), manic (Goodwin and Jamison, 1990), the behavior of division sample (Carlsson, 1978 of relating to unusually of dopamine; Snyder, 1976), ADHD (Shaywitz etc., 1976), conduct disorder or attack (Rofeness etc., 1986; Valzelli, 1981; King, 1986), alcohol abuse (Blum etc., 1990) and stutter.In addition, reported haloperidol, a kind of DRD2 receptor antagonist, effective (Murray etc., 1977 in the treatment of some stutters; Prins etc., 1980).Though the most normal participation compelling sex behavior of 5-hydroxy tryptamine energy mechanism, also be considered unusually (Austin etc., 1991 of dopamine; Delgado etc., 1990).Unusual path in thalamus, basal nuclei and the frontal lobe participates in obsession, and (Baxter etc. 1992; Rauch etc., 1994; Modell etc., 1989) and particularly dopamine is a kind of major neurotransmitter involved in striatum and frontal lobe.

The defective of maincenter noradrenaline energy mechanism usually participates in the etiology of attention deficit hyperactivity disorder.Verified, compare with the ADHD child who does not have cognitive defect, have read and the ADHD child of other cognitive defect in plasma norepinephrine (NA) significantly increase (Halperin etc., YEAR).It is relevant that they infer that the ADHD+ cognitive defect notices that with influence top/temporal lobe the NA of maincenter lacks of proper care.Because contiguous audition of these brain regions and Language Processing district are so this may cause symbiotic cognitive disorder.From clinical angle, with common remarkable improvement the (Hunt etc., 1985 that symptom takes place, clonidine treatment back; Comings etc., 1990), the effect of this prompting NA in some ADHD at least.Clonidine is a kind of presynaptic a ₂-noradrenergic receptor antagonist, it causes norepinephrine to be released into the inhibition (Starke etc., 1974) of synapse.

By inference, NA and locus coeruleus (LC) in awakening of the importance of attention and sleeplessness, play a role (Aston-Jones etc., 1984).By inference, to the stress tolerance of task and good performance with the low basis of catecholamine or nervous level and spirit stress the time higher acute release relevant (Dienstbier, 1989).May be opposite in ADHD, have the NA baseline that increases intense and stimulative and stress process in the reduction of catecholamine discharge (Pliszka etc., 1996).For detecting the hypothesis that the NA defective participates in ADHD, carried out the excretory research of NA metabolite in many CSF, blood plasma and the urine (3-methoxyl group-4-hydroxy phenyl glycerol (MHPG)).Some studies show that ADHD patient has MHPG excretion rate (Oades, 1987 that are lower than contrast; Shekim etc., 1983; Shekim, Dekirmenjian and Chapel, 1997; Yu-cum and Yu-feng, 1984) and studies show that of other do not change (Rapoport etc., 1978; Zametkin etc., 1985) or the increase (Khan and Dekirmenjian, 1981) that transforms of NA.Reported and compared with contrast, in the ADHD main body, epinephrine level obviously lower (Hanna etc., 1996; Klinteberg and Magnusson, 1989; Pliszka etc., 1994), perhaps show a kind of torpid reaction (Girardi etc., 1995) to glucose uptake.Norepinephrine is converted into epinephrine by the Phenylethanolamine N-methyltransferase by the PNMT gene code.

The model of a kind of ADHD comes tetanic property inhibition locus coeruleus NA neuron based on adrenergic obstacle.It is reported that the d-amphetamine and the desmethylimipramine that all are generally used for the ADHD treatment cause the excretory remarkable reduction of MHPG (Mefford and Potter, 1989; Shekim etc., 1979).Yet, be used for the most frequently used medicine methylphenidate of ADHD treatment and do not cause the excretory reduction of MHPG (Zametkin etc., 1985), for example Fenfluramine (Donnelly etc., 1989) is invalid in treatment ADHD and other reduces the excretory medicine of MHPG.These observe with the participation of the ADHD that has several types and various neurotransmitters and gene consistent.

Play a role (Posner and Peterson, 1990 of the imbalance at NA and the adrenergic alpha-2 receptor LC place by attention system after the cortex of top/temporal lobe in the ADHD of some forms by inference; Pliszka, 1996), and the ADHD of another kind of form is because the dopaminergic defective of the main influence prefrontal lobe attention system that impulsion property is relevant with carrying out obstacle together.Have been found that several dopaminergic genes, for example dopamine D ₂Receptor (DRD2) (Comings etc., 1991), dopamine D ₄Receptor (DRD4) (Lahoste etc., 1996) and dopamine transporter (DAT1) (Cook etc., 1995; Comings etc., 1996; Gill etc., 1997; Waldmaqn etc., 1996) gene is relevant with ADHD.

It is reported that the boy's blood plasma MHPG level with ADHD and reading disorder is significantly higher than boy (Haperin etc., 1993 that only have ADHD; Halperin etc., 1997).In one research of back, they have also confirmed remarkable negative correlation between blood plasma MHPG level and WISC-R speech IQ and reading, spelling and the arithmetic problem estimated by WRAT-R (the wide region attainment test of revision).This difference with show that the ADHD with cognitive disorder is previous other people research (August and Garfinkel, 1989 of a kind of different subtype of ADHD; McGee etc., 1989; Pennington etc., 1993) unanimity.Also show, tend to a kind of optionally attention deficit (Posner and Peterson, 1990 with the relevant attention deficit type of top defective; Dykman etc., 1979; Richards etc., 1990).

By inference, the ADHD+ cognitive disorder is because a kind of NA Metabolic disorder that relates to the LC of adrenergic alpha-2 receptor, and mainly influence cortex of parietal lobe back attention system (Halperin etc., YEAR).Because these brain region adjacent auditions and Language Processing zone, this may cause symbiotic cognitive disorder.Because ADHD is a kind of polygenes unusual (Comings etc., 1996), therefore suppose that these are that pure form is wrong, and most individuality may have two types gene.The defective that studies show that NA and adrenergic alpha-2 receptor in the primates in the prefrontal lobe cognitive defect, also play a role (Arnsten, 1997).

Multiple research has pointed out dopamine receptor to participate in addictive behavior.The cocaine patient shows reduction on their neuron activity level, this is consistent (Volkow etc., 1993) with accepting the reduction of dopamine ability.The neuron that has the D2 dopamine receptor in the rat of morphine addiction shows as and reduces 25%, and they accept the ability (Nestler etc., 1996) of dopamine in a small amount from contiguous neuron to lose major part.Rely on the minimizing of observed D2 receptor in the main body at opioid, this prompting main body before they begin Drug abuse just has low D2 receptor by inference, and this reduction may make them be easier to oneself take medicine (Wang etc., 1997).

Though it is complicated participating in the biological neurotransmitter of recompense system, but known at least three kinds of other neurotransmitteies participate in the site, a few place of brain: 5-hydroxy tryptamine at hypothalamus, endorphins (opioid peptides) at veutro Gai Qu and volt nuclear and inhibitory nerve mediator GABA in black substance, veutro Gai Qu and volt nuclear (Stein and Belluzzi, 1986; Blum and Kozlowski, 1990).What is interesting is that glucoreceptor is an important connection between serotonergic system and the opioid peptides in the hypothalamus.Another recompense path is included in the release to norepinephrine of the nerve fiber that risen by locus coeruleus in the Hippocampus.

Have opium can and dopaminergic system on dissecting and the evidence that interrelates on the function, this shows that endogenous opiate can system have effect in the effect that mediates ethanol and the other medicines pair brain dopaminergic path relevant with recompense.The damage influence of dopaminergic the path preceding former A activity of endorphins (Morris etc., 1988 in dopamine antagonist and the brain; Normand etc., 1988).Behavioristics, pharmacology and neuro chemistry studies show that opium energy and dopaminergic system have effect (Blum etc., 1976a, b in the effect that strengthens ethanol and other medicines abuse; Blum, 1982a; Blum, 1977; Blum, 1973; Koob and Bloom, 1988; Weiss etc., 1993).Zooscopy shows that opioid receptor agonist increases the preference to ethanol, and the antagonist of these receptors then reduces consumption (Blum, 1975 of ethanol; Le etc., 1993).In addition, to the animal and human alcoholic studies show that opiate receptor antagonist in reducing the positive reinforcement effect (positive reinforcing effects) of alcohol consumption be effectively (O ' Malley, 1992; Swift etc., 1994; Blum, 1975; Volpicelli etc., 1992).In addition, the increase of alcohol induced brain dopamine level is blocked (Widdowson and Holman, 1992 by opiate receptor antagonist Allylnoroxymorphone and naltrexone in animal; Benjamin etc., 1993).The summary of being done by Gianoukalis and de Waele (1994) is supported the effect of endogenous opiate-like sub stance and Drug abuse (ethanol) recently.

In the normal person, these neurotransmitteies stimulate and complex reaction between a kind of excitement or suppress co-operation in the cascade reaction, cause a kind of good feel, final recompense.In the cascade reaction principle of recompense, the destruction of these cell-cell interactions causes negative feeling.The long-term abuse that comprises the genetic abnormality of some polymorphism, long-term anxiety or psychoactive drug (comprising glucose) all can cause the self-continuous-mode (Blum, 1991) of unusual addiction in the animal and human.

The effect of pharmacology (bromocryptine, amfebutamone and the nor-apomorphine of N-propyl group) part is by the dopamine D 2 genotype decision of individuality.The A1 carrier pharmacology of DRD2 gene goes up the reaction of D2 agonist stronger.Research shown with the direct microinjection of the nor-apomorphine of D2 agonist N-propyl group to as the volt nuclear of rat in the remarkable symptom behind the inhibition animal refraining opium type material, and dopamine itself suppresses withdrawal symptom (Harris and Aston-Jones, 1994 of ethanol; Blum etc., 1976b).In this respect, dopamine/endogenous opioid peptides interactional evidence in other place of volt nuclear and brain is arranged, and may be the reduction (Pothos etc., 1991) that the overstimulation of opioid peptides system is caused the dopamine function by exogenous opioid.When comparing with normal non-alcohol preference rat, the alcohol preference rat has less 5-hydroxy tryptamine neuron, has the endorphins of higher level in hypothalamus (because seldom being released), at Fu Hezhong low-density D2 receptor (McBride etc., 1995 in more GABA neuron and some zone in limbic system arranged in hypothalamus; Smith etc., 1997; With McBride etc., 1997).

Verified amino acid precursor and D-phenylalanine (a kind of active material of the inhibition enzymatic lysis raising endorphins (U.S. Patent number 4 that passes through that gives some neurotransmitter (5-hydroxy tryptamine and dopamine) as proband of clinical experiment to SUD or carbohydrate gluttony, 761,429 and 5,189,064) in the time of), finds to reduce addiction, reduces nervous sickness rate, reduces relapse rate and increase restorative possibility.

Many laboratorys are followed the trail of the contact between some gene and various actions obstacle, comprise that d2 dopamine receptor allele is with the relation between the mandatory addictive behavior of many impulsions.Except that have research by the SPECT scanning technique show in the Tourette impaired patients dopamine transporter site increase (Malison etc., 1995; Tiihonen etc., 1995), for same DAT ₁10/10 allele or D β HB ₁The polymorphism that allele is associated is expressed and is known little.It is main that ADHD, tourette's syndrome (TS), conduct disorder, ODD, reading difficulty, learning disorder, stutter, drug dependence and alcoholism all show as the male.(Comings etc. are 1996a) with clinical genetics research (Comings, 1994b for the molecule genetics research of DRD2, D β H, DAT; 1994c; 1995b; Bierderman etc., 1991; Comings and Comings, 1987) show that these are obstacles relevant on the etiology.The someone advocates that the defective of neurotransmitter participates in alcoholism (Blum, 1991).The gene studies that participates in the neurological path is described below.

Androgen receptor gene it is reported that the specific sudden change of androgen receptor (AR) gene causes the androgen insensitivity symdrome (Gottlieb etc., 1977) of wide range of types.In addition, cause two cover polymorphism trinucleotide repeats sequences of amino acids bundle in the albumen, CAG (Edwards etc., 1992) and GGC (Sleddens etc., 1993; Sleddens etc., 1992), be present in first exon of AR gene.Show that the CAG trinucleotide repeats sequence 43 to 65 times, causes the chain Duchenne-Arandisease of X (La Spada etc., 1991) when highly expanding.The length of repetitive sequence is 11 to 31 times (Edwards etc., 1992) in normal population.

The allele of the non-height amplification of little satellite that exists in normal population and moonlet may be brought into play directly effect in the adjusting of gene.This is based on such observation, and promptly most of short placed in-line repetition is correlated with (Schroth etc., 1992) with the formation of Z-DNA, and Z-DNA repeats to participate in various aspects (Rich etc., 1984 of Gene regulation; Hamada etc., 1982; Wolff etc., 1996).Because the amount of formed Z-DNA is extremely sensitive (Schroth etc., 1992) for multiple length, thus prompting repeat allelic size itself may be with phenotypic effect relevant (Comings, 1997).

Some researchs (Olweus etc., 1988; Mattsson etc., 1980; Schiavi etc., 1984; Kreuz and Rose, 1972) but be not all research (Bradford and McClean, 1984; Schaal etc., 1998) point out aggressive behavior relevant with the plasma testosterone level.Add in the main body of ADHD the sick condition of normally a kind of symbiosis of aggression obstacle (Comings, 1995 at TS; Stewart etc., 1981; Biederman and Sprich, 1991) and between TS and ADHD, have high level symbiosis (Comings and Comings, 1984,1990; Knell and Comings, 1993).

Dopamine D ₁Acceptor gene (DRD1) is in contrast and suffer from the order-checking of carrying out the DRD1 gene among schizophrenia, manic depression and the crapulent patient and do not identify phenotype and the exons mutation (Jensen, the 1993k that produce effect on the association study of schizophrenia and TD; Gelertner etc., 1993k).D1 receptor in cortex of frontal lobe may in memory, play a role (Comings etc., 1997k; Williams etc., 1995k).Reported D in rat ₁And D ₂The adverse effect (Self etc., 1996) to cocaine searching behavior of receptor stimulating agent.TD proband, smoker and pathological gambling person meet negative heterosis, and wherein the most consistent difference is the relative reduction and the 11 and 22 homozygous increases (Comings etc., 1996) of 12 heterozygote frequencies of Dde1 polymorphism.On the contrary, positive hybrid vigor is present in the DRD2 gene, and its 12 heterozygotes of quantitatively marking are for the highest, and 11 and 22 homozygotes are minimum.Though the result for ADHD is not remarkable in the DRD1 site separately, detect the negative heterosis on the DRD1 gene and/or the just heterotic existence on the DRD2 gene have significant additive effect (Comings etc., 1997k).

Research before the d2 dopamine receptor gene (DRD2) shows that in the individuality of ADHD, TS, CD and SUD, D2A1 allele exists probability obviously to increase (Comings etc., 1991).Because these obstacles are feature with the stress of difference all, and a lot of diagnostic criterias for PTSD have many symptoms with the ADHD unanimity, Vietnam time-expired soldier of country re-adjustment research (Kulka etc., 1990) has reported the childhood period of PTSD and ADHD and CD concordance to have significant dependency between the symptom history.From addiction therapy, once in Vietnam experiences the people of fierce war condition, screened posttraumatic stress disorder (PTSD), concurrent present PTSD have dependency between the allelic individuality of D2A1 with carrying (Comings etc., 1996k).

Detected relatedness between the TaqA1 allele of ADHD and DRD2 gene (Comings etc., 1991k).The stimulant methylphenidate increases regional flow, and it reduces blood flow in other people.The metabolic change of frontal lobe, temporal lobe and cerebellum is relevant with the density of D2 receptor---and it is big more that the high more blood flow of density increases.Methylphenidate reduces the metabolic activity in the ganglion basal.These results are with this identical of views, it thinks that the genetic defect in the dopamine metabolism that causes dopaminergic state in limbic system and frontal lobe causes that the compensatory of dopaminergic activity increases in ganglion basal, and methylphenidate by associating its through the dopamine activity that the inhibition of dopamine transporter improved the brain dopaminergic nerve (Volkow etc., 1996k) and in ganglion basal dopaminergic activity reduce subsequently and ganglion basal in the minimizing of blood flow reverse this phenomenon.These researchs are also with other people unanimity as a result, show between CSF level and have positive correlation (Castellanos etc. the reaction of methylphenidate and HVA, 1996k), HVA is a kind of metabolite of dopamine, and its level in CSF is relevant with the density of D2 receptor.Although D2 receptor deficiency in cerebellum, methylphenidate still increase metabolism (Volkow etc., the 1996k of cerebellum; Hall etc., 1994k).This with more and more show evidence that cerebellum plays a significant role in attention, study and memory be consistent (Leiner etc., 1989k).Reported the dependency between A1 genotype and regional flow.TaqI D2A1 carrier shows in the cortex of shell (lentiform nucleus), the volt gyrus of nuclear, frontal lobe and temporal lobe and middle prefrontal lobe, pillow temporal lobe and socket of the eye relative glucose metabolism and significantly is lower than those and has the genotypic individuality of A22 (Nobel etc., 1997).Taq D2 A1 carrier in ganglion basal, have significantly reduced d2 dopamine receptor Bmax (Nobel etc., 1991k).Endorphins increase with the blood flow of methylphenidate similar area and therefore may participate in a kind of dopaminergic mechanism (Blum etc., 1985k).Have disengaging (detachment), social isolation and lack the remarkable reduction that detects dopamine receptor density in the individuality of best of friends (Farde etc., 1997k).

Though the DRD2 gene pleiomorphism is relevant with many mental maladjustment, between finding among the drug abuser who is kept in detention, do not have dependency (Smith etc., 1993) with certain psychosis.The report of dependency changes greatly between alcoholism and DRD2 allele, finds dependency (Smith etc., 1992 between D2A1 allele and many material/drug dependence; Noble etc., 1993; O ' Hara etc., 1993; Comings etc., 1994; U.S. Patent number 5,210,016 and 5,500,343).Behind the dependency of finding first between DRD2 A1 and severe alcoholism (Blum etc., 1990), several seminar all can not repeat out this observed result.Prompting has two possible reasons: the first, and not enough for ethanol, medicine and tobacco abuse contrast screening; The second, sample error (Blum etc., 1997 of disease chronic nature and seriousness evaluation aspect; Bolos etc., 1990; Gelertner etc., 1991; Schwab etc., 1991; Turner etc., 1992; Cook etc., 1992; Goldman etc., 1992; Goldman etc., 1993; Suarez etc., 1994).

Dopamine plays regulator (LeMoal and the Simon of multiple different behaviors, 1991), and many researchs have reported at allele and cocaine addiction, many substance abuses, smoking, attention deficit hyperactivity disorder (ADHD), tourette's syndrome, vision-perceptual disturbance, behavior disorder, posttraumatic stress disorder, the pathological gambling of DRD2 gene and have forced to have significant dependency (Blum etc., 1995 between the feed; Blum etc., 1996).Though these dependencys are arranged, order-checking research does not find to explain any sudden change of these discoveries in exon.If the non-exons mutation that plays a role in regulating the DRD2 function of the same the unknown of D2A1 allele connects unbalance then can explain these discoveries (Comings etc., 1991).In addition, the type of having reported crapulent seriousness and used contrast is important determiner (Noble etc., 1994 of DRD2A1 allele with the alcoholism dependency; Geijer etc., 1994; Parsian etc., 1991; Blum etc., 1990; Lawford etc., 1997).

Utilization repeats polymorphism at the Taq1 in DRD2 site " A " RFLP and a kind of little satellite and suffers from the compatriot who carries out in the crapulent family at pilosity linkage analysis is shown, in this site with develop between the tendentiousness of severe alcoholism and have significant dependency.Do not find corresponding sudden change in the DRD2 gene, this effect may be from the closely linked site (Cook etc., 1996) beyond the DRD2 gene itself.The verified point mutation (Finch etc., 1995) in alcohol dependence patient's DRD2 gene the 8th exon, and other people are reported in the coding region of DRD2 gene and do not have structural mutation (Gejman etc., 1993).

Have been found that DRD2 gene A 1 allele is relevant with many behaviors, comprise severe alcoholism, many substance depilatories, crack (crack)/cocaine addiction, smoking, pathological gambling, the outbreak of shortage major depression and carbohydrate gluttony, perhaps be summarised as DSM-IV matter utilization obstacle (Blum etc., 1996e; Blum etc., 1995b; Comings etc., 1996c).Compare the division sample that MCMI-II estimates/avoidances group scale significant correlation (Corbisiero etc., 1991) with other II (Anix II) diagnosis group (antisocial, autophilia, paranoid) with alcohol abuse.Have and represent the patient group of the MCMI-II rising of dividing sample/avoidance quality to tend to keep treatment less day, and recur morning (Fals-Stewart, 1992).Division sample/avoidance group's high score is relevant with the patient (Kranzler and Satel, 1994) that male alcoholic (Matano etc., 1994) who does not have patience and cocaine rely on.Find division sample/elusive behavior (comprising low-level sensation) and consume more ethanol and have higher MAST scoring (Ohannessian and Hesselbrock, 1995) than patient with high level sensation; And in experimenter with severe gluttony obstacle avoidance personality significant correlation (Yanovski etc., 1993).

In healthy individual and alcoholic, show the molecular hybrid advantage at dopamine receptor gene place.Healthier volunteer comprises the HVA of dopamine, the 5-HIAA of 5-hydroxy tryptamine and the MHPG of norepinephrine with monoamine metabolite level in DRD2 TaqI A1A2 and the genotypic cerebrospinal fluid of B1B2.The result shows 1,1+1, and 2 homozygotes had the significant difference of statistics to 1,2 o'clock, and were analyzing 1, and 1+1 does not then have when 2 pairs 2,2 genotype and 1 pair 2 allele.TaqI B1B2 polymorphism draws result (Jonsson etc., 1996) much at one.On the contrary, in Finland and U.S. alcoholic, detect CSF HVA level and DRD2 TaqI A1/A2 polymorphism, when detecting 1 pair 2 allele, do not find dependency, and 1,1+1 does not have (Goldman etc., 1992) during 2 pairs 2,2 genotype yet.

By the CSF level of HVA relatively show the hybrid vigor at DRD2 gene place be utilize the DRD2 genotype of TaqI polymorphism (Jonsson etc., 1996k).Be used for TD experimenter's absent minded grade form, 12 heterozygotes show as the highest absent minded scoring experimenter, and they are 12 heterozygotes (Jonsson etc., 1996) with CSF HVA of floor level.Observe the HVA of top level in 11 homozygotes, 22 homozygotes are medium.Some researchs, be not all studies show that in the child who suffers from ADHD and TD (Shaywitz etc., 1979k) the CSFHVA level obviously lower (Cohen etc., 1979k).Between electrophysiological abnormality and DRD2 A1 allele, find to have significant dependency (Blum etc., 1994k).These can observe (Comings etc., 1991k unusually in ADHD experimenter and crapulent child; Noble etc., 1994k).

Have positive correlation (Comings etc., 1991 between the TaqA1 that it is reported at the DRD2 gene and attention-deficient obstacle (ADHD) and the Tourette disease; Comings etc., 1996a), and other people find do not have dependency (Sunohara etc., 1996) with the ADHD proband.The 48bp variant that the ADHD proband shows with the D4 gene has significant dependency, and same DRD2, DRD3 or serotonin transporter gene then do not have.7 times of DRD4 are repeated allele and obviously occur more continually in having the child of ADHD.Have 7 times of the D4 acceptor gene repeat allele with novel the pursuit (with get excited, probe into, impetuousness, irritability, temper is suddenly made an uproar and words and deeds wantonly are feature) have evidence (Epstein etc., 1996 of dependency; Benjamin etc., 1996).It is relevant with opposite state to rely among the proband DRD2 A1 allele at cocaine: low novelty is pursued, its with consider carefully, stubborn, ascetic, temper, avoidance and the depression of shrinking back with reinforcement are feature (Compton etc., 1996) slowly.Molecule genetics research has been found that D ₂Dopamine receptor (DRD2) A1 allele is with the dependency between alcoholism and drug dependence (Blum etc., 1990).Shown with those and do not had the allelic (A1 of A1 ^-) compare, carrying A1 allele (A1 ⁺) the experimenter in have the central dopamine of reduction can function (Nobel etc., 1997).Though the many DRD2 of studies show that genes have significant effect in many serious cases at present, be responsible for crapulent gene still unknown (Noble, 1993; Blum etc., 1995).

(Comings and Comings 1987b) and addiction, impulsive behavior, comprise mandatory feed, gambling and smoking be associated (Self etc., 1996 with the same compulsion of DRD2 gene; Ogilvie etc., 1996; Blum etc., 1995b; Blum etc., 1996e).Before be reported among the experimenter different these behaviors according to this with DRD2 gene-correlation (Comings etc., 1993a with TS group; Noble etc., 1994d; Blum etc., 1996a; Comings etc., 1996c; Noble etc., 1994c; Noble, 1993; Comings etc., 1996e).

D2 dopamine receptor significantly is less than non-alcoholic in the alcoholic, and with the natural law irrelevant (Volkow etc., 1997) behind the last alcoholism.The DRD2 receptor to the ratio of transport protein amount in non-alcoholic apparently higher than the alcoholic.Compare with non-alcoholic, the alcoholic shows D2 receptor (postsynaptic sign) but not the remarkable reduction of DA transport protein amount (presynaptic sign).Because the D2 receptor in the striatum mainly is arranged in the GABA cell, these results provide the unusual evidence of dopaminergic seen in the GABA participation alcoholic.

Dopamine D ₃Acceptor gene (DRD3) disappearance DRD3 gene knock out (knockout) mice significantly than they have the brood mice of normal DRD3 gene active (Williams etc., 1995k).Have been found that the schizophrenia reported the negative heterosis of DRD3 site (Crocq etc., 1992k).TD (Comings etc., 1993j) and pathological gambling (Comings etc., 1996) in DRD3MscI 12 heterozygositys significantly reduce.

Dopamine D ₄Acceptor gene (DRD4) is at dopamine D ₄In the acceptor gene (DRD4), reported in coding is responsible for being bonded to the DNA of proteic the 3rd the endochylema ring of guanylic acid to have a 48bp and 16 the multiple polymorphisms of aminoacid (Van Tol etc., 1992k; Lichter etc., 1993k).Repeat 2 to 11 times in this DNA zone, and modal allele is 2,4 and 7 repetitions.Inhibition performance a kind of to the dopamine blunt reaction of 7 allele by adenosine cyclase in the cell (Asghari etc., 1995k).Independently study for two of the normal subjects and shown in 7 allelic existence and novel the pursuit between (novelty seeking) (a kind of) to have dependency (Benjamin etc., 1996k with the relevant character of impulsion; Ebstein etc., 1996k).There is a research not find this dependency (Malhotra etc., 1996).An ADHD child study report of comparing with contrast, with contrast compare more ADHD child carry at least one 7 allele (LaHoste etc., 1996k).Reported dependency between 7 allele of DRD4 gene in TD (Grice etc., 1996k).Other work in this field be not equivocal (Spielman etc., 1993k).

Dopamine transporter gene (DAT1) is being derived from the DAT1 genetic marker frequency of capsule transport protein site among different the white Americans of different European countries and is showing suitable inhomogeneity, but the commaterial abuse does not have tangible dependency (Uhl etc., 1993; Persico etc., 1993).The allelic distribution of DAT1 VNTR can not be distinguished any substance abuse person or spiritual activating agent misuser's control sample (Persico etc., 1996), yet observes with Japanese alcoholic relevant (Muramatsu and Higuchi, 1995).The DAT1 gene also be indicated in force with the addiction obstacle in play a role.Because a kind of model of action of cocaine is to suppress dopamine transporter function (Riz etc., 1992; Ritz etc., 1990),, and comprise that other of parkinson disease (Uhl, 1990) and tourette's syndrome (Singer etc., 1991) is unusual so it participates in drug dependence biology.

Utilize the SPECT scanning technique verified in tourette's syndrome the dopamine transporter site increase (Malison etc., 1995), and compare with nonviolent alcoholic, the dopamine transporter acceptor site significantly increases (Tiihonen etc., 1995) in the violence alcoholic.TS experimenter's sample after death research report, the dopamine uptake number of loci increases in striatum, points out or has more that the DAT1 molecule or the dopamine teleneuron quantity of more number increase (Singer etc., 1991).This be widely used chemical compound methylphenidate in ADHD treatment (Volkow etc., 1995k) and the action site of Dexedrine.These analeptic suppress transport process, cause the increase of synapse dopamine.Reported same contrast ratio, and significantly the increasing of dopamine transporter protein level in TD experimenter's the striatum (Maison etc., 1995k).The DAT1 knock-out mice, much it more moving in the space that limits, to its studies show that the brain dopamine level increases by 500, D2 receptor down-regulated, D2 function of receptors de-connect and body size reduce by 57% (Giros etc., 1996k).Still do not know whether more rare DAT1 repetition allele is correlated with the increase or the reduction of DAT1 molecular amounts.

(Cook etc. 1995k) and in the variation of the behavior in the Tourette obstacle (TD) (Comings etc., 1996) have reported with the dependency between 10 allele of DAT1 gene in the ADHD/ADD case history.Remarkable increase in lonely experimenter shows TS and solitarily is that the hereditism is correlated with and relates to research (Burd etc., 1987 of several groups of similar genes with some; Comings and Comings, 1991b; Sverd, 1991) be consistent.

Non-alcoholic's contrast with 93 ethnic group couplings is compared, show to give up at 93 and observe the advantage that repeats allele VNTR polymorphisms at 9 of DAT1 gene 3 ' untranslated region among outbreak or the alcoholic that goes mad and significantly increase (Sander etc., 1997).In the drug dependence experimenter, detected 5 ' UTR 40bp repetition polymorphism in the DAT1 gene, and found that comparing any 3 ' UTR repetition gene frequency with normal control does not have marked difference (Persico etc., 1993).Found that 9/10 genotype is relevant with " pathologic violence " teenager; And it is relevant with 9/9 genotype in the alcohol dependence of giving up outbreak or going mad.Be reported in multiple 9 allele of DAT1 gene 40bp with the dependency between the inductive paranoid of cocaine (Gelernter etc., 1994a).

Dopamine-D β H is one of metabolic main enzyme of dopamine, and the catalysis dopamine is to the conversion of norepinephrine (NE).In zooscopy, the active inhibition of D β H causes the reduction of noradrenaline levels, and the inhibition that it discharges tyrosine hydroxylase causes the excessive generation of dopamine.The same hyperkinesia of the latter, aggressivity, self-excitation and stereotyping movable relevant (Randrup and Scheel-Kruger, 1966; Shekin etc., 1983k).This enzyme that studies show that of blood D β H enzyme level is pursued (sensation seeking) (Kuperman etc., 1988k in sensation; Comings etc., 1996), ADHD and conduct disorder (Rogeness etc., 1982k; Rogeness etc., 1989k) effect in.

In the past with the active disorder of dopamine (D β H) with the childhood period CD and alcoholism be associated (Pliszka etc., 1991).By inference, come to the surface (externalizing) obstacle such as CD are relevant with increasing of the reduction of norepinephrine energy function and dopaminergic function, and this is a pair of condition (Quay, 1986) that may only be caused by D β H defective.The diagnosis frequency that other people are reported in CD among the boy of the emotional disturbance with low plasma D BD level increases.Yet Bowden etc., out-patient in 1988 discover that the probability that D β H level is low in suffering from the ADHD child of CD is than the ADHD child who does not have CD much bigger (Rongeness etc., 1987; Pliszka etc., 1988; Bowden etc., 1988; Comings etc., 1996).On the contrary, in out-patient's research of young detention center, there be not the dependency of discovery between CD and plasma D β H.Umberkomen etc. (1981) have shown the dependency between low D β H level and sensation pursuit behavior.

Comprise that the detection of CSF D β H level among major depression, bipolar affective disorder and the schizoid patient finds that unique significant dependency is between low CSFD β H and bipolar affective disorder (Lerner etc., 1978) having multiple mental maladjustment.Contact research between D β H locus and schizophrenia (Aschauer and Meszaros, 1994), alcoholism, depression, manic depression and tourette's syndrome (Comings etc., 1986) is negative.Yet some sib-pair analysises show between abo blood group and D β H and some mental maladjustment such as depression and alcoholism weak related (Wilson etc., 1992).

Contact research between D β H locus and schizophrenia, alcoholism, depression, manic depression and tourette's syndrome is negative (Aschauer and Meszaros, 1994; Comings etc., 1986).Between D β HTaqIB1 allele and pathologic SAB, do not find dependency (Blum etc., 1997).It is reported that TaqI B1/B2 polymorphism is relevant with the contrast of getting rid of medicine, ethanol and tobacco abuse through screening.Yet the B1 allele of dopamine gene is also with TD proband and ADHD proband relevant (Comings etc., 1996).

Fructus Cannabis ester receptor (CB1) more may be the second order effect of the adjusting effect in the dopamine metabolism by anandaide and Fructus Cannabis ester receptor though Fructus Cannabis ester receptor may be primary with the contact of recompense path.This CB1 receptor that coexists is consistent with the similarity between DRD2 receptor result.Similar with the CD1 gene, hereditism's variant of DRD2 gene with the contact between many substance abuses (O ' Hara etc., 1993; Smith etc., 1992; Noble etc., 1993; Comings etc., 1994) than being more repeatably with the contact between alcoholism itself.An explanation of these observations is dopaminergic-Fructus Cannabis recompense paths, and particularly cocaine and amphetamine are activated than by ethanol activated many (DiChiara and Imperato, 1988) by medicine.

The recurrence of the known activation initiation cocaine pursuit behavior of center edge dopamine system in the drug dependence animal model.This priming effect is by dopamine D ₂Agonist is strengthened, but by dopamine D ₁Agonist suppresses (Self etc., 1996).In this, anandamide causes D in the striatum ₁And D ₂The ability that the receptor ratio reduces may be the reason that explanation CB1 variant acts in drug dependence.

Monoamine oxidase, MAO after testing the relevant Fnu4H1 polymorphism of the T → C variant of 1460 site that coexist of MAO-A cDNA and the relevant EcoRV polymorphism (Hotamisligil and Breakefield, 1994) of T → G variant of 941 site that coexist.Because they all relate to the displacement in the 3rd base of a codon, so they are irrelevant with amino acid replacement.They have detected active 40 cell lines of the known MAO-A of having.All cell lines of carrying Fnu4H1 C variant all also carry EcoRV G variant.When based on low when higher MAO-A activity is divided into two groups with sample, be present in 25% the cell line more rare Fnu4H1 C or+allele (inventor's 2 allele) is with greater activity group significant correlation.Lin etc. (1994) have reported in manic depression with the relevant (Lin etc. of lower MAO level, 1994), more general MAOAFnu4H1 T or 1 allele (Lin etc., 1994) significantly increase, and (1995) such as Craddock etc. (1995) and Nothen fail to confirm this point.

Vanyukov etc. (1993) utilize CA to repeat polymorphism and have detected the MAOA gene and contrasted compare (Black etc., 1991) with 31 male and 78 women in 23 male and 34 women alcoholics.In young substance abuse person in the male among (P=0.17) rather than the women (P=0.8) exist one with higher molecular weight allele (＞dependency trend between 115bp), and＞allele of 115bp is with there being weak dependency (P=0.03) trend between the age that begins.Tivol etc. (1996) have measured the MAO-A enzymatic activity recently and have shown＞40 contrast male's of 100 times of variations exon sequences.There is significant conservative at coded sequence.Only find 5 kinds of polymorphisms.Wherein, 4 kinds relate to the 3rd codon site and aminoacid sequence does not have change.Another kind is a lys → arg displacement.

The gene mapping of nicotine receptor gene code CHRNA4 gene is formed (Steinlein etc., 1996) in chromosome 20q13.2-13.3 (Steinlein etc., 1994) and by 6 exons above the genomic DNA of 17kb.In the Australian pedigree of an expansion, find to stride Ser248Phe missense mutation in the film district 2 with autosomal dominant frontal lobe epilepsy at night (ADNFLE) relevant (Steinlein etc., 1995) at the CHRNA4 gene.The insertion of in Norway's pedigree, finding three nucleotide (GCT) in M2 domain C-terminal coding region with autosomal dominant frontal lobe epilepsy at night relevant (Steinlein etc., 1997b).Two kinds of brain function other is unusual, benign neonatal familial convulsions (Leppert etc., 1989; Malafosse etc., 1992) and low-voltage EEG (Steinlein etc., 1992) also the zone with the CHRNA4 site is relevant.D20S19, the site of a height polymorphism is with causing all three kinds these unusual genes closely linked (Steinlein etc., 1996).

Weiland and Steinlein have reported the dinucleotide VNTR polymorphism (Weiland and Steinlein, 1996) of the height polymorphism in first intron of a kind of CHRNA4 of being positioned gene.Single base pair polymorphism (Steinlein etc., 1995 have also been reported; Phillips and Mulley, 1997; Guipponi etc., 1997; Steinlein etc., 1997a).Utilize three kinds of single base pair polymorphisms, Steinlein etc. (1997a) find that the CHRNA4 gene is with not having dependency between frightened obstacle.Utilization is with ADNFLE relevant Ser248Phe missense mutation and 4 kinds of reticent polymorphisms, Steinlein etc. have reported with contrast (.027) and have compared that the allelic frequency moderate of the T of CfoI 595 polymorphisms increases (.085) in the PGE common the childhood period.

Little/moonlet polymorphism is found with the relevant behavior phenotype research of little/moonlet polymorphism at different neural spiritual candidate genes place, little or little inter-satellite short or longer allele and following gene with various quantitative behavioral traits has significant dependency: MAOA, MAOB, HTR1A, DAT1, DRD4, HRAS, HTT, OB, CNR1, GABRA3, GABRB3, FRAXA and NO (Comings etc., 1996k; Comings etc., 19961; Comings etc., 1996m; Johnson etc., 1997; Comings etc., 1998; Grade etc., 1997).Significant phenotype behavior effect and DAT1 (Cook, 1995; Gelernter etc., 1994), DRD4 (Benjamin etc., 1996; Ebstein etc., 1996; Grice etc., 1996; Lahoste etc., 1996), HRAS (Herault etc., 1993; Eggers etc., 1995; Thelu etc., 1993), HTT (Ogilvie etc., 1996; Lesch etc., 1996), INS (Bennett etc., 1955; Kennedy etc., 1995; Pugliese etc., 1997; Vafiadis etc., 1997) relevant with the allele of the specific size of DBH (Wei etc., 1997) the identical polymorphism of gene.These researchs also are not precluded within the important function that exists single base pair to change in the subgroup of these length variants (face and Grice etc. as follows, 1996; Lichter etc., 1993; Krontiris etc., 1985).

At the unusual (Caskey etc. of multiple neurological, 1992) comprise fragile X mental retardation, Huntington Chorea (Huntington Chorea united research group, 1993), myotonia atrophica, Ken Nidishi disease, Friedreich ataxia (Campuzano etc., 1996) and the evidence that exists long triplet to repeat to participate in other (Caskey etc., 1992).At least these in unusual 5 relate to that GAG repeats in the gene that produces poly glutamine bundle in the aminoacid sequence of gene outcome separately.

Obesity-related gene research does not in the past identify any sudden change (Ezzel etc., 1995 of people OB gene in hundreds of obese individuals; Hamilton etc., 1995; Considine etc., 1996b).Yet, early-stage Study (Comings, 1996b; Comings etc., 1996c) shown participate in the unusual sudden change of polygenes may be beyond exon, and the repetition of polymorphism dinucleotide itself may regulated their institutes play a role in the expression of gene (Krontiris etc., 1993; Green and Krontiris, 1993; Trepicchio and Krontiris, 1992; Trepicchio and Krontiris, 1993; Bennett etc., 1955; Kennedy etc., 1995).

The TaqI polymorphism (APOE-D) of apolipoprotein gene is found with obese subjects and relevant between APO-D and fasting insulin.This work shows that the APO-D polymorphism may be hereditism's mark (Vijayaraghavan etc., 1994) of fat and hyperinsulinemia.

The functional variety of this gene of 5-hydroxy tryptamine gene may be the viewed reason that 5-hydroxy tryptamine and tryptophan raise simultaneously or reduce in various obstacles.4 kinds of different polymorphisms of people TDO2 gene have been identified.Correlation research shows that one or more same TS, the ADHD of these polymorphisms and drug dependence have significant dependency.Intron 6G-T variant with the horizontal significant correlation of platelet 5-hydroxy tryptamine (Comings etc., 1996a).

Polygenes is analyzed the joint-detection of dopamine d 4 receptor gene (DRD4), Fructus Cannabis ester acceptor gene (CNR1) and GABAB3 acceptor gene (GABRB3) and has been explained 25% (Saucier etc., 1996 that intravenous pharmacy abuse character changes; Comings etc. 1997; Johnson etc., 1997).According to observations, the Body Mass Index of the test interpretation 22.8% of OB and DRD2 gene changes, and shows that many genetics factors influence body weight, and the dependency by the determined congeniality symptom of single detection only be a less ratio reason (Comings etc., 1996b).Shown each polymorphism at three dopaminergic gene places: the TaqIB1 of the TaqIA1 of d2 dopamine receptor (DRD2), dopamine (D β H) and the same TS of multiple 10/10 genotype of 40bp, ADHD and the CD of dopamine transporter (DAT1) gene have significant dependency (Comings, 1996).

The effect of neurotransmitter and amino acid precursor participates in the gene of dysautonomia except being considered to, after deliberation neurotransmitter and medicine producing and alleviating effect in some spiritual character.In human body, shown that the preceding dopaminergic activity of central volume participates in human cognitive (Weinberger etc., 1988).Parkinsonian and may be in the schizophrenic, in cognitive process, activate and have before the volume dopaminergic function clinical sign (Weinberger etc., 1988k).The conversion of animal deutocerebrum chemical substance verified the load of precursor aminoacid or systematicness and directly variation (Blum etc., the 1996a of neurotransmitter levels after central nervous system's administration; Blum etc., 1996b).Zooscopy shows that NE and dopamine participate in relating to the corelation behaviour of attention widely of searching and exploration activity, distractibility, response rate, differentiation power and attention conversion.In a word, animal and human's research prompting dopamine and NE play a role in the processing in the early stage and late period of information respectively (Sara etc., 1994k).Several neurotransmitteies, particularly the dopamine that in brain function and mood regulation, plays a crucial role, 5-hydroxy tryptamine, norepinephrine, GABA, glutamine and opioid peptide can acutely be influenced by the cyclical level of their precursor aminophenol nutrient (Wurtman, 1981k).Data prompting amino acid precursor and endorphine enzyme inhibitor provide important effect (Blum etc., 1987a for the rehabilitation of ethanol, cocaine and food addiction; Blum etc., 1987b; Blum etc., 1987c; Blum etc., 1989b; Blum etc., 1990; Strandberg etc., 1996).

The innerv critical function of cerebral cortex catecholamine may be the control attention.What is interesting is especially from network structure and throw to corticocerebral catecholamine, promptly the dopamine neuron in midbrain veutro lid district and in last pons the NE neuron of locus coeruleus.Have been found that acetylcholine (ACH) and dopaminergic system (DA) all are crucial for keeping accurate cognitive performance.A series of the discovering of these aspects of detection cognitive function that disclosed by radiation arm labyrinth, these two kinds of neurotransmitter systems are with a kind of mode of complexity interact (Levin etc., 1995).Select the accuracy defective by blocking-up muscarine-or nicotine-ACH receptor-inducible.By being reversed by the dopamine receptor-blocking agent haloperidol with the inductive selection accuracy defective of scopolamine blocking-up M-ChR.Specificity DAD1 blocker SCH23390 also has this effect, and specificity D ₂The receptor blocking agent raclopride does not but have.Use D ₂The antagonist raclopride can be seen this effect, and SCH23390 does not then have with the D1 antagonist.Find selective d ₂Agonist LY1771555 reverses the selection accuracy defective that is caused by mecamylamine, shows D ₂Receptor participates among the effect of nicotine to cognitive function.These selective ds A treatment effectiveness in reversing cognitive defect be since the ACH activation not enough (Levin etc., 1990k).

More and more evidences shows that 5-hydroxy tryptamine may be regulated cholinergic function in several zones of mammal brain and these 5-hydroxy tryptamine energy/cholinergic interaction influence is cognitive.Be not all by inference by 5-hydroxy tryptamine can and the parallel processing of cholinergic system inductive palinmnesis confusion can change owing to the 5-hydroxy tryptamine of cholinergic system.The cognitive competence of a body is not only from the interaction between several neurotransmitteies, for example as the DA in the brain structure of Hippocampus or cortex, and from of the influence of other general function to memory, this may relate to the different brain substrate of the classical relevant substrates of those and memory function (for example attention, awakening consciousness accuracy etc.) (Cassel etc., 1995k).

In addition, determine that the external adjusting of Hippocampus θ depends on the co-activating of cholinergic and septal area, the middle level input of GABA energy.The cholinergic projection provides afferent excited power for Hippocampus θ-Kai cell, and septal area GABA can throw by suppressing the aggregate level that Hippocampus GABA energy relay cell (Hippocampus θ-pass cell) reduces inhibition.Generation for Hippocampus θ territory and cytoactive must have these two kinds active existence.Whether the balance between cholinergic and GABA can systems may determine Hippocampus synchronism (θ) or asynchrony that (Symthe etc., 1992) take place.

Other neurotransmitter such as norepinephrine (NE) may also play a role in learning and memory.The neuroregulation characteristic of NE shows that locus coeruleus cortex (LC) NE is incident upon in attention and the memory process and plays an important role.For example, in the target sensory system, discharge the gate of NE and tunning effect and can promote selective attention (Sara etc., 1994) at the crucial moment that changes to related stimulus.Other studies show that stress or physiological challenge process in the activatory the possibility of result of LC be that release by DA and NE increases or keeps awakening (Page etc., 1994).

It is reported, and the effect of the neuronic discharge of NE LC prompting LC system in regulating state of attention or insomnia in behavior rat and monkey (Aston-Jones etc., 1991k).Other supports the effect that LC increases persistent erection of the penis in the journey in short-term or the long time journey that promote complicated sensory stimuli reaction for the research of NE in the dentate gyrus, and with the effect unanimity of LC in memory and attention process (Harley etc., 1988k).External source gives or the NE of endogenous release can start one of dentate gyrus journey or long time journey strengthen (LTP) in short-term.

Studied and understood neurokinin Substance P (SP) and its N-and C-terminal fragment effect for memory, reinforcement and cerebral metabolism.Studies show that SP promotes memory and invigoration effect is arranged when administered peripherally.Yet, the most important thing is to find these effects as if by different SP sequential codings, because the SP1-7 hypermnesis of N-end, and C-seven peptides of SP and six peptide sequences are proved to be to have come invigoration effect with the equimolar dosage of SP.These different behavior effects sexually revise parallel with active selectivity of DA and site-specific, because SP and its C-end but not the terminal DA that increases in the volt nuclear (Nac) of N-then do not have in neostriatum.These results show that the invigoration effect of SP that periphery is given may be by the mediation of its C-end sequence, and this effect may coexist DA among the NAC active relevant (Huston etc., 1991k).

As for dopaminergic activity, former research has shown bromocryptine (bromocriptine), and a kind of dopamine-receptor stimulant can play useful (Kimberg etc., 1997) for vision-space working memory function in the normal experimenter.Find all that in the damage of carrying out with monkey and single unit record research and the neural video research in the mankind this memory form (having kept short interval aspect some of its moderate stimulation) is also with prefrontal cortex function closely link to each other (Goldman etc., 1987k; Jonidas etc., 1993).Also reported in the release rate of bromocryptine in genotypic situation reduction alcoholic and had selectivity positive effect (Lawford etc., 1995) according to d2 dopamine receptor.In addition, verified dopamine antagonist in the monkey of carrying out memory tasks for neuron active direct effect period of delay (Williams etc., 1995k).Phentermine, a kind of dopamine releasing agent participates in losing weight (Weintramb etc., 1992).

In addition, influence vision-space working memory in the control of the pharmacology of human and animal's deutocerebrum dopamine concentration, afterwards an effect is positioned prefrontal cortex.Yet dopamine agonist is still known little for the mankind's effect.By inference bromocryptine may by its to the cortex dopamine receptor and be projeced into receptor under the cortex in the zone of neopallium effect and to having influence (Kimberg etc., 1997) with the relevant cognitive function of cortex before the volume.They find that bromocryptine depends on experimenter's working memory ability to young normal subjects's effect.High ability experimenter is poor to drug reaction, and low ability person then improves.A kind of experience contact between the working memory system of these result's proof dopamine mediations in the mankind and higher cognitive function.Also show DRD2 A1 allele equally also relevant with vision-spatial memory defective (Berman etc., 1995k).

A double-blind study proves alcoholic's a kind of D2 agonist bromocryptine or a kind of placebo that carries A1 allele (A1/A1 and A1/A2 genotype) or only carry A2 allele (A2/A2), has reduced serious hope and anxiety in the A1 carrier with the bromocryptine treatment.The proportion of goods damageds with in the A1 carrier of placebo treatment for the highest.Bromocryptine to the A1 carrier act on it through the treatment in six weeks the time much strong.Dopamine D 2 agonist bromocryptine can improve high-grade cognitive function.

In animal, utilize advanced techniques, comprise that microdialysis detects, the verified change (Hernandez etc., 1988) of neurotransmitter output behind load precursor aminoacid.In addition, systematicness or directly the central nervous system give the change (Blum etc., 1972) of the behavior that in animal, confirmed behind the precursor aminoacid.Though some L-aminoacid is the precursor of neurotransmitter and neuromodulator, their racemate, D-aminoacid also have biologic activity.Particularly, D-phenylalanine, D-leucine, other D-aminoacid and some metabolite (for example hydrocinnamic acid) reduction are for degraded (Blum etc., 1977 of the opioid peptide of the adjusting key of mental state and behavior; Della Bella etc., 1980).

In some individualities, scientist described a kind of phenylalanine defective (PHD) (Lou, 1994k).In this, two in phenylalanine and the tyrosine composition dopamine biosynthesis are opened the beginning step, and dopamine is the metabolic precursor thereof of NE.The extracellular concentration of phenylalanine synthesizes by the reduction dopamine in PHD influences cerebral function.Shown that it induces EEG to slow down and prolong the carrying out time of neuropsychology test.Tyrosine concentration reduces in PHD among the CNS, and this may point out because competition suppresses, and for example passes through blood brain barrier, and is used for the substrate deficiency of the synthetic tyrosine of catecholamine.In experimentation, shown that synthetic the and release of dopamine can be influenced by the increase of tyrosine amount.The shortening of the extra diet of three kinds of dosage tyrosine picked-up (160mg/kg/24h) induced reaction time and reduce transmutability in PHD, and reported that in a double blinding cross-over experiment a similar dosage induces the improvement of psychology test, lower dosage does not then have.

Have endorphine enzyme and suppress the treatment (United States Patent (USP) 5,189, No. 064) that the amino acid whose associating of active precursor can be used for the cocaine dependence.The acute application of known cocaine can improve brain electricity physiological function obstacle some aspect (Maurer etc., 1988k).Chronic cocaine abuse change attention processing procedure (Noldy etc., 1990k).The acute application of known cocaine can improve some aspect (Jonsson etc., 1996) of brain electricity physiological function obstacle.Yet, contradiction be that chronic cocaine abuse changes notes processing procedure (Braverman and Blum, 1996).Though still have dispute, shown that the attention process is to depend on (Lyoo etc., 1996) that biogenic amine is regulated.

Obesity and nervous function are defined as obesity above ideal body weight 20% or more usually.Attempt multiple slimming method, comprised balanced diet low in calories, " fashion " diet, behavior adjustment, medicine (being D-phenflouramine, phenteramine etc.), surgery, the associating of hunger, the wiring of Hubei Province portion and these methods fully.The short-term solution that these method major parts are problems, and be of short duration effectively, some in addition may bring serious risk (Lockwood and Amatruda, 1984).Even proved in a short time to lose weight, but body weight is recovered after the scheme that loses weight is paused usually again.Though American's obesity of about 28% thinks that extensively obesity is a kind of food addiction, a kind of beauty treatment but not state (Kral etc., 1989 that a kind of oneself of healthy indication forces; Weintraub and Bray, 1989).

From recently for some reasons of obesity and treat the research that the difficulty of this state carries out and produced a kind of understanding.Verified strong hereditism basis (Bouchard, 1989 of obesity in than the twins study in horse American Indian; Stunkard etc., 1990).Obesity is that a kind of cause of disease differs and very general obstacle, and it has hereditism and environment composition.The macroscopic view of various foods select and familial matter utilization obstacle (SUD) between relation put down in writing by document, and neuro chemistry research has been supported by ethanol, nicotine, cocaine and carbohydrate common point (Nobel, 1998 by the dopaminergic system reinforcement; DiChiara, 1988).In this respect, obesity and SUD all can be considered to appetite and force.Some genes for example d2 dopamine receptor (DRD2) and dopamine transporter (DAT1) gene may be not only for obesity (Noble etc., 1994; Comings etc., 1993; Blum etc. are 1995a) and for total SUD and other spiritual mental maladjustment (Noble etc., 1994; Smith etc., 1992; Comings, 1994; Blum etc., 1995b; Comings etc., 1996; Cook etc., 1995) be a risk factor.In addition, hope is lighted in the clone and the order-checking of mice ob gene and people OB analog thereof, and the defective of this gene may be brought into play significantly effect in the reason of human obesity disease, and leptin, its gene outcome may useful in treatment (Zhang etc., 1994; Peileymounter etc., 1995).Though hereditism's effect can work separately, hereditism's situation is only set the state (promptly following the obvious increase of body weight when increase is ingested) that heredity-environmental interaction chance is provided in most of case.For the people with this hereditism's unsafe condition, obesity is lifelong state, equally with other chronic disease needs to carry out long-term treatment.

Specific reasons to the uncontrollable absorption behavior of ethanol, medicine and food (particularly carbohydrate) is also understood fully.Yet, be clear that these appetite compulsions are products of heredodiathesis and environmental damage factor.A large amount of research has shown opioid, opioid peptides, CCK-8, glycogen, DA and insulin interaction (Morley and Levine, 1988 in the selectivity picked-up of glucose utilization and carbohydrate; Moore etc., 1982; Morley etc., 1985; Riviere and Bueno, 1987).The dopamine (DA), norepinephrine (NE), epinephrine (EPI) and the 5-hydroxy tryptamine (5-HT) that comprise monoamines in the main neurotransmitter that participates in dietary behavior; Inhibitory nerve mediator γ-An Jidingsuan (GABA); And multiple neuropeptide for example pancreatic polypeptide, opioid peptides, releasing factor and various intestinal brain peptide (summary is seen Cooper etc., 1988; Gosnell, 1987; Bouchard, 1994).Coordinative role in center edge recompense system is controlled the extensive evidence (Leibowitz and Hor, 1982) of normal trophic behavior for many brain monoamine compounds and neuropeptide.The analysis of cerebrospinal fluid prompting is with particular disorder (Kaye etc., 1985 of the relevant cerebral nerve chemical functional of pattern of ingesting unusually in the human and animal; Kaye etc., 1984).

One to the mistake trencherman studies confirm that absorption contains a kind of dietary supplement PHENCAL of amino acid precursor ^TMThe experimenter of variant on average alleviated 27lbs in 90 days, and only alleviated 10lbs (Blum, 1990) in matched group.Find PHENCAL ^TMOr other similar neurotrophic agents helps rehabilitation (Blum etc., 1988c that alcoholic, multiple medicines thing misuser, heroin misuser and cocaine rely on individuality; Blum and Trachtenberg, 1988; Cold, 1996), and then point out a kind of co-therapy mode (Blum etc., 1996 for these multiple substance addictions; Blum etc., 1997).

The effect nicotine of nicotine also discharges dopamine, but also finds that nicotine improves memory (DiChiara etc., 1988) in rat, monkey and people's multiple test.Nicotine is reduced in rat in a kind of dose-dependent mode and distinguishes incorrect response (Geller etc., 1970) in the behavior.This effect is similar with chlordiazepoxide, but can not be by stimulant caffeine simulation (Geller etc., 1970).Proved that it is effectively that nicotine that natural gum or skin paste form suffers from the treatment of twitching among the experimenter of tourette's syndrome for some, and reported that smoking improves attention, awakening, study and memory (Wesnes and Warburton, 1984; Warburton, 1992; Balfour and Fagerstrom, 1996) and symptom (Coger etc., 1996 of improving ADHD; Conners etc., 1996; Levin etc., 1996).

The double-blind study of having reported a placebo determines to utilize effect (Levime etc., 1996 of nicotine in the adult's who suffers from ADHD treatment; Conners etc., 1996).Among 17 experimenters, 6 is that smoker and 11 are the non-smokers.All all meet the DSM-IV standard of AD in adults HD.The plaster that utilizes percutaneous with medicine being that 7mg/ days and non-smoker are that 21mg/ days dosage gives to the smoker.With equilibrated order active and about 1 week of placebo plaster are arranged.Nicotine causes the significantly overall improvement of clinical global impression (CGI) scoring.This effect even yet be significant when only considering the non smoker, this prompting only is not because the alleviation that regular smoking is given up.Nicotine causes as the remarkable increase of the vigor of being measured by emotional state table (POMS) test and in the overall significantly shortening that continues the response time in the performance testing.On Inattention (inattention) index, significant reduction is arranged also.The accuracy that the nicotine raising time is estimated and the variability of time response estimator curve of reducing.Because smoking is obviously than the adult of those no ADHD more general (Conners etc., 1996) in suffering from the adult of ADHD.

The nicotine system may be important with this effect of interaction partners of dopamine system.Utilization in radiation arm labyrinth, win the rat of transfer (win-shift) working memory task carried out nicotinic agonist and antagonist with the interactional a series of researchs of dopamine system (Levin and Rose, 1995k).The working memory defective that is caused by the nicotinic antagonists mecamylamine is by D1/D1DA antagonist haloperidol and specificity D ₂The antagonist raclopride is strengthened.On the contrary, the inductive defective of mecamylamine giving altogether reverse by D2/D3 agonist quinpirole.About the working memory behavior in radiation arm labyrinth, nicotine also has tangible interaction with the dopamine medicine.The dopamine agonist pergolide is not selected accuracy by itself improving the radiation arm.Nicotine is effective in reversing this defective.With respect to single wherein a kind of medicine of using, when giving jointly with nicotine, D2/D3 agonist quinpirole improves RAM and selects accuracy.Mecamylamine effectively weakens the working memory function (Noble etc., 1998) of radiating in the arm to the acute local infusion of midbrain dopamine nuclear.

A kind of inorganic matter (Jeejeehboy etc., 1977 during the effect trivalent chromium of chromic salts (CrP and CrN) to normal insulin function key; Schwartz etc., 1959).Some and be not that studies show that before all are replenished risk factor (Abraham etc., 1992 that chromium may advantageously change coronary artery disease (CAD) and non-insulin-dependent diabetes mellitus (NIDDM); Anderson etc., 1991; Donaldson etc., 1985; Glinsmann etc., 1966; Kaats etc., 1991; Levine etc., 1968; Page etc., 1991; Press etc., 1990; Roeback etc., 1991).Chromium is considered to cause by its strengthening effect to insulin (Offenbacher etc., 1988) of these changes.

Zooscopy is supported CrP can reduce insulin resistance and is improved the argument (Liarn etc. of organization, 1993), a human body discovers that the positive of organization when carrying out CrP replenishes changes (Hasten etc., 1992), another has reported positive findings, though organization does not have to change significantly on the statistics (Hallmark etc., 1993), and the positive change (Clancey etc., 1994) of the 3rd organization when finding not that any CrP of carrying out replenishes.Having shown that CrP replenishes improves organization, particularly reduces unnecessary body fat (Page etc., 1992).Yet, observed in the past the work that replenishes chromium simultaneously and carry out athletic training and be limited to effect to body weight and composition, the mutual contradiction of result (Clancy etc., 1994; Evans etc., 1993; Hallmark etc., 1996; Hasten etc., 1992).

Though the effect on about chromic salts (pyridine carboxylic acid salt and nicotinate) organization and TBW being alleviated is still controversial (Abraham etc., 1992; Anderson, 1995; Hallmark etc., 1993; Clancey etc., 1994; Bulbulian etc., 1996), but as if some reports be supported in the positive change (Kaats etc., 1996) of organization in the human body.On the contrary, (Grant etc., 1997; Bulbulian etc., 1996) reported in human body, have or do not have under the exercise situation with chromium picolinate and weight increase then shows positive effect (Kaats etc., 1992) for nicotinate in identical colony.

Chromium picolinate (CrP) is to use, studies and the chromium compound of promotion dynamics maximum, also may be feasible but in vitro study shows nicotinic acid chromium losing weight and changing in the field of organization.Research has in the past shown the additional reduction fat mass of chromium picolinate and has increased fat-free quality (Kaats etc., 1991; Page etc., 1991).Exercise research has in the past shown the increase (Stefanick, 1993) of fat-free quality equally.Though to studies show that of young male (Evans, 1989) and women (Hasten etc., 1992), unite and take exercise and the additional change that improves the organization that takes place when performing physical exercise of chromium picolinate, this discovery also is not proved (Clancy etc., 1994; Hallmark etc., 1996).It is reported, nicotinate (CrN) may in addition than pyridine carboxylic acid salt more important (Grant etc., 1997).

The stricture of vagina of the nutritional supplementation neurotransmitter effect in the behavior treating dysfunction disorderly may constitute basis (Blum etc., the 1996e of multiple spirit and behavior disorder; Peisico and Uhl, 1997; Noble etc., 1991).Particularly, the unusual adjusting of dopamine, 5-hydroxy tryptamine, norepinephrine, gamma aminobutyric acid (GABA), glutamine and opioid peptides is considered in assuetude disturbance, particularly those relate to performance key effect (Pohjalainen etc., 1996) in ethanol and the cocaine abuse.Thereby these observations provide opportunity for the nutraceutical absorption of selectivity can sway the emotion and then influence this design of behavior.Though the nutrition strategy uses (Grandy etc., 1989) in the past, the confirmation of effectiveness is very limited.Shown that associating amino acid precursor and endorphine enzyme inhibitor are for the rehabilitation of the RDS behavior that comprises ethanol, cocaine and gluttony from some have a significant effect (Noble etc., 1993; Noble etc., 1994; Blum etc., 1994; Balldin etc., 1993; Duffy etc., 1994; The sick community of Americanism task groups, 1991, United States Patent (USP) 5,189, No. 064).

The polygenes analysis of psychosis related gene and other polygenic character are by inference, psychotic behavior has identical gene, in case and the balance of dopamine-5-hydroxy tryptamine and other neurotransmitteies is upset, cerebral disorder can cause different widely behavior (Comings, 1990a as a result; Comings and Comings, 1991a; Winokur etc., 1970; Comings, 1994b; Comings, 1995b).Other people support the personality character may have neuro chemistry and such supposition (Cloninger, 1983 of hereditism's substrate that the different hereditism's transmutabilities by in dopamine and the transhipment of other neurotransmitter is mediated; Benjamin etc., 1996; Epstein etc., 1996; Cloninger, 1991).(Comings etc. are 1996a) with clinical genetics research (Comings, 1994b for the molecule genetics research of DRD2, D β H, DAT; Comings, 1994c; Comings, 1995b; Biederman etc., 1991; Comings and Comings, 1987) show that ADHD, tourette's syndrome, behavior disorder, ODD, dyslexia, learning disorder, stutter, drug dependence and alcoholism are pedigree obstacles relevant on the etiology, have male's advantage.

In the past twenty years, the most gene of these obstacles has all been identified, has been located, has been cloned and order-checking.Owing to still have the number of this genoid to be identified to reduce, so there is increasing interest in more common polygenes obstacle.It has been generally acknowledged that and identify that the gene that participates in these obstacles is with much more difficult.This difficulty is fully showed by mental disorder.Though to manic depressive disorder, schizophrenia, tourette's syndrome, terrified obstacle, unsociable and eccentric and other (the two-phase obstacle may make an exception) many chain researchs (Risch and Botstein, 1996) are arranged, almost do not have multiple discovery.Many is by utilizing LOD score analysis (lod score analysis), other linkage analysis form or haplotype relative risk technology based on family to attempt single-gene list disease model is used to polygenes obstacle service (Falk and Rubinstein, 1987) for the effort of finding gene in the complex barrier.At present, be used for identifying that right full genome screening is formed by suffering from the compatriot for the most frequently used method of complex barrier gene.Analyze chain nonparametric technique (Weeks and Lane, 1988) and be suitable for complicated heredity (but seeing Greenberg etc., 1996).Yet, when a given gene is explained less than 8% variation, must detect a large amount of parent-sons to or born of the same parents to (Carey and Williamson, 1991).

Recognize to have only correlational study may have ability (Risch and Merikangas, 1996 of evaluation more and more to the less gene of the percentage ratio contribution of a given polygenic character variation; Collins etc., 1997).Correlational study by relatively in the proband who seriously suffers from and between the anosis contrast that matches of irrelevant fully ethnic group the frequency of mutant candidate gene can differentiate these little effect (Weeks and Lathrop, 1995; Comings, 1996; Owen and McGuffin, 1993).DRD2, D β H and DAT gene (Comings etc. have been detected, 1996j), DRD1 and DRD2 gene (Comings etc., 1997a), OB and DRD2 gene (Comings etc., 1996d) and the additive effect of other gene associating gene in TS, ADHD, behavior disorder, stutter and corelation behaviour.In the TS syndrome, the effect of having found to identify three dopamine genes (DRD2, D β H and DAT1) is by to quite a large amount of TS experimenters, their relatives and the detection of contrast and determination, this shows that TS is polygenic inheritance with relevant obstacle, and each gene only is responsible for sub-fraction (Comings etc., 1996a in any behavior scoring change; Comings, 1996b; Comings etc., 1996d; Comings, 1996c).

Most of mental disorder be polygenic (Comings, 1996b) and also each gene is responsible for less than 10%, usually less than the change of 5% given behavior variable.In both research, increased the intensity of dependency by the additive effect that detects an above gene.One of main obstruction of correlation research extensive use be many candidate genes place of cloning and checking order or near lack available suitable polymorphism (Comings, 1994).Yet, even when the linkage technology of this technology of use or classics, find in research subsequently, can not be repeated usually (Egeland etc., 1987 from one group of investigator's the positive; Kelsoe etc., 1989; Blum etc., 1990; Bolos etc., 1990).Because colony's layering, this technology can also produce false positive, yet by a large amount of experimenters being utilized haplotype relative risk step (Falk and Rubinstein, 1987) it can be reduced to minimum.Whether possible the small-scale of these effects and the difficulty in repetition have caused a kind of about the gene that differentiate to participate in the polygenes obstacle downbeat mood (Moldin, 1997).Summary of the invention

Only just there are 18,000,000 alcoholic, 28,000,000 alcoholics' child, 6,000,000 cocainomaniacs, 14,900,000 other material of abuse persons, 25,000,000 nicotine addiction persons, 54,000,000 overweight at least 20% people, three and one-half million to suffer from the school age population of ADHD or tourette's syndrome and about 3,700,000 compulsive gambling person in the U.S..Inventors of the present invention believe the allelic genotype of measuring other gene of being correlated with the psychology obstacle among people's DRD2 gene and the present invention, are actually towards the first step to destructive problem rational therapy in the society.

The present invention at first provides a kind of compositions that is used at experimenter's treatment recompense deficit syndrome (RDS).In some aspects, said composition comprises a kind of of following compositions at least: opium destroys the material of enzymatic degradation of at least a inhibition neuropeptide acyl opium of amount of suppression, and it is aminoacid, peptide and analog thereof or derivant; At least a precursors of neurotransmitters of the synthetic promotion amount of a kind of neurotransmitter, it is a kind of dopamine precursor such as L-Tyr, L-Phe and L-DOPA, a kind of 5-hydroxy tryptamine precursor such as L-Trp and 5-hydroxyryptophan or a kind of gamma aminobutyric acid (GABA) precursor such as L-glutaminate, L-glutamic acid and L-glutamate, Glu; A kind of chromium picolinate (chromium picolinate) of tryptophan concentration raising amount or nicotinic acid chromium (chromium nicotinate); A kind of chemical compound that discharges endorphins, it is, but is not limited to a kind of peptide, preferred a kind of amino acid whose peptide of D-that contains; Perhaps a kind of at least a chemical compound of opium antagonism amount in δ, μ, κ, σ or epsilon receptor place blocking-up opium effect.Except top list especially, further described endorphine enzyme inhibitor, precursors of neurotransmitters, opium in this application and destroyed the type of mortifier, opiate antagonist and/or chromium compound, and forgive in the present invention.The present invention some preferred aspect in, compositions is used to prevention or reduces the unwanted body weight of experimenter.In some others of the present invention, compositions preferably is used in the treatment of attention deficit hyperactivity disorder, attention process or memory.In the present embodiment, for attention deficit hyperactivity disorder, attention deficit hyperactivity disorder (ADHD) attention process or memory, compositions more preferably comprises at least a Rhodila of being selected from of the synthetic promotion amount of neurotransmitter or the synthetic material that promotes of neurotransmitter of huperzine (hubazine).Used " derivant " can refer to a kind of chemical compound of chemical modification in this, and " analog " refers to a kind of different chemical compounds that have similar characteristic or structure with the chemical compound that is compared.

Aspect some, said composition can be used for the treatment of these all RDS corelation behaviours of announcing of the present invention.The RDS behavior is those with the unbalance relevant behaviors of chemistry, and this chemistry is unbalance to show as one or more has the relevant behavior disorder of happiness with individual to anxiety, indignation or to a kind of material serious hope.The RDS behavior comprises family history and the obesity of alcoholism, SUD, smoking, BMI or obesity, pathological gambling, carbohydrate gluttony, axle 11 diagnosis, SAB, ADD/ADHD, CD, TS, SUD, is described in this.

The present invention also provides the method for a kind of experimenter's of treatment RDS behavior, and these RDS behaviors include but not limited to irritability syndrome after SUD, obesity, smoking, tourette's syndrome, ADHD, schizophrenia/elusive behavior, attack, the wound, PMS or tobacco abuse.The RDS behavior is not to be confined to these obstacles especially, and the inferior obstacle of many types is also contained by these conditions.For example, attention deficit hyperactivity disorder (ADHD) can show as ethanol, medicine, forces coercive action, the problem concerning study in the learning disorder, reading problem, gambling, manic symptoms, phobia, frightened outbreak, opposition challenge behavior, conduct disorder, primary school, smoking, sexual behaviour, division sample, somatization, depression, sleep disorder, extensively anxiety, stutter and tic disorder.As a part of the present invention, all these behaviors, and relevant described in this or all be included in the RDS behavior with other behavior of the gene-correlation that participates in RDS related neural path with the RDS behavior.In addition, in this used for as the many clinical term of the multiple particular obstacle of RDS obstacle at " DSM-IV ^TMThe quick reference of diagnostic criteria " (Quick Referenceto the Diagnostic Criteria From DSM-IV ^TM) APA,American Psychiatric Association, the Washington D.C. can be found in 1994,358 pages.In this list of references, can find the particular obstacle of definition, and they are at DSM-IV ^TMIn numbering comprise: anxiety disorder comprises that anxiety disorder 293.89 due to the agoraphobia 300.22, special phobia 300.29, social phobia 300.23, obsession 300.3, posttraumatic stress disorder 309.81, acute stress disorder 308.3, generalized anxiety disorder 300.02, overanxious disorder of childhood 300.02, body situation (indicating) of the panic disorder 300.01 of not accompanying agoraphobia, the panic disorder 300.21 of companion's agoraphobia, no panic disorder history, material cause anxiety disorder 293.89, anxiety disorder (unreceipted) 300.00; How moving attention deficit and vandalism obstacle comprise the moving obstacle mixed type 314.01 of the outstanding how moving-impulsive type 314.01 of the moving obstacle of attention deficit/outstanding Inattention type 314.00 of obstacle, attention deficit/how, attention deficit/how, attention deficit/how moving obstacle (unreceipted) 314.9, conduct disorder 312.8, disobey sexual disorders 313.81, disruptive behavior disorder (unreceipted) 312.9; The two-phase obstacle comprises two-phase I type obstacle 296.0x, 296.40,296.4x, 296.6x, 296.5x and 296.7, two-phase II type obstacle 296.89, ring disposition thread obstacle 301.13, two-phase obstacle (unreceipted) 296.80; Depressive disorder comprises repeatedly major depressive disorder 296.3, dysthymic disorder 300.4, the depressive disorder (unreceipted) 311 of outbreak, the major depressive disorder 296.2 of single outbreak; Eating disorders comprises bulimia nervosa 307.51, anorexia nervosa 307.1, eating disorders (unreceipted) 307.50; Impulse control disorder comprises intermittent explosive disorder 312.34, kleptomania 312.32, pyromania 312.33, pathological gambling 312.31, trichotillomania 312.39, impulse control disorder (unreceipted) 312.30; Personality disorder comprises antisocial personality disorder 301.7, avoidant personality disorder 301.82, obsessive-compulsive personality disorder 301.4, schizoid personality disorder 301.20; Schizophrenia comprises intolerance style 295.30, disorderly type 295.10, catatonic type 295.20, not typing 295.90, residual type 295.60, Schizoaffective mental disorder 295.70, division sample mental disorder 295.40; Sleep disorder, comprise former sleep disorder, for example comprise former insomnia 307.42, former excessively sleep 307.44, narcolepsy 347, the circadian rhythm obstacle 307.45 of sleep, the parahypnosis of sleep disorder (unreceipted) 307.47, comprise nightmare disorder 307.47, sleep and shy obstacle 307.46, sleep walking 307.46, the sleep relevant issues of unusual (unreceipted) 307.47 of sleep relevant issues unusually, the sleep disorder relevant with other mental disorders-it comprises and [axle I or axle II obstacle] relevant insomnia 307.42, with [axle I or axle II obstacle] relevant excessive sleep 307.44, comprise the sleep disorder 780.xx that body situation (indicating) causes, other sleep disorder of the sleep disorder 780.xx that material causes; The substance abuse obstacle comprises that the ethanol associated disorders for example accompanies psychotic disorders 291.5 due to the ethanol of vain hope, alcohol abuse 305.00, alcoholism 303.00, ethanol gives up 291.8, alcoholism delirium 291.0, ethanol is given up delirium 291.0, persistency dementia 291.2 due to the ethanol, persistency amnestic disorder 291.1 due to the ethanol, alcohol dependence 303.90, psychotic disorders 291.3 due to the ethanol of companion's hallucination, affective disorder 291.8 due to the ethanol, anxiety disorder 291.8 due to the ethanol, sexual dysfunction 291.8 due to the ethanol, sleep disorder 291.8 due to the ethanol, ethanol associated disorders (unreceipted) 291.9, alcoholism 303.00, ethanol gives up 291.8; The nicotine associated disorders, it comprises nicotine dependence 305.10, nicotine withdrawal 292.0, nicotine associated disorders (unreceipted) 292.9; The amfetamine associated disorders, it comprises that amfetamine relies on 304.40, amfetamine abuse 305.70, poisoning by amphetamine 292.89, amfetamine gives up 292.0, poisoning by amphetamine delirium 292.81, psychotic disorders 292.11 due to the amfetamine of companion's vain hope, psychotic disorders 292.12 due to the amfetamine of companion's hallucination, affective disorder 292.84 due to the amfetamine, anxiety disorder 292.89 due to the amfetamine, sexual dysfunction 292.89 due to the amfetamine, sleep disorder 292.89 due to the amfetamine, amfetamine associated disorders (unreceipted) 292.9, poisoning by amphetamine 292.89, amfetamine gives up 292.0; The Fructus Cannabis associated disorders, it comprises, and Fructus Cannabis relies on 304.30, anxiety disorder 292.89, Fructus Cannabis associated disorders (unreceipted) 292.9, cannabism 292.89 due to psychotic disorders 292.12, the Fructus Cannabis due to the Fructus Cannabis of psychotic disorders 292.11, companion's hallucination due to the Fructus Cannabis of cannabis abuse 305.20, cannabism 292.89, cannabism delirium 292.81, companion's vain hope; The cocaine associated disorders, it comprises that cocaine relies on 304.20, cocaine abuse 305.60, cocaine poisoning 292.89, cocaine gives up 292.0, cocaine poisoning delirium 292.81, psychotic disorders 292.11 due to the cocaine of companion's vain hope, psychotic disorders 292.12 due to the cocaine of companion's hallucination, affective disorder 292.84 due to the cocaine, anxiety disorder 292.89 due to the cocaine, sexual dysfunction 292.89 due to the cocaine, sleep disorder 292.89 due to the cocaine, cocaine associated disorders (unreceipted) 292.9, cocaine poisoning 292.89, cocaine gives up 292.0; The hallucinogen abuse obstacle, it comprises hallucinogen dependence 304.50, hallucinogen abuse 305.30, hallucinogen intoxication 292.89, hallucinogen gives up 292.0, hallucinogen intoxication delirium 292.81, the hallucinogen caused by mental disorder 292.11 of companion's vain hope, the hallucinogen caused by mental disorder 292.12 of companion's hallucination, affective disorder 292.84 due to the hallucinogen, anxiety disorder 292.89 due to the hallucinogen, sexual dysfunction 292.89 due to the hallucinogen, sleep disorder 292.89 due to the hallucinogen, hallucinogen associated disorders (unreceipted) 292.9, hallucinogen intoxication 292.89, hallucinogen persistency sensory disturbance (flashback) 292.89; The inhalation (inhalatio) associated disorders, it comprises, and inhalation (inhalatio) relies on 304.60, inhalation (inhalatio) abuse 305.90, inhalation (inhalatio) are poisoned 292.89, due to the inhalation (inhalatio) of inhalation (inhalatio) poisoning delirium 292.81, companion's vain hope due to the inhalation (inhalatio) of psychotic disorders 292.11, companion's hallucination due to psychotic disorders 292.12, the inhalation (inhalatio) anxiety disorder 292.89, inhalation (inhalatio) associated disorders (unreceipted) 292.9, inhalation (inhalatio) poison 292.89; The opioid associated disorders, it comprises, and opioid relies on 304.00, opioid abuse 305.50, opioid are poisoned 292.89, due to the opioid of opioid poisoning delirium 292.81, companion's vain hope due to the opioid of psychotic disorders 292.11, companion's hallucination due to psychotic disorders 292.12, the opioid anxiety disorder 292.89, opioid associated disorders (unreceipted) 292.9, opioid poison 292.89, refraining opium type material 292.0; Many materials associated disorders, it comprises many substance depilatories 304.80; Tic disorder, it comprises Tourette obstacle 307.23, chronic motor or vocal tic disorder 307.22, the property a crossed tic disorder 307.21, tic disorder (unreceipted) 307.20, stutter 307.0, autistic disorder 299.00 and somatization obstacle 300.81.In addition, other RDS obstacles be defined as with by known to those of skill in the art those, for example novel the pursuit is defined in (Clonigen etc., 1993).If definition especially in this, other obstacle comprises general abbreviation with by in common knowledge identical of those skilled in the art.

Of the present invention aspect some in, giving the amount of the various top chemical compounds of putting forward that are used for RDS behavior or treating dysfunction every day can be about 1, about 2, about 4, about 5, about 6, about 7, about 8, about 9, about 10, about 11, about 12, about 13, about 14, about 15, about 16, about 17, about 18, about 19, about 20, about 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, about 50, about 55, about 60, about 65, about 70, about 80, about 90, about 100, about 110, about 120, about 130, about 140, about 150, about 160, about 170, about 180, about 190, about 200, about 220, about 240, about 260, about 280, about 300, about 325, about 350, about 375, about 400, about 425, about 450, about 475, about 500, about 550, about 600, about 650, about 700, about 725, about 750, about 775, about 800, about 825, about 850, about 875, about 900, about 925, about 950, about 975, about 1,000, about 1,100, about 1,200, about 1,300, about 1,400, about 1,500, about 1,600, about 1,700, about 1,800, about 1,900, about 2,000, about 2,100, about 2,200, about 2,300, about 2,400, about 2,500, about 2,600, about 2,700, about 2,800, about 2,900, about 3,000, about 3,250, about 3,500, about 3,750, about 4,000, about 4,250, about 4,500, about 4,750, about 5,000, about 5,250, about 5,500, about 5,750, about 6,000, about 6,250, about 6,500, about 6,750, about 7,000, about 7,250, about 7,500, about 7,750, about 8,000, about 8,250, about 8,500, about 8,750, about 9,000, about 9,250, about 9,500, about 9,750, about 10,000, about 11,000, about 12,000, about 13,000, about 14,000, about 15,000, about 16,000, about 17,000, about 18,000, about 19,000, about 20,000, about 21,000, about 22,000, about 23,000, about 24,000, about 25,000, about 26,000, about 27,000, about 28,000, about 29,000, about 30,000mg or more.In addition, though do not list especially, all amounts in the particular range all can be used and be contained by the present invention in the above.For example, about 4,751, about 4,752, about 4,753mg etc. though do not list especially in the sentence in front, can use in the present invention.

In certain embodiments of the invention, wherein the RDS behavior is an obesity, and the preferable range of compositions and composition are to give 460mg DL-phenylalanine, 25mg L-tryptophan, 25mg L-glutaminate and 5mg 2-methyl-3-hydroxy-4-formyl-5-hydroxymethylpyridine .-5 '-phosphoric acid every day.This embodiment other preferred aspect in, utilize announce in this or method known to those skilled in the art come test subject to have the family history that chemicals rely on to determine whether the experimenter, wherein this family history point out the probability height of successfully treating.In the others of this embodiment, this treatment suppresses gluttony.In others of the present invention, this treatment suppresses addiction.In aspect preferred, compositions contains chromic salts.

Described in this, utilizing a kind of molecular Biological Detection to come test subject to detect a kind of is a part of the present invention with RDS or the relevant allele of psychology behavior, and the allelic existence of a kind of like this diagnostic is that the experimenter more may be to the indicant of the compositions generation positive reaction that is used for the treatment of of being announced in this.In aspect this embodiment is preferred, the method for utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps tests by a kind of molecular Biological Detection, detects existence at least a in the following allele: D ₂TaqI A1, B1, C1 or exon ^6-7Haplotype, HTR2A-C allele isozygoty, the 0B-1875 dinucleotide repeats polymorphism＜the allelic homozygosity of 208BP, No. 2 chromosome microsatellite polymorphisms of people, APO-D-TaqI 2.2 or 2.7BP or 0B gene D7S1875, the allele of being mentioned above wherein detecting prompting improves the probability for the treatment of successfully reaction.This embodiment other preferred aspect in, compositions comprises the nicotinic acid chromium of effective quantity, and detects the allelic existence of experimenter DRD2 A1, wherein the allelic existence prompting of DRD2 A1 improves the probability of therapeutic response.This embodiment other preferred aspect in, compositions comprises the chromium picolinate of effective quantity, and detects the allelic existence of experimenter DRD2A2, wherein the allelic existence prompting of DRD2 A1 improves the probability of therapeutic response.

In one embodiment of the invention, wherein the RDS behavior is a tobacco abuse, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, at least a existence: D1 (Dde A1's isozygotys) in the allele below detecting, D2 (Taq A1), D4 (VNTR2), D5 (dinucleotide 13 allele, scope 135-159BP), DAT1 VNTR (10/10), D β H (TaqI B1 allele), the allele of being mentioned above wherein detecting prompting improves the probability of the successful reaction of treatment.

In one embodiment of the invention, wherein the RDS behavior further comprises unsociable and eccentric disease, tourette's syndrome or ADHD, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, at least a existence: D1 (Dde A1's isozygotys) in the allele below detecting, D2 (Taq A1), D4 (VNTR2), D5 (dinucleotide 13 allele, scope 135-159BP), DAT1 VNTR (10/10), D β H (TaqI B1 allele), MAOA (X), the allele of being mentioned above wherein detecting prompting improves the probability for the treatment of successfully reaction.In aspect some of this embodiment is preferred, compositions comprises Rhodila or huperzine.

In one embodiment of the invention, wherein the RDS behavior is a pathological gambling, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect at least a existence: D1 (Dde A1's isozygotys), D2 (Taq A1, B1, C1) in the following allele, the allele of being mentioned above wherein detecting prompting improves the probability for the treatment of successfully reaction.

In one embodiment of the invention, wherein the RDS behavior further comprises pathologic violence, division sample/avoidance (SAB), attacks, gets angry, is hostile to or posttraumatic stress disorder, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test, detect at least a existence: D2 (TaqA1, B1, C1, exon in the following allele by a kind of molecular Biological Detection ^6-7), DAT1 (VNTR 10/10), mNOSIa-≤201BP isozygoty, the allele prompting of being mentioned above wherein detecting improves the probability for the treatment of successfully reaction.

In one embodiment of the invention, wherein the RDS behavior is PMS, and the method for utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps tests by a kind of molecular Biological Detection, detects DAT1 (VNTR 10/10), D ₂TaqI A1, B1, C1, exon ^6-7At least a existence in the allele of haplotype allele or DRD1, DRD2, DRD4, HTT, HTRIA, TDO2, D β H, MAO, COMT, GABRAB, GABRB3, PENk, ADRA2A or ADRA2C gene, the allele of being mentioned above wherein detecting prompting may raising property to what treat successfully reaction.

The present invention and then a kind of definite experimenter is provided the method to hereditism's quality of at least a RDS behavior, it is by detecting from including but not limited to that at least one allele carries out in DRD1, DRD2, DRD3, DRD4, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, HTR1D β, HTR2A, HTR2C, IFN-, CD8A or the PS1 gene, and wherein this equipotential gene is diagnostic for a kind of RDS behavior.In one embodiment of the invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the existence of at least one MAOA allele VNTR polymorphism, wherein this allelic existence is manic for comprising, OCD, the property, sleep, primary school's behavior, gambling, study, Inattention, ADHD, ADDR, impulsion, MDE, CD, how moving, phobia, the behavior of division sample, extensive anxiety, somatizationization, medicine, intravenous pharmacy, read, ODD, twitch, the experimenter of the RDS behavioral genetics quality of ethanol or tobacco abuse is diagnostic.

In another embodiment of the present invention, the method for utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps tests by a kind of molecular Biological Detection, detects at least one DRD ₂Gene A ₁Allele, DAT ₁Gene, VNTR 10/10 allele or D β H gene B ₁Allelic existence, wherein this allelic existence is diagnostic for the experimenter of the RDS behavioral genetics quality that comprises division sample or avoidance.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, the multiple existence of (AAT) n triplet that detection is accelerated in the CNR1 gene, wherein this allelic existence is diagnostic for the experimenter of the RDS behavioral genetics quality that comprises drug dependence.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, the D7S1873 that detection is accelerated in the OB gene, D7S1875, D7S514 or the multiple existence of D7S680 dinucleotide, wherein this allelic existence is for comprising obesity, anxiety, depressed, psychosis, be hostile to, paranoia's sample imagination, force, the symptom summation, general SI, the novel pursuit, whole summation, nervousness, and the experimenter of careful RDS behavioral genetics quality is diagnostic.In this embodiment, preferred allele is the D7S1875 dinucleotide repetition of length greater than 225bp, and this equipotential gene all exists in two copies of CNR1 gene.In this embodiment, also preferred another allele that detects is the D of DRD2 gene ₂A1 allele.In this embodiment, also preferred RDS behavior is an obesity.

In another embodiment of the present invention, the method for utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps tests by a kind of molecular Biological Detection, detects the D of DRD2 gene ₂The allelic existence of A1, wherein this allelic existence for comprise tourette's syndrome, manic symptoms, disobedience, property, ADHD-R, division sample, ADHD, twitch, major depression, conduct, stutter, force, the experimenter of the RDS behavioral genetics quality of somatization symptom, alcohol abuse, study and sleeping problems is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the allelic existence of Taq A1 of D β H gene, wherein this allelic existence for comprise tourette's syndrome, ADHD, smoking, study, primary school, ADHD-R, disobedience, twitch, manic, ethanol, reading, drug dependence, sleep, stutter, force, the experimenter of the RDS behavioral genetics quality of somatizationization and major depression is diagnostic.In this embodiment, preferred allele is the Taq B1 allele and the Taq A1 allele of D β H gene, and the RDS behavior is a tourette's syndrome.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect 10 allelic existence of DAT1 gene, wherein this allelic existence for tourette's syndrome, autism, somatizationization, ethanol, ADHD-R, major depression, terrified, force, the experimenter of the heredodiathesis of extensive anxiety, manic, disobedience, property, reading and ADHD is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test, detect 10 allele, the Taq A1 allele of D β H gene or the D of DRD2 gene of DAT1 gene by a kind of molecular Biological Detection ₂The allelic existence of A1, wherein this allelic existence for ADHD, stutter, ADHD-R, disobedience, twitch, conduct, force, manic, ethanol, extensively the experimenter of the heredodiathesis of anxiety, terrified, division sample, sleep, property, medicine and major depression is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the allelic existence of DdeI of DRD1 gene, wherein this allelic existence for ethanol, smoking, force feed, twitch, gambling, medicine, reading, shopping, disobedience, major depression outbreak, divide sample, ADHD, conduct disorder, force and the experimenter of manic heredodiathesis is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the TaqI A1 and the allelic existence of TaqI A2 of DRD2 gene, wherein those allelic existence for disobedience, conduct disorder, gluttony, smoking, gambling, ADHD, force, the experimenter of the heredodiathesis of manic and ethanol is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect 11 or 22 genotypic existence of DRD1 gene, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis of tourette's syndrome, smoking and gambling.In this embodiment, the preferred allele that detects is two copy/genome DRD1 gene Dde1 allele.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect 11 genotypic existence of DRD1 gene, wherein this allelic existence for disobedience behavior, conduct disorder, force feed, smoking, gambling, ADHD, manic, stutter, force and the experimenter that divides the heredodiathesis of sample is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the allelic existence of DdeI of DRD1 gene, wherein this allelic existence for gambling, smoking, force feed, disobedience, major depression outbreak, ADHD, conduct disorder, division sample, force, the experimenter of the heredodiathesis of manic and ethanol is diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect 11 or 22 genotypic existence of DRD1 gene, wherein this allelic existence for ethanol, smoking, to force the experimenter of the heredodiathesis of feed, twitch, gambling, medicine, reading, shopping, gambling and primary school's problem be diagnostic.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect tryptophan 2, the existence of the intron 6G → A polymorphism of 3 dioxygenases, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis of tourette's syndrome.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect tryptophan 2, the existence of the intron 6G → T polymorphism of 3 dioxygenases, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis of ADHD, alcohol dependence, drug dependence, pathological gambling.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect tryptophan 2, the existence of the intron 6DGGE polymorphism of 3 dioxygenases, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis of ADHD, alcohol dependence, drug dependence, pathological gambling and depression.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, (＜181bp) the existence of polymorphism, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis of drug dependence to the low base pair allele of detection ADRA2C dinucleotide repetition polymorphism.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, the existence of the high base equity position gene pleiomorphism of two 〉=183bp of detection ADRA2C dinucleotide repetition polymorphism, wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis that comprises alcohol abuse.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect (PS-1) two homoeoallelic existence of polymorphism of senilism element-1 (presenilin-1), wherein this allelic existence is diagnostic for the experimenter of the heredodiathesis that comprises ethanol and tobacco abuse.In this embodiment, the preferred allele that detects be the PENK gene repeat two homology allele of polymorphism greater than the CA dinucleotide of 80bp.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the allelic existence of short GGC of AR gene, wherein this allelic existence is diagnostic for the experimenter who comprises CD, ODD or the heredodiathesiies of moving more.

In another embodiment of the present invention, the method of utilizing in this to be announced is carried out hereditism's test to the experimenter, perhaps test by a kind of molecular Biological Detection, detect the allelic existence of DRD2, wherein this allelic existence is diagnostic for the experimenter who comprises the heredodiathesis that crapulent type B behavior, cocaine addiction or RDS demonstrate,prove earlier.

In the present invention and then provide a kind of and determine the method for the heredodiathesis of polygenic character is comprised that detection is from least one the relevant allele in DRD1, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, BABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, DRD3, DRD4, HTR1D β, HTR2A, HTR2C, IFN-, CD8A or the PS1 gene.In one embodiment of the invention, polygenic character is ADHD, and the same DRD1 of allele, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, BABRB3, CNR1, CNRA4, NMDAR1, PENK, AR or CRF gene-correlation.In another embodiment, polygenic character is the shortage of ADHD susceptibility, and the same DRD3 of allele, DRD4, HTR1D β, HTR2A, HTR2C, IFN-, CD8A or PS1 gene-correlation.

In another embodiment of the present invention, polygenic character is OOD, and the same DRD1 of allele, DRD2, DRD3, DAT1, HTT, HTR1A, HTR2A, HTR2C, DBH, ADRA2A, ADRA2C, MAOA, GABRA3, GABRB3, CNR1, CHRNA4, NMDAR1, PENK, AR or CD8A gene-correlation.

In another embodiment of the present invention, polygenic character is twitch, and the same DRD1 of allele, DRD5, HTR1A, HTR1D β, HTR2C, TDO2, DBH, ADR2C, COMT, GABRA3, CNR1 or CHRNA4 gene-correlation.

In another embodiment of the present invention, polygenic character is LD, and the same DRD1 of allele, HTR2C, TDO2, DBH, ADR2A, ADR2C, MAOA, CNR1 or CNRA4 gene-correlation.

In another embodiment of the present invention, polygenic character is LDL, and the same HTT of allele, OXYR, DRD2 or PS1 gene-correlation.

In another embodiment of the present invention, polygenic character is long-lived, and the same PS1 of allele, OXYR or APOE gene-correlation.

The present invention provides a kind of method of setting up diagnostic polygenes detection method in addition, comprises evaluation character to be studied, sets up and wait to study measuring of the serious Cheng Du of character; Select at least a may be to the contributive candidate gene of described character, identify at least a polymorphism relevant with candidate gene, the allele pattern of polymorphism is associated with described measuring, and compares the dependency of allele pattern candidate gene and character dependency.To add together with the positively related allele pattern of character, form the polygenes detection method that a kind of polygenic character susceptibility to the experimenter has diagnostic significance.To form with the allele pattern addition of character negative correlation and a kind of the experimenter be lacked the diagnostic polygenes detection method of polygenic character susceptibility, this also is a part of the present invention.

In one embodiment of the invention, candidate gene includes, but are not limited to DRD1, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, BABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, DRD3, DRD4, HTR1D β, HTR2A, HTR2C, IFN-, CD8A or PS1 gene.

In another embodiment of the present invention, polygenic character includes, but are not limited to ADHD, ADHD shortage, ODD, CD, LD, twitch, drug dependence/dependence, smoking, osteoarthritis, cholesterol levels rising, the LDL level raises or longevity.

In one embodiment of the invention, detect to detecting the allele of at least one and DRD1, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, BABRB3, CNR1, CNRA4, NMDAR1, PENK, AR or CRF gene-correlation for the polygenes of ADHD.

In another embodiment of the present invention, the polygenes that lacks for ADHD detects and comprises the allele that detects at least one and DRD3, DRD4, HTR1D β, HTR2A, HTR2C, IFN-, CD8A or PS1 gene-correlation.

In one embodiment of the invention, detect for the polygenes of OOD and comprise the allele that detects at least one and DRD1, DRD2, DRD3, DAT1, HTT, HTR1A, HTR2A, HTR2C, DBH, ADRA2A, ADRA2C, MAOA, GABRA3, GABRB3, CNR1, CHRNA4, NMDAR1, PENK, AR or CD8A gene-correlation.

In another embodiment of the present invention, detect for the polygenes of twitching and to comprise the allele that detects at least one and DRD1, DRD5, HTR1A, HTR1D β, HTR2C, TDO2, DBH, ADR2C, COMT, GABRA3, CNR1 or CHRNA4 gene-correlation.

In one embodiment of the invention, detect for the polygenes of LD and comprise the allele that detects at least one and DRD1, HTR2C, TDO2, DBH, ADR2A, ADR2C, MAOA, CNR1 or CNRA4 gene-correlation.

In another embodiment of the present invention, the polygenes that raises for the LDL level detects and comprises the allele that detects at least one and HTT, OXYR, DRD2 or PS1 gene-correlation.

In one embodiment of the invention, detect for the life-prolonging polygenes and comprise the allele that detects at least one and PS1, OXYR or APOE gene-correlation.

In another embodiment of the present invention, detect for the polygenes of osteoarthritis and also to comprise the allele that detects at least one and COL2A1, COL2A1, COL2A1, COL9A1, COL9A1, AGC1, IGF1, IGF1, IGF1R, IGF1R, IGF2, IGF2R, TGFB1, TGFB2, IL1A, IL1B, IL1R1, IL1RN, MMP9, TIMP1 or vitamin D3 gene-correlation.

When using in this article, " a kind of " or " one " or " individual " should be understood to mean a kind of or more than one.The accompanying drawing summary

Following accompanying drawing constitutes the part of this description and is used to further specify particular aspects of the present invention.The present invention can be by being able to better understanding with reference to a secondary or how secondary accompanying drawing with to the detailed description to particular herein.

Fig. 1. " super contrast (normally) " and serious " RDS " proband's DRD2 gene.The linear trend that is used for each group shown in the comparison is analyzed p＜0.000001.Computer sorting group:

Group I (C1): to alcoholism, the substance abuse obstacle, many substance depilatories, chemistry relies on and fat family history, nicotine dependence (smoking) BMI surpasses 25, carbohydrate gluttony, autism, Tourette, ADHD, Axis II, pathological gambling and posttraumatic stress disorder are carefully estimated.The inventor utilizes the DSMIV criterion to estimate the substance abuse obstacle.(N＝11)

Group II (C2): the identical exclusion standard of organizing I except Axis II is included.(N＝6)

Group III (C3): except the identical exclusion standard of organizing I and II substance abuse obstacle and the fat positive family history is included.(N＝20)

Group IV (C4): except cigarette smoking (nicotine dependence), organize I, II, the identical exclusion standard of III is included.(N＝21)

Group V (C5): except benzodiazepine abuse/dependence, organize I, II, III, the identical exclusion standard of IV is included.(N＝31)

Group VI (C6): except substance abuse obstacle (being ethanol and cocaine), organize I, II, III, IV, the identical exclusion standard of V is included.(N＝74)

Group VII (C7): except BMI organizes I above 25, II, III, IV, V, the identical exclusion standard of VI is included.(N＝140)

Group VIII (C8): except the BMI of symbiosis substance abuse obstacle (abuse of alcohol and cocaine) surpass 25 organize I, II, III, IV, V, VI, the identical exclusion standard of VII is included.(N＝31)

Group IX (C9): except the BMI of the many substance depilatories of symbiosis (being ethanol and cocaine) surpass 25 organize I, II, III, IV, V, VI, VII, the identical exclusion standard of VIII is included.(N＝11)

In addition, in order to carry out statistics relatively, the present invention also comprised by document in advance gene type 18.5% D2A1 prevalence rate (Blum etc. are arranged, 1990, Blum etc., 1991, Noble etc., 1993, Parsian etc., 1991, Comings etc., 1991, Smith etc., 1992, Amedeo etc., 1993) healthy white man (L1) Men's and Women's that adds up to 286 (N=286) (only be used to screen ethanol and drug abuse person and screen the nicotine misuser in some cases).In addition, present invention includes the experimenter (L2) of sum 714 (N=714), they only are used to screen ethanol or many substance depilatories (Blum etc., 1990, Parsian etc. by document, 1991, Comings etc., 1993, Noble etc., 1994, Amedeo etc., 1993, Comings etc., 1994, Noble etc., 1993, Schwab etc., 1991, Uhl etc., 1992, O ' Hara etc., 1993, Uhl etc., 1992), the D2A1 prevalence rate is 25.9.In addition, the present invention has also comprised the experimenter (L3) of sum 980 (N=980), presses document (Bolo etc., 1990, Grandy etc., 1989, Gelernter etc., Uhl 1992, Goldman etc., and Finns 1994, Nothen etc., Noble etc., 1994, Jonsson etc., 1993, Hedebrand etc., 1993, O ' Hara etc., 1993) they have 32.9 D2A1 prevalence rate (contrast of unknown situation).

Fig. 2 .Phencal ^TMEffect to the body weight reduction.This figure explanation is at biennial PHENCAL later on ^TMWith non-PHENCAL ^TMThe weight ratio of group.Last research in 2 years and do not use PHENCAL ^TM52.8% the comparing of matched group (n=117), use PHENCAL ^TMAverage overweight 23.5% (p＜0.0001) of experimenter (n=130).

Fig. 3 .MAOA VNTR polymorphic allele distribution (allele sum=768).

Fig. 4. increase the adding up property effect of the number of adding up property genetic mutation gradually to the ADHD scoring.It shown a kind of from have only 4 or 5 variant genes 1.0 to 25.0 the carrying out property increase trend that 15 variant genes are arranged.P value＜10 of the linear X 2 test of carrying out property of ADHD scoring increase ^-8

Fig. 5. the adding up property and the subtracting property effect of all 29 gene pairs ADHD scorings, and the adding up property effect of adding up property gene only.The r of each gene of scoring is at random specified in the open squares representative of bottom ²Value, it matches with the scoring frequency of observing.r ²Adding up property of the carrying out property effect of value is represented by the square of sky, only uses positive relevant adding up property effect to represent by the square that an x is arranged.What the point on the line came labelling by x is adding up property gene, on the line what o'clock come labelling by a solid dot is adding up property and subtracting property gene.Use the final r of adding up property and subtracting property gene simultaneously ²Be .0001.Only use the final r of adding up property random gene ²Be .0004.Neither one is significant.In addition, though in PENK gene at random substituting r ²Up to .008, but it rolls back .0004 after the last gene of adding up property at random (CD8A) adds.

Fig. 6. all 29 genes are the adding up property of (n=166) and subtracting property effect in sample in two.Some genes are adding up property subtracting property again in two groups, some genes in one group be adding up property and another the group in subtracting property.The data of solid dot representative group I, filled squares representative group II data.

Fig. 7. the variant gene distributed number that does not have the experimenter of DSM-IV ADHD symptom to compare with the experimenter of the DSM-IV standard of those ADHD.

Fig. 8. four gene HTT, OXYR, DRD2 and PS1 are to the adding up property effect of cholesterol and LDL blood levels.This figure illustrates that the MAA technical appraisement goes out four genes, when combining with 154 experimenters, provides p=2.0 * 10 ^-4Remarkable result.Filled circles is the data of cholesterol, and open circles is the data of low density lipoprotein, LDL.The description of illustrative embodiment is used for the polygenes of RDS and the diagnosis of other polygenic characters

The present inventor believe many psychology obstacles by common biology substrate interrelate, " rigid line " system in the brain provides enjoyment in the process of some behavior of recompense.The present inventor proposes in the present invention, and the congenital chemical imbalance that changes volt nuclear or other recompense district, edge intercellular signal conduction may be with anxiety, indignation or the addiction to material (as ethanol) that can relax negative feeling is replaced individual happiness feel.This chemical imbalance show as one or more behavior disorders under the vocabulary of having created " recompense deficiency symptoms " (Blum etc., 1996a).

In recompense deficiency symptoms or RDS, hereditism's defective best appreciated of recompense approach is the polygenes obstacle, and the hereditism detects needs to detect a plurality of genes.Susceptibility that the present invention has identified RDS and dependency: dopaminergic gene DRD1, DRD2, DRD3, DRD4, DRD5, dopamine transporter gene (DAT1) including, but not limited to following gene; 5-hydroxy tryptamine gene HTT, HTRA, HTRDb, HTRA, HTRC, tryptophan 2,3-hydroxylase (TD02); The noradrenaline plain gene, D β H, ADRAA, ADRAC, NT, catecholamine metabolism gene, MAOA, COMT, GAGA gene, GABRAA, GABRAB, Fructus Cannabis ester acceptor gene, CNR, nicotine cholinergic, CHRNA; The nmda receptor gene, NMDAR, enkephalin gene, PENK; Androgen receptor gene, AR, IFN-gene, INFG, CDA; The senilism element-, PS-; The CRF gene, CRF, ob gene (OB), leptin receptor gene; 5-hydroxy tryptamine HTR1A acceptor gene, 5-hydroxytryptamine receptor (5HT2R) gene, catechol (catachol)-O methyl-transferring enzyme (COMT) gene, neuron nitric oxide synthase gene (nNOS1a), apolipoprotein-D (APO-D) and uncoupling protein (UCP1 and UCP2).

The unusual importance of the character of Fu Za RDS and corelation behaviour and many environmental factorss has been eliminated the probability that in fact a specific gene or environmental factors play 100% decisive action.Though believe that " recompense cascade pattern " causes the RDS behavior when suffering damage generally, the present inventor points out modestly, though a more than gene may be to being responsible at the certain proportion of the total variation of the inferior character of a kind of RDS, it is may relevant RDS behavior with another few or it doesn't matter.

Improved genotyping technique has made that the applied genetics method is drawn gene map in the commercial possibility that becomes in the etiology of common human diseases.The numerous disease gene identifies by the linkage analysis method, and this method is with random marker site being divided into from detecting in the kinsfolk of disease character.Major part in them belongs to the single-gene Mendel disease gene (Weeks and Lathrop) with simple inheritance mode.Now, human inheritance scholar hereditism for example hypertension, diabetes, heart disease, multiple sclerosis, arthritis and the RDS behavior such as the obesity of multi-factor disease that beginning one's study.Multi-factor disease is by interacting and cause with the interactional a plurality of genes of environmental factors, producing the gradient to disease genetic quality.The interaction of upper degree and type or these genes influences strongly uses the chance that the linkage analysis method detects gene.Even without upper, if keep hereditism's heterogeneity, several different loci cause character independently, and opportunity of success may reduce.Disease such as RDS for complexity, it is can be not very effective that tradition LOD scoring is analyzed, because there is the single principal disease site (special hereditary pattern is arranged) that can explain most of hereditary variation in its supposition, now known is not such concerning RDS, so correlational study will be more effective and will be better method.

Prior art is for example trying to find out alcoholism itself or resemble TS and during the gene of the RDS behavior of ADHD, important mistaken ideas are that most of worker comprises the inventor, all concentrate on the individual gene method, therefore be merely able to identify the sub-fraction of all variations, as DRD2 allele.For example for TS, groundwork also is the chain research that utilization has the autosomal dominant inheritance, AD pattern that lowers penetrance, is not correlational study, though got rid of almost hundred-percent genome at present, this method is not effective.The present inventor has illustrated that RDS is the polygenes obstacles from parents' gene.When using the chain research of Lod scoring in the polygenes obstacle in a reality, in labelling, can cause many mistakes, even so that, also can obtain minus lod scoring in fact participating in the gene that phenotype limits.In addition, when obstacle is polygenic inheritance, about got rid of the argument of specific gene effect by linkage analysis, as in the DRD2 gene, no longer valid (Devor etc., 1994; Gelernter etc., 1990).

Among the present invention, an individual gene with the polygenes group is referred to as a polygenes sometimes.In the past ten years, the inventor on inspection the latent effect of dozens of gene of many behaviors and neurological's phenotype.Based on the calculating of correlation coefficient, the inventor finds, does not consider the level of significance, and specific polymorphism explains that the percentage ratio of QTV variation is between 0.5% to 2.5%.Because specific allele is to character low effect level like this, correlational study may be unique effective ways of identifying the polygenes effect.Because polygenes explains the percentage ratio of phenotype and reduce, the difficulty of identifying this effect increases and the experimenter's that must study quantity also increases.A criticism to correlational study is, if contrast is to come from different races or ethnic group, the appearance of gene frequency or difference can produce wrong result in these colonies.This can eliminate by haplotype relative risk technology in theory, concerning two times of types (diabolic) sign, when gene frequency 0.1 when between 0.2, changing, and the gene of research only causes that when being lower than 20% variation, the ability of this technology is severely limited.At last, refuting reason for one of correlational study method supporter (especially for DRD2 research) is, careful screening contrast is to get rid of many relevant RDS behavior (SUD, TS, ADHD, CD, SAB, and other) failure and the condition of inspection can cause obtaining incoherent false supposition (Blum etc., 1995; Blum etc., 1997).

Yet several meta analyses of available data have illustrated that this being correlated with is strong (Cloinger, 1991; Gorwood etc., 1994; Noble an Blum K, 1993; Pato etc., 1993; Cook and Gurling, 1994; Uhl etc., 1993; Blum etc., 1995; Blum etc., 1997).The key factor that research DRD2A1 allele is relevant with alcoholism (it also is suitable for other RDS behavior) is to use the comparative control type.The binding analysis of former used contrast has been illustrated that A1 allele has than remarkable high prevalence rate (Uhl etc., 1993 in the contrast of carrying out alcoholism and other related factors evaluation in the contrast that does not have evaluation; Noble etc., 1994a).This result is more outstanding in nearest research, Neiswanger etc., and 1995 and Hill and Nyswander, 1997, they find the strong relatedness between A1 allele and the alcoholism by using " super normal " contrast.This seminar concludes that so the matched group of evaluation was than errors of sampling or the prior explanation of colony's layering to former observed deflection the possibility of result.

Super contrast.The inventor is at Priceton, 184 proband have been estimated in New Jersey neuropsychiatry and medical science clinic, they are modestly near contrast, so each individuality has all been got rid of many RDS behaviors and (comprised alcoholism, SUD, smoking, BMI or obesity, pathological gambling, carbohydrate gluttony, axis 11 diagnosis, SAB, ADD/ADHD, CD, TS, SUD family history, and obesity), carry out the TaqID2A1 gene type then, find to organize differently, among standard compliant 30 people, have only one to carry DRD2A1 allele or have only 3.3% (seeing Fig. 1 .) with 32.9% the allelic white man of screening of DRD2A1 that carries.In fact when all groups all compared, they found, compared with the contrast of fine evaluation, and the gradual percentage ratio (%) of many substance depilatories of symbiosis group (seriously) A1+ allele prevalence rate increases obviously wants high.(X2＝78.7，df＝1，p＜0.000001)。

The application's a important embodiment comprises separately or the method for some genetic mutations of joint-detection, separately the effect of diagnosis RDS behavior is carried out based on them.The inventor believes by the associating that utilizes gene and detects above-mentioned specific polymorphism or actual sudden change, can identify the risk individuality more accurately, accuracy is higher when promptly having only a gene detected like that such as the DNA detection of the dopamine receptor gene of the patent suggestion of announcing in the past.In order to be illustrated more clearly in this potentiality, infer that some genes participate in the recompense path.Although being described, neurotransmitter participates in the recompense path here, but the related gene that the nomological validity of the polymorphism reported and genetic defect is arranged in substance abuse and other impulsion, addictive behavior, the inventor advises that these genes and many other unidentified genes will form complete RDS differential mode figure (allomorphic map).It is as a kind of basis, can think carrying out along with time and Geng Duo research, the gene that influences other neurotransmitteies will join among this polygenes group, and other polygenic psychiatric disturbances (especially for " recompense " of the present invention and serial behavior) application present technique also will be decoded.In fact there is a kind of like this probability, many chromosomal focis and specific markers or gene will be found in the near future, be used to detect the MULTIPLEX GENESCAN that is defined as the relevant many associated genes of the impulsion of recompense deficiency symptoms-force-addictive behavior and other polygenic characters with the inventor but the objective of the invention is to develop ^TMIn addition, the present inventor has developed the diagnostic method of the polygenic character that comprises RDS, is called as the association that adds up (MAA) technology more.

The step of multiple adding up property association (MAA) technology.The application of certain embodiments explanation MAA technology in making up RDS associated disorders diagnostic analysis.Yet, some embodiment that the inventor proposes be with psychiatric disturbance irrelevant and illustrate that the MAA technology extends to all polygenes obstacle and polygenic character.Therefore, the inventor imagines the MAA technology becomes method to all polygenes obstacles.The polygenes obstacle is considered to the additive effect owing to many genes, and each gene can only cause the sub-fraction of phenotypic variation alone.They present different degree in whole individualities.The polygenes obstacle has influenced 1% to 20% of population than general of single-gene obstacle more.The part example is a hypertension, obesity, most of psychiatric disturbances, multiple sclerosis, lupus erythematosus, osteoporosis, coronary artery disease, rheumatoid arthritis, osteoarthritis, body weight, height, blood pressure, at the age (life-span), psychology character and any other are to a certain extent by the character that surpasses the decision of a gene or allele.How lecture the MAA technology below implements.

This MAA technology has the additional features of following uniqueness.At first, it enlarges the gene dosage that can detect significantly to thousands of.It is by using correlation coefficient (r), variation ratio (r ²) replaced character itself that all quantitative traits or two divergent character are placed same measuring.It introduces such notion, and when individual gene adds fashionablely, they may be (r and the r of adding up property ²Raise) or (r and the r of subtracting property ²Reduce).It utilizes this rising and reduces identifies those genes that work in obstacle or character because their adding up property and in obstacle or character inoperative those genes because their subtracting property.Only utilize adding up property gene to assess the percent of total r of the variation that causes by the adding up property gene of identifying ²Analyze again.Adding up property effect by assessing a plurality of genes rather than assess gene in the mode of next gene, the MAA technology when identify participating in the gene of polygenes obstacle than for example lod scoring of the method for individually assessing gene, blood relation to, haplotype relevant risk (Falk and Rubinstein, 1987) and genetic unbalance check (Spielman and Ewens, 1996) more effective.P value in the correlational study of MAA technology demonstration one next gene whether participates in the polygenes obstacle with a gene or character is seldom relevant.The MAA technology can be described as having following steps, and some steps are that this technology is distinctive.

Step 1. first step is to identify polygenes obstacle or the character that will study, as attention deficit hyperactivity disorder (ADHD), depression, cholesterol levels, body weight, height, life-span, blood pressure, multiple sclerosis, perhaps any other polygenes or supposition are polygenic obstacle or character.Except as otherwise noted, adopt existing diagnosis meet method such as DIS (diagnosis meet program) (Robins etc., 1991) or SCID (Williams etc., 1992) use the DSM standard, carrying out known or inferring is the psychiatric diagnosis of polygenic psychology character individuality.The inventor imagines up-to-date DSM version and can be used to carry out such diagnosis, but old version still can use.

Second step of step 2. is to set up measuring of assessment polygenes obstacle seriousness.This may be quantitative trait or two distortion amounts (QT or DV).For example, research during blood pressure quantity to measure may be diastolic blood pressure, can use the height of foot or centimetre expression during research height, measuring when research is fat may be body weight or BMI, measuring when research is depressed may be depressed positive DSM-IV standard number, or the like.Two divergent character also can be used.For example, if study the experimenter of 200 contrasts and 200 multiple sclerosiss, contrast scoring may be 0 and the scoring of multiple sclerosis is 1.The population size that the scoring of character, phenotype or QTV is referred to character.For example, if I am body weight 200Ibs, the scoring of my body weight will be 200.If I have level is 250 cholesterol, my cholesterol scoring will be 250, or the like.It is relevant with minimum, medium or the highest phenotypic effect according to which kind of genotype that gene is marked, and will mark 0,1 or 2 to be assigned to genotype.

The 3rd step of step 3. is to identify the candidate gene that will detect.As an example, the application is ADHD, disobey obstacle, conduct disorder, learning disorder, ethanol problem have been selected 29 candidate genes, and they are to comprising dopamine, 5-hydroxy tryptamine, norepinephrine, GABA, and the gene that works of the adjusting of other neurotransmitter.Those of ordinary skill in the art can recognize the gene that may work to polygenic character potentially.At first select those that gene of metabolism or physiology relation may be arranged with the character of assessment, help to be chosen in a candidate gene or one group of gene of using in the MAA technology.Gene or the allele describing or find in the available scientific literature in public library or from the such computerized data bank of the gene bank (Genebank) of NCBI expect that all being used for the MAA technology as candidate gene diagnoses or prognostic analysis specific trait.Those skilled in the art can recognize other genetics information resource, comprise individual knowledge or do not deliver the knowledge that maybe can not openly obtain, they can be used as the candidate gene of identifying most of polygenic character and obstacle, and such resource can be used for the practice of MAA method.

An example that relates to the candidate gene of the polygenic character as height or osteoporosis susceptibility relates to the gene of bone and/or formation, growth and/or adjusting.The character that another one can be estimated is fat, and candidate gene will comprise that these genes add OB gene, OB acceptor gene, npy gene and neuropeptide Y receptor gene.For the character as blood pressure, reasonably candidate gene will be that those relate to the metabolic gene of norepinephrine, epinephrine, steroid and renin.

In an embodiment of the present invention, selected many suppositions to relate to the gene of polygenic character.Particularly importantly infer the gene that relates to the RDS behavior.The standard of selection gene comprises bibliographical information, and they participate in one or more RDS behaviors or other purpose polygenic character, and/or inventor's experimental data (this can describe hereinafter).The inventor expects that the dependency of gene or polymorphism and certain character can be used for the diagnostic analysis of this character, and whether is applicable to that for certain gene and/or its special polymorphism the MAA diagnostic analysis of purpose polygenic character provides guidance.For this gene or polymorphism, character is not necessarily related with RDS.

The DRD1 gene.This specific gene is compared with contrast in inventor's preliminary study not to be had relatedly with serious excessive drinking proband, like this, is that other people is taken aback to the additional discovery of large sample more.Studies show that their more vast scales (Comings etc., 1997) than any one independent gene explanation serial behavior variation are found in the inspection of DRD1 and two dopamine receptor genes of DRD2.The D1 receptor gene polymorphism may be more relevant with serious excessive drinking with many substance abuses ratios.

Dopaminergic gene, violence and division sample/elusive behavior.Discovery in addition is supported in complicated personality disorder as the notion of polygenic inheritance is arranged in " pathologic violence " and the division sample/elusive behavior (SAB).The inventor finds between DRD2A1 allele and " pathologic violence " in the teenager proband strong correlation is arranged, and also finds the similar dependency of 10 allele of dopamine transporter gene (DAT1).Found the strong correlation of DRD2A1 allele and SAB, D β H gene is not found relevant, though find DAT1 gene and SAB more weak dependency (Blum etc., 1997) is arranged.

The allelic inspection of D β H may be the exact method of assessment D β H gene latent effect in the behavior of men obstacle.This is the research first of supporting DRD2TaqIA1 allele and SAB and PV strong correlation.When these complex characters did not show simple Meng Deer mode of inheritance, expectability did not cause simple hereditism's answer by individual gene.The relation of dopamine gene and pathologic violence will be further described in specific embodiment 17.

The d2 dopamine receptor gene.With polymorphism mark chain unbalance in the sudden change of inferring that is positioned at gene 3 ' and 5 ' noncoding region that keeps, may influence transcribing and/or the stability of mRNA and so influence the quantity of DRD2 receptor of DRD2.This is based on following practical work: find and the stronger dependency that is marked with that is positioned at flanking region rather than exon point mutation; Be reported in and do not have sudden change in " the A1 labelling " the DRD2 allele exon (except the exon 8); And be reported in do not have Kd to change and when the A1 labelling is arranged DRD2 Bmax reduce people such as (, 1991) Noble.This may help to explain with polymorphism TaqID2 and lack dependency.Lacking dependency with TaqID2 conforms to the population genetics analysis, although latter explanation TaqI A and B have intensive linkage disequilibrium each other, 3 ' the flank labelling of A1 presents the imbalance less with TaqI D, therefore may expect the weak dependency that it and analeptic likes people such as (, 1994) Suarez.

For complicated problems more, may be because the molecular hybrid advantage that Comings (co-inventor) proposes for the explanation of failure in some researchs.This occur in when genetic polymorphism be that the experimenter of heterozygosis is that the experimenter of isozygotying is when having obviously higher (positive hybrid vigor) or lower (negative hybrid vigor) average than one of two allele on quantitative trait parameter (QTV).

Positron emission tomograph unit (PET) research has illustrated that the brain district glucose metabolism antidotal alcoholic and abuse cocaine person descends.In this research, use ¹⁸The F-deoxyglucose has A1 ⁺And A1 ^-Measured regional glucose metabolism among the experimenter of the non-ethanol/non-drug abuse of allelic health.Comprise lentiform nucleus, volt nuclear, frontal convolution and temporal convolution and prefrontal inboard, pillow-temporo and orbital cortex in many brains zone, average relative glucose metabolic rate (GMR) is at A1 ⁺Compare A1 in the group ^-Obviously low in the group.At A1 ⁺The relative GMR of group descends and also is found in motor speech area, preceding Reil's island, Hippocampus and material black substance.Yet at A1 ⁺Part brain zone presents the relative GMR of increase in the group.

The evidence that has several explanation dopamine metabolic deficiencies in stress obstacle etiology, to act on.In the brain of A1 allele carrier, the d2 dopamine receptor of lesser amt may be converted into the dopaminergic activity of reduced levels in the part that brain relates to recompense.The A1 carrier may not can find that from the A2 carrier stimulation of gratification obtains competent recompense.This may be converted into the lasting addiction of A1 carrier or the behavior of sensation seeking.Since known dopaminergic reduce stress or addiction, the A1 carrier may turn to other material or movablely discharge the dopamine of greater number to obtain temporary transient alleviation.Ethanol, cocaine, Fructus Cannabis, nicotine and carbohydrate (as chocolate) can both cause the release of dopamine in the brain and bring respite to addiction.These materials can use separately, unite and use or use interchangeably.Accordingly, the dependency of DRD2 gene and smoking we (people such as Comings, 1996b) and others (people such as Noble, 1994; People such as Lerman, 1997) obtain proof in the research.

Dopamine-.Because D β H is arranged in sympathetic nerve terminal and is released into circulation at the dispose procedure of norepinephrine, the gene of therefore controlling it may be positioned on the chromosomal foci rather than D β H itself.Therefore, hereditism's labelling and ADHD in D β H site, CD, the correlation research of alcoholism and other RDS corelation behaviour may be negative.On the other hand, if the serum levels of D β H establish owing to a series of environmental factorss are common, with D β H hereditism labelling correlation research may the effect in these obstacles provides more accurate evaluation to D β H gene than blood levels.

These are found and other discovery illustrates that together the polymorphism of D β H gene may only work in specific RDS behavior, by predicting that about the blood levels of D β H or its activated product (dopamine or norepinephrine) hereditism result is difficult in the past.

Dopamine-in drug dependence and other character.The active inhibition of D β H causes the excessive generation (Randrup and Scheel-Kruger, 1996) with many moving, attack, dopamine that self stimulation is relevant with stereotyped movement.This shows D β H may act in human aggressive behavior, ADHD and conduct disorder (CD).In the affective disorder boy of low plasma D β H level CD diagnosis frequency increase existing report (O ' people such as Connel, 1992; People such as Rogeness, 1984; People such as Rogeness, 1986; People such as Rogeness, 1988; People such as Rogeness, 1987).Some researchs have shown in low D β H level and some personality character as the scoring of turning up (Roy and Brockington, 1987) in the Eysenck personality inventory with feel searching (people such as Ballenger, 1983; People such as Umberkoman-Wita, 1981) relation between, and in a research, plasma D β H level and sensation are sought between scoring positive correlation (Folatein and Rutter, 1977).

Another aspect of the present invention relate to the correlation research of dopamine-and show D β H dinucleotide repeat between polymorphism and the drug dependence pattern dependency first (people such as Comings, 1996a).

Find D β H dinucleotide repeat polymorphism more than the 175bp or below double mode allele distributions is arranged.With experimenter's gene type be low (≤174bp) or high (≤176bp) isozygoty perhaps heterozygosis.Usually, finding to have the patient that high base isozygotys uses in the intravenous pharmacy that ASI had greater amount drug use, longer cocaine to use the amphetamine intravenous injection of history, higher frequency, higher frequency in the past.These subjects reported more frequent father and mother's alcoholism and the sexual abuse history of Childhood.The genotypic patient of high bp divides relevant with the lower assessment of " oneself accepts, and imparts rudimentary knowledge to beginners second characteristic and oneself's guidance " (Self-Acceptance, Enlighted SecondNature, and Self-Derectiveness).

As DRD2 A1 allele, D β H B1 allele and variables A DHD, obsession, manic, it is maximally related disobeying and sleeping.Difference comprises that the dependency of DRD2 A1 allele and schizophrenia, property, conduct and stutter variable is bigger, and D β H B1 allele and study, reading and school's problem have stronger dependency.With opposite in the DRD2 research of tending to be listed in the bottom, it is noticeable especially that the variable relevant with school's behavior such as reading, study and primary school studied the high trend of middle grade at D β H.This may be relevant with the effect of D β H in memory.

Tourette syndrome (TS) may be RDS the most complicated can the identification form in a kind of.Because all behaviors relevant with the active inhibition of D β H are common (Comings and Comings, 1984 in Tourette syndrome (TS) patient; Comings and Comings, 1987b; Knell and Comings, 1993; Comings, 1990), may between D β H Taq B polymorphism (d ' people such as Amato, 1989) and TS, manner obstacle, attention deficit hyperactivity disorder (ADHD), there be dependency.

The eliminating of DAT1 gene in morbid obesity.The allelic existence indication of DRD2 A1 is not only for also risk increase for other addictive behavior of obesity, and BMI surpass 25 itself (not having macroscopic view to select the features of [carbohydrate gluttony] or symbiosis SUD) for the allelic dependency of DRD2A1 be not competent standard people such as (, 1996) Blum.

Both estimated work that DRD2 A1 also estimates the morbid obesity field of DAT1 gene show in the woman 34% or more body fat and in the man 28% or more body fat only relevant and have nothing to do with the DAT1 gene with DRD2A1 allele (p＜O.0001).The highest body weight appears in the individuality with DRD2A1/A1-DAT1-10/10 haplotype, illustrates that the D2 acceptor gene has much bigger distribution in the distribution of two genes in morbid obesity.

Fructus Cannabis ester acceptor gene.The tendentiousness of intravenous drug (IV) drug use may be subjected to hereditism and Effect of Environmental.In order to explore these genes-cultural etiology factor, assessment has been done in the contrast that 77 the non-spanish white people of male material misusers and 70 races match.Patient is by the medication of addiction severity index; All proband are by " home environment scale " and " childhood period experience application form " medication, and hold gene type to dopaminergic, Fructus Cannabis and GABA energy gene.It is higher that the present inventor finds that genotype only contains the prevalence rate of the allelic experimenter's medium-sized vein of low-molecular-weight drug utilization of the high molecular allele of CB1 (Fructus Cannabis) acceptor gene and GABRB3 gene.Trinucleotide repeats sequence of CB1 gene, at least 9 kinds of allele are arranged, the present inventor finds that evidence shows, its effect to phenotype can not be carried out the straight line prediction from allelic relative weight, and the interaction between they and intravenous pharmacy utilize is more complex.Several environmental variables (comprise in the home environment scale a kind of) are correlated with the susceptibility of intravenous pharmacy utilization after removing the variation relevant with genetic variance, although these environmental variables itself may be under the influence of identified gene not.

Electrophysiology unusually with substance use disorders (SUD): related with d2 dopamine receptor and Fructus Cannabis ester acceptor gene.Corresponding to the contrast of " super contrast " and many documents, the obvious dependency between serious substance use disorders (SUD) and the DRD2A1 allele is observed (people such as Blum, 1997; People such as Hill, 1997).The amplitude of waking the P300 ripple of related potential (ERP) up descends and be associated with ethanol and drug dependence incubation period (Braverman, people such as R., 1990).The preclinical obvious prolongation of P300 and three risk factor relevant (1) parents' SUD; (2) chemistry relies on (relying on as cocaine) and (3) carbohydrate gluttony.The family history of decline of P300 amplitude and alcoholism and SUD is relevant, but does not have related with DRD2A1 allele.

In this application, the inventor provides in decline of frontal lobe P300 amplitude and Fructus Cannabis ester acceptor gene (CB1) 〉=5 and has repeated obvious relevant evidence between allele homozygosity.Relevant in order to assess whether same genotype with ethanol or drug dependence, CB1 is carried out in 98 experimenters coming from ATU and 69 contrasts repeat allelic gene type.All experimenters are right and wrong Spain white man.Use " addiction severity index ", the diagnosis plan of meeting " and MAST-R ATU experimenter is assessed.Only use MAST-R from contrast, to get rid of medicine and alcohol abuse/dependence.The result shows that 5 repeat allelic isozygotying and many different types of drugs dependence (cocaine, amphetamine, Fructus Cannabis), the time limit with hallucinogenic agent, inhalation (inhalatio), heroin, amphetamine, cocaine and barbiturates use, use and overdose with intravenous pharmacy, and the legal issue relevant with drug dependence (drugs are accused, drugs are determined a crime, driven violating the regulations and weapon, attack and the charge of intentional vandalism) has tangible dependency.By contrast, there is not tangible dependency with the variable relevant with alcohol dependence.This may be because contrast and/or serious crapulent evaluations deficiency, particularly for RDS phenotype completely.Yet these results are consistent with the research that explanation Fructus Cannabis ester receptor works in recompense path and the metabolism of adjustment dopamine.

The present invention identified BEAM (BEAM) unusual and and DRD2 genotype dependency between obvious relation.The inventor believes that this important demonstration test as the RDS behavior disposition of diagnosis genetic induction has commercial value.Suggestion relate to method that above-mentioned people's dopamine receptor gene A1 allele and Fructus Cannabis ester acceptor gene (CNR1) detect follow the mind map of standard (be Nicolett ( ^TN)).

In addition, the linear trend of weighting has disclosed with the DRD2A2 genotype and has compared with Fructus Cannabis SUD, and event related potential significantly worsened effect (p＜0.0001) under DRD2A1 allele existed.Duncan ' s Range check explanation is compared with the DRD2A2 contrast, has worsened EP ' S significantly with or without the allelic SUD of DRD2A1.These results show the effect of DRD2A1 allele in the non-behavior Pathophysiology phenotype that relates to brain function and potential addiction tendency.

The 5-hydroxy tryptamine gene.The metabolic defective of 5-hydroxy tryptamine, and blood 5-hydroxy tryptamine and tryptophan levels are unusual, in many psychiatric disturbances report are arranged.5-hydroxy tryptamine 2,3-dioxygenase (TDO2) are that the decomposition 5-hydroxy tryptamine is the rate-limiting enzyme of N-formoxyl kynurenin.The inventor attempts to determine whether the hereditary variant of 5-hydroxy tryptamine HTR1A gene is relevant with the phenotypic expression of TS or its symbiosis behavior.Between rare allele (shorter and longer allele) and ADHD, CD, disobedience obstacle (ODD), twitch, property and other behavior scorings, tangible dependency is arranged.Contribution to these scorings is a spot of, only causes the variation of 2-4%.The effect of HTR1A and DRD2 gene is an additivity, causes the variation of ADHD, CD and ODD scoring 5.1% to 5.4% jointly.These results are that the lifting manipulation of polygenes obstacle is consistent with these, and part is owing to influence the probability convergence of 5-hydroxy tryptamine and the metabolic genetic mutation of dopamine, and environmental factors.Repetitive sequence itself may be in polygenic inheritance works in the generation of important function equipotential polymorphism variant.

Utilization " meet of Axis II personality disorder structure " and " addiction severity index " (ASI) and " Buss-Durkey hostility scale " (BHDS) the possible dependency of the T/C polymorphism of 5HT-2 acceptor gene has been done assessment.In the patient's sample, 22 genotype are relevant with depressed (p＜0.05) with border personality disorder (p＜0.05).This be marked among the ASI with spend in medicine on money quantity (p＜0.05) and rape history (p＜0.05) and shoplifting/relevant to the destruction (p＜0.05) of art.In BHDS, 22 genotype are relevant with the scoring that attack (p＜0.01) and non-direct hostility (p＜0.05) subscale raise in man's contrast.In woman's contrast, 5HT-2R gene and non-direct hostility (p＜0.05), negativism (p＜0.05), oral hostility (p＜0.005) and guilty sense (p＜0.05) and total hostility scoring (p＜0.01) are relevant, but relevant polarity has been reversed (11 genotype are relevant with higher value in all scorings).The sex of genotype dependency reverses and shows that it is and the interactional complicated gene of gonadal hormone.

Estrogen receptor, aromatic series enzyme site and arginine vasopressin gene and conduct disorder.Because the female hormone receptor gene knock-out mice shows offensive behavior people such as (, 1996) Ogawa, may be relevant with conduct disorder to the multiple research of this gene dinucleotide.Two other related genes are people such as (, 1991) Polymeropoulos and spermine acid vassopressin (AVP) gene (Summar, 1992) in aromatic series enzyme (CYP19) site.

Nicotine (nicotine) acceptor gene.The nicotine receptor of prefrontal cortex participates in the reaction task that postpones, and muscarinic receptor more participates in the overall work memory people such as (, 1995) Granon.Many researchs have shown the tight interaction between nicotine and dopamine.With other dependence producing drug one time-out, nicotine is the same with other addictive drugs to cause increase (Dichiara and Imperato, 1988 that middle edge and volt nucleus neuron dopamine discharge; People such as Corrigall, 1994; People such as Pontiefi, 1996) and intensive oneself take medicine (Corrigall and Coen, 1989; Corrigall and Coen, 1991).Yet tolerance increases sharply along with repeating to take medicine (people such as Lapin, 1989).Nicotine suppresses the absorption of dopamine not as most dopamine uptake inhibitors, and it has only 50% inhibition people such as (, 1991) Irenwasser.The influence that studies show that of dopamine uptake inhibitors and nicotine receptor agonist and inhibitor taken in dopamine is by nAChR mediation people such as (, 1991) Irenwasser.

Neuronal nitric oxide synzyme (NOS) gene.The nitrous oxide synthetase gene recent findings is relevant with the aggression of mice.With respect to the brood mice of non-ob/ob, the research of ob/ob mice has been illustrated the increase of nitric oxide synthetase level.Research to the NOS knock out mice makes the effect of nitric oxide in attack and sexual behaviour more outstanding.The ob/ob mice has tangible noradrenaline levels increase and at bow type funnel tangible dopamine level decline (Oltman, 1983) is arranged at paraventricular nucleus and lateral hypothalamus.

Checked that neuronal nitric oxide synthase gene (nNOS1a) dinucleotide repeats the dependency of polymorphism.The degree of being hostile to, diagnostic characteristic and the personality character of the contrast that 67 the non-spanish white people of male material misusers are conformed to 68 age races are assessed.Patient's sample is assessed with the meet of AXIS-II personality disorder structure, addiction severity index (ASI) and Cloniger temper and personality investigation (TCI).The allele distributions of patient and contrast is different (p=0.056) reluctantly, the patient who has high molecular allele to isozygoty.In AXIS-II meets, the more frequent diagnostic criteria that meets schizophrenia (p＜0.05) and border (p＜0.05) personality disorder of the patient that high molecular allele isozygotys.In ASI, this genotype is relevant with the increase of following scoring: showed violent behavior (p＜0.005) in the past 30 days, forge history (p＜0.05), theft history (p＜0.005), use ethanol (p＜0.005) in the past 30 days, inhalation (inhalatio) service life (p＜0.0075), medicine detoxifcation (detoxes) number of times (p＜0.05) experiences the natural law of ethanol (p＜0.005) and medicine (p＜0.005) problem in the month before.This genotype also relevant with the friend with smaller amounts (p＜0.04) has still less friendship relevant (p＜0.0005) with their friend, with the more heterogeneous pass of marriage number of times (p＜0.05).In TCI, this genotype is with following relevant: impulsion increases (p＜0.01), and the attachment scoring reduces (p＜0.05), relies on (p＜0.02), and recompense relies on (p＜0.05).Purposiveness (p＜0.01), the oneself instructs (p＜0.05), and emotion moves into (p＜0.05), helps (p＜0.02) sincere conscience (p＜0.02), and collaborative (p＜0.05).

Monoamine oxidase, MAO gene (MAO).MAO is a main enzyme being responsible for the degraded of brain synapse neurotransmitter.Can suppress the obvious improvement that the active medicine of MAO obtains emotion and other behaviors by using.Many researchs have shown that the MAO level is with following related: alcoholism (people such as Wiberg, 1977; People such as Gottfries, 1975; People such as Devor, 1994; 09); Schizophrenia (people such as Wyatt, 1979); Depressed (people such as Sherif, 1991; People such as Pandey, 1992); Manic depressive disorder (people such as Pandey, 1980); Commit suiside (people such as Gottfries, 1975; People such as Sherif, 1991; People such as Buchsbaum, 1976; People such as Buchsbaum, 1977; Meltzer and Arora, 1986); ADHD has another name called ADDH (people such as Skekim, 1982); And take a risk, seek to feel or the personality character of externalization (people such as Vonknorring, 1991; People such as Buchsbaum, 1976; People such as Schooler, 1978; People such as Skekim, 1989; People such as Vonknorring, 1984).Yet other research but can not find with these character in one or more relevant (Mann and Stanley, 1984; People such as Propping, 1981; People such as Tabakoff, 1988).Utilization enzyme level (people such as Wiberg, 1977; People such as Gottfries, 1975; People such as Devor, 1994; People such as Vonknorring, 1991) and the former effect of MAOA gene in substance abuse that studies show that of hereditism's variation people such as (, 1993) Vanyukov.

Except dopaminergic system and Fructus Cannabis ester receptor, proved that monoamine oxidase, MAO (MAO) also plays significant effect in RDS.The present invention also provides the first association of monoamine oxidase, MAO gene in the Tourette syndrome.Outstanding in the male of the soluble ADHA of the allelic genetic defect of the MAOA of X-linkage or MAOB, TS and relevant obstacle.Three allele that repeat polymorphism, two and of MAOB gene of MAOA have done assessment in 351 patients Ts, relatives and contrasts.Each experimenter has finished an Investigation on structural table, and 23 the varying number character relevant with school's problem with behavior, study are estimated.The longer base pair allele of MAOVNTR and MAOB polymorphism and the shorter base pair allele of MAO CA-1 polymorphism have the higher assessment of significant trend and ADHD, stutter, manic, depressed, conduct and problem concerning study to divide relevant.The most significant result is that the CA-1 of MAOA gene repeats for ADHD (p=0.005), major depression (p=0.005) and stutter (p=0.007).And the regression coefficient of seven kinds of behaviors is significant＜0.01, the R of sex-linked MAOA gene ²Or the percentage ratio of variation has contribution to a series of behaviors, and the degree that relates to is not enough to explain the degree that the male of TS, ADHD or CD gives prominence to.The hypothesis that the polygenic inheritance mechanism of result and TS obstacles and moonlet itself may work in Gene regulation is corresponding to.

OB in the obesity, people's the 2nd chromosome, uncoupling protein 2 and APO-D gene.Colony studies show that the women has the fat genetic load of higher degree than the male, and have such possibility: genetics factor more likely participates in obesity in youngster than the old people greater than 50 years old, incidence rate fat in greater than 50 years old old people increases, account for crowd's significant proportion, and acquired factor may more important (people such as Stunkard, 1986) as longer sedentary life style.

Relating to a fat important protein product is the serum albumin leptin.It synthetic is by the OB Gene Handling and be considered to working people such as (, 1995) Maffei aspect the adjusting of body fat.Human body leptin level and total obesity-related height of individual related people such as (, 1996) Considine have been found.Microsatellite polymorphism, D2S1788 is found to be positioned at by lod scoring and shows with the obviously chain chromosome 22 p21 of Serum Leptin Levels and go up people such as (, 1997) Comuzzie.This locus has caused 47% serum leptin level variation, and contains several fat candidate genes, comprises that glucokinase regulates albumen (GCKR) and proopiocortin (POMC) people such as (, 1997) Comuzzie.A potential mechanism about pomc gene is that it is the precursor of thyroliberin (ACTH), and thyroliberin acts on the generation that adrenal cortex causes glucocorticoid.Yet might reason out pomc gene also works as the opium propeptide.

The mitochondrial protein of uncoupling protein (UCP1) by name is in play an important role aspect heat production and the transformation heat (Nicholls and Locke, 1984).Relevant with glucose metabolism (Himms-Hagen, 1990) are formed with body temperature, body in this path.Yet the brown adipose tissue that contains UCP1 does not participate in inhabiting the human body weight adjusting of pining for property environment.UCP-2 has 59% aminoacid identical with UCP-1, have with the corresponding to characteristic of effect in diabetes and obesity (people such as Fleury, REFERENCE).When yeast obtains to express, UCP-2 has stronger effect to mitochondrial membrane potential than UCP-1.The UCP-2 wide expression is included in the abundant tissue of macrophage in adult soma, and the edible reaction of fat is raised in white adipose.

The present invention combines the polygenes analysis that all these is had the ob gene of many different physiological mechanisms.These differences may be considered adding up property effect rather than synergism, cause more accurate pre-diagnostic test based on DNA.In a sample, be embodiment preferred, rather than use any one gene in conjunction with DRD2, OB, the 2nd chromosome, UCP-2 and APO-D gene.In addition, invention determines that MO proband is not according to BMI, but determines according to the percentage ratio of body fat: the women 34%, and the male 28%.

The polygenes method of SUD.The relative effect that to assess experimenter's many heterogeneic allelic distinct advantages on the same group be each gene can compare by different quantitative variations.In addition, the bigger exquisiteness and the accuracy of analysis of environmental activity given in genotypic evaluation, because the effect of gene can be separated with the reason of environment.

The scoring of ten scales of Moos ' home environment scale (FES) is used as the index of examined person's childhood and adult phase home environment.Utilization SPSS (SPSS, " dependency and part correlation software " analytical data Inc).

The stepwise regression analysis that starts from using the marker gene type as seven locus of forecasting index to carry out to the analysis of every kind of medicine variable.Require the p value to be lower than 0.05 during variable substitution product equation.After each regression analysis, get rid of variable with minimum predictable gene-correlation.The result has shown the corresponding correlation coefficient of different genes and environmental factors in table 1.

Table 1

To different pharmaceutical and alcohol abuse variable, specific gene type prediction effect

Multiple regression analysis progressively

	The intravenous pharmacy user	Hallucinogenic agent uses a year number (Yrs)	Fructus Cannabis is used a year number (Yrs)	Amphetamine uses a year number (Yrs)	Cocaine uses a year number (Yrs)	The serious ranking of alcohol abuse	The serious ranking of drug dependence
								N	??100 ??.25** ??.26** ??.28**	??100 ??.19* ??.20** ??.27**	??118 ??.19* ??.27**	?94 ?.23* ?.23*	128 .26*	130 .24**	128 .17* .37**
Genotype
	DRD1 1
GABRB3 2
	CNR1 3
DRD4 4
	DβH 5

	The intravenous pharmacy user	Hallucinogenic agent uses a year number (Yrs)	Fructus Cannabis is used a year number (Yrs)	Amphetamine uses a year number (Yrs)	Cocaine uses a year number (Yrs)	The serious ranking of alcohol abuse	The serious ranking of drug dependence
								Amount to	???.46 ???.21 ???.0002	????.46 ????.21 ????.0001	??.32 ??.10 ??.002	??.38 ??.14 ??.003	??.26 ??.07 ??.013	??.24 ??.06 ??.007	??.41 ??.17 ??＜0.0001
??R
	??r 2
??p
	The FES environmental variable, significant hereditary forecasting index separates (Partialed Out)
??ICO 6									???-.3.1* ???-.21* ???-.23*	????-.2.2** ????-.12 ????-.16	??-.2.5** ??-.26** ??-.19*	??-.2.3** ??-.11 ??-.22*	??-.15 ??-.19* ??-.27**	??-.31* ??-.21* ??-.35**	??-.16 ??-.17 ??-.21*
	??ARO 7
??C 8
	??*＜0.5?? 4DRD4 ± 7 an allele * *＜0.01 D β H Taq B1 heterozygosis vs. isozygotys 1 DRD1 11 genotype 6The intelligence of ICO=FES and culture are directed 2GABRB3＜188 bases 6The positive entertainment oriented of ARO=FES is to allele 3CNR 〉=5/ 〉=5 genotype 8Family's cohesion of C=FES

The 4th step of step 4. is one or more polymorphisms of evaluation and each gene-correlation.These may be single base pair restriction fragment length polymorphisms (RFLPs), or dinucleotide, trinucleotide or other repetition polymorphisms, repeat as variable series connection, or any other labelling of locus.Except polymorphism disclosed herein, hereinbefore about the description of the method for identifying candidate gene, this polymorphism and detection method can obtain in the former at an easy rate work of delivering or not delivering as previous.In addition, if suspecting a gene works in the purpose polygenic character, but after consulting document and genetic database, do not have the polymorphism that in the MAA technology, to use at present, just can adopt the polymorphism of genetic analysis with the gene of determining to be applied to the MAA technology.Except technology described herein, this analysis is usually used in the literature and is described.According to actual conditions, can use the screening method of the sudden change in accurate detection genomic DNA, cDNA or the RNA sample.

The present invention relates to detection, diagnosis, prognosis and the treatment of RDS disease, and utilization MAA technology is to detection, diagnosis, prognosis and the treatment of polygenic character.The allele labelling that polygenic character is worked adding up property or the effect of subtracting property is also disclosed, it is the nucleotide sequence form of separating and identifies and detect the new labelling that is used for the MAA analysis from individuality method.These labellings are indexs of the polygenic character of analyzing, also are that body presents the diagnosis index of the potential probability of specific trait one by one.

Those skilled in the art will recognize that nucleotide sequence disclosed herein, reach available in public database (for example finding in NCBI), can use the MAA technology from the available nucleotide sequence of the scientific literature of delivering, they will be used for many application of detection, diagnosis, prognosis and the treatment of RDS or other polygenic character like this.The example of these application comprises within the scope of the present invention, use one or more labellings of special primer amplification polygenic character, detect the labelling of polygenic character, as hybridizing with oligonucleotide or nucleic probe, isolating nucleic acid is inserted in the carrier, and the RNA that comes from carrier expresses.

A plurality of genes of check behavior disorder and other polygenic characters are feasible and do not need new technology.Polymorphism that relates to and variant have two types, 1) single base changes and causes restriction fragment length polymorphism (RFLP) and 2) short series connection repeats polymorphism (STR) [particularly two and trinucleotide repeat].These methods of checking are in a large number had nothing in common with each other for every type.

RELP.Applied Biosystem (applying biological system), a branch of Perkin-Elmer company has developed a kind of new technology and instrument, is used to carry out the PCR of single base RFLP type genetic polymorphism ^TMFast detecting.This instrument, promptly the Prism of applying biological system 7200 sequence detection systems (TaqMan) can carry out the polygenes detection.This method utilization standard primer electrophoresis contains the DNA part of restricted enzyme pleomorphism site.The unique aspect of this technology be the design two weak points oligonucleotide primers, one with allele in one match, another and another allele matches.Fluorescent dye is connected to an end of each primer, and the cancellation dyestuff is connected to the other end.If primer cooperates correct, when archaeal dna polymerase arrived the antisense oligonucleotide primer object location, primer can be degraded to nucleotide when polymerase passed through.The strongest fluorescence is sent in the release of quencher and dyestuff.Yet if oligonucleotide primers mismatches, it can come off from the site and cancellation continues rather than nuclease degradation.Dual wavelength reading to plate can be distinguished 11,12,22 genotype.The overall process of reading 96 sample result needs the time to be lower than 15 minutes, and the result is transfused to computer and analyzes and preserve.This technology, the PCR that under computerized work station helps, sets up ^TMReaction can detect hundreds of different RFLP in one day.

STR. use the PCR that sets up STR with the same work station of above using ^TMReaction.Difference is that the STR primer itself is by different fluorochrome labels.Can accurately identify the allele that has only two base differences to the 373DNA of the applying biological system sequenator of second kind of dyestuff of sample labelling.Each all detects by carrying out laser scanning at different wavelength.For example, PCR ^TMPrimer by fluorescence HEX Amidite (the applying biological system, the Foster city, CA) or other fluorochrome label.The PCR of 10 times of dilutions of 2 μ l ^TMROX 500 standards that add 2.5 μ l deionized formamides and 0.5 μ l in the product, 92 ℃ of degeneration 2 minutes join in 6% the denaturing polyacrylamide gel of AB 373 dna sequencing instrument then.Gel electrophoresis 5 hours under constant 25W.Use inside ROX 500 standards of each swimming lane gel is carried out laser scanning and analysis.Peak value is discerned based on the color fragment of base pair length decision by Genotyper (1.1 version).

In history, many diverse ways are used for the test point sudden change, comprise denaturing gradient gel electrophoresis (" DGGE "), restriction endonuclease polymorphism analysis, chemistry and enzyme action method, and other method.The more common method of using comprises directly to PCR at present ^TMThe purpose sequencing fragment (seeing below) and the single-strand conformation polymorphism analysis (" SSCP ") of amplification.

The method of another screening point mutation is based on the RNA enzyme to RNA/DNA and the right enzyme action of RNA/RNA heterodimer base mismatch.When using in this article, vocabulary " mispairing " is defined as one or more unpaired or wrong paired nucleotide in double-stranded RNA/RNA, RNA/DNA or the DNA/DNA molecule.This definition comprises by insertion/deletion mutation like this, and the mispairing that causes of single and a plurality of base point mutation.

U.S. Patent No. 4,946,773 have described the restriction analysis of a kind of RNA enzyme A mispairing, comprise the annealing of single stranded DNA or RNA test specimen and rna probe, and with RNA enzyme A the nucleic acid dimer are handled subsequently.Behind the endonuclease reaction of RNA enzyme, inactivation, enzyme action product heat denatured and the electrophoretic analysis on denaturing polyacrylamide gel of RNA enzyme by proteolysis degraded and organic extraction.According to the single stranded product that the RNA enzyme A of molecular size electrophoretic separation handled, compare to detect mispairing with the contrast dimer of same treatment.The sample that contains the more small fragment (enzyme action product) that does not have in the contrast is positive.

Present available RNA enzyme mispairing restriction analysis comprises those according to U.S. Patent No. 4,946, and 773 analyses of implementing need the radiolabeled rna probe of utilization.Myers and Maniatis are in U.S. Patent No. 4,946, and 773 have described the detection to base-pair mismatch with RNA enzyme A.Other researcher has been described the application of a kind of escherichia coli enzyme (RNA enzyme I) in mispairing is analyzed.Because RNA enzyme I has wideer enzyme action specificity than RNA enzyme A, if can find to lower the composition of non-specific enzyme action increase to the enzyme action of mispairing, RNA enzyme I is applied to base-pair mismatch to detect ideal enzyme.The RNA enzyme I that is used for the mispairing detection has description at the document of Promega Biotech.Promega sells a kind of test kit that contains RNA enzyme I on market, their document shows, if the level of this enzyme is enough high, and its in can four known mispairing of enzyme action three.

The RNA enzyme protection is analyzed first Application in the end that detects and show special mRNA in the solution.This analysis depends on can be easily by the radioactive mark RNA probe of in vitro transcription generation with the complementary high specific acitivity of purpose mRNA.At first, the plate of touching of in vitro transcription is the recombiant plasmid that contains phage promoter.Probe mixes to hybridize their complementary fragments with total cell RNA, handles with the RNA enzyme then and decomposes the superfluous not rna probe of hybridization.Same, the same with initial expection, the RNA enzyme of use is special to single stranded RNA, so the double-chain probe of having hybridized can be protected and do not degrade.After the RNA enzyme-deactivating was removed, the probe (quantitatively proportional with the quantity of purpose mRNA) of recovery protection was also analyzed on polyacrylamide gel.

The analysis of RNA enzyme protection also is fit to the detection of single base mutation.In the RNA of the type enzyme A mispairing restriction analysis, radiolabeled rna probe that the wild-type sequence in vitro transcription obtains and the complementary target fragment hybridization that comes from sample survey.Though can use target RNA (endogenous mRNA) once in a while, the target that detects comprises DNA usually, and (existing genomic DNA also has by the DNA of clonal expansion on plasmid or PCR ^TMThe DNA of amplification).If difference on single nucleotide (or more) sequence is arranged at the probe of hybridization with between the target fragment, cause the disconnection of (" mispairing ") Watson-Crick hydrogen bond in this position, this can be identified and be opened by strand specific ribonucleic acid enzyme action in some cases.Up to now, RNA enzyme A almost is unique enzyme that is used for the single base mismatch enzyme action, though that RNA enzyme I also finds recently to the enzyme action of mispairing is effective.Recently also relevant for the report that detects single base mismatch with MutS albumen and other DNA repairase.Other methods of nucleic acid and sudden change that detect are described in this article.

Nucleic acid. as what describe herein, one aspect of the present invention is 29 known genes, their polymorphic allele is the sign of polygenic character, comprises the sign that resembles ADHD, disobeys the such polygenic character of obstacle, conduct disorder, learning disorder, ethanol, cholesterol and LDL.

In one specific embodiment, the nucleotide sequence that discloses herein will can be used as hybridization probe or amplimer.For example, these nucleic acid may be used for the diagnostic assessment of tissue sample, or clone's full-length cDNA or genomic clone correspondingly.In specific embodiment, these probes and primer are oligonucleotide fragment.These fragments should long enough to be used for that RNA or DNA tissue sample are carried out special hybridization.Typical sequence will be a 10-20 nucleotide, but also may be longer.Longer sequence as 40,50,100,500, is better choice until total length to specific embodiment.

Be selected from any gene that can be used for diagnosis of the present invention and Therapeutic Method and have approximately 10,15,17,20,30,40,50,60, the nucleic acid molecules of 75 or 100 or 500 continuous nucleotides can use.Also be fine with complementary molecule of above-mentioned sequence and the molecule that under the height stringent condition, combines these sequences.These probes are useful in many crossing schemes, for example dna molecule hybridize and RNA molecular hybridization.In some instances, probe can be used for not influencing with a plurality of target sequence hybridization the ability of their efficient diagnosis polygenic characters.

Can be around disclosed nucleotide sequence or with the polymorphism of marking various probes of sequential design and primer on every side, so long as gene disclosed herein or joined gene in 29 these groups of gene described herein afterwards.Expection can be founded new cohort with other gene be used to assess different polygenic characters, in the MAA technology, use any other gene or preferably gene pleiomorphism also be a part of the present invention.Primer can be any length, is typically a long 10-20 base.By setting the numerical value of sequence, for example, it is 2 that first residue is 1, the second, or the like, the algorithm that can define all primers is: n is to n+y

N is that y is that the length of primer deducts one (9 to 19) from 1 integer to the last numeral of sequence, and n+y can not surpass the last numeral of sequence.Therefore, for 10 bases, probe is with respect to base 1 to 10,2 to 11,3 to 12 ... or the like.For 15 bases, probe is with respect to base 1 to 15,2 to 16,3 to 17 ... or the like.For 20 bases, probe is with respect to base 1 to 20,2 to 21,3 to 22 ... or the like.

In the embodiment of some, wish that a plurality of probes of utilization are used for the hybridization of single sample.Utilization length is the duplex molecule that the probe of 14 to 100 nucleotide can form stable selectivity.The length of complementary series is normally preferred above the molecule of 20 bases, so that increase the stability and the selectivity of hybridization, and therefore improves the quality and the degree of the special hybrid molecule of acquisition.People prefer designing the nucleic acid molecules of 20 to 30 nucleotide usually, and are perhaps longer in needs.This fragment prepares easily, and for example direct sequence synthetic by chemical method or that will select is inserted in the recombiant plasmid and be carried out recombinant production.

Correspondingly, nucleotide sequence of the present invention can be used for according to their character optionally forming duplex molecule or tissue-derived DNA of amplification or the primer of RNA being provided with the complementary portion of gene or RNA.According to the usage of expection, will use different hybridization conditions to obtain probe to target sequence selectivity in various degree.

When the higher selectivity of needs, can take strict relatively condition, promptly select low relatively salt and/or high temperature conditions, for example NaCl from about 50 ℃ to 70 ℃ about simultaneously 0.02M to about 0.10M.Such height stringent condition allow hardly (if any) probe and touch plate or the purpose chain between mispairing, and be particularly suitable for separating special gene or detect special mRNA and transcribe.It has been generally acknowledged that condition can be controlled more strictly by the Methanamide that adding is accelerated.

For specific usage, for example, carry out aminoacid by positional mutation and replace, thinking need be than the stringent condition of low degree.Under these conditions, even probe sequence and target chain are not correctly complementary, hybridization also can take place, but in the mispairing of one or more sites.Can make condition more not strict by increasing salinity and reducing temperature.For example, moderate stringent condition can be the NaCl from the about simultaneously 0.1M of about 37 ℃ to 55 ℃ temperature to about 0.25M, and can be that temperature is from changing the salt of about simultaneously 0.15M to about 0.9M about 20 ℃ to 55 ℃ than the stringent condition of low degree.Therefore, hybridization conditions can be controlled at an easy rate, and normally select according to the result who wants.

In other embodiments, hybridization can obtain in following condition: 50mMTris-HCl (pH 8.3), 75mM KCl, 3mM MgCl ₂, the 10mM dithiothreitol, DTT arrives under about 37 ℃ temperature conditions at about 20 ℃.Another hybridization conditions of utilization will comprise about 10mM Tris-HCl (pH 8.3), 50mM KCl, 1.5 μ M MgCl ₂, from about 20 ℃ under the temperature conditions of about 37 ℃ of variations.

In certain embodiments, nucleotide sequence best incorporated proper implements of the present invention uses, such as the label that detects hybridization.Many suitable marking toolses are known in the literature, comprise fluorescence, radioactivity, enzyme or other aglucon such as affinity element/biotin, and they can be detected.In preferred embodiments, can use fluorescent labeling or enzyme labelling such as urease, alkali phosphatase or peroxidase to replace radioactivity or other is to the disadvantageous reagent of environment.When using enzyme labelling, the colorimetric indication substrate is known, can be used to provide the visible or observable detection method of spectrophotometer of human eye, to identify and the specific hybridization that contains the sample of complementary nucleic acid.

Generally, can expect that hybridization probe described herein both can be as the reagent of liquid hybridization, as at PCR ^TMIn detect the expression of corresponding gene, also can be used for using the scheme of solid phase.In relating to the scheme of solid phase, test DNA (RNA) is adsorbed or is fixed on the substrate or surface of selection.Fixed single-chain nucleic acid is hybridized under the condition that requires with selected probe then.The selection of condition according to specific situation based on needed specific criteria (for example, according to the content of G+C, the type of target nucleic acid, the source of nucleic acid, the size of hybridization probe, or the like).After the hybridization surface washing is removed the probe molecule of non-specific binding, by marker detection hybridization, even quantitatively.

Amplification and PCR ^TMThe nucleic acid-templated methodology according to standard of amplification usefulness is separated people such as (, 1989) Sambrook from the cell of biological sample.Nucleic acid can be genomic DNA or sub-fraction or whole cell RNA.When using RNA, may wish to change RNA into complementary DNA.In one embodiment, RNA is whole cell RNAs and the template that directly is used as amplification.

Contact with isolating nucleic acid under the condition of selective cross with the paired primer of gene corresponding nucleic acids selective cross in the polygenic character.Ding Yi speech " primer " means and can start the synthetic any nucleic acid of newborn nucleic acid in the mode that template relies in this article.Typically, primer is the oligonucleotide of long ten to 20 or 30 base pairs, but longer sequence also can be used.Primer can be the form of two strands or strand, though the form of strand is better.

In case after the hybridization, nucleic acid-primer complex contacts the nucleic acid that is beneficial to the template dependence with one or more enzymes synthetic.Carry out the amplification of many wheels, also be known as circulation, up to the amplified production that produces sufficient amount.

Next step is the detection of amplified production.In specific usage, detection can be macroscopic mode.In addition, detection can be the non-direct evaluation of product, by system's (Affymax technology) of chemiluminescence, the radiolabeled radioactivity flicker of being mixed or fluorescent labeling or utilization electronics or thermal pulse signal.

The method that many templates rely on to be used for the increasing flag sequence of certain sample.One of the most known method is that the polymerase chain reaction (is called PCR ^TM), it is in U.S. Patent No. 4,683, have a detailed description in 195,4,683,202 and 4,800,159, each all integral body be incorporated herein by reference.

In simple terms, at PCR ^TMIn, prepare two primers, with the regional complementarity of two reverse complemental chains of flag sequence.In reactant mixture, add excessive triphosphoric acid DNA (deoxyribonucleic acid) nuclear, archaeal dna polymerase such as Taq polymerase.If flag sequence is arranged in the sample, primer will be attached on the sign, and polymerase extends primer by adding new nucleotide along flag sequence.By improving or reduce the temperature of reactant mixture, the primer of extension will separate forming new product from indicating, excessive primer will be attached to sign upward and on the product, and this process is able to repetition.

Reverse transcription PCR ^TMAmplification method is used for the quantity of the mRNA of amplification is carried out quantitatively.Reverse transcription RNA is well-known and people such as Sambrook to the method for cDNA, description is arranged in 1989.The alternative method of reverse transcription is utilized the dependent archaeal dna polymerase of heat-staple RNA.These methods have description in WO90/07641, the application on the 21st of nineteen ninety December is incorporated herein by reference.It is well-known in the literature that polymerase chain reaction method is learned.

The other method of amplification is ligase chain reaction (" LCR "), and it is open in EPA No.320 308, and the latter is incorporated herein by reference in full.In LCR, it is right to prepare two complementary probes, and when target sequence was arranged, on the reverse complemental chain that every pair will be attached to target, so they adjoined mutually.When ligase is arranged, two probes will be to being connected to form single unit.Circulation by temperature resembles at PCR ^TMIn, close-connected unit separates then from target sequence and is used for the right connection of excess probe as " target sequence ".United States Patent (USP) 4,883,750 have described the bonding probes similar to LCR to the method to target sequence.

The Qbeta replicative enzyme of describing in PCT application No.PCT/US87/00880 (being incorporated herein by reference) also can be as another amplification method among the present invention.In the method, target sequence has the replicability RNA sequence in complementary district to join in the sample under the situation of RNA polymerase is arranged.Polymerase will duplicate this replicability sequence, and it is detected subsequently.

A kind of homothermic amplification method, wherein restricted enzyme and ligase are used to increase contain the target molecule of nucleotide 5 '-[α-sulfur]-triphosphoric acid on chains of restriction site, also can be used for the present invention's nucleic acid amplification.

Strand displacement amplification (SDA) is the method for another constant-temperature amplification nucleic acid, and it comprises a plurality of circulation strand displacements with synthetic, i.e. nick translation.A similar methods is called and repairs chain reaction (RCR), comprises that several probes anneal in the zone that is used for increasing, and is to repair reaction then, and wherein four bases only occur two.Two other base can be used as biotin derivative and adds so that detect.Similar methods is used in SDA.The purpose specific sequence also can react (CPR) with circle probe and detect.In CPR, has the probe and the hybridization of the DNA in the sample of the intermediate sequence of 3 ' and 5 ' sequence of non-specific DNA and special RNA.After the hybridization, reactant is handled with RNA enzyme H, discharges behind enzyme action as the probe product of distinctiveness product.Initial template and the annealing of another circle probe also repeat this reaction.

Also have other amplification method to be described at GB application NO.2 202 328 with in PCT application No.PCT/US89/01025, each document all is incorporated herein by reference, and they also can be used for the present invention.In previous application, " modification " primer can be used for PCR ^TMIt is synthetic that sample, template and enzyme rely on.Primer can be by capturing group (as biotin) and/or detection moiety (as enzyme) labelling is modified.In one application of back, add excessive label probe in the sample.When having target sequence, probe in conjunction with and by catalytic decomposition.After decomposing, target sequence is able to complete release and by the excessive probes combination.The existence of the decomposition indication target sequence of label probe.

Other nucleic acid amplification method comprises based on the amplification system of transcribing (TAS), and it comprises amplification (NASBA) based on nucleotide sequence and 3SR people such as (, PCT applies for WO88/10315, is incorporated herein by reference in full) Gingeras.In NASBA, phenol/chloroform extracting that can be by standard, the thermal denaturation of clinical sample, handle with DNA isolation and RNA or extract RNA with lysis buffer and microcentrifugation post and prepare nucleic acid and be used for amplification through guanidine hydrochloride.These amplification techniques comprise the annealing that contains target distinguished sequence primer.After the polymerization, the DNA/RNA heterozygote is decomposed by RNA enzyme H, and the while double chain DNA molecule is thermal denaturation once more.In each case, single stranded DNA becomes two strands completely by the adding of second target special primer, carries out polymerization then.Double chain DNA molecule is transcribed by many times via RNA polymerase such as T7 or SP6 subsequently.In the constant temperature circular response, RNA is inverted record for single stranded DNA, changes double-stranded DNA then into, repeatedly transcribes under the effect of the RNA polymerase as T7 or SP6 so subsequently.The product that obtains no matter be incomplete or completely, has all shown the target-specific sequence.

People such as Davey, EPA No.329 822 (be incorporated herein by reference in full) discloses a kind of nucleic acid amplification method, comprises the synthesizing single-stranded RNA in cyclicity ground (" ssRNA "), ssDNA and double-stranded DNA (dsDNA), this method can be used for the present invention.SsRNA is the template of article one oligonucleotide primers, and it can be prolonged by reverse transcription (archaeal dna polymerase that RNA relies on).RNA removes from the DNA:RNA two strands that forms by the effect of RNA enzyme H (RNA enzyme H, special two strands at RNA and DNA or RNA formation) then.The ssDNA that obtains is the template of second primer, and it comprises the sequence with rna polymerase promoter (for example t7 rna polymerase) of template homologous sequence 5 '.This primer extends under the effect of archaeal dna polymerase (for example big " Klenow " fragment of Escherichia coli polymerase I), produced double-stranded DNA (" dsDNA ") molecule, its have with two primers between the identical sequence of initial RNA, promoter sequence is at one end arranged in addition.By suitable RNA polymerase, this promoter can be used to prepare many RNA copies of DNA.These copies can reenter circulation and increase fast.Select suitable enzyme, this amplification can be carried out under constant temperature and need not in each circulation enzyme-added.Because the circulation characteristics of this method, the sequence of beginning both can be that DNA also can be RNA.

People such as Miller, PCT Shen feelings WO89/06700 (be incorporated herein by reference in full) finds a kind of nucleic acid amplification scheme, it is based on promoter/primer sequence and purpose single stranded DNA (" ssDNA ") hybridization, the many RNA that transcribe this sequence subsequently copy.This scheme is not circulative, does not promptly produce new template from the rna transcription body that obtains.Other amplification method comprises " RACE " and " unidirectional PCR ^TM" (Frohman, 1990, be incorporated herein by reference).

Two sections (or more) oligonucleotide connect when existing based on the nucleic acid in " two-oligonucleotide " sequence that contains generation, thereby and the method for this two-oligonucleotide that increases, also can be used for amplification step of the present invention.

After any amplification, all amplified production should be separated with assessment whether special amplification has taken place from template and excessive primer.In one embodiment, amplified production separates (people such as Sambrook, 1989) with the method for standard by agarose, agarose-acrylamide or polyacrylamide.

In addition, chromatographic technique also can be used for separating.Have many kinds of chromatography can be used for the present invention: absorption, distribution, ion exchange and molecular sieve, and the technology of many specializations comprises post, paper, thin layer and gas chromatography.

The product of amplification must develop the color to conclude the amplification of flag sequence.Typical coloration method comprises with ethidium bromide and glue is dyeed and observes under ultraviolet.In addition, if amplified production with radioactivity or fluorescently-labeled nucleotide labelling intactly, amplified production can exposure X-ray sheet or is separated the back and observe under suitable exciting light.

Colour developing that can be indirect in one embodiment.After amplified production separated, the nucleic probe of labelling contacted with the flag sequence of amplification.Probe preferably combines with chromophore but also can carry out radioactive label.In another embodiment, probe is connected with binding partners, as antibody or biotin, but has detection moiety in conjunction with another right member.

In one embodiment, detection is to carry out by dna molecule hybridize with the hybridization of label probe.The technology of dna molecule hybridize is well known to those skilled in the art, can find in the molecule manipulation reference book of many standards.See people such as Sambrook, 1989.Amplified production can separate simply by gel electrophoresis.Gel contacts with film then, as nitrocellulose filter, carries out the transfer and the non-covalent combination of nucleic acid.With caudacoria be connected with chromophoric and can hatch with the probe of purpose amplified production hybridization.By detecting with film exposure X-ray sheet or with the emission of ions checkout gear.

An embodiment of preceding method is in U.S. Patent No. 5,279, is described (being introduced into this paper as a reference) in 721, and it discloses a kind of nucleic acid electrophoresis and transfer device and method of automatization.This device need not the outside just can carry out electrophoresis and hybridization to the operation of gel, and it is the ideal implementation method of the present invention.

The imitate essential all substances and the reagent of genetic marker of one or more polygenic characters in the product of detection of biological can be assemblied in the test kit together.This generally includes the primer of the specificity marker of choosing in advance.Can also comprise the different polymerases that are suitable for amplification of nucleic acid (RT, Taq, or the like), DNA (deoxyribonucleic acid) and buffer, to provide amplification essential reaction system.

This test kit generally comprises the right different vessels of primer that fills every kind of reagent and enzyme and every kind of sign by rights.The preferred primer of amplification of nucleic acid comes out to be used to the sequence that increases and list hereinafter to selected: SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ IDNO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, SEQ IDNO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ IDNO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ IDNO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ IDNO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ IDNO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ IDNO:32, SEQ ID NO:33 and SEQ ID NO:34.

In another embodiment, this test kit comprises the specific hybridization probe of the gene that relates to polygenic character, corresponding to hereinafter planting the sequence of listing: SEQ ID NO:1, SEQ IDNO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ IDNO:l1, SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ IDNO:15, SEQ ID NO:16, SEQ ID NO:17, SEQ ID NO:18, SEQ IDNO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ IDNO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ IDNO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ IDNO:31, SEQ ID NO:32, SEQ ID NO:33 and SEQ ID NO:34.The inventor expect known can with polymorphism allele effectively any primer of hybridization can be used for this test kit, suspect that this polymorphism allele has diagnostic significance in the methods of the invention, particularly in the MAA technology.This test kit comprises the different vessels that fills every kind of reagent and enzyme and every kind of used probe of sign hybridization usually by rights.

The dna fragmentation of coding specific gene can import in the recombinant host cell in order to express ad hoc structure albumen or to regulate albumen.In addition, by using gene engineering, the part of selected gene or derivant also can be used.Comprising the upstream region of control band such as promoter region can be separated and be used to express selected gene.

Should be appreciated that the present invention is not limited only to concrete probe disclosed herein, special expection comprises and the nucleotide sequence of disclosed sequence hybridization or the functional similar sequences of these sequences at least.For example, one section incomplete sequence can be used to identify gene or its source full-length gene group DNA or the cDNA clone of structurally associated.Those skilled in the art know that foundation can be as the method for the cDNA and the genomic library of the target of above-mentioned probe people such as (, 1989) Sambrook.

For the application in nucleic acid fragment insertion carrier of the present invention such as plasmid, cosmid or the virus; these fragments can combine with other DNA sequence; for example promoter, polyadenylic acid signal, restriction enzyme site, multiple clone site, other coded sequence and similar sequence, their total length can have sizable variation like this.The expection almost nucleic acid fragment of any length can be used, but simple in order to prepare the use of recombinating in operating with DNA, total length preferably is restricted.

The neomethodology that detects a plurality of genes in a DNA sample is because many genes of expectation and their pleomorphism site need be used to diagnose RDS and corelation behaviour, and the inventor has proposed polygenes method for screening (GENESCREEN ^TM).This may use new dna technique, as the gene chip of Affymetrix development.Putting it briefly, is exactly that photoactivation compound bag is by the glass dummy slider.These chemical substances contain prevent DNA base and chip and mutually between bonded protection against light sensitivity group.Yet when these chemical substances of irradiate light, the blocking group concurrent biochemical association reaction that is inactivated.Can allow " face shield " in specific region rather than other zone of irradiate light chip by using, the DNA base can be incorporated into institute's favored area of chip.Each the new base that adds all has bonded blocking group, so this method can repeatedly be connected to a base on another.Use this method, a large amount of different sequence dna probes can be synthetic simultaneously on one 1/2 inch chip.

400,000 probes that make up 20 DNA bases of any group leader only need 80 chemical steps.It is impossible utilizing the probe of the synthetic different sequence similar number of traditional dna synthesizer in reasonable time.These instruments are synthesising probing needle rather than synthetic with the used extensive parallel mode of Affymetrix in a continuous manner.In this DNA analysis fast, sample DNA at first joins in the probe array on the chip then with the fluorescence labels labelling.If sample finds complementary probe on chip, it will be in conjunction with getting on.If there is not complementary strand, it will be from eluting on the chip (dna fragmentation that complementary base is arranged is to say that they are complementary: for example, Segment A TTTGCGC (SEQ ID NO:1) will be in conjunction with the complementary fragment TAAACGCG that contains following sequence (SEQ ID NO:2)).The sequence of each probe and position are known, so which probe scanner can combine by judgement sample.Because the sequence of probe is known on the chip, the sequence of sample DNA is just known, because their sequence is complementary.Use gene chip not need messenger RNA is changed into cDNA and can detect messenger RNA and cDNA in large quantities.

Yet for the analysis of proposing among the present invention, the additive method that depends on the DNA instrument may be enough to carry out commercialization.For example, expection uses mass spectrography to carry out gene type.As using biochip technology simultaneously to the substitute technology of many gene types, Sequenom (SanDiego, CA) adopted the attached effect of the auxiliary laser desorption of substrate/ionizing flight time mass spectrum (MALDI-TOF) to carry out a large amount of gene types (people such as Little, 1997 that single base pair and short series connection repeat polymorphism; Braun, Little, Koster, 1997; People such as Braun, 1997).It is finished by following steps.At first, with one in the biotin labeling primer, PCR ^TMAmplification polymorphism zone (people such as Jurinke, 1997).In second step, the DNA of amplification is fixed on the streptavidin pearl people such as (, 1997) Jurinke.In the 3rd step, be close to the hybridization (Braun, Little, Koster, 1997) of a primer of pleomorphism site.The 4th step is when the ddNTP that dNTP is arranged and do not occur with the deoxidation form exists, with archaeal dna polymerase extend past pleomorphism site.When according to the sequence appropriate design, this can only cause the adding (Braun, Little, Koster, 1997) of a small amount of extra base.In the 5th step, DNA processes then to remove untapped nucleotide and salt.In the 6th step, remove and utilize the voltage pipet to transfer on the silica gel (O ' people such as Donnell, 1997) short primer+pleomorphism site by degeneration.In the 7th step, the quality of short primer+pleomorphism site is measured (people such as Li, 1996 by postponing to extract the MALDI-TOF mass spectrography; People such as Tang, 1995).The quality that single base pair in the sequence and series connection repeat to make a variation by them is easy to judge.This last step is very fast, and each is analyzed only needed for 5 seconds.All these steps all are machine automatizatioies.This technology has the potentiality of carrying out 20,000 gene types every day.

This technology is split hair fast, and is applicable to any polymorphism.With respect to biochip technology, its remarkable advantage is that it is more accurate, can either identify that single base pair can identify that also short series connection repeats polymorphism, and in seconds add or remove the polymorphism of check with minimum cost.

The different genes that polygenes test kit-genescreen test kit proposes according to the application, table 2 describes in detail based on GENECHIP ^TMThe possible gene obstacle test kit of notion.

Table 2 obstacle gene alcoholism D1, D2, D4, DAT1, TDO2, nNOS1a drug dependence (comprising that intravenous pharmacy uses) D1, D2, D4, CNR1, GABAB3, TDO2, HT-2R,

MAOA (X), COMT, nNOS1a, DOMC, preceding endorphins analeptic is given up depressed D2 substance abuse obstacle (SUD) D1, CNR1

SUD subclass: alcoholism D2, GABAB3

SUD subclass: many substance abuses D4, HT-2R

SUD subclass: intravenous pharmacy abuse DAT1, MAOA (X)

The SUD subclass: analeptic is given up COMT, nNosla

The SUD subclass: analeptic is given up: TDO2

Depressed seriousness

The SUD subclass: ethanol is given up DAT1 (9/9 allele) and is forced D1, TDO2

Force subclass: pathological gambling D1, D1, TDO2

Force subclass: property is forced fat D2, OB, DAT1, APOD, UCP2, CHROME-2

Fat subclass: female body APOD, CHROME-2

Fat (＞34%)

Fat subclass: male body DAT1, OB

Fat (＞28%)

Fat subclass: BMI＞25

Fat subclass: carbohydrate gluttony table 2 (continuing) obstacle gene attention deficit 5HT-transport protein

Attention deficit subclass: autism D1, DAT1

Attention deficit subclass: Tourette D1, D2, TDO2, D β H, MAOA (X), HTR1A

Syndrome

Attention deficit subclass: ADHD HTR1A, D2, DAT1, D β H, TDO2

Attention deficit subclass: ADD MAOA (X) cigarette smoking D1, D2

Cigarette smoking subclass: smoking every day

The bag number

Cigarette smoking subclass: beginning

Cigarette smoking subclass: the smoking time limit

Cigarette smoking subclass: recurrence personality disorder DAT1, D2

Personality disorder subclass: pathologic violence D2, DAT1, D4, CNR1

The personality disorder subclass: the division sample is avoided D2, DAT1

Personality disorder subclass: P300 D2, CNR1

Personality disorder subclass: AER/VER D2

Personality disorder subclass: stress D2 after the wound

Personality disorder subclass: the low novel D2 that seeks

Personality disorder subclass: the high novel D4 that seeks

Personality disorder subclass: defence type D2

Inventor's suggestion, in order to identify the excessive risk crowd of RDS safely, these all genes must be checked.When finding other gene, expectation is also tested to them.Because have only several genes to constitute the check list, the original plan of check comprises PCR ^TMTechnology.。When these other gene discoveries, expect that they also join GENECHIP ^TM(Affirmatrix, Santa Clara, CA).

In the 5th step of step 5.,, determine a scoring to genotype based on which kind of genotype of explanation and maximum quantity relevant which kind of genotype of scoring and the relevant independent studies of minimum quantity scoring.Determine that to gene scoring is based on application ANOVA, assess the height of its average Kellgren scoring of each genotype in every kind of polymorphism.These researchs are carried out in one group of experimenter, and are irrelevant with the experimenter among the MAA.For example, for dopamine D ₂The Taq I A1/A2 polymorphism of acceptor gene, DRD2, all researchs shown 1 allele with a series of impulsions, force, addictive behavior is relevant, and some study the heterozygous genes types 12 that illustrate, divide relevantly with higher assessment, 11 is relevant with minimum scoring with 22 genotype.Therefore, by utilization Taq I A1/A2 polymorphism, genotype 22 or 11 scoring=0 in the DRD2 gene, the scoring of genotype 12=2.If it is relevant with minimum scoring in the quantitative trait that independent studies shows 22 genotype, 12 is relevant with medium scoring in the quantitative trait, and 11 divide relevantly with higher assessment, and scoring should be 22=0,12=1 and 11=2.

When using dinucleotide and trinucleotide polymorphism,, also allele might be divided into short and long allele even many allele are arranged.If long allele is relevant with maximum phenotypic effect, scoring should be SS=0, SL=1 and LL=2, and wherein S is short allele, L is the long gene that waits.

Step 6. is set up polygenes or the PG variable fabricated by constantly adding the scoring of each gene.For example, if the scoring of first gene is that the scoring of 2, the second genes is 1 in example 1, then PG scoring variable is 3.1 to n example will be assessed by regression analysis for the correlation coefficient r of quantitative trait variable scoring.More preferably use the off-the-self statistical procedure.The percentage ratio of variation is r ²Then, for 1 to n example, add the gene scoring of the 3rd gene and repeat this process.Draw final result.

Step 7. is carried out the single argument regression analysis of PG to QT or DV.Be an example that can be generalized to any amount character or two distortion amounts below.Before the analysis beginning, the hypothesis variable (PG) of gene scoring sets=0.Regression analysis is carried out in the scoring that adds each other gene then, adds the scoring of second gene, repeats regression analysis.Continuing this process all adds up to all check genes.For example, PG is set at 0 at first.Scoring as fruit gene 1 equals 2, and the PG value then adds 2, or PG=PG+2.Carry out the recurrence of PG then to QT or DV.Scoring as fruit gene 2 equals 1, and the PG value then adds 1, or PG=PG+1.Perhaps, equal 2 as the scoring of fruit gene 2, the PG value then adds 2, or PG=PG+2.After the scoring of gene 2 adds, carry out PG once more to the regression analysis of PG to QT or DV.

The method for optimizing of an analytical data is with all data input statistical package such as SPSS.Utilize the value of the DRD2 gene of describing in the 5th step in the above, following exemplary algorithm will be opened and set up to the syntax file of SPSS program.Compute PG=0.If (DRD1 eq2) PG=PG+2.If (DRD2 eq2) PG=PG+2.If (DRD3 eq2) PG=PG+2.If (DRD4 eq2) PG=PG+2.If (DRD5 eq1) PG=PG+1.If (DRD5 eq2) PG=PG+2.If (DAT1 eq1) PG=PG+1.If (DAT1 eq2) PG=PG+2. regression variable=ADHD PG/ dependence=ADHD/ method=input PG

The PG variable is set to 0.Because DRD1, DRD2, DRD3, DRD4 gene are counted 0 or 2 fen, if the experimenter carries DRD1 DdeI 11 genotype and/or DRD2TaqI 12 genotype and/or DRD3 MscI 11 or 22 genotype and/or DRD446,47 or 77 genotype, they only need the PG single line to increase by 2 fens.On the contrary, DRD5 and DAT1 gene are counted 1 or 2 fen, and they just need the two-wire code.When adding each gene, carry out the regression analysis of PG score to QTV (ADHD scoring).

Step 8. drawing result.The result is drawn into figure, and the gene that carrying out property adds up is positioned at X-axis, r ²Be positioned at Y-axis.Shown in the lines among Fig. 5: its expression is to adding up property and the subtracting property gene of ADHD.

This process of gene redundancy of step 9. an adding up property of use.The result as shown in Figure 6, its expression adding up property gene.

The treatment of RDS corelation behaviour

Some anti-addiction material is by strengthening the function of endogenous or exogenous nerve polypeptide or neurotransmitter, as the release of dopamine at Fu Hezhong, and suppresses serious hope to the sense of euphoria, thereby obviously reduces unusual addiction sexual behaviour, claims " RDS " behavior again.The material that can cause the sense of euphoria comprises ethanol, cocaine, Opiate, nicotine, glucose or other carbohydrate, and sexual behaviour, gambling, attack, violence etc., but is not limited in this.There is common hereditary basis in the RDS behavior, and with in-limbic system in the capacity supply of neurotransmitter relevant.Final recompense approach comprises the dopaminergic activation of acceptor site (D1, D2, D3, D4, D5 and various hypotype), and the density of these receptors by their separately gene and responsible dopamine is synthetic, storage, release and the decision of metabolic gene.

The present invention's suggestion is used enkephalinase inhibitor and a kind of enkephalin releasing agent such as kyotorphin (tyrosine-arginine) or its stable analog tyrosine-D-arginine occupying for a long time with promotion D2 receptor by uniting, make it to take place hypertrophy or just regulating (Fitz etc., 1994), thus strengthen the release of synapse dopamine.

The present invention relates to the viewpoint of polygenic inheritance and has proposed with a plurality of gene locuses as the notion of differentiating RDS group of people at high risk's determiner on the part degree.Do not find the foundation of single-gene mechanism in the quantitative trait loci research in diverse each the strain mice of inherited character.Detect the reaction of 15 kinds of mices to ethanol, morphine, metamfetamine, find the reaction difference of each strain to above-mentioned 3 kinds of materials, prompting is by the susceptibility of different genes decision to different addiction materials.These find that it is not independent phenomenon that further prompting is subjected to the sensitivity to ethanol of effect gene, and on the contrary, it has specificity (Crabbe etc., 1994) to the response variable of being studied.

Therefore one aspect of the present invention is above-mentioned specific gene and suspection other gene type relevant with RDS behavior or other polygenic features, obtains targeted therapy RDS dependency behavior or the important haplotype information of other polygenic features.When a certain specific genetic variance was found relevant with certain behavior disorder, the inventor can utilize this knowledge to help the clinician to adopt medicine to carry out hereditary targeted therapy.The present invention adopts gene type and amino acid precursor to introduce, suppresses enkephalinase, induces the composition as anti-addiction such as enkephalin release, anesthesia antagonism and chromium picolinate or nicotinic acid chromium etc., increases the favourable release of dopamine at Fu Hezhong.

In the present invention, any one gene alteration that inventor's proposition relates to various developed by molecule in the recompense cascaded loop (may reach more than 100) all can make individual RDS of generation and corelation behaviour, even ADHD and violence.Although above-mentioned may be true, be difficult to prediction at present and be specifically related to which gene, propose to adopt the MAA technology up to the present invention.Yet the effect evidence of D2 dopamine receptor gene is fully at least, and the inventor thinks that the D2 dopamine receptor gene represented main " the recompense gene " that plays an important role on RDS and corelation behaviour.Preferred version of the present invention is TaqA1 DRD2 allele (A1, B1, C1, haplotype polymorphism between the exon 6-7) and the consequence of targeted therapy connect that (attention is concentrated, and prevents that body weight from ging up, and stops smoking, reduce the carbohydrate gluttony, reduce number AMA time, prevent many substance depilatories, reduce violent behavior and other).The inventor also will provide some gene type examples at above-mentioned specific gene, and they can produce optimum response to certain class medicine.

On neurotransmitter recompense cascade model, the domination of hypothalamus 5 one hydroxytryptamine serotonergic neurons also activates the met-enkephalin serotonergic neuron, and the latter suppresses γ-An Jidingsuan (GABA) neuron, and the latter activates the DA neuron of tegmentum veutro subsequently.The DA neuron projects Cluster A1 (CA1) cell cluster of Acb and Hippocampus subsequently, and the DA neurotransmitter is as main recompense substrate (Stein and Beluzzi, 1978) there.

Volt is examined and enkephalin plays an important role in this complicated loop (Heidreder etc., 1988).Local with inducing DA behind the enkephalinase inhibitor in striatal release, point out this adjusting to carry out (Chesselet etc., 1981) by stimulating the δ receptor.In fact, kyotorphin also may protect cholecystokinin-8 (CCK-8) not by brain peptide enzymatic degradation.Control desire nerve polypeptide and DA that this is important are present in Fu Hezhong together, and and CCK-8, DA and endogenous opioids polypeptide between close getting in touch (Koob, 1992) arranged.

Neurotransmitteies such as 5 one hydroxytryptamines (5-HT), DA, norepinephrine (NE) and enkephalin are proved to be the picked-up that can reduce the sweet food thing.Therefore special design compositions of the present invention, by giving amino acid precursor, comprise 1-tryptophan (5-HT precursor), 1-phenylalanine (DA and NE precursor) and enkephalinase inhibitor d-phenylalanine are to increase these food inhibitory nerve mediators (Blum etc., 1986).

The inventor studies to establish the effect of these reagent classifications the Lewis rat.As if the Lewis rat has the tendency of many substance abuses, and is used as the important animal model of RDS behavioral study.We adopt the various dose of 3 kinds of reagent (D-phenylalanine [DPA], naltrexone [NTX], tyrosine-D-arginine [TA]) and combination to treat to ethanol cocaine, nicotine, the selection voluntarily of Fructus Cannabis or carbohydrate in this.Compare with any 2 kinds of combination of agents, three kinds of agent combination are the most effective (seeing Table 3) together.

Table 3

Uniting and independent utility of anti-RDS reagent, minimum from being up to

The DPA+NTX+TA+ chromic salts

The DPA+NTX+ chromic salts

The DPA+TA+ chromic salts

The NTX+TA+ chromic salts

The DPA+ chromic salts

The NTX+ chromic salts

The TA+ chromic salts

Chromic salts

The DPA=D-phenylalanine; The NTX=naltrexone; TA=tyrosine-D-arginine

The compositions of treatment RDS: although various obstacle can range RDS and invade polyamines energy system, a kind of medicine can not be treated all patients that suffer from RDS similarly.A large amount of other systems also can be invaded, and therefore must treat it simultaneously for the curative effect that obtains comprehensive satisfaction.The disease that relates to medicine or substance abuse especially needs individualized treatment.Although therefore a total therapeutic scheme comprises the amino acid precursor of enkephalinase inhibitor, enkephalin releasing agent and dopaminergic system to patient's effective dose, total also need adopt the therapeutic effect of other composition to strengthen.

Some treatment reagent is praised highly by " D2 genetic recipient defect theory ".The level that has confirmed DRD2A1 carrier DRD2 receptor is very low.Patient need rely on these receptors to tackle the society of today fully under tension pressure.An important aspect of the present invention provides some amino acid precursor, trace meter such as chromium picolinate and/or nicotinic acid chromium, enkephalinase inhibitor, narcotic antagonist and the enkephalin releasing agent of therapeutic dose (alone or in combination), finally cause discharging to induce the D2 hypertrophy, especially in the D2A1 carrier naturally of dopamine.This total viewpoint obtains check already in cocaine addiction.The TROPAMINE that is called by first inventor development ^TMPreparation research and adopt in patient.Through comparing TROPAMINE with other medication ^TMThe major advantage (Halikas etc., 1993) that has detoxifcation and keep giving up.See Table 4.

Table 4 generally is used for the treatment of the ordering of the medication effectiveness of cocaine abuse

Detoxication is given up and is kept the antidote patient and count internist's number and render a service and to keep the medicine patient and count internist's number effect Tropamine 190 17 4.00 Tropamine 150 17 3.50Buspiran 11 2 4.00 clonidines 15 2 3.00 nortriptylines 100 3 4.00 desmethylimipramines 8,824 306 2.84 phenobarbitals 2,250 10 4.00 imipramine 2,940 129 2.64 benzodiazepine classes, 155 3 3.33 fluoxetines 2,386 145 2.61 clonidines, 412 15 3.33 L-tyrosine, 350 9 2.60 LIYANNINGs, 510 14 3.25 carbamazepines 1,384 86 2.57 stable 2,710 18 3.23 bromocriptines 5,256 132 2.56L-tyrosinase 15s, 10 9 3.13 amantadines 6,527 119 2.39 bromocriptines 15,511 262 2.95 L-tryptophans 6,247 110 2.20 amantadines 19,189 225 2.69 Neuroieptics 1,494 54 2.19 desmethylimipramines 10,352 287 2.65 naltrexones 1,255 40 2.15

Detoxication is given up and is kept the antidote patient and count internist's number effect and keep the medicine patient and count internist's number effect imipramine 2,885 122 2.48 phenobarbitals, 100 3 NR L-tryptophans, 15,112 198 2.33 " other drugs " 853 47 2.64

Mixture fluoxetine 1, the mixture of 527 111 2.25Bupranorphine, 148 19 2.00 naltrexones, 817 31 1.68 Mazindol 11 2 1.00 " other drug " 8,007 119 3.11

Sum 79,760 468 2.43 sums 37,166 381 2.57

An ingredient of this scheme be earlier patient by gene type, provide suitable cocktail treatment by his/her genotype then.Unusual for D2, the inventor has developed a suitable cocktail scheme, and description is arranged in the text.Table 5 has been summed up the application that the specific gene type has been confirmed the various compositionss of effective the present invention, table 6-16 has listed treating the effectively preferred component of these compositionss of various obstacles, and table 17-19 has shown how some element influences stimulus object (cocaine etc.), opiates and the inductive recompense of sedative hypnotics.

Table 5 polygene diagnosis and anti-addiction compositions: the RDS behavior effectiveness study type genes of interest relevant that directed prevention and the therapeutic combination that treatment is selected are received treatment with the reaction of improvement

(SI)/allele Alcotrol ^TMThe anti-addiction of substance abuse obstacle, anxiety reduces, and recurrence DBPC-inpatient DI be (the allelic Dde homozygosity of A1

Stress calmness-hypnotic especially and reduce excessively, antagonism doctor advice rate frequency increases)

With (reducing health and D as ethanol, opioid, (AMA) ₂(Taq A1, B1, exon ^6-7Doubly single

Barbiturates) the BESS score have improved-type, C1)

DBPC-outpatient DATI VNTR (10/10)

CNRI (VNTR＜5bp repeats to isozygoty)

MOAA(x)-VNTR＜335bp

*

NNOSIa-≤201BP allele isozygotys

DBH-TaqI B1 allele

COMT-108 valine allele

TDO2 intron 6 (G → A) and/or (G →

T)

D ₄The RDS behavior effectiveness study type genes of interest relevant that VNTR 2 or 7 therapeutic combinations of selecting are received treatment with the reaction of improvement

(SI)/allele C ocotrol ^TMThe substance abuse obstacle stresses and Alcotrol especially ^TMIdentical, add DBPC-inpatient D ₂TaqI A1, B1, C1 or exon ^6-7

Psychoanaleptics abuse cocaine thirsts for reducing haplotype, D1 (the allelic Dde of A1

The frequency of (cocaine, metamfetamine) homozygosity increases)

DBPC-outpatient Comt-108-valine allele, D3Alcotox ²Calmness-hypnotic is given up and (is comprised that wine benzodiazepine class demand reduces inpatient OT DAT1 VNTR (9/9)

Essence, opioid, barbital are given up to tremble and are alleviated, and depression alleviates A1, B1, C1 or exon with canonical solution poison comparison D2 TaqI ^6-7

Class) haplotype PHENCAL ^TMIt is fat that (the carbohydrate gluttony loses weight, and the gluttony incident reduces DBPC-outpatient D ₂TaqI A1, B1, C1 or exon ^6-7

Anorexia, bulimia nerovsa) the positive change formed of health, anti-haplotype

Ending fat-reducing back body weight bounce-back HTR2A-C allele isozygotys

The OB-1875 dinucleotide repeats polymorphism

＜208BP allele isozygotys

No. 2 chromosome microsatellite polymorphisms of people,

D2S1788

UCP-2 (polymorphism is undetermined)

APO-D-TaqI2.2 or 2.7BP

The RDS behavior effectiveness study type genes of interest relevant that the therapeutic combination that leptin receptor (polymorphism the unknown) is selected is received treatment with the reaction of improvement

(SI)/allele Nicarest ^TMCigarette smoking smoking stops, and abandons easily (inhaling DBPC-outpatient D1 (Dde A1 isozygotys)

Cigarette), prevent from heavily to inhale D2 (TaqI A1)

D4(VNTR?2)

D5 (dinucleotide 13 allele, model

Enclose 135-159BP)

DAT1?VNTR(10/10)

D β H (TaqI B1 allele) HyperGen ^TMAutism, Tourette, the ability of ADHD job task strengthens, TOVA DBPC-outpatient D1 (Dde A1 isozygotys)

Score is improved, Cull/Blum ADHD D2 (TaqI A1)

Score is improved, and hyperactivity reduces D4 (VNTR 2)

D5 (dinucleotide 13 allele, model

Enclose 135-159BP)

DAT1?VNTR(10/10)

D β H (TaqI B1 allele)

The RDS behavior effectiveness study type genes of interest relevant that the therapeutic combination that MAOA (X) selects is received treatment with the reaction of improvement

(SI)/allele Gambotrol ^TMPathological gambling spends to reduce, and bets that frequency reduces OT outpatient D1 (Dde A1 isozygotys)

Give up D2 (TaqI A1, B1, C1) violent behavior of Tempaminc pathologic easily, division sample/violent behavior outburst reduces hostility OT outpatient D2 (TaqI A1, B1, C1, exon ^6-7)

Avoid disease (SAB), aggressivity reduces, defence type normalization, DAT1 (VNTR 10/10)

Angry, hostility stress anxiety reduce after the wound, and SAB reduces mNOSIa-≤201BP allele and isozygotys

Obstacle PMX ^TMPMS pain and cramp reduce, an OT outpatient and Alcotrol ^TMAnd Cocotrol ^TM

Alleviation of pain is few, and whole mental state is improved identical

Endorphins/dynorphin before the POMC-/Orphan

Opiate receptor

δ, σ, lonely, μ, κ, ε ¹Prescription sees Table 6 described to 17 ²Substituting composition limits as follows: D-phenylalanine 500mg/ capsule, 6 of every days; Tyrosine-D-arginine, every capsule 1mg, 6 of every days; Naltrexone, every capsule 50mg, 1 to 3 of every day. ^*Isozygoty HTR1A-TC polymorphism HTR2A-C allele (isozygotying) abbreviation of GABRB3-dinucleotide repetition 〉=185bp: ADHD=attention deficit hyperactivity disorder DPBC=double blinding, placebo PMS=premenstrual syndrome POME=Propiomelancortin OT=open trial

Table 6

Kantroll ^TMComposition basic or core capsule mg/ capsule core or " target " capsule biotin 0.050 core calcium 35.000 core chromium nicotinate chromiums 0.033 core pyridine carboxylic acid chromium 0.033 core copper 0.033 core 5-hydroxyryptophan＞.5 core DL-phenylalanine 460.000 core folic acid 0.030 core iodine 0.025 core iron 1.000 core Glus 25.000 core magnesium 2.500 core methionines 50.000 core nicotinic acid (not reddening) 10.000 core pantothenic acid 0.330 core selenium 0.012 core vitamin A IU 277.710 core vitamin B-2 0.800 core vitamin B-1,0.375 core vitamin B-6,1.000 cores (P-5 phosphoric acid) vitamin B-12s 0.005 core vitamin C 100.000 core vitamin E IU 5.000 core zinc 1.500 core L-BETAINs 10.000 fat Ginko B 25.000 obesities; 150.000 gambling of focus TYR; Exciting. anxiety; Nicotine; Cocaine; Fat ornithine aspartate 10.000 fat kola nuts (caffeine) 20.000 fat L-arginine 10.000 fat pyroglutamate yellow chamomile * 25.000 nicotine taurine * 25.000 excitements, anxiety valerian * 10.000 nicotine extract of willow bark * 60.00 PMS symptoms

Annotate: for basic recompense deficiency symptoms behavior total in various symptoms, the inventor recommends to take " core " neurologic agent (Neutraceutical), every day three times (before the meal).If the patient has the addiction/impulsion/compulsion of a series of stubbornnesses, or obvious serious addiction/impulsion/compulsion, the inventor recommends to take " core " neurologic agent, adds that every day, suitable number of times was taken suitable additional neurologic agent.

For obesity, the patient should take white capsule when taking " core " nerve under physician guidance, early should take before the meal

Orange capsule.

For nicotine dependence, the patient should be at the blue capsule of hora somni sumendus.

Table 7

PHENCAL ^TMThe every capsule of composition (in milligram) DL-phenylalanine 500.000L-glutamine 15.000L-tyrosine 25.0005-hydroxytryptophane 2.5mg kola nut (caffeine) 20.000L-carnitine 10.000 pyridine carboxylic acid chromium or 0.033 nicotinate chromium calcium, 35.000 iron, 1.000 magnesium, 2.500 zinc, 2.500 biotins, 0.050 pantothenic acid, 0.330 iodine, 0.025 bronze medal, 0.333 selenium, 0.012 vitamin C, 13.300 vitamin B-1s 0.330

Table 7 (continuing)

PHENCAL ^TMThe every capsule of composition (in milligram) vitamin B-2 0.500 nicotinic acid (not reddening) 3.330 vitamin E IU 5.000 vitamin B-6s (P-5 phosphoric acid) 0.330 folic acid 0.066 vitamin B-12 0.001Ginko B 25.000

Table 8

Tropagen ^TMThe every capsule of composition, (in milligram) DL-phenylalanine 250.000L-glutamine 50.000L-tyrosine 150.0005-hydroxytryptophane 2.5mg pyridine carboxylic acid chromium or nicotinate chromium 0.020 calcium 25.000 iron 1.500 methionines 50.000 magnesium 2.500 zinc 5.000 pantothenic acid 1.500 vitamin Cs 100.000 vitamin B-1s 1.670 vitamin B-2s 2.500 nicotinic acid, (not reddening) 16.700 vitamin B-6s, (P-5 phosphoric acid) 3.330 folic acid 0.670 vitamin B-12 0.670

Table 9

Alcotrol ^TMThe every capsule of composition (in milligram) DL-phenylalanine 480.000L-glutamine 25.0005-hydroxytryptophane 2.5mg pyridine carboxylic acid chromium or nicotinate chromium 0.020 calcium 25.000 iron 1.500 magnesium 2.500 zinc 1.500 biotins 0.050 pantothenic acid 15.000 vitamin A I.U.333.300 copper 0.333

0.013 vitamin C 13.300 vitamin B-1 2.417 vitamin B-2s, 0.850 nicotinic acid (not reddening) 3.300 vitamin E I.U.5.000 vitamin B-6s (P-5 phosphoric acid) 3.000 folic acid 0.670 vitamin B-12 0.670

Table 10

Nicarest ^TMThe every capsule of composition, (in milligram) DL-phenylalanine 500.000L-glutamine 50.0005-hydroxytryptophane 2.5mgL-tyrosine 125.000 pyridine carboxylic acid chromium 0.033 calcium 50.000 iron 1.000 magnesium 2.500 zinc 2.500 biotins 0.050 pantothenic acid 0.330 iodine 0.025 bronze medal 0.333 vitamin C 100.00 vitamin B-1s 2.417 vitamin B-2s 0.850 nicotinic acid, (not reddening) 3.330 vitamin B-6s, (P-5 phosphoric acid) 3.000 folic acid 0.670 vitamin B-12 0.005 yellow chamomile 25.000* radix valerianae 10.000** h.d.

Table 11

PMX ^TM

The every capsule of composition, (in milligram) DL-phenylalanine 375.000L-glutamine 50.0005-hydroxytryptophane 2.5mg pyridine carboxylic acid chromium 0.020 calcium 50.000 magnesium 25.000 iron 1.500 zinc 1.500 biotins 0.050 pantothenic acid 1.250 vitamin A I.U.277.710 vitamin Cs 30.000 vitamin B-1s 0.375 vitamin B-2 0.800 nicotinic acid, (not reddening) 10.000 vitamin B-6s, (P-5 phosphoric acid) 1.000 folic acid 0.030 vitamin B-12 0.005 yellow chamomile 25.00 extract of willow bark 25.00

Table 12

HyperGen ^TMThe every capsule of composition (in milligram) DL-phenylalanine 400.000L-glutamine 25.000L-tyrosine 150.0005-hydroxytryptophane 3.5mg methionine 50.000 pyridine carboxylic acid chromium 0.020 calcium 25.000 iron 1.000 magnesium 3.500 biotins 0.050 pantothenic acid 15.000 vitamin C 13.300 vitamin B-1 0.375 vitamin B-2,0.800 nicotinic acid (not reddening), 10.000 vitamin B-6s (P-5 phosphoric acid) 3.330 folic acid 0.030 vitamin B-12,0.001 Ha Maling (pharmaline) 50 huperzines (huberzine) 150ugGinko B 40.000

Table 13

Stress-X ^TMThe every capsule of composition; (in milligram) DL-phenylalanine 400.000 vitamin C 100.000L-glutamy 35.0005-hydroxytryptophane 3.5mg calcium 25.000 pantothenic acid 15.000 nicotinic acid; (not reddening) 3.300 vitamin B-6s; (P-5 phosphoric acid) 3.000 magnesium 2.500 vitamin B-1s 2.417 iron 1.500 zinc 1.500 vitamin B-2s 0.850 folic acid 0.670 biotin 0.050 pyridine carboxylic acid chromium 0.020 vitamin B-12 0.005 vitamin A I.U.333.300 vitamin E 5.000I.U. taurine 0.005 radix valerianae 25.000

Table 14

Tempamine ^TM

The every capsule of composition, (in milligram) biotin 0.050 calcium 25.000 pyridine carboxylic acid chromium 0.020DL-phenylalanines 400.000 folic acid 0.670 iron 1.500L-glutamine 35.0005-hydroxytryptophane 2.5000L-tyrosine 100.000 magnesium 2.500 nicotinic acid, (not reddening) 3.300 pantothenic acid 15.000 vitamin A I.U.333.300 vitamin B-1s 2.417 vitamin B-2s 0.850 vitamin B-6, (P-5 phosphoric acid) 3.000 vitamin B-12s 0.005 vitamin C 100.000 vitamin E I.U.5.000 zinc 1.500

Table 15

Aggression

The every capsule of composition, (in milligram) DL-phenylalanine 460.000L-glutamy 25.000L-tyrosine 100.0005-hydroxytryptophane 2.5mg pyridine carboxylic acid chromium 0.020 calcium 25.000 iron 1.500 magnesium 2.500 zinc 1.500 biotins 0.050 pantothenic acid 15.000 vitamin A I.U.333.300 vitamin Cs 100.000 vitamin B-1s 2.417 vitamin B-2s 0.850 nicotinic acid, (not reddening) 3.300 vitamin E I.U.5.000 vitamin B-6s, (P-5 phosphoric acid) 3.000 folic acid 0.670 vitamin B-12 0.005

Table 16

Gambotrol ^TM

The every capsule of composition (in milligram) calcium 25.000 pyridine carboxylic acid chromium 0.020DL-phenylalanines 400.000 folic acid 0.670 iron 1.500L-glutamine 35.000L-tyrosine 100.0005-hydroxytryptophane 2.5mg magnesium 2.500 methionines, 50.000 nicotinic acid (not reddening) 6.700 pantothenic acid 1.500 vitamin B-1s, 1.670 vitamin B-2s, 2.500 vitamin B-6s (P-5 phosphoric acid) 3.330 vitamin B-12s 0.005 vitamin C 100.000 zinc 2.500

Table 17

Cocaine and amphetamine recompense

Example encourages the intracranial self administration of medication to the intracranial oneself electric stimulation of hypothalamic outside growth encourage foot lid district veutro that influences of recompense

Prefrontal cortex middle part (cocaine) encourages

Volt nuclear (amphetamine) encourages vein not to be had the 5-HT of change receptor antagonist from administration norepinephrine receptor antagonist and encourages the M-opiate receptor antagonist and do not have the D1 of change and D2 dopamine-receptor antagonist and suppress norepinephrine denervation (6-HAD) and do not have the 5-HT of change denervation (the two hydroxytryptamines of 5,7-) and encourage dopamine denervation (6-HAD)

Volt nuclear suppresses

Foot lid district veutro suppresses

The prefrontal cortex middle part does not have change

Table 18

The opiates recompense

The example recompense effect encephalic oneself electric stimulation of hypothalamic outside encourages encephalic and encourages the hypothalamus outside from administration volt nuclear and encourage pin lid district veutro and encourage intravenous and suppress Δ-receptor antagonist from administration opiate receptor antagonist M-receptor antagonist and do not have the K-of change receptor antagonist and do not have the dopamine-receptor antagonist mixing resultant dopamine denervation of change (6-HAD) volt nuclear and do not have and change

Table 19

Tranquilizer/somnifacient recompense

The example recompense effect intracranial oneself electric stimulation of hypothalamic outside encourages intracranial self administration of medication foot abdomen lid veutro and encourages per os self administration of medication GABA _AReceptor antagonist suppresses GABA _AReceptor stimulating agent encourages opiate receptor antagonist and suppresses the inhibition of dopamine-receptor antagonist inhibition 6-HT receptor stimulating agent inhibition norepinephrine synthetic inhibitor

Therapeutic Method: treat in the present invention and to comprise the D-phenylalanine in the preparation, D-leucine and any D-aminoacid (seeing Table 6-16) that contains hydrocinnamic acid.Said preparation also comprises brain coffee enzyme inhibitor in addition, as some protein synthesis inhibitor, and as bacitracin, bestatin and puromycin; Polypeptide amino acid such as the D type monamino acid that dissociates, the dipeptides or the tripeptides of D type essential amino acids; The sulfydryl benzyl amino acid (2-[sulfydryl-3-phenyl-propiono]-the L-leucine); The carboxyalkyl methyl ester (N-[(R, S)-2-carbethoxyl group-3-phenylpropanol]-the L-leucine); And unrelated compounds such as quinalbarbitone (secobarbital) on other the structure in a large number, pyrophosphate, O-phenanthroline, phosphamide (phosphamidon), Z-leucine-NHOH and Z-glycine-NHOH.

In addition, the inventor thinks that they can improve the reaction of patient to treatment very significantly by the enkephalin releasing agent further is provided.Therefore enkephalin releasing agent tyrosine-arginine and tyrosine-D-arginine also are included in the treatment preparation.

There are a mechanism that influenced by psychoanaleptics stimulus object and non-psychoanaleptics stimulus object in a large amount of allele and multiple impulsion, associated cue mandatory and assuetude disturbance, can cause that dopamine (DA) preferentially is discharged into volt nuclear (Acb) middle part.This machine-processed hereditary basis comprises at least and contains dopaminergic gene and various regulatory enzyme (D1-D5, DAT1, D β H, MAOA, polymorphism COMT); Therefore, change neurotransmitter in the recompense cascade (comprise 5-hydroxy tryptamine, enkephalin, GABA, composition DA) to material and action obstacle should be useful effect.Substance abuse and behavior disorder comprise: ethanol, cocaine, nicotine, glucose, Fructus Cannabis, opiates and opiate derivative, gambling, how moving property is forced,, long-term violent behavior and stress disorders, and with the relevant symptom of premenstrual syndrome (PMS).

In order to induce the release of dopamine at volt nuclear place, finally also having related to opium can be active.Therefore wish to unite and use the enkephalinase inhibitor and the enkephalin releasing agent that prevent the enkephalin degraded active to strengthen the opium energy to greatest extent.

The pith of this scheme is first gene type, and provides suitable cocktail treatment according to his/her genotype.Developed suitable HAART about the unusual inventor of D2, and stated in the text.

Synthetic agonist is not ideal treatment reagent.A certain specific agonist can act on several receptors, or the similar receptor on the different cell, and is not only a certain receptor or the cell that we wish.As can produce toleration (by the receptor number with to the change of medicine affinity) to medicine, also can produce and toleration antagonist in the same old way.More special in bromocryptine (bromocryptine) or metamfetamine, itself can produce drug dependence.Bromocryptine can be by Rhesus Macacus self-administration (Woolverton etc., 1984) as everybody knows.

In contrast, D-phenylalanine (DPA) does not have toleration.According to the research of S.Ehrenpreis, do not produce toleration and dependency (Chicago medical college) by 500mg/kg/ days dosed administration for a long time mice.

The reasonability of this combination also is just to work when short releasing agent has only existence to discharge object, they can not be treated DOPA and lack, in fact, the inventor worries that short releasing agent can increase the weight of the long-term lacking of dopamine, from this point, what precursor adopted is the adjusting approach of comparison nature, and only precursor just is converted into neurotransmitter whenever necessary, and organism distributes its product as required then.

Prior art adopted DPA that ethanol and cocaine dependent are carried out anti-addiction therapy.The data of Blum etc. shows: in 90 days open trials that reduce about body weight, they have proposed DPA and 1-tryptophan, the 1-glutamine, and ' phosphoric acid is united use to 2-methyl-3-hydroxy-4-formyl-5-hydroxymethylpyridine .-5.In this research, only reduce by 10 pounds with the collator and compare, the supplement therapy group has on average reduced by 26 pounds.In addition, test group only 18.2% reduction is no more than 15 pounds, and matched group is 81.82%, and these results show that the overweight whose body weight reduction of using this product is 2.7 times of the person of not using.

The inventor finds to go on a diet in the program at 247 influencial families, the low-calorie in 2 years of people of diagnosing a disease, group that replenish to take in DPA, other aminoacid and trace meter with take core vitamin group and compare, show: 1) comprise masculinity and femininity, 2 times reduction is all arranged on overweight percentage ratio.2) women's addiction reduction by 70%, the male reduces by 63%.3) women's gluttony behavior reduces by 66%.Man's surname reduces by 41%.In the seminar of going on a diet, body weight has only been gone up them in 14.7% of the body weight that loses of going on a diet therebetween, and matched group then recovered 41.7%.5) the logarithm regression model shows, supplement therapy, women, morbid obesity and fat family history are the important predictors of weight increase after 2 years.

These data show, use DPA separately or unite use with other amino acid precursor, trace meter such as chromium picolinate or nicotinic acid chromium, during 2 years, special at the prevention weight increase or eradicate body weight and can produce dramatic effect aspect rising again, up to today, the inventor does not also find have this series products can call oneself so great discovery on market, comprise and Redux ^(Interneuron, Cambridge, quality) or Fastin  (Smith/Kline Beacham, Philadelphia, Pennsylvania).

In addition, also having carried out some is difficult to reduce body weight and interrupts the principle research of the body weight after the fat-reducing back keeps alleviating about the overweight people, these studies confirm that body is how " to defend " a specific body weight, so-called " set point ", in the face of the calculated energy expenditure of fat-reducing reduces, body by increase hunger sensation, reduce oxygen consumption, gastronintestinal system more effectively absorbs nutrition, thyroid hormone secretion keeps " set point ", this point had had description (Keesey, 1989).In addition, body weight reduces, and the influence that health care or Drug therapy etc. are regulated disappears, cause the stable perhaps more piece trencherman of body weight to be described as the phenomenon of " platform ", even body weight go up, from this point replacement therapy, not only can prevent body weight, as if can also overcome platform effect.

List is with fenfluramine and unite use phentermine and PHENCAL ^TMComparison: the result who uses the d-fenfluramine is that retainable body weight reduces, and returns to normal level although food is freely taken in.Up to today metabolic rate after the test of humans and animals does not all prove administration can increase.And when using fenfluramine, as if when energy expenditure, it can both strengthen energy expenditure, and this medicine can both strengthen the heat effect of food to humans and animals.The ability that its is strengthened energy expenditure can not produce toleration, to it in the effect aspect the apositia then different (Levitsky and Troiano, 1992).

For novel diet products, most important treatment effect is to prevent that the body weight that reduces from ging up, in fact, it is the purpose of fat-reducing strategy that the persistent body weight of advising Turner reduces, and according to the evidence of the present parties concerned, as if the d-fenfluramine can both play the effect that reduces body weight and not produce toleration reaching in the year, toleration is that restriction uses other drug to grow the problem that course of therapy should disease.Although the d-fenfluramine impels those that dissatisfied patient who reacts of strategy that other reduce body weight is reduced body weight by serotonergic system, the follow-up investigation prompting of carrying out subsequently to the maximum duration of d-fenfluramine, to serious adiposis patient, want to keep body weight to reduce and answer continued treatment.With regard to the rise that prevents body weight, the d-fenfluramine seemingly works by the 13-25% that reduces caloric intake every day, and it also can reduce the serious hope to rich food.Viewpoint of the present invention is: use PHENCAL ^TMThe effect that prescription can reach far surpasses the therapeutic effect that d-fenfluramine and other analeptic etc. have reached, in addition be better than by first of FDA's approval be used for slimming medicine the therapeutic effect that should reach.The inventor believes, adopts amino acid precursor to introduce and the enkephalinase inhibition, is the Therapeutic Method that up to the present state-of-the-art prevention body weight is gone up.

The preferred therapeutic scheme: the basic scheme of treatment RDS should comprise following a kind of material at least, uses separately or unites use.This table comprises the main component (concrete prescription sees Table 6-16) of running after fame and being used for the treatment of with medicine or neurologic agent:

Table 20

The effective dosage ranges that the composition of therapeutic combination is considered

D-phenylalanine 16 to 5000mg

D1-phenylalanine-3,4-quinone 2 to 10,000mg

Naltrexone 1 to 1000mg

Tyrosine-arginine 15ug to IS mg or

Tyrosine-D-arginine (at same dosage range)

Chromium picolinate 10ug is to 10000ug

Chromium nicotinic acid chromium 10ug is to 10000ug

L-carnitine 1 to 200mg

L-tyrosine 9 to 90,000mg

L-glutaminate 3 to 30,000mg

L-tyrosinase 15 to 5,000mg

5-hydroxyl-tyrosine 0.5 to 500mg

L-arginine pyroglutamate 1 to 1000mg

Hepa-merz 1 to 1000mg

D-leucine 16 to 5000mg

DL-leucine 32 to 10,000mg

Hydroxycinnamic acid 1 to 100mg

Theanine (Taiyo Intl) 1 to 1000mg

Huperzine 1.5 to 1500ug

Vitamin B6 (pyridoxol HCI) 1 to 1000mg

Rhodioia rosea extract 5 to 500mg

(Pharmline)

Expect that other enkephalinase inhibitor also is useful (patent before seeing, United States Patent (USP) the 5th, 189, No. 064, the 4th, 761, No. 429, Canadian Patent the 1st, 321, No. 146 in these Therapeutic Method.Here be used as list of references and quote).

Carbohydrate gluttony behavior or obesity chemical compound: in this, the aminoacid that the front was narrated is introduced with enkephalin and is suppressed to influence many major issues relevant with body weight, the most important thing is to prevent body weight rise after 2 years.

The detailed protocol of anti-SUD: Alcotrol ^TMAnd Cocotrol ^TMUse according to gene type.Be used in improved preparation on the original patent basis, in an open trial, estimated amino acid precursor introducing and enkephalinase and be suppressed at the effect that prevents ethanol and cocaine dependence proband recurrence.280 routine patients have been observed in this research between 2 years, the standard of recurrence is the evaluation according to the expert who qualifies: whether the proband recovers regularly to use ethanol and cocaine.Patient is divided into 2 groups: group A=is the one-A-Day vitamin only, and B=is to alcohol addiction Alcotrol for group ^TMA kind of, to cocaine addiction Cocotrol ^TMA kind of.

After 2 years, matched group has 78 people recurrence in this research, and relapse rate 86% does not have 1 example recurrence with 189 patients of aminoacid group, and relapse rate is 0.Results suggest is preventing that very positive effect is arranged in patient's SUD recurrence, and this effect has increased typical treatment in the resulting result of patient on one's body in fact, and they accept 12 traditional steps, near rehabilitation.

Alcoholism and cocaine abuse person's classification: proband's gene type is A type and Type B.Researcher empirical tests the alcoholic is divided into the notion of A type and Type B, and find now this notion to research cocaine abuse effective equally.Hypotype is meant to have the individual further classification of one or more common traits and the system of research.Classification can be understood risk factors such as gene, personality and environment better in the complex interactions that causes on the alcoholism to alcoholic's hypotype, and influenced by these risk factors and the bounce-back that takes place.

Alcohol abuse is more serious than A type the Type B alcoholic.As if the Type B alcoholic has several characteristic: it is more relevant with inherited genetic factors than A type person; More often occur in the male; Type B has more impulsion property and tendency has serious alcohol abuse family history; They have more Childhood conduct disorder, more serious alcohol dependence, multiple medicines thing abuse and mental disorder, especially antisocial personality.

For the cocaine abuse person, certain damage factors, as alcoholism or drug dependence family history, pursue sensory behavior and the childhood period conduct disorder etc. be more prone to cause its more serious Type B cocaine to rely on.Therefore, the cocaine abuse person (A type) that other does not have above-mentioned feature may mainly be subjected to the influence of society, environment rather than heredity, mental state or spiritual influence and produces dependency.

Can find the importance of 3 kinds of main features on one's body Type B cocaine abuse person, as risk factor before sick (the substance abuse family history, the childhood period conduct disorder and ADHD, pursue the behavior of sensation and the age of beginning Drug abuse); The variable of substance abuse is (as the number of times of cocaine use, a large amount of times of using cocaine, the symptom that cocaine relies on, the symptom of alcohol dependence, the multiple medicines thing uses, medical science and social consequence) and spiritual problem (as the seriousness (Ball etc., 1995) of depressive symptom and antisocial personality obstacle and above-mentioned spiritual problem.

In addition, in the score of pursuing aspects such as sensation, attack, crime, violence and social adaptation weaken, Type B person mark is higher than the A type.The former uses amount, number of times and the persistent period of cocaine also to be higher than A type cocaine user.The Type B person also more is prone to the use side effects of pharmaceutical drugs, as lose consciousness, violence (and other), and to give up painful ratio higher to alleviate to use more the multiple medicines thing in them.It is young more that age of Type B cocaine abuse person of addiction symptom is regularly used, uses, occurs for the first time every day for the 1st time in the 1st use, the 1st excessive use, the 1st time.

Two kinds of hypotypes use cocaine and the 1st appearance to rely on the time span of symptom at the 1st time; Use approach such as nasil, smoking or injection; The tactful number that control is used; The aspects such as persistent period of giving up forbidden drug or ethanol in the past do not have notable difference.What is interesting is that most of participation experimenter is divided into Type B, but in the hospitalization patient, the quantity of A type and Type B is almost equal.In the experimenter of outpatient and participation treatment, the 75%th, the A type.

One aspect of the present invention is that the type B behavior determination flag of new molecular gene diagnostic techniques and establishment is in the past combined, and is ethanol and cocaine abuse person typing more accurately.The parameter that new genetic discovery is relevant with Type B astoundingly match (Ball etc., 1995).The report of nineteen ninety-fives such as a tree name Ball, most Type B parameters are relevant with DRD2 type gene, and evidence is from the polymorphism research (seeing Table 20-A) of some relevant non-Spain white man d2 dopamine receptor gene gene types.

Table 20-A

Cocaine abuse person's genotype classification

Reason polygenes 53% Noble of category-B DRD2A1 list of references abuse problem etc., risk factor family history before more p of 1993 sex male＜0.05 are sick (father and mother p＜0.016 Noble etc. at least, 1993

One of be alcoholic)

ADHD p＜0.0001 Comings etc., factor regular use cocaine p＜0.030 Noble etc. for the first time childhood period of 1991,1993

Impermissible behaviour before (CD)

Have 3 risk factor: FH+ p＜0.007 Noble etc., 1993

Strong cocaine uses use p＜0.015 of the strong cocaine of CD substance abuse variable

(the intravenous average % time

, free alkali, and crack)

From using for the first time cocaine to p＜0.033 Noble etc., 1993

The equispaced week number that uses next time

Table 20-A (continuing)

The repeatedly medication of category-B DRD2A1 list of references substance abuse (every kind of medicine p＜0.0005 Comings etc., 1994 seriousness spend above 25 dollars) weekly

(comprise cocaine and its at all medicines

Its analeptic is except that opiates) on

All spend more money p＜0.01 Comings etc., 1994 initial age A1 carrier=23.2 years old p＜0.026 Comings etc., 1994

The A2 carrier=the psychopathology order of severity was higher in 26.7 years old, had more antisociality Comings etc., 1994

The childhood period violence p＜0.002

Adolescence violence p＜0.0001 Blum etc., 1996

Violent crime p＜0.011 Comings etc., the high impulsion property of 1994 personalities is sought sensation

(pathological gambling) p＜1 * 10 ^-8Comings etc., 1994

Division sample/avoidance disease p＜0.006 Blum etc., 1997

(indifferently with separate)

ADHD ADHD proband's subject matter is and can not focuses one's attention on.What the inventor was recent studies show that one of them is called Kantroll ^TMThe a series of electric physiology result of recipe-imposed (seeing Table 6A)

Known to the inventor, this is that the first relevant people eats the be correlated with report of ability (ERP) of a kind of special ispol influence incident relevant with cognitive performance every day.Cognitive ER is produced by two kinds of computerized vision attention projects in normal youth volunteer's test, three-dimensional location project (SOT) and accidental persistence performance project (CCPT).Each was tried to take in before amino acid precursor 28-30 days and the back is detected under the situation that is subjected to my control fully.After 2 kinds of projects were mixed, (the p＜.009) and the quickening of cognitive process speed (p＜.015) appearred strengthening significantly on the statistics in ERP composition P300 amplitude.The enhancing of function of nervous system and RDS patient's (as substance use disorders, attention deficit hyperactivity disorder, carbohydrate gluttony) the promotion rehabilitation after replenishing aminoacid is consistent in the viewed normal control in this research.

This is operated among the ADHD proband and does not also finish.But along with a series of ADHD cases many moving reports that alleviate with the improvement of learning capacity after replenishing aminoacid, its reasonability also is tangible.The inventor has data to show, and is pointed as this paper front, and DRD2A1 allele and P300 and AER and VER are unusually also relevant.Be also shown in Comings about the work of Fructus Cannabis to the P300 wavelength affects.All these make and adopt special formulation (HyperGen ^TM) and be that enkephalinase inhibitor (dl-phenylalanine) treatment necessitates at least.Utilize this principle inventor also will to obtain relevant DRD2A allele and be called the data that concerns between the standard computer ADHD test of TOVA.

Attention is concentrated obstacle.One aspect of the present invention is that the treatment attention is concentrated obstacle and other RDS related syndromes.The inventor is to be subjected to neurotransmitter to control and can treat by the fact that specific amino acid precursor is regulated certain special neurotransmitter that normal brain cognitive function is responsible for according to the attention centralized procedure.Should be to the understanding of the electrophysiological function of brain owing to the biogenesis aspect that relates to the Chemical Regulation material that attention concentrates.

Cognitive competence infringement (detecting by the P300 wavelength) that a nearest problem of being concerned about is the alcoholism child and patient's ADD/ADHD attention are concentrated the ability reduction.About this point, support such notion on evidence, be many destructive youngster's phases and adolescence behavior disorder-comprise ADHD, Tourette syndrome, learning disorder, substance abuse, disobey obstacle and conduct disorder and be interrelated behavior-a part, it has very strong genetic constitution, is polygenic inheritance, share a series of genes that influence dopamine and 5-hydroxy tryptamine and other neurotransmitter, and from father and mother both sides.Concentrate the observation prompting of relevant attention deficit hyperactivity disorder: it preferably is divided into by behavior and a kind ofly self regulates or carry out the obstacle of function.Neural imaging on the anatomy is discovered, relevant regulating loop comprises prefrontal cortex and ganglion basal, they are regulated by arranging from the dopaminergic nerve of midbrain, and discharge the stimulant adjusting that activates dopamine receptor by agonist activity or dopamine.

The invention provides and can strengthen the neurotransmitter control action, cause discharging naturally of dopamine, overcoming PHD and occupying d2 dopamine receptor by the synapse dopamine stimulates its outgrowth compositions (comprise and use phenylalanine).

5-hydroxy tryptamine precursor 5-hydroxy tryptamine (5-HTP) is a CNS mediator, and it is present in intestinal chromaffin system and the platelet.The biosynthesis of this neurochemical is by earlier L-tryptophan hydroxyl being changed into 5-hydroxy tryptamine acid, then latter's decarboxylation formed 5-hydroxy tryptamine.Hydroxylating (rate-limiting step) is finished by tryptophan hydroxylase.Decarboxylation is finished by ubiquitous L-aromatic acid decarboxylase, this enzyme require pyridoxal 5-phosphate fellowship.

5-hydroxy tryptamine is metabolized to the 5-hydroxyindoleacetic acid by monoamine oxidase, MAO.This metabolite passes through homaluria subsequently.Mesencephalic centre 5-hydroxy tryptamine mechanism plays an important role in some neuroendocrine mechanism of control mood, behavior, locomotor activity, feed and hypothalamic control hunger, thermoregulation, sleep, hallucinatory state.

The life-time service cocaine can reduce the concentration of 5-hydroxy tryptamine and metabolite thereof.Cocaine obviously reduces the absorption of 5-hydroxy tryptamine precursor tryptophan, therefore reduces the synthetic of 5-hydroxy tryptamine.Can activity (Reith etc., 1985) thereby the activity of tryptophan hydroxylase that also reduces cocaine reduces 5-hydroxy tryptamine.

Enkephalin and endorphins content in hypothalamus rather than brain, have been increased simultaneously with the drug treating rat (fenfluramine, PCPA or 5-7-DHT) that consumes 5-hydroxy tryptamine.Because do not have mRNA and proenkephalin precursor (PE) or popiomelanocortin content to change, the prompting 5-hydroxy tryptamine can conduct the utilization of regulating the opiates polypeptide rather than influence it and synthesize.

Following hypothesis is supported in this discovery, i.e. the release of enkephalin minimizing causes the active reduction of dopamine, shows as depressive state.After aggravating activities, some behavior disorder such as pain, depression, sleep disordered can the generation.Replenishing 5-hydroxy tryptamine with the L-tryptophan can conduct and help the mental state of recovering positive.Having been found that provides tryptophan in diet, as replenishing precursor there is definite influence (Moir and Eccleston, 1968) in the metabolism of brain 5-hydroxy tryptamine and related compound.The content of cerebrum 5-hydroxytriptamine depends on the level (Fernstrom and Wurtman, 1971) of blood plasma tryptophan.Feed to lack tryptophan food, the brain 5-hydroxy tryptamine level of mice is very low, and the L-tryptophan can make its recovery.If the L-tryptophan is expelled in the blood, brain tryptophan and 5-hydroxy tryptamine level can raise 9 times and 2 times respectively.To having the department of neurology patient of depressed and insomnia defeated simultaneously, can make 6 times (Gillman etc., 1981) of cortex tryptophan levels rising with tryptophan.

8 healthy males are carried out the double blinding crossing research (Lieberman etc., 1983) of tryptophan (50mg/kg), tyrosine (100mg/kg) or placebo, and tryptophan rather than tyrosine can obviously ease the pain.Other studies show that the toleration (Hosobuchi etc., 1980) that tryptophan can alleviate clinical pain (Seltzer etc., 1983), prevention of migraine (Poloni etc., 1974) and reverse analgesic.Thereby tryptophan by 5-hydroxy tryptamine can the activation release that strengthens dopamine reach pain relieving.

Be different from tyrosine hydroxylase, under the normal physiological situation, tryptophan hydroxylase is not saturated, that is to say and has not given play to maximum effect.Therefore, the activity of tryptophan hydroxylase is subjected to the influence of L-tryptophan significantly.The quantity of operational free L-tryptophan is determined by many factors, comprises the uptake rate of concentration and its brain and the presynaptic terminal of circulation L-tryptophan in the blood plasma.The inventor expects that employing L-tryptophan or 5-HTP can recover by the destructive serotonergic system of cocaine.

5HTP effectively treats reagent.The speed that the L-tryptophan enters brain depends in the blood plasma freely the ratio with bonded tryptophan, and this ratio is subjected to that neural aminoacid, insulin and drug concentrations influence in the blood, and protein binding site and tryptophan in these materials competition blood plasma are taken in the site.In addition, 5HTP not only by the 5HT neuron also by other neuronal uptakes.Therefore synthetic the increasing of 5HT is not limited in the 5-hydroxy tryptamine neuron.

The 5-HT synthetase inhibitors comprise irreversible tryptophan hydroxylase inhibitor (the DL-fenclonine, 6-fluorine tryptophan, L-propyldoracetamide) and 5HTP decarboxylase inhibitor (carbidopa and 1-methyl-5HTP).5-hydroxy tryptamine mode with exocytosis under the influence of action potential and medicine is discharged into synaptic space.Cocaine, (+)-amphetamine, metamfetamine, fenfluramine, can promote the release of 5HTP to chlorine amphetamine, clomipramine and amitriptyline.

What propose at present has 3 kinds of 5HT acceptor types (5HT-1 ,-2 ,-3).The 5HT receptor stimulating agent comprises LSD, quipazine, N, N-dimethyl-tryptamines (DMT).The antagonist of 5HT receptor comprises Cyproheptadine, methysergide, LSD, 2-bromo-CDS (BOL), ketanserin, xylamidine tosilate, cinanserin and 1-(-)-cocaine.The active transport mechanism that the inactivation of 5HT relates to the energy dependence of high-affinity is used for the 5HT of synaptic space is sent back to presynaptic neuron.

The material that suppresses neuronal uptake 5HT comprises three ring anti-depressant (imipramine, desipramine (desimipramine), amitriptyline, clomipramine, fluvoxamine, fenfluramine (a kind of fenisorex)) and cocaines.The 5HT of any non-storage form will be changed into metabolite by MAO, yet if MAO is suppressed, 5-hydroxy tryptamine will be metabolized to N-methyl or N-N-dimethyl product by O-methyl-transferring enzyme (COMT).

The reinforcing agent and the releasing agent of the reinforcing agent of opiates polypeptide/releasing agent opiates polypeptide.One aspect of the present invention is to use the material that suppresses the degraded of opiates nerve polypeptide.These opioids promote the synthetic of dopamine and discharge.Can increase striatum DA biosynthesis and metabolic speed for the animal opioid, this effect is positioned at the special opiate receptor that black striatal dopaminergic do not hold and is regulated (Clouet etc., 1970; Biggio etc., 1978; Regiawi, 1990).Give beta-endorphins and enkephalin for a long time and can produce dopaminergic toleration (Iwatsubo etc., 1975; Arden etc., 1972).Postsynaptic DA receptor responsive unusually (Schwartz etc., 1978) in tolerific animal.

Cocaine also influences opium can neural activity.After rat contacts cocaine for a long time, can observe the bonded change of naloxone of dose dependent.The density of opiate receptor obviously reduces on several brain structures, but increases in the hypothalamus outside, and this shows that the combination of opiates is subjected to the influence (Hammer etc., 1978) in " recompense maincenter " rather than other zone especially.In this external another research, naloxone has been blocked the thresholding (Bain and Korwetsky, 1987) of brain recompense maincenter reduction cocaine effect effectively.As if cocaine influences the analgesic effect (Misra etc., 1987) of some opiates preparation in addition.The inventor believes that the invigoration effect of cocaine can be partly by the opiate system adjusting of brain recompense maincenter, and the latter changes by long-term contact cocaine.

It is found that anesthetics can be used for various " opiate receptors ".Endogenous opiate (EO) is contained in brain and other nervous tissue.In brain, also find relevant pentapeptide, methionine and leucine enkephalin (Hughes etc., 1975).Enkephalin activates δ and μ receptor, and the β endorphins activates epsilon receptor.The endocrinologist shows that having been recognized a kind of B-lipotrophin (B-LPH) of doing pituitary hormone contains 5 amino acid whose MET-enkephalin sequences and be hydrolyzed into activated opioid beta-endorphins (Li etc., 1976).

The known EO chemistry family that has 3 kinds of separate sources, difference in functionality at least of present inventor is although all polypeptide all contain the sequence of tyrosine-glycine-glycine-phenylalanine-X at the N end.Endorphins family comprises big precursor, opium prohormone (pro-opiocortin), and B-LPH and beta-endorphins, second EO family is enkephalin family.Met-enkephalin and leucine enkephalin are all come the big polypeptide precursor of self-contained 2 kinds of sequences.Respectively there are 1 or 2 basic amino acids to be connected to six peptides and seven peptides and a kind of seven peptides of enkephalin c-terminus: the intermediate (Hexum etc., 1980) that [Met] enkephalin-Arg-Phe seemingly exists naturally.The 3rd family is kappa agonist such as dynorphin 1-13 and 1-17.The effect of these CNS composition antagonism morphines.Dynorphin may change the change (κ is to δ) that inferior endorphins form (E5) can cause receptor affinity into as precursor, the formation N end of leucine enkephalin, illustrates that may there be new regulating action in the enzyme of regulating ligand expression.

Polypeptide from each family seems all not only as neurotransmitter but also as neuro hormone.5 peptide enkephalins are positioned at the not tip of nerve, discharge from neuron behind the irriate.Leucine and met-enkephalin are discharged into the blood from adrenal medulla, are used as neuro hormone.Beta-endorphin is discharged into the blood from hypophysis cerebri, as the neurotransmitter (Bloom etc., 1978) of brain zones of different.Endorphins and enkephalin all produce the reaction of biochemistry or medicine reason, comprise toleration, dependency and sexual repression, give EO with humans and animals after the caused reacting phase of narcotic analgesics seemingly.Endogenous opioid is the same with anesthetics to be the member of " opium " family.The enzyme of degraded enkephalin (E5) is referred to as " enkephalinase ".

Contain the multiple metabolic peptidase of pentapeptide enkephalin (E5) that makes in the known tissue at present.The enzyme that acts on similar enkephalinase comprises soluble and the bonded amino peptidase of graininess (Hersh, 1981), other act on glycine 3-phenylalanine-4 position as peptidyl dipeptidase or metal inscribe peptidase (Benuck etc., 1982; Schwartz etc., 1980).Metallocarboxypeptidase A makes enkephalin be cracked into the terminal dipeptides of tyrosine-glycine-glycine-C and methionine-phenylalanine or Ile-Phe.With mainly be responsible for making the biogenic amine of its inactivation different at target site by single enzyme, the degraded of enkephalin needs multiple enzyme, although the seemingly main enkephalinase of metal inscribe peptidase.Following scheme shows and the relevant enzyme site of action of E5 degraded.

A strategy that prevents the E5 degraded provides its substitute.For stoping of the degraded of various histaminases to enkephalin, must carry out several chemical modifications, these comprise: a) N end tyrosine is modified, and makes the tyrosine modification analog of methionine can prevent degraded (Coy and Kastin, 1980); B) 2 introduce of the effect of 1 D-aminoacid in the position with the blocking-up amino peptidase; And/or c) modifies or introduce 1 D-aminoacid at position 3-5 and act on the effect of the enzyme of glycine 3-phenylalanine 4 keys with blocking-up peptidyl dipeptidase or other.Other analog comprises D-alanine-enkephalinase amide or FK33-824, as MU agonist, enkephalin-arginine-phenylalanine as delta agonists and dynorphin 1-13 or 1-17 as kappa agonist (Wisler etc., 1981).

Whether the E5 agonist of not clear these candidates has the problem aspect potential addiction tendency, toleration and other toxicology when clinical use up till now, many these modifieds, to the probability of the substitute addiction of antienzyme, significantly reduced their clinical practice.

Second kind also is preferably to increase enkephalin in vivo or the active strategy of enkephalin is to use specific enzyme inhibitor, some enkephalin segment (glycine-glycine-phenylalanine-methionine or glycine-glycine-Phe-Leu 2: PN: WO2004057976 PAGE: 32 claimed protein, phenylalanine-methionine, Phe-Leu 2: PN: WO2004057976 PAGE: 32 claimed protein), can be used as the inhibitor of enkephalin, and enkephalin form that may be bigger itself just has inhibition.

In the present invention; " enkephalinase inhibitor " is including but not limited to D-phenylalanine (DPA); DL-phenylalanine (DLPA); hydrocinnamic acid and D-aminoacid such as D-leucine; expect that other enkephalin inhibitor also has special significance; they are selected from some protein synthesis inhibitor (bacitracin, bestatin, and puromycin); polypeptide amino acid (free D-type monamino acid; the dipeptides and the tripeptides of D-type essential amino acids, the sulfydryl benzyl amino acid (as 2-{ sulfydryl-3-hydrocinnamoyl }-the L-leucine), the carboxyalkyl methyl ester is (as N-{ (R; S)-2-carbethoxyl group-3-{ phenyl-propanol }-the l-leucine }; benzomorphans-enkephalin, and unrelated compounds such as quinalbarbitone on other structure, pyrophosphate; the O-phenanthroline; phosphamide, Z-leucine-NHOH, Z-glycine-NHOH.Dipeptides such as D-phenylalanine-D-leucine and D-phenylalanine-D-methionine, with polypeptide such as L-tyrosine-glycine-glycine-D-phenylalanine-D-leucine, with L-tyrosine-glycine-glycine-D-phenylalanine-D-methionine, and the D-phenylalanine, D-leucine and hydrocinnamic acid.

Proved that the D-phenylalanine can suppress hydroxyl PEPA A (Hartruck and Lipscomb, 1971), and proved that recently it also has pain relieving (Ehrenpreis etc., 1978; Della Bella etc., 1979) and antidepressant effect (Beckmann etc., 1977).

For estimating the effectiveness of D-phenylalanine as enkephalinase inhibitor, it can obviously reduce the degraded (Gail etc., 1983) of oligopeptide (D-alanine 2-D-leucine) enkephalin (DAPLA) and tyrosine-D-alanine-glycine-phenylalanine (TAAGP) on the mice intestinal mucosa through evidence.Yet, the D-phenylalanine suppresses the activity very low (Amsterdam etc., 1983) from the enkephalinase A of Medulla Bovis seu Bubali and B in vitro tests, what is interesting is, after only adding 1 aminoacid formation dipeptides D-phenylalanine-tyrosine, just can increase significantly and suppress to render a service.

The D-phenylalanine can suppress the degraded of enkephalin and beta-endorphins, and it is better than the enzyme of catalysis B-enkephalin for the effect of the enzyme of catalysis enkephalin decomposition.Its activity or tissue-specific, at hypothalamus, 80% enkephalinase is suppressed, and endorphine enzyme is 5%; In cortex, enkephalinase is 60%, and endorphine enzyme only is 18%; At striatum, brain coffee enzyme is 78%, and endorphine enzyme is 10%; At spinal cord, enkephalinase is 84%, and endorphine enzyme is 40% (Ehrenpreis etc., 1981).Other studies show that the injection DPA after after 90 minutes, methionine one enkephalin has increased by three times in that CNS is actual, and after 6 days, also maintain higher level (Balagot etc., 1983), after the increase of mouse brain Met-enkephalin was found in the injection hydrocinnamic acid similarly, it was a kind of known D-phenylalanine metabolite.

Another aspect of the present invention is that enkephalinase inhibitor and enkephalin releasing agent are combined, and the theoretical basis of doing like this is that the inventor can obviously strengthen the effect (as δ or μ) of enkephalin at separately opiate receptor position.For reaching this purpose, the inventor preferably uses tyrosine-arginine dipeptides (kyotorphin) and its stable analog tyrosine-D-arginine, and the latter has analgesic effect, and increases intracellular Ca2+ in the synaptosome at Mus brain striatum position.These materials are the Met-enkephalin releasing agent seemingly, its model of action the unknown (Ueda etc., 1986).

In order not only to suppress enkephalinase but also to increase the release of neuron enkephalin, the consumption that is called material (tyrosine-arginine) every day of kyotorphin should be between 15ug-15mg.Can replace it as enkephalin releasing agent (Tajima etc., 1980 with the more stable analog (tyrosine-D-arginine) of dosage between 15ug-15mg every day; Ueda etc., 1986).Therefore, a kind of enkephalin releasing agent can with the combination of brain coffee enzyme inhibitor, in synapse, reach high-caliber enkephalin neurokyme activity, with the release of further increase neuron dopamine.This can be used as a kind of mode of " replacement therapy ", reduces " addiction " and other RDS behavior as described herein to cocaine.This treatment will be the most effective in behind the cocaine detoxification 12 months.

The precursor of γ-An Jidingsuan (GABA).GABA is a kind of inhibitory nerve mediator (Gessa etc., 1985) that the control dopamine discharges.As if it reduce the epilepsy after the ethanol withdrawal.The main path of synthetic γ-An Jidingsuan (GABA) is with the decarboxylation of L-glutamic acid by glutamate decarboxylase (GAD).The same with other amino acid decarboxylases, this enzyme require vitamin B6 (pyridoxal 5-phosphate) is as cofactor.GAD exists only in the cytoplasm of synapse GABA teleneuron.The synthetic key of control GABA is GAD, and it also is the synthetic rate-limiting step of GABA.GABA suppresses to influence the activity of GAD by end-product.Be in saturatedly in the concentration of presynaptic neuron L-glutamic acid, therefore, the material concentration of increase can not influence the aggregate velocity of GABA, so the exogenous L-of giving glutamic acid can not obviously increase neurotransmitter GABA, unless the horizontal abnormality of L-glutamic acid is low.There is test to show, continues to take the content that glutamine can significantly increase (500ng/kg, every day) glutamate, Glu in the brain, GABA and taurine for the inductive adult albinism mice of different ethanol by drinking-water in 10 days.Glutamine is the effective intermedium that ammonia is transported out brain, the catabolism of different aminoacids in the tissue that therefore can affect the nerves greatly.Behind deaminizating, glutamine can become the precursor of glutamate, Glu, thereby is the precursor (Thawki etc., 1983) of GABA.

Have 2 kinds of GABA receptors at least: the GABA-A receptor is very sensitive to the competitive retardation of bicuculline and Radix Cocculi Orbiculati picrotoxin or Radix Cocculi Orbiculati picrotoxinin, and these receptors are positioned on the post-synaptic structures, regulates typical GABA inhibitory action.And the GABA-B receptor is positioned at the presynaptic end, and is insensitive to the retardation of bicuculline.Not only scalable GABA is in the release of CNS for the GABA-B receptor, and scalable NE is from the release at some position of sympathetic nervous system.

The someone some dysfunction such as dyskinesia, HuntingtonShi chorea, insane disease, alcoholism etc. is clinically proposed may be relevant with the GABA system.The GABA receptor is regulated by the albumen of a kind of crying " GABA regulin " all to the change of the barbiturates binding site of the benzodiazepine binding site of the affinity of GABA, benzodiazepine and barbiturates.It is the same that the GTP relevant with the adenyl cyclase associated receptor regulates albumen, and the activity of GABA regulin is determined by phosphorylation.

GABA is usually relevant with the inhibitory neuron of the substantia gelatinosa [of Rolando at hypothalamus, Hippocampus, brain ganglion basal, posterior horn of spinal cord and retina place.The aixs cylinder path that in the CNS some is long also can be active relevant with GABA through assert.

Gaba agonist comprise acetic acid imidazoles, sulfonic acid 3-aminopropane and THIP (4,5,6,7-tetrahydrochysene-isoxazoles also-the pure and mild 5-aminomethyl-3-hydroxyisoxazole of in agaricfly, finding of [415-C]-pyridine-3-(3-hydroxyl-5-amino-methyl isoxazole).The GABA antagonist comprises bicuculline, Radix Cocculi Orbiculati picrotoxin, Radix Cocculi Orbiculati picrotoxinin and benzylpcnicillin.

The dependent shooting system of sodium that has a high-affinity in presynaptic GABA teleneuron and neuroglia composition makes its deactivation thereby the GABA that discharges removed from ECS.The inhibitor that GABA takes in comprises: neuron absorption type: as DAB and suitable-2, the amino thiacyclohexane carboxylic acid of 3-; Neuroglia absorption type: as the B-alanine; Other absorption type: nipecotic acid, benzodiazepine class and, neuroleptics and tricyclics.

GABA discharge and with acceptor interaction after can come back to presynaptic neuron, be a recycling neurotransmitter therefore.GABA carries out metabolism and conversion at teleneuron and glial tissue by enzyme, as become succinic acid semialdehyde by mitochondrion GABA aminotransferase (GABA-T) effect in the presence of the oxo glutamic acid of A-.The succinic acid that forms enters tricarboxylic acids (Krebs) circulation.GABA-T needs pyridoxal 5-phosphate as cofactor.Succinic acid semialdehyde is oxidized into succinic acid by succinic semialdehyde dehydrogenase rapidly, and succinate dehydrogenase also will use NAD and NADH as cofactor.Consider this point, the inventor adds the promoter of pyridoxal 5-phosphate as the oxidation-reduction path in prescription.

From this point, the animal and human, can increase GABA concentration for following GABA-T inhibitor, as ethanolamine-P-phosphoric acid, γ acetenyl GABA, γ vinyl GABA, gabcuculline, hydrazinepropionic acid, two-N-propyl-acetic acid sodium (valproate sodium), amino glycolic (vitamin B6 inhibitor), L-glutaminate (Bloom, 1985).

The precursor catecholamines dopamine (DA) of catecholamine (dopamine, norepinephrine), norepinephrine (NE) and epinephrine (E) all are neurotransmitteies.Catecholamine is the chemical compound that has 2 adjacent hydroxyls (OH) on phenyl ring.In vivo, this class material is aromatic amino acid L-tyrosine hydroxyl to be turned to L-3,4 dihydroxyphenylalanines (L-DOPA) and synthetic by tyrosine hydroxylase.L-tyrosine is taken in the plain energy of adrenalectomy teleneuron effectively.The L-phenylalanine is precursor (Blum and Kozlowski, 1990 of L-tyrosine; Schwartz etc., 1992).

Tyrosine hydroxylase is positioned at the cytoplasm of nor-parathyrine serotonergic neuron, is the synthetic rate-limiting enzyme of NE.Studies show that in a large number reduced form pteridine cofactor, molecular oxygen and ferrous ion are essential to activity.In endochylema, the L-DOPA is by being DA with pyridoxal 5-phosphate (vitamin B6) as the L one AADC decarboxylation of cofactor.Dopamine is initiatively taken in the granular storage vesicle, under the effect of DOPA-B hydroxylase the DA hydroxylating is formed norepinephrine (NE).This enzyme require copper, molecular oxygen and ascorbic acid are as cofactor.In some neuron of CNS, NE further is transformed into epinephrine (E) by phenylethanolamine N transmethylase.

The activity of tyrosine hydroxylase is influenced by following factors: " end-product " suppresses, and caused by NE concentration increase in the teleneuron, and the speed that L-tyrosine is converted into the L-DOPA reduces; The increase of sympathetic activity in the CNS, thus the synthetic of NE increased; The synthetic increase of NE of Angiotensin II mediation; Agonist of adrenoreceptor (as clonidine) and blocker (as phentolamine) change the rate of release of NE by the adrenoreceptor that is positioned at the presynaptic teleneuron.

The inhibitor of NE synzyme comprises: methyl-P-tyrosine (inhibition tyrosine hydroxylase), carbidopa (being suppressed at the AADC in the outer tissue of CNS), with the dithiocarbonic acids diethylester, the FAI63 and the sulfur (inhibitor of dopamine-B-hydroxylase) that relieves the effect of alcohol

NE is stored in a plurality of regions of anatomy of teleneuron with the form of multiple storage complex.It is particulate composite that a kind of NE stores type, is positioned at the vesicle of noradrenergic nerve tip.Particulate composite comprises dopamine-B-hydroxylase and Mg++, Zn++ and Cu++ by being attached to the NE on the ATP and being referred to as several albumen of doing chromograin and forming.

To be an active transport process in DA and the NE absorption storage vesicle, need ATP to provide energy and Mg++ to activate the dependent ATP enzyme of Mg++, this Mg++ is dependent to be taken in DA and NE the process of storing vesicle and DA and NE and passes neuron membrane to enter neuronic process be separate different, and the latter is Na ⁺/ K ⁺-ATP enzyme is dependent.

The stability of NE-ATP-albumen-ion storage complex can be destroyed by some chemical compound as the Mg++ intercalating agent.The magnesium deficiency that long-term cocaine abuse person occurs sometimes may be relevant therewith.In this regard, give the answer that cocaine can increase NE for a long time.

NE is to finish by the exocytosis of a Ca-dependent from the release of teleneuron.Vesicle film and plasmalemma merge, by NE, and ATP, the Vesicle-Containing that dopamine-B hydroxylase and chromograin are formed is discharged into synaptic space.But a known control NE need rely on the presynaptic receptor of the teleneuron that is positioned at release NE to the mechanism of the availability of postsynaptic receptor.A shooting system that is positioned at the presynaptic teleneuron can make NE leave synaptic space, thereby NE is terminated in the effect of synaptic space.There are two kinds of neuronal NE to take in mode: mode I, mode II.

Mode I is an energy dependence, need be by the ATP of the dependent ATP enzymatic degradation of sodium.This is the process of a high-affinity, that is to say the low concentration NE that can remove synaptic space effectively.The neuron shooting system is transported to teleneuron with NE, is stored in the vesicle at these most of NE, and the inhibitor of this process comprises cocaine, tricyclic antidepressants, amphetamine and tyramine.

Mode II relates to the gathering of NE in the non-neuron tissue.Because the NE blood plasma high level that medulliadrenal stimulation or intravenous injection catecholamine cause, can be absorbed by non-nervous tissue such as liver, muscle, connective tissue.NE or other catecholamine material can be diffused into blood circulation again, and more are to be destroyed by monoamine oxidase, MAO (MAO) and catecholamine-O-transmethylase (COMT) in cell.

MAO is present in all and contains in the mitochondrial tissue, and is combined on the adventitia.In the active tissue of liver, brain, nerve, muscle and all metabolism MAO is arranged all, it is c with the NE oxidative deamination, the 4-dihydroxy-mandelic acid, the latter methylated by O-subsequently (by COMT) be 3-methoxyl group-4-hydroxyl mandelic acid.MAO is actually one group of isomerase with different tissues distribution, substrate, inhibitor and physical characteristic.For example, it is substrate that MAO A likes with NE and 5-HT, can be suppressed by the clorgiline selectivity.It is substrate that MAO B likes with olopamine and phenethylamine, can be suppressed by deprenyl (selegiline) selectivity.Other MAO inhibitor of knowing comprises: iproniazid, niaguitil (nialamide), tranclypromine, and phenelzine.

It is found that to have a large amount of COMT in the hepatocyte, in CNS, COMT does not act on and is taken in and the E and the NE of inactivation again by neuron.The inhibitor pyrogallic acid works by the dependent Methyl transporters process of retardance COMT, and this transfer relates to arrives NE with a Methyl transporters from S-adenosine-L-methionine, on the catechol ring 3 ' hydroxy position of E and DA.Dopamine is the precursor of E and NE, and plays an important role in CNS and more autonomic neuroganglions.

The DA of high concentration suppresses tyrosine hydroxylase by end-product inhibition in the neuron, thereby reduces the synthetic of DA.In addition, the synthetic rate-limiting step of DA is that tyrosine under tyrosine hydroxylase is done is to the conversion of L-DOPA.Under normal circumstances, tyrosine hydroxylase is saturated by L-tyrosine fully, and therefore, the tyrosine level can not be accelerated the synthetic speed of DA in the increase circulation.Yet, when DA amount reduces and tyrosine hydroxylase for example is subjected to the influence of cocaine and when being destroyed, this fact will change.

The L-DOPA is initiatively taken in the CNS neuron and is changed into DA at this.After the treatment of L-DOPA, DA is synthetic and that store, and amount obviously increases.Compare with dopaminergic system, after the treatment of L-DOPA, NE synthesizes only a small amount of increase.

Dopamine is stored in to be stored in the granule, is combined with each other at this catecholamine and chromograin, bivalent metal ion and ATP.DA is considered to be discharged into synaptic space by exocytosis.As NE, this is the process of a Ca-dependent, and produces when the action potential that arrives teleneuron or medicine are reacted.Following material can increase DA and discharge: cocaine, (+)-An Feitaming, metamfetamine, tyramine, amantadine, m-phenmetrazine, phentermine and nomefensine.Except causing that DA discharges, these chemical compounds also suppress the reuptake of neuron to DA to some extent.

After DA was released to synaptic space, its effect was stopped by neuronic reuptake system, and this system is a high-affinity, energy dependence, active transport process, and is similar with the NE system of once describing.MAO and COMT respectively DA are converted into 3-4-dihydroxyphenyl acetic acid (DOPAC) and 4-hydroxy-3-methoxy-.alpha.-toluic acid. (HVA, 3-methoxyl group-4-hydroxyl-phenylacetic acid) is responsible for.Cocaine is by prolonging the effect of DA at synaptic space to the retardance of DA being taken in again the presynaptic teleneuron.

The rising of brain tyrosine level causes the synthetic increase of brain L-DOPA, and the metabolism subsequently of L-DOPA is a dopamine.After giving tyrosine, the synthetic and release of dopamine increases.Tyrosine in the food can increase the answer and the release of dopamine and norepinephrine, but does not increase catecholamine levels.Thereby pressure, cold and some drugs can cause the neural level that increases the catecholamine that makes teleneuron that consumes to reduce.

The L-phenylalanine is an essential amino acids, and it also is dopamine and the synthetic precursor of norepinephrine neurotransmitter.By detecting metabolite HVA, DOPAC and the MHPH of these neurotransmitteies, find that they obviously change when aggravating activities and physical fatigue.The L-phenylalanine can cause dopamine to exhaust the back in order to replacement at cocaine abuse, or recovers dopamine with L-cheese ammonia cheese or L-DOPA and store.

Adopt these precursors to can be used as or influence that dopamine, norepinephrine, epinephrine are taken in again or suitably the replenishing of each stage of the reagent of degrading treatment with dopaminergic releasing agent, blocker, agonist or antagonist.Yet, the more important thing is to comprise that whole dopaminergic activities synthetic and that discharge are subjected to the adjusting (as enkephalin and endorphins) of some opiates polypeptide on some degree.Central is given the opiates polypeptide all can cause the blood plasma catechlolamine level at humans and animals rising (Clouet, 1982).In fact, retardance presynaptic dopaminergic receptor can strengthen the release of beta-endorphins, shows the interactive relationship of a uniqueness.The chemical compound that can be used as precursor comprises L-tyrosine, L-phenylalanine, pharmaline.

Rhodiola Rhosea extract (pharmaline) .Rhodiola Rhosea or golden root are a kind of herbaceos perennials of Crassulaceae, are grown in the arctic and area, Alps.In Montes Altai, thing Berli Asia, Tien-sdhein and the Far East Area, people successfully grasp the cultivation technique of Rhodiola Rhosea.Promptly can again can be from plant propagation (Polozhy etc., 1985 from seed; Saratikov and Krasnov, 1987).Its rhizome contains phenolic compounds, wherein the most important thing is p-tyrosol (tyrasol) and its glucosides rhodioside (salidroside), and it has determined the biological activity (Saratikav etc., 1968) of Rhodiola Rhosea preparation.Rhodiola has stimulation and causes adaptive character, and it is considered to improve is engaged in blue-collar ability, lessens fatigue, and the recovery time after the shortening muscle over loading, makes cardiovascular kinetic energy normalization.In high strength muscular work process, prevent losing of micurgic phosphoric acid in brain and the muscle by optimizing oxidative phosphorylation process, stablize the activity of lipid in muscle, the metabolic index of enhances skeletal flesh (activation) to the aminacyl-t-RNA-synzyme, increase the content of RNA, increase the especially blood supply of brain (Saratikov etc., 1968 of muscle; Saratikov, 1974).Rhodiola can strengthen attention scope, memory, improve mental work and physical work.With this active relevant brain region is thalamus cortex and PH (Marina etc., 1973).Rhodiola is also relevant with other multiple effects: prevent hyperglycemia and hypoglycemia, leukocytosis and minimizing, erythrocytosis and minimizing, hypoxia, the protection action of the heart stress and be played in minimizing.The effect of stress regulating of Rhodiola makes the effect of hypophysis-adrenal system and opium energy system normalization relevant with it.People also find the antitumor tolerance of Rhodiola energy enhancing body, obviously suppress the growth of experimental tumor, reduce the frequency of neoplasm metastasis, prolong the life cycle of tumor recurrence animal, reduce the consequence (Dementyeva and Yaremenko, 1983) of spontaneous tumor.Show on evidence that also Rhodiola can alleviate neurosis, to resisting fatigue (Saratikov, 1977).

The rhodioside (SAL) of rhodioside (a kind of Rhodiola extract) the 30mg/kg inductive animal brain NE of sulfur that can prevent to relieve the effect of alcohol reduces.Rhodioside influences brain NE by the activity that suppresses COMT and MAO, rhodioside does not reduce the ability that catecholamine and 5-hydroxy tryptamine precursor see through blood brain barrier (BBB), this characteristic makes it to the present composition, and the scheme of especially treating attention disorders is very useful.(contain rhodioside, the aglycone tyrosol rosavin), can increase the concentration of DOPA in the brain neopallium, dopamine (DA) and 5-HT, reduces NE level in the caudatum to give 0.2mg/kgrhodosine in not having the mice that hinders.Other has the people to prove the subcutaneous injection rhodioside after 30 minutes, and epinephrine (EPI) and DOPA level there is no change.When giving 30mg/kg dosage, NE content reduces 26%, and 5-HT content reduces 15%; When giving the dosage of 100mg/kg, the concentration of NE, DA, 5-HT reduces 20%, 28%, 23% respectively.In the test of taking L-DOPA (50mg/kg) and 5-HTP (100mg/kg) to mice, show, compare, take rhodioside (30mg/kg) and can make exogenous DOPA of animal and 5-hydroxy tryptamine raise 26% and 13% respectively with saline-DOPA-5-HT contrast.These data disclose said preparation has increased the permeability of blood brain barrier to the catecholamine precursor.In addition, Petkov (1981) studies show that, rhodioside has reduced the activity of MAO, has suppressed the activity of COMT, and the catecholamine that therefore slows down methylates and the deactivation of oxidative deamination mode with O-.Further have test to show: rhodioside does not change the activity of 5-HTP decarboxylase, and therefore, it does not influence synthetic to 5-hydroxy tryptamine by 5-HTP, but by slight inhibition MAO, slows down the biotransformation of amine.Obviously, relate to the increase of uniting 5-hydroxy tryptamine in the research midbrain that gives 5-HTP and rhodioside, strengthen blood brain barrier by rhodioside the permeability of 5-HTP is controlled.

Document has shown some interactions between Rhodiola and the neurotransmitter kinetics, conclude having of getting up to have reported: dopamine is in the minimizing of Fu Hezhong, the increase of NE in Hippocampus, in the increase of hypothalamus 5-HT, and the agonist-like activity of cholinoceptor.Some mechanism of same-action is accepted about various neurotransmitteies rise not on cognitive question in neuroscience.Choline can neuromechanism be the fixed basis of mnemonic trace.The brain noradrenergic system strengthens the positive reinforcement effect, and 5-hydroxy tryptamine can mechanism relate to the consolidation of memory process more.

Someone is by using several effects (Petkov etc., 1986) that have the positive escape method research Rhodiola of negativity and positive reinforcement effect to rat.Employing has the labyrinth method that negativity (punitive) is strengthened, and can improve learning and memory after finding to give every rat Rhodiola extract 0.1ml.In memory test, use same dosage to treat 10 days continuously, can obviously improve longterm memory.Employing has " stair " method of positive reinforcement (food), gives every rat 0.1ml Rhodiola extract, has also obtained favourable effect in training process.On the contrary, adopt other method, learning and memory is not produced actual effect for every rat 0.01ml Rhodiola extract, prove that this alcohol-water puies forward the discordance of thing.

The albinism rat is used to study meclofenoxate and the Rhodiola effect (Lazatova etc., 1986) to convulsibility galvanic shock memory impairment.With the dosage peritoneal injection meclofenoxate of 100mg/kg body weight 5 days, after training the 3rd, the 24th hour, through keeping evidence: it can prevent the caused retrograde amnesia of convulsibility galvanic shock.On the contrary, although the Rhodiola extract can improve learning and memory in other experiment, per os takes Rhodiola extract 0.10ml, not generation effect in continuous 10 days every rat.

The huperzine huperzine is a kind of chemical compound that belongs to acetylcholinesteraseinhibitors inhibitors, and it can suppress to destroy the enzyme of acetylcholine, and acetylcholine is a kind of important neurotransmitter or brain chemical substance, it is believed that learning and memory very important.Huperzine is naturally occurring chemical compound, is separated to from Lycopodium clavatum Huperzine Serrala the earliest.Be used as Chinese medicine, carried out limited clinical trial as the treatment amnemonic medicine relevant recently in China with the age.Results suggest can be improved learning and memory in some patient.Yet the result of this hint property is not confirmed by clinical trial.This natural materials is considered in this patent in the desired compositions to influence the attention centralized procedure.The recommendation using dosage that is used to strengthen man memory is 150ug/ days (the therapeutic scope of medication is 1.50 to 1500mcg/ days).

Huperzine A compares external the effect of the memory impairment that brought out by scopolamine and the effect of E2020 and tacrine (tacrine), use be operation of radial labyrinth and acetylcholine esterase inhibition method.Scopolamine (0.2mg/kg) has damaged spatial memory significantly in rat.Huperzine A (oral 0.1-0.4mg/kg) and E2020 (0.5-1.0mg/kg is oral) and tacrine (1.0-2.0mg/kg is oral) can lack reverse to the memory that these scopolamines bring out.Huperzine A, E2020 and the tacrine of being determined by colorimetry is to butyrylcholine esterase: the ratio of acetylcholinesterase is respectively 884.57,489.05 and 0.80.The result has proved that huperzine A is to have optionally acetylcholinesteraseinhibitors inhibitors most, and, improved the memory shortage of bringing out more significantly than E2020 or tacrine by scopolamine, this shows that it may be a kind of medicament likely (Cheng etc., 1996) that can be used for alzheimer's disease patient's cognitive impairment clinical treatment.

Huperzine A is a kind of new effective reversible selectivity acetylcholinesterase (AchE) inhibitor, and people expect that it is more outstanding than other acetylcholinesteraseinhibitors inhibitors of the treatment that is used at present to remember shortage in alzheimer's disease patient.After measured huperzine A influence (people such as Zhi, 1995) that the rat of being handled by AF64A-is showed in radial labyrinth.(every limit 2nmol, i.c.v.) ability to mouse execution space working memory operation has caused obvious impairment to AF64A.The damage of this behavior and choline acetyltransterase (ChAT) active remarkable minimizing in Hippocampus is relevant.Huperzine A (0.4-0.5mg/kg, lumbar injection) has improved the memory that is caused by AF64A significantly and has lacked.These results hint AF64A be a kind of by changing the Hippocampus cholinergic function useful reagent of destruction work memory process, and such damage can be improved ..1995 such as () Zhi effectively by huperzine A.

Huperazon ^TMMain ingredient be the proprietary extract of Lycopodium clavatum Huperzia serrata, be used for the treatment of alzheimer's disease.Studies show that China carries out: the active substance huperzine A of this extract is the likely new therapy that is used for alzheimer's disease.Other the huperzine A that studies show that is a kind of good acetylcholinesteraseinhibitors inhibitors (AchE), has the half-life in the outstanding penetration of CNS and the noticeable body.The double blinding clinical trial of carrying out in China proves: huperzine A is safe, and is effective to the long-term treatment of alzheimer's dementia.Except having the acetylcholinesteraseinhibitors inhibitors activity, nearest discovery shows that huperzine A also has other neuroprotective sexual function: huperzine A can suppress the cytotoxicity by glutamate induction in the culture of the newborn Hippocampus of rat and cerebellar neuron; Huperzine A promotes the branch of the dendron in the neuron culture.

The characteristics of alzheimer's disease are neuronic unusual and degenerations, and these neuron normal activities and survival ability are to rely on acetylcholine and acetylcholinesterase.These neurocytes that are positioned at the forebrain basilar part are also with relevant as Parkinsonian other sacred diseases.Huperzine A is to compare Cognex on activity ^With effective inhibitor of the outstanding acetylcholinesterase of E2020, Cogex ^Be first medicine that is approved for treatment alzheimer's disease in the U.S., E2020 be recently by Eisai Pharmaceuticals permit medicine.In addition.Huperzine A has shown in tissue culture and can neuroprotective unit to have avoided the death that caused by excitatory amino acid glutamic acid.Because huperzine A has dual pharmacological activity, Huperazon ^TMJust the treatment for attention deficit and old memory shortage provides a kind of uniqueness important activity.Studies show that of the toxicology of huperzine A and effect, even administration of human therapeutic dose 50-100 dosage doubly, it does not have toxicity yet.When extract is 2ug/kg at dosage, can onset 6 hours and do not have noticeable side effect.

In alzheimer's disease, use the Weschler scale to measure, 160 above patients' double blind control result of study shows: only every day twice 150ug dosage (3-5ug/kg) just can make the patient obtain significant improvement.In the patient evaluation that is undertaken by its caretaker, huperzine A and placebo are compared, use 11 patients of placebo to report that aspect sanity improvement is arranged, and 26 use huperzine A patients report has this improvement, the patient of 8 use placebo is proved to be and has improved memory, and the patient of 16 use huperzine As has improved memory, and, the patient of 1 use placebo is proved to be has linguistic progress, and the patient of 8 use huperzine As has this progress.

In the comparison that the memory of carrying out between the patient of patient who uses huperzine A and use piracetam improves, verified: the patient of the patient of 50% use piracetam and 85% use huperzine A has improved memory, the patient who has also confirmed to improve significantly memory has 30% in using the piracetam patient, in using the huperzine A patient, have 70%, and, only there is the patient of 15% use huperzine A to be proved to be the improvement that does not have memory, uses the patient of piracetam to have 50% not to be proved to be and to improve.

Huperzine A has 2 important features can be used for being different from the experimental compound of Cognex  and E2020 and other researchs.Huperzine A has very high specificity to brain acetylcholinesterase (AChE) rather than the acetylcholinesterase in the health other places.The lower toxicity that it is believed that this selectivity and extract has relation.In addition, different with E2020 with 2 kinds of medicine Cognex  that are approved for treatment alzheimer's disease, huperzine A has been proved to be shortage to being present in receptor among the CNS such as the combination of M-ChR M1 and M2, and this combination can cause side effect.

The active persistent period of huperzine A (3 hours) will be grown than Cognex  (2 hours) and physostigmine (30 minutes).In the behavioral study about the learning and memory reinforcement that carries out in animal, the difference concerning memory and study between effective extract doses and the non-toxic effect dosage (from toxicity research) is 30-100 times.These data have hinted that effectively huperzine A can effectively treat alzheimer's disease and have only MIN side effect.

Chromium in chromic salts (as pyridine carboxylic acid salt, the nicotinate etc.) food is the nutrient of needed by human, and its value aspect human nutrition has had final conclusion.To the interest of chromium from a kind of view, promptly because chromium is a kind of essential trace quantity mineral and the cofactor of insulin, so by the stiffening effect to insulin action, it can work in glucose, lipid and aminoacid metabolism.The support of this words is according to being that people observe the chromium deficiency result and cause the infringement of glucose tolerance, insulin resistant, blood glucose to raise and the disease of type ii diabetes; In addition, the chromium of an amount of physiologically active can reduce the demand (.1996 such as Kaats) of insulin in the human body.

State-run academy of science classifies chromium as trace mineral, and recommends to take in every day 50～200ug.But, the most reliable research report: in the American (also similar concerning other country), this intake be time good dosage-only be woman minimal requirement 40%, man's 60%.Human research more than 25 points out: replenish chromium the people who lives is at home had useful effect, comprise and improve glucose, insulin and lipid levels, improve the glucose tolerance of damaged; Reduce the cholesterol levels that the adult has raise; Improve insulin and hypoglycemia patient's situation (Mertz, 1992).

In order to increase the bioavailability of chromium, several researchs have proposed to use pyridine carboxylic acid, and it is the derivatives that the tryptophan natural metabolism produces.It seems that pyridine carboxylic acid can combine in intestinal and blood with trace metal ion, promotes the collection and the utilization (Evens and Bowman, 1992) of trace mineral.

Because the lipidosis of health it seems that part is to be regulated by insulin, insulin utilizes the improvement of situation should cause the minimizing of lipidosis.Because insulin instructs aminoacid to enter muscle cell, so the effect of improving insulin can be woven with definite influence to muscle groups; In case aminoacid enters muscle cell, they are assembled by the influence of insulin pair cell hereditary material DNA and ribonucleic acid becomes protein.This effect of chromium is very important to the present invention, because can reduce some aminoacid such as valine and leucic competition like this, thereby makes the amount of tryptophan increase (Wurtman, 1982).Insulin also makes the proteinic disintegrate of health or decomposition slack-off, and by network effect, increase can be used to build the proteinic amount of tissue.Chromium can impel fat-free matter (FFM) to be maintained to potentiality or increase.The someone advises: if CrP can reduce insulin resistant, it just can improve health and constitute, because insulin resistant or shortage cause leading to the glucose and the damage of amino acid whose passage of muscle cell, increased the decomposition of muscle protein, and the potentiality that strengthened the acceleration lipid deposition that insulin deficit had (people such as Kaates, 1996).Other document shows: insulin resistant helps the obese patient and stablizes body fat, promptly is on the fat level; Play " set point " effect, the prevention body weight further increases (Eckel, 1992).In general, although zooscopy has been supported this opinion ..1993 such as () Liarn, human body has been discovered the variation of determining that additional CrP causes people such as (, 1992) Hasten on health is formed; Another report also is sure, although from the statistics angle, the composition of health does not significantly change people such as (, 1993) Hallmark; And the 3rd report fails to find that the CrP supplementary function can produce any definite variation (people such as Clancey, 1994) aspect the health composition.These report that controversial essence is: the study subject sample size that most human research use is very little, and, study subject often just enters motion or adjusting program subsequently, thereby has increased the demand of chromium, and the amount that can provide in these researchs quantitatively has been provided.

The previous observation chromium supplementary function and the coefficient work of training are limited in body wt and constitute go up on the observation on effect, and the result is mutual contradiction (people such as Clancy, 1994; People such as Evans, 1989; People such as Evans, 1993; Hallmark etc., 1996.Hasten etc. 1992).

The pyridine carboxylic acid salt of chromium is to use, studies and promote maximum chromium compounds, still, in vitro study work suggestion nicotinic acid chromium may lose weight and the variation of health formation aspect be great-hearted.About this point, the very recent work of inventor discloses nicotinate may be more important than pyridine carboxylic acid salt ..1997 such as () Grant.Here these data of Ti Chuing are as the illustration of the serviceability of nicotinic acid chromium, and are added in the basic compositions in the specific embodiments of present patent application.

Pharmaceutical composition waterborne compositions of the present invention comprises all cpds of the effective dose of the relevant obstacle of the RDS of being used for the treatment of disclosed by the invention, these obstacles comprise obesity, ADHD, Tourette syndrome, PMS, smoking and other behavior of setting forth hereinto, these compound dissolutions or be dispersed in pharmaceutically acceptable carrier or the aqueous medium.Phrase " pharmacy or pharmacology go up acceptable " is meant such molecular entity and composition, and promptly it does not produce disadvantageous, hypersensitive or other undesirable reactions when giving the suitable administration of animal or human.

" pharmaceutically acceptable carrier " comprises any one and all solvents, disperse medium, coating thing, antibacterium and antifungal, isotonic agent or delay absorbable preparation or the like as used herein.Such medium and reagent are known by those skilled in the art to the using method of pharmaceutically active substance.Unless incompatible with active constituents of medicine, traditional sucrose or reagent all can be used in the therapeutic combination.The active component of complementarity also can be mixed in the pharmaceutical composition.When taking to the people, said preparation must meet in Food and Drug Administration's biological preparation standard the standard about aseptic, pyrogen, Generally Recognized as safe and purification.

Biologic material need be through large tracts of land dialysis removing unwanted small-molecular weight molecule, and/or carry out lyophilization on demand and be formulated in the carrier of wanting preparing at any time.Reactive compound generally is made into the dosage form of parenterai administration, for example passes through through intravenous, intramuscular, subcutaneous, local even Intraabdominal injection administration.According to disclosure of the present invention, the preparation that contains the waterborne compositions of active component has been that those skilled in the art are well-known.Typically, such compositions can be made into the solution of liquid or the injection of suspension form; Also can be prepared into and before injection, add the suitable solid form for preparing solution or suspension in the relevant liquid; In addition, preparation also can be an emulsion.

The medicament forms that is suitable for injecting use comprises: aseptic aqueous solution or dispersion; The preparation that comprises Oleum sesami, Oleum Arachidis hypogaeae semen or aqueous solution of propylene glycol; Be used for preparing the sterile powder of injectable sterile solution or dispersion then and there.In whole examples, dosage form must be aseptic, must be that fluid is to be easy to pass through syringe needle.It must keep stable under manufacturing and condition of storage, must be kept under the condition of microbial contaminations such as avoiding class bacteroid and fungus.

Go up the reactive compound of acceptable salt form as free alkali or pharmacology, can be in water with surfactant for example hydroxypropyl cellulose suitably mix and make solution.In addition, also can in glycerol, liquid macrogol and their mixture and oil, prepare dispersion.Under common storage and service condition, comprise in these preparations that antiseptic is to prevent the growth of microorganism.

The chemical compound of the RDS of being used for the treatment of associated disorders of the present invention can be made into the composition of neutral form or salt form.Pharmaceutically acceptable salt comprises acid-addition salts (forming with proteinic free amine group group), with mineral acid for example hydrochloric acid or phosphoric acid, perhaps with organic acid such as acetic acid, oxalic acid, tartaric acid, mandelic acid or the like formed salt.In addition, also can be by the formed salt of free carboxy group: sodium hydroxide, potassium, ammonium, calcium or hydrated ferric oxide. and organic base such as 2-aminopropane., trimethylamine, histidine, procaine or the like from inorganic base, for example.As active component, quote 4,608,251 with peptide therapeutics as a reference here; 4,599,230; 4,596,792 and 4,578, No. 770 United States Patent (USP)s, its technology can be used.

Carrier can be a kind of solvent or disperse medium, comprises for example water, ethanol, polyhydric alcohol (for example, glycerol, Polyethylene Glycol of propylene glycol and liquid or the like) and their suitable mixture and vegetable oil.Suitable flowability is kept with following method, for example by using coating thing such as lecithin; By keeping the particulate size in the decentralized photo and passing through to use surfactant.Prevention to microbial activities can be used various antibacteriums and antifungal preparation, for example parabens, methaform, phenol, sorbic acid, thimerosal or the like.Under many circumstances, preferably contain etc. and to ooze preparation such as sugar or sodium chloride.The absorption delay effect of Injectable composition can produce by the reagent that use to postpone absorbs in said composition such as aluminum monostearate and gelatin.

The preparation process of sterile injectable solution is: reactive compound is joined in the appropriate solvent of aequum, mix with the composition of above-named various needs, then filter sterilization.In general, the preparation process of dispersion is, various active component through sterilization processing joined comprise in basic disperse medium and above-mentioned those other the required composition sterile carrier enumerated.For the sterilized powder that is used for preparing sterile injectable solution, preferred manufacturing procedure is vacuum drying and Freeze Drying Technique, and the generation active component adds the powder from other required composition of above-mentioned filtration sterilization solution.Also the expection preparation is used for the denseer solution of direct injection, and anticipation is used DMSO as solvent, can cause the infiltration that is exceedingly fast, with active agent delivery to a less zone of high concentration.

As for the preparation of medicine, solution will be with the form administration that conforms to dosage form, and dosage should be effective in treatment.Medicine is formulated into various dosage forms so that administration: for example can be the injectable solution form of setting forth above, but also can be drug release capsules or the like.

As for the parenteral aspect of aqueous solution, for example, solution should suitably be cushioned in the case of necessary, and diluent at first will reach etc. with competent glucose or normal saline and ooze.These special aqueous solutions are particularly useful for intravenous, intramuscular, subcutaneous and intraperitoneal administration.About this point, the aseptic aqueous medium that will use is well-known according to of the present invention disclosing those skilled in the art.For example: the medicine of certain dosage can be dispersed in the isoosmotic NaCI solution of 1ml, the subcutaneous injection that perhaps is added into 1000ml is with in the liquid, perhaps injected (referring to " Lei Mingdun pharmaceutical science " the 15th edition, 1035-1038 page or leaf and 1570-1580 page or leaf) in specified injection site.According to the condition of the study subject of receiving treatment, the variation of some necessity can take place in administration.Under any circumstance, the people of responsible treatment will determine individual's suitable dose.

May be formulated in the mixture of treatment usefulness at this active component of being set forth, content is about 0.0001～1.0 milligram, or about 0.001～0.1 milligram, perhaps about 0.1～1.0 milligram even about 10 milligrams/dosage.In addition, also can take a plurality of dosage.

Be used for the parenteral form except chemical compound is mixed with, beyond the dosage form as intravenous or intramuscular injection, the acceptable form of other pharmaceutics comprises: for example: the tablet of oral administration or other solid form; The liposome form; Capsule form by time release; And the form of any other current use, comprise ointment.

In addition, in the present invention, solution or spray, aerosol or the inhalant that also can use per nasal to suck.The solution that per nasal sucks is aqueous solution normally, is to prepare to splash into or spray delivery through nasal meatus.The solution that per nasal sucks is produced similarly to the secretion of nose aspect a lot, and like this, normal cilia activity is maintained.Thereby the aqueous solution that per nasal sucks is normally isoosmotic, and is slightly cushioned, to keep 5.5～6.5 pH value.In addition, if necessary, be similar in those the antimicrobial antiseptic that use in ophthalmic preparation and suitable medicine stabilizing agent be also included within.The nasal preparation of various commercializations is known, comprises for example antibiotic and the antihistaminic that is used for prevention of asthma.

Other dosage form that is suitable for other administering modes comprises vaginal suppository and pessulum.In addition, rectum holder or suppository also can be used.Suppository is the solid dosage forms of various weight and shape, inserts rectum, vagina or urethra after adding medicine usually.After the insertion, the suppository softness is got up, is melted or is dissolved among the intracavity fluid.In general, for suppository, traditional binding agent and carrier can comprise: for example poly alkylene glycol or triglyceride; Such suppository can be formed by the mixture of the active component that contains 0.5%-10%, is preferably the active component that contains 1%-2%.

Comprise excipient commonly used in the oral formulations, for example, pharmaceutical grade mannitol, lactose, starch, magnesium stearate, saccharin sodium, cellulose, magnesium carbonate or the like.These compositionss adopt solution, suspension, tablet, pill, capsule, sustained release forms, perhaps form of powder.In certain embodiments, combination of oral medication will comprise a kind of inert diluent or absorbable consumption carrier, perhaps, they can be enclosed in the hard or soft gelatine capsule, perhaps, they can be pressed into tablet, and perhaps, they can directly be incorporated among the food of diet.To the administration of oral therapeutic drug, reactive compound can combine with excipient, with edible tablet, and buccal tablet, lozenge, capsule, elixir, suspension, syrup, forms such as wafer are used.Such compositions and preparation should comprise at least 0.1% reactive compound.Certainly, the percentage ratio of composition and preparation can be changed, be more conveniently from Unit Weight 2% to about 75%, perhaps more suitably be between 25-60%.There is the appropriate amount of the reactive compound in the therapeutic combination should be able to obtain suitable dosage this.

Tablet, lozenge, pill, capsule etc. can comprise following composition: binding agent such as tragakanta, acacin, corn starch or gelatin; Excipient such as dicalcium phosphate; Disintegrating agent such as corn starch, potato starch, alginic acid or the like; Lubricant such as magnesium stearate; And sweeting agent, resemble sucrose, lactose or glucide; Perhaps flavoring agent such as Herba Menthae, oil of wintergreen or Fructus Pruni pseudocerasi flavoring, they can be added into.When dosage unit form when being capsular, it not only can comprise the material of the above-mentioned type, can also comprise a kind of carrier of liquid phase.Various other materials also can be used as the coating thing or exist for the physical form that changes dosage unit.For example: tablet, pill or capsule can be by Lac, sugar or the two quilts that wraps.The elixir syrup can comprise reactive compound, and with sucrose as sweeting agent, methyl parahydroxybenzoate and propyl ester also comprise dyestuff and flavoring agent as antiseptic, as Fructus Pruni pseudocerasi and mandarin orange flavor.

Test kit treatment test kit of the present invention comprises mentioned reagent or the chemical compound that one or more are used for the treatment of RDS and relevant behavior.Such test kit generally will comprise in proper container: a kind of pharmaceutically acceptable formulation forms or aforesaid any pharmaceutically acceptable formulation forms.Test kit can have single container (packing), and perhaps, it can have different separately container (packing) to each chemical compound.

When the component of test kit is a form with one or more liquid solutions when being provided, liquid solution will be a kind of aqueous solution, especially preferred aseptic aqueous solution.In addition, the treatment chemical compound () that is used for the RDS associated disorders also can be formulated into and be injectable compositions.In this case, container itself may be exactly syringe, pipette or like instrument, and the composition for preparing can be applied to certain part of health by it, injects in the animal body, perhaps mixes with other ingredient of test kit and uses.

But, the component of test kit also can be used as dry powder and is provided.When reagent or composition were provided as dry powder, dry powder can the reconstruct by adding suitable solvent.In addition, can imagine: solvent also can be provided with another kind of package means.

The following example provides in order to prove the preferred embodiments of the invention.Those skilled in the art will recognize that the technology representative that in the following example, discloses by inventor's those technology that find, that in practice of the present invention, go on well, and can think that this is the optimal way of this engineering practice.But,, those skilled in the art will recognize that: under the situation that does not break away from spirit of the present invention and scope, can also carry out many changes to disclosed specific embodiments and still obtain similar or close result according to disclosure of the present invention.

Embodiment 1

Familial hyperphagia patient is loaded aminoacid and suppresses the enkephalinase introduction

The inventor thinks that RSD lacks the reaction that is produced to one or more neurotransmitteies.Attempting to alleviate the unbalance of neurotransmitter by medicine-receptor activation will only be to substituting that recompense (award) lacks, and will only produce temporary transient peacefulness sense.For this consideration, the inventor shows: by taking precursor aminoacid and enkephalinase inhibitor, using the neurotrophy material to repair the brain chemical substance and lack, is (Blum etc., the 1988a that important facilitation is arranged for the recovery from illness of some uncontrollable trophic behavior (as SUD); Blum etc., 1988b; Brown etc., 1990).Whether inventor's decision is to loading precursor aminoacid and suppressing enkephalin and can assess this problem of potentiation of having kept that loses weight of outpatient, and the time limit is 2 years.Method:

The selection of study subject and program: the study subject of this research is that (Behavioral Medicine Group Clinicin Sacramento California) carries out 247 influencial families that a kind of utmost point low-calorie replenishes the fasting programmed treatment people that diagnoses a disease in the behavioral medicine group clinic that California Sa lattice sliding doors is prevented stock prices from falling.Study subjects use Optifast  as trophism fasting product, reduce to target weight up to their body weight to differ in 15% the scope, perhaps when the patient selects no longer to continue fasting till.The inventor thinks: other dietary amount recipes of under internist's care, leaving can replace Optifast  and same effectively.Standard city resident's life insurance (Standard Metropolitan Life Insurance) height/body weight form is used to determine ideal body weight.All study subjects are all taken core vitamin (Centrum vitamins) during whole research.Each study subject all agrees to participate in research after fully being informed, according to a fat research agreement, this current draft also obtains the approval of " behavioral medicine doctor organizes clinic meeting examination board " and " being positioned at the health science center examination board of association of University of Texas of San Antonio ".

Whether to take PHENCALTM about the patient ^TMDecision after patient's fasting finishes, make.Patient and those main suits that those complaints obtain the highest gluttony score are resisting some carbohydrate, feel the patient of difficulty the most during as the temptation of foods such as confection (sweet food), bread, Citrus, pasta, and those are considered to more refractory aspect weight reduction or actually reach the inconvenient patient in target weight aspect.Complain that maximum patients can not control maybe that the patient of feed is selected to come out to take PHENCAL ^TM(seminar; N=130).Show to such an extent that be easy to if keep treatment, just do not provide PHENCAL such patient ^TM(matched group; N=117).For the selected study subject that comes out, PHENCAL ^TMTherapy when the end of fasting, begin and keeping treatment during continue to carry out, up to the end of the researchs in 2 years.PHENCAL ^TMBeing the supplement of a seed amino acid and vitamin, also is specific PHENCAL ^TMThe predecessor, by 1899 Co., Ltds (San Antonio, Texas) development with produce, sell by Weider nutriment group (salt lake city, the Utah State).The study subject of picking out is taken six PHENCA every day ^TMCapsule, each capsule contain 460 milligrams of DL-phenylalanine, 25 milligrams of L-tryptophans, 25 milligrams of L-glutamy, 5 milligrams of 2-methyl-3-hydroxy-4-formyl-5-hydroxymethylpyridine .-5 '-phosphate esters, 33 microgram chromium picolinates and 10 milligrams of L-carnitinies.

Patients are before participating in an educational class and will weigh in weekly afterwards.Each patient will participate in the study of this course, the medical science person in charge and a registered dietitian by this plan bear teaching, lesson structure comprises following knowledge: about obesity and its social popularity, the prevention of its potential probability, it is to the influence of personal health and welfare, its treatment, the responsibility of family, the probability of genetic disorder, the usage of trophism supplement, the effect of suitable nutriment in diet, high protein, low fat and notion extremely low or the carbohydrate diet, the effect of motion, or the like.In addition, also to monitor weekly patients' psychology situation and medical condition.The monitoring of psychology situation is according to the degree graduation score of patient to addiction, stability of mood and the gluttony of food.To not take food with feeling well even the expressions in 5 fens on the food addiction scale are uncontrollable fully yet.1 submeter is shown with the feed hope of feeling of guilt, and promptly he knows that he is met, and should not be on the physiology the food addiction.Stability of mood is by with similar point system record, and expression in 5 fens is that emotion is very unstable, and represents MIN emotional lability in 1 minute.What the gluttony index write down is to take food weekly to surpass the number of times of being satiated with food a little.Laboratory hemanalysis and urine analysis carry out every other week, so that relevant medical information to be provided.Blood sample analysis detects is chemical substance and SMAC-complete blood count in the blood, and on the other hand, urine analysis mainly is to detect ketone and/or glucose.Educational class is to carry out weekly one hour, and what course was emphasized is the variation of nutrition, motion, behavior and principle how to handle stress, loses weight and to keep treatment over a long time supporting.

The study subject demographic characteristics: 247 study subjects, 84% is the woman.All study subject all is white people, and the mean age is 40 years old.When entering this programmed treatment, study subject average overweight 74%.

Take PHENCAL ^TM130 research study subjects do not having marked difference with 117 contrast study subjects aspect the family history (seeing Table 20-B) of age, ideal body weight, beginning body weight, overweight percentage ratio, food addiction, anxious state of mind, gluttony or obesity.But they are then having any different aspect the family history of chemicals product dependences (CD+).In the study subject in the seminar 65% has the chemicals product to rely on the family history of (CD+), then has only 30% in the contrast study subject.(p＜0.005)。Complain that the more or out of control a bit study subject of the body weight during weight maintenance just can be selected into seminar because have only those, so these data hint the study subject of those CD+ and have more complaint, and can show more difficultly during whole weight maintenance treatment.

Table 20-B

It is right to be tried

Resemble the sequence number age (year) the overweight % OB% of women % ideal body weight % CD%

Sum 247 39.8 84.2 130.2 74.3 68.1 52.3

The grouping number

PhenCal organizes 130 38.9 83.8 129.1 74.4 64.5 65.4 non-PhenCal and organizes 117 40.7 84.6 131.4 74.3 71.8 39.3 study subject features: each digitized representation meansigma methods.% is overweight=[body weight-ideal body weight during beginning]/ideal body weight; The %OB=report has the percentage ratio of fat family history group.The %CD=report has the chemicals product to rely on the percentage ratio of family history group.Phencal ^TMIt is the product that is positioned at the Weider nutritional companies of 1899 responsibility company limiteies of Texas San Antonio and salt lake city, the Utah State.

Sex difference.In this research, average overweight 76% when 208 woman begin, and 39 men average overweight 66%.Standard city resident's life insurance height/body weight table is used to determine ideal body weight.About woman of 3/4ths (73%) is a morbid obesity, (promptly overweight at least 50% or more), and by comparison, man's morbid obesity number ratio is about half (49%).This in the works the ratio of woman and man's number surpass 5 to 1.In the interview that when beginning plan carries out, have to surpass 90% woman and reported and crave for food, on the other hand, have among the man to be slightly less than 80% people and to have reported this serious hope.Aspect gluttony, suitable evident difference is arranged also between the men and women.80% woman has reported gluttony, and only has 64% people that this kind performance is arranged in the man.

Patient's family history aspect.When this plan of entering, each study subject has been inquired that all relevant chemicals product rely on and fat family history.Be slightly more than 70% woman and 56% man and reported to have fat family history.Have among the patient of fat family history at these, the number (accounting for 73%) that fat mother is arranged approximately is the twice of number (accounting for 38%) that fat father is arranged, has those philtrums of fat family history to have 11% people to report that its father and mother are all fat.Almost respectively there is the people of half all to report the family history that the chemicals product rely among woman and the man.But in this case, mainly be that its father has this feature.Having the chemicals product to rely in the study subject of family history, totally have 86% people to report that their father is chemicals product addicts, is chemicals product addicts and have only mother of 31% people.The study subject report of about morbid obesity of 2/3rds (63%) (overweight at least 50%) has fat mother, and has 60% morbid obesity person to report that their father is the chemicals product addict of certain form.Because the man only accounts for 16% of total number of persons, so the statement of any relevant family history subgroup can only be at most preliminary.The result

What lose weight keeps.247 study subjects are alleviating 68.4 pounds through weight average after the fasting in average 20.0 weeks.Seminar is last fasting, and is beginning to take PHENCAL ^TMDifferent with matched group before, but difference is not remarkable.Seminar is overweight 22% when fasting finishes, and matched group is overweight 32%.If do not consider this species diversity, the body weight the when fasting of statistical analysis proof finishes did not influence losing weight in 2 years.When research finished in 2 years, take PHENCAL ^TMStudy subject average overweight 23.5%, do not take PHENCAL ^TMMatched group average overweight 52.8%, (p＜0.0001) [seeing Fig. 2].Study subject after 2 years in the seminar has only recovered 14.7% of body weight that they lose, and by comparison, the contrast study subject has recovered them and lost 41.7% (p＜0.0001) of body weight.

Influence in the heredity:, find that all take PHENCAL according to family history divide into groups comparison study subject and contrast crowd (OB+/CD+, OB+/CD-, OB-/CD+ and OB-/CD-group) ^TMStudy subject after 2 years than any contrast subgroup all still less overweight (p＜0.0001).With regard to this example, OB is not meant gene, and is meant fat family history.All had patient's group of OB+ family history all to organize overweight more (p＜0.05) than the patient that the OB-family history is arranged after 2 years.But study subject that the chemicals product rely on family history is arranged to PHENCAL ^TMwell reply aspect and other people do not have significant difference on the statistics.

Heredity and sex: see that on the whole all have the male group of family history to compare with women's group, the recovery extent of its body weight is light (p＜0.0001) significantly all.In the best group of situation (OB-/CD+), after the researchs in 2 years finish, the body weight that the man does not virtually completely have recovery to cut during fasting.Because the ratio of woman and man's number surpasses 5 to 1 in this research, and, the woman aspect several characteristic and the man different, the inventor determines further to identify woman's characteristics.70% woman has reported the family history of OB+, and woman's report of 54% has the family history of CD+.Only 12% woman report had not both had the family history of CD-not have the family history of OB-yet.There is the woman of woman's average specific OB-/CD-family history of OB+ and CD+ family history to weigh 58%.And the woman of OB+/CD+ is overweight maximum; Overweight that occupy next is the woman of OB+/CD-, follows woman by OB-/CD+, and that minimum is the OB-/CD-woman.Similarly situation appears at and thirsts for it was reported that the OB+/CD+ woman is showing the most outstandingly aspect food addiction, gluttony and the being happy and angry uncertainly in food, gluttony, feed and the stability of mood score.

To food addiction and gluttony: patient's psychology state and the tracked supervision of medical condition.The psychology state according to clinically to the grade classification scale of food addiction, emotional stability and gluttony by monitoring record qualitatively.To not take food with feeling well even the expressions in 5 fens on the food addiction record are uncontrollable fully yet.1 submeter is shown with the feed hope of feeling of guilt, and promptly he knows that he is met, and should not be on the physiology the food addiction.Stability of mood is by with similar point system record, and expression in 5 fens is that emotion is very unstable, and represents MIN emotional lability in 1 minute.What the gluttony index write down is to take food weekly to surpass the number of times of being satiated with food a little.The patient assesses by inquiry this experimental dietetic compliance.

When research finishes, take PHENCAL ^TMStudy subject compare with matched group, the former food addiction is reduced to 1/3rd (p＜0.0001 at least).In the contrast crowd, the food addiction does not reduce fully.With do not take PHENCAL ^TMThe contrast crowd compare, the number of times of gluttony incident is being taken PNENCAL ^TMStudy subject in be to have reduced significantly.When this research began, study subject was reported the gluttony incident weekly 10.9 times, and when this research finished, study subject was only reported the gluttony incident weekly 2.9 times.In contrast to this, when the beginning of this research, do not take PHENCAL ^TMMatched group report the gluttony incident weekly 8.3 times, when this research finished, matched group was reported the gluttony incident equally weekly 8.3 times, this shows significant change does not take place.After 2 years,, take PHENCAL aspect the addiction and gluttony incident of food ^TMGroup is compared with matched group, and the former is reduced to 1/3rd.

Multiple regression and variance analysis progressively multiple regression are used to check the significance that returns the predictor that increases percentage ratio in the later body weight over 2 years of the beginning of treatment procedure.Whether this predictor is divided into feminine gender (0) or positive (1), thereby shows whether the patient is morbid obesity, in the hardship that stands gluttony and food addiction, the family history that whether has the chemicals product to rely on, whether whether fat family history is arranged, be the women, whether takes PHENCAL ^TMProgressively (SPSS, version 6.13 (SPSS Inc., Chicago, Illinois) selects to use PHENCAL to option program ^TMTreatment, women, morbid obesity and fat family history are as the significance predictor of weight increase after 2 years.The family history that gluttony behavior, addictive behavior and chemicals product rely on is not the predictor of significance on the statistics.The block mold of the predictor of selecting is significance (p＜0.0001), explained 39.8% of weight increase degree of variation in two years by these predictors.The most influential predictor is PHENCAL in this predictor group ^TM, then be morbid obesity, women and fat family history (seeing Table 20-C).Further analyze to have compared and taking PHENCAL ^TMBetween group and the contrast core group in the gluttony score of 2 years experimental sessions front and back.Also used a kind of two-way analysis of variance method, a factor is PHENCAL ^TMFactor between the group of treatment, another is about repeating to estimate factor between the gluttony score before and after the treatment phase in 2 years, analyzing and find to have significant interaction (p＜0.001) therebetween.Also used paired t-check respectively to taking PHENCAL ^TMThe variation of the gluttony score of group and matched group is checked.From the statistics angle, matched group does not have detectable variation.And take PHENCAL ^TMGroup then has tangible reduction on the gluttony score.

Table 20-C

The standard error β flexible strategy T P of variable slope (B) B

Morbid obesity 0.111 0.050 0.130 2.4 0.180

Gluttony score 0.023 0.003 0.420 7.70＜0.001 is used PhenCal (being) 0.327 0.003 0.426-7.80＜0.001

(constant) 0.151 0.045 3.34＜0.001 discussed

The data of the 2 terms open trial research of here showing shows: neurotrophy Substance P HENCAL ^TMCan in known carbohydrate glutton, suppress unusual feed behavior, and prevent the body weight recovering to have alleviated.The inventor thinks PHENCAL ^TMObvious useful effect can be construed as precursor aminoacid and the combined effect of enkephalinase inhibitory action in the result in the recompense loop of center edge system.A mechanism accurately can't be provided for the effect of this neurotrophy mixture of substances inventor at this moment, can not point out accurately that the combination of any composition or composition suppresses the gluttony behavior to carbohydrate in can present inventor's research most effectively.

Neurotransmitter 5-HT, DA, NE and enkephalin have been proved to be absorption (Leibowitz, 1985 that can reduce sweet food; Leibowitz etc., 1982; Kaye etc., 1984; Riviere etc., 1987; Blum etc., 1990).Thereby, PHENCAL ^TMBe specifically designed by loading precursor aminoacid, comprise 1-tryptophan (5-HT precursor), 1-phenylalanine (DA and NE precursor), thereby increase the inhibitory nerve mediator in these foods, and increase enkephalinase inhibitor d-phenylalanine, so that enkephalin raises (Blum etc., 1986).Observed PHENCAL in these researchs ^TMA predicable mechanism of effect comprises to the restitution of some insufficient monoamine such as 5-MT, NE, EPI with to the restitution of neuropeptide methionine-enkephalin and CCK-8.All these are considered to be subjected to feed (carbohydrate) property material (Frohman, 1983 of glucose or genetic influence; Fullerton etc., 1985; Matsumura etc., 1984).

Compare PHENCAL with the man that identical family history is arranged ^TMCan bring out best effect to the woman that OB+ and CD+ family history are arranged, this point is very worth gazes at.Perhaps, this species diversity has relation with the discovery of (1996b) such as nearest Comings, they notice: only in the women, the hereditary variant of the hereditary variant of human obesity disease (OB) and human dopamine D2 (DRD2) gene account for body-mass index (BMI) variation up to 22.8%.Interaction between OB in adiposis patient and the DRD2 gene may relate to combining of leptin and OB receptor, and this combination has activated a kind of Wright's nerve mediator or neuropeptide, thereby behavior and appetite and metabolism are exerted an influence.Known that about this point the Ob/ob mouse also shows the obvious minimizing of dopamine level (Oltmans, 1983) in arc funnel.According to this work, the inventor thinks: as in the past referred, it is similar that glucose relies on (for example: cocaine, ethanol, heroin) in conjunction with the chemicals product with other.

In 2 years, take PHENCAL ^TMAminoacid absorption group with do not take in PHENCAL ^TMCore vitamin group compare, demonstrate: 1) though the former the group in the men and women, its overweight percentage ratio all reduces twice; 2) woman's addiction minimizing 70%, and man's addiction minimizing 63%; 3) woman's gluttony behavior reduces 66%, and man's gluttony behavior reduces 41%;

Embodiment 2

VNTR allele is related with Tourette syndrome and drug dependence

Tourette's syndrome is the syndrome of neural spirituality obstacle, it is characterized in that chronic motion and pronunciation twitch, and far-ranging some corelation behaviour, comprise abuse, the depression of ethanol and medicine and force, attention deficit hyperactivity disorder, conduct, sleep, study, property and anxiety disorder (Comings, 1987c; Comings, 1990, Comings and Comings1993; Comings 1995d).Tourette's syndrome is assumed to be that usually being is Gts and hand down by heredity (Comings etc., 1984, Pauls and Leckman as a kind of autosomal dominant inheritance, AD character, 1986), whole genome has in fact been investigated in chain research, but does not find (Fog, 1985 of existing of Gts gene; Tsui, 1994).Yet, verified recently conduct and disobey Gts in the impaired patients and the genetic load of ADHD gene have highly significant increase (Comings, 1995a).Hereditism's family about the Tourette syndrome studies show that: this disease may relate to a plurality of genes, is lower than 12/1000ths (Comings etc., 1984) unless suffer from the danger of this disease in life.The frequency that school age, tourette's syndrome took place among the boy accounts for 1/90th of whole tourette's syndromes, and possible tourette's syndrome accounts for 1/40th (Comings etc., 1990).Similarly the result also can see .1994 such as () Kurlan in other research, and the frequency of tourette's syndrome or chronic Gilles de la Tourette's syndrome is confirmed as 2.6% (Zohar etc., 1992) among the Israel soldier.In nearer family research, find: as the whole behavior spectrum (Comingss relevant with tourette's syndrome, 1990, Comings, when 1995d) all being included in the research range, the evidence that a lot of families provide shows that these genes are from father and mother both sides' heredity, this supports that tourette's syndrome is polygenes obstacle (Comings, 1990; Comings, Comings, 1993; Comings, 1994b; Comings, 1995b; Kurlan etc., 1994).

Consider that the wide region of tourette's syndrome associated disorders and Gts gene are evidence (Kurlan etc., 1994 from father and mother both sides' heredity; Comings, 1990), this prompting tourette's syndrome is a kind of multigenic disease, and these genes (MAOACX, DBH, DRD2, DATI or the like) relate to the metabolism of dopamine, 5-hydroxy tryptamine, norepinephrine and other neurotransmitter, the variation that each gene provides only accounts for 1% to 10% (Comings, 1996b of total variation; Comings, 1995a; Comings, 1995b; Comings, 1996a).Method

Group I: tourette's syndrome group.

57 collators have been comprised in the object of study, 229 tourette's syndrome proband, most of people among the latter suffer from multiple relevant behavior disorder (Comings1990), and 90 ill and not ill tourette's syndrome proband's family members.Whole study subjects are the white people of right and wrong Spain system all.The aspect feature of other of mean age, diagnosis typing (tourette's syndrome or chronic exercise are twitched) and object of study be described elsewhere (Comings etc., 1996a).

The behavior score.Each tourette's syndrome collator, proband or family members are required to fill in a questionnaire based on diagnosis meet flow sheet (Robins etc., 1981) or DSM-III-R (1987) standard.This provides a framework review to the psychiatric symptom in the broad range.These symptoms are divided into 27 different behavior groups, comprise ADHD, substance abuse, mental state, anxiety, school's performance, stutter, twitch and other classes.In other articles, be described in detail (Comings 1995a about the problem of how to give these behavior score; Comings 1994a; Comings 1994b:Comings1995b; Comings etc., 1996a:Robins etc., 1981:Comings 1995c).Two kinds of behavior point systems are used to ADHD is assessed.First kind is called ADHD, and it is that whether minimum half variable exists and assess in 22 kinds of ADHD variablees of a series according to DSM-III and DSM-III-R standard.Second kind is called ADHD-R, it according to DSM-III-R diagnostic criteria assess.Untapped QTV is Inattention, impulsion and hyperkinesia before three kinds.They are three kinds can produce the little branch of ADHD score by accumulation.The QTV abbreviation comprises: conduct disorder (CD), disobey obstacle (ODD) (Comings 1995a) and major depression outbreak (MDE) (1995 c Comings) symptom.

The reason of verifying the behavior of falling ill altogether is: once observing some gene in the past may be more close with the relation of some specific illness behavior of existing in the tourette's syndrome, rather than with diagnosis itself more close .1996a such as () Comings.This part questionnaire and not meaning that provides DSM-III-R or DSM-IV diagnosis, and provides a very structurized method that produces the QTV in different behaviors field.The advantage of continuous trait is that they can provide wideer diagnostic area to the seriousness of the state of an illness, rather than carries out either-or diagnosis.Some other colleague (Gadow and Sprafkin, 1994; Grayson and Carlson 1991) the operating position of the operating position of such questionnaire with the same structurized questionnaire that is used to speak face to face treatment compared, proved that this part is based on accuracy, practicality and sensitivity to the questionnaire of the approach of symptom evaluation.Use the review of the study subject of this part questionnaire to demonstrate them to good hundreds of name and correctly reflected in the past the information that will just can obtain by personal interview.

Group II: substance abuse group.

Patient group is made of the honky of 120 non-Spain systems.

Assessment.All study subject is all used " test of Michigan alcoholism seriousness " (Davis etc. 1987) (self management's questionnaire of one 24 project, through having comprised medicine thing abuse (MAST-R) after the revision), by " clinical interview program (DSM-III-R version) (Robins etc.; 1981) " (being used for the diagnosis whether substance depilatory sexual disorders exists) of clinician's management, with assess by " addiction severity index the 5th edition (Hodgins; and Guebaly, 1992) (ASI) " (being used for the assessment that ethanol and medicine thing use variable) of clinician management.

The inventor has utilized ASI's " medicine thing/alcohol abuse " and " legal status " part.Use to specific material is assessed, inquired the problem of lifelong (for many years) operating position of relevant following material, as:reach drunk ethanol and use the use of heroin, other Opium medicine/analgesic, barbiturates, other calmness/sleep peacefully/tranquillizer, cocaine, amphetamine, Fructus Cannabis, psychedelic drug and inhalant.To above-mentioned every kind of material, study subject will be asked the problem of relevant route of administration when relevant.That selectable approach has is oral, per nasal, suction smog and intravenous injection.The continuous variable of # intravenous injection medication is by the whole numerals through the different pharmaceutical of intravenous injection (IV) being added up and being calculated.The variable of intravenous injection medication is a kind of dichotomic variable, 0 the representative not used intravenous drug, the problem relevant with medicine that the experimenter of used one or more intravenous injection medicines then be designated as≤1. is asked comprises: and " did you have how many times Alcoholic DT? overdose? " " did you have the alcohol problem that how many days run in the past 30 days? the medicine problem? " " you in the past 30 days 1 li how much has been spent at alcohol? on medicine? " in addition, also be asked about the various legal problem of medicine and alcohol abuse: " how many times that has in life at you is accused of driving when intoxicated? " " you the arrested that how many times arranged in life because medicine is accused and prosecuted? " these charges finally have how many times to be determined a crime? " May use from 0 during to any numeral when answering, " 0 " is just counted " 0 " and is divided, and any other numeral is counted " 1 " and divided.Those use relevant problem to be added up to the total ethanol score of calculating with ethanol, and those problems relevant with drug use are added up to the total medicine score of calculating.The interviewer judges according to the assessment to the seriousness of problem the patient whether treat ethanol and/or drug dependence by needs, the evaluation scope from 0 (not needing treatment) to 9 (needs are treated to handle life-threatening situation).

The substance abuse matched group.The collator of substance abuse group is the contrast that is independent of the tourette's syndrome patient.They are made up of two groups.First group is the white race student's (30.1 years old mean age) from the non-Spain of 45 the older male system of the state university in California that is positioned at the San Bernardino.Test is excluded those people that remarkable substance abuse problem arranged based on MAST-R.Form by twin father for second group from twin children's family study project of Minnesota.Because these people have this common ground of twin children of 11 years old or 17 years old and found out from the population of Quanzhou and decide that they compare with the university students according to them, more can represent a kind of combination at random of whole colonies of social economy and education colony.Although all the score of collator on the substance abuse variable all is negative, because the result of substance abuse assessment is also unavailable to twin children (father) collator, in fact some people's score may be positive.But, because this is the intersection region at random in a prevailing state of agricultural, so the inventor supposes that the false negative numeral is very little in this group.

PCR ^TMPolymorphism.Utilized MAOA VNTR polymorphism (Hinds etc., 1992) in the experiment.The polymorphism of this complexity is made of a kind of little satellite of GT that is directly adjacent to the new 23-bp VNTR of a kind of imperfect two strands motif, its allele dinucleotide repeat with the multiple numeral of VNTR on different.From whole blood, extract DNA with standardization program.Target DNA is through PCR ^TMAmplification (Mullis etc., 1986).For labelling PCR ^TMProduct has used each 0.1uM of every kind of primer (seeing Table 20-D) of fluorescence HEX or FAM Amidite (Applied Biosystems, Inc., Foster city, California) labelling in reaction.2ul is through the PCR of 10 times of dilutions ^TMProduct is added into deionization Methanamide and 0.5ul standard ROX 500 (Applied Biosystems, Inc. of 2.5ul, the Foster city, the California) in,, loads on 6% polyacrylamide gel in the applying biological system 373 type dna sequencing instrument 92 ℃ of degeneration 2 minutes.Gel electrophoresis 5 hours under the condition of 1100 volts of voltages and firm power 30W.Gel is analyzed with inner ROX 500 standards through laser scanning.According to color fragment, with Genotyper (version 1.1) (Applied Biosystems, Inc.) identification peak value by the decision of base pair length.The complete information of each sample that obtains from each gel file all is printed, and the also submitted analysis of data of compilation.

The allele group.May the hypothesis relevant in order to verify the allelic length of MAOA with the effect of phenotype, allele has been divided into 4 groups (seeing the result).They are marked as 1 to 4, and are promptly the longest from being short to most, have formed MAOA genotype variable.Only in the tourette's syndrome group, just there is the women to participate in.Have only the homozygote of those given allele groups just to be included in the analysis.

Statistical analysis.For Tourette syndrome group, variance analysis is used to check the degree of correlation of each QTV to 4 different allele groups.The linear equation analysis is used to check 4 average recruitments of significance progression between the allele group.Used the statistical package of SPSS (SPSS Inc., Chicago, Illinois).For the linear equation analysis, the subcommand multinomial is set as 1.Multivariate analysis of variance is used to determine when whole variablees are all verified simultaneously, and whether any one QTV significance.Polynary linear regression analysis is used as second approach, determines when whole variablees when all the while is verified, and whether any one QTV significance.The MAOA genotype is set to the subordinate variable, and 27 QTV are progressively imported as variable independently.

Card side.Above-mentioned studies show that: the longest allele group has the highest mean concerning most of QTV.Between 4 groups of tourette's syndrome, compared≤the potential progressivity reduction of 335bp allele group frequency, the symptom of these 4 groups gradually reduces: the tourette's syndrome proband of ADMD, the tourette's syndrome proband who does not have ADHD has the relatives of tourette's syndrome and does not have the relatives of tourette's syndrome.

Multivariate analysis of variance.To the substance abuse group, multivariate analysis of variance is used to determine between 4 MAOA allele groups and 2 summary variablees whether have significant association, and these two variablees are ethanol and medicine score.Variance analysis is used to check the ethanol of 4 allele groups and the mean of medicine score.

Linear X 2 test.Linear X 2 test is used to check between 3 groups≤the potential progression recruitment of 335bp group frequency.These three groups are: matched group, (no ATU) substance abuse person of no behavior organizes and has substance abuse person's group of behavior (ATU is arranged).In why no ATU group is included in is in order to get rid of a kind of probability: promptly owing to the common morbidity of certain different behavior, the frequency of this allele group in substance abuse person may increase.In order to help to get rid of this factor, the frequency among the substance abuse person with certain behavior in this allele group is at least than high by 20% the substance abuse person who does not have this behavior.Owing to suppose: these allelic frequencies will little by little increase in these 3 groups, so use linear X 2 test statistical method.

Regression analysis.Largest percentage for the variance of determining variable that can be illustrated with the MAOA gene, relevant with medicine, the inventor has used regression analysis, wherein those carry≤the allelic study subject of 335bp counted 1 fen, those carry≤the allelic study subject of 335bp counted 2 fens.Because this is to get in touch chi-square variable the most closely with the MAOA gene, it just is used as drug dependence variable (contrast=1, no ATU actor counts 2, has the ATU actor to count 3).The result

The allele group.The allelic distribution of two groups is represented with Fig. 3.Because this is complicated VNTR, allele does not fall into odd number or even number base pair one model clearly.The result is the form that is produced by gene type (Genotyper) program.Between bp 316 and 323bp, do not have allele, produced like this＜320 and＞2 clearly main groups of 320bp.But the effect of phenotype may the hypothesis relevant with base length be checked in order to allow, bigger 323-339bp group allele is divided into three subgroups, be respectively: the allele subgroup that is shorter than the 320-333 of main peak, the main peak allele subgroup of 334bp, be longer than main peak≤335bp allele subgroup.219 men and 156 woman are arranged in the tourette's syndrome group, and adding up to has 375 study subjects.In the woman, 88 people are allozygotes.After they are deducted, be left 287 study subjects in this research, wherein 36 is the collator.In this final group, in the frequency distribution of 4 allele groups, woman and man do not have significant difference.

The tourette's syndrome group.The The results of analysis of variance of each QTV of 4 allele groups is shown in table 20-D.Conventional The results of analysis of variance shows below F-ratio and p value.The F-ratio that linear equation is analyzed is at F ₂Show subscript below the row ¹Represent that these values are to have＜0.05 significance.At F ₂In the row, QTV lines up regularly according to the F-ratio that successively decreases.By the α value be set at≤0.05 Tukey test determines, mean significantly is lower than≤those allele groups of 335bp group represent with asterisk.To say for 24 remaining QTVs except stutter, shopping and terrified (being given minimum F-ratio), those carry≤and the mean of 335 allelic study subjects is the highest.

Statistical analysis.The result of the multivariate analysis of variance of whole 27 QTV (has significance p=0.023, gambling (p=0.025) and manic (p=0.025) to sex (p=0.012), problem concerning study.Whole 27 QTV check with multiple regression analysis progressively simultaneously that variable primary school problem (p=0.012) and gambling (p=0.038) have significance.According to r ²Value, MAOA gene have only illustrated 3.9% of these QTV variations.Use chi-square analysis to find: carry≤percentage ratio of 335 allelic study subjects has significant progressivity to reduce, promptly from tourette's syndrome proband that ADHD is arranged (24%, n=125); To the proband of the tourette's syndrome that does not have ADHD (20.0%, n=50); To the proband family members that tourette's syndrome is arranged (12.5%, n=16); To the proband family members of no tourette's syndrome (5.6%, n=56) (p=0.003).

The substance abuse group.The collator is according to the ATU study subject.For the collator of 160 merging, the distribution of 4 allele groups is as follows:＜320-34.4%, 320-333-38.1%, 334-335-21.3% ,≤335-6.3%.For 120 ATU study subjects, frequency is as follows:＜320-39.2%, 320-333-18.3%, 334-20.8% ,≤335-21.7%.There is significant difference, x ²=22.17, p=0.00006; Frequency and two matched groups of≤335bp group have comparability, and San Bemardino group is 8.9%, and twin father and mother's group is 5.2% (x ²=0.744, p=0.38).

Multivariate analysis of variance.Ethanol is pointed out with the multivariate analysis of variance of medicine score: when the two all with MAOA gene and VNTR (the parameter series connection repeats) when having significance related, compare (p=0.012) with the ethanol score, the significance of medicine score bigger (p=0.001) (table 21).In addition, the result of the multivariate analysis of variance of merging also is significant (p=0.007).This numeral of 257 people (N) is littler than 160 collator+120 ATU actors' total or 280 people's number because only 97 have the study subject of ATU behavior to finish ASI.On the contrary, DIS has been finished in the affirmation of all 120 people that the ATU behavior arranged DSM diagnosis that all is ethanol and/or drug dependence.

Variance analysis.The variance analysis that shows two score of each allele group mean all uses table 22 to represent.As for the tourette's syndrome group, the highest mean is present in≤335bp allele group in.For the medicine score, according to Tukey test, 3 other the allele group obviously than≤335bp organizes low.

Card side: whether have superiority relatedly in order to determine the MAO gene, 14 variablees relevant with the use of this material are verified with certain type substance abuse.Among the collator≤frequency in the frequency of 335bp allele group and the ATU of no behavior (the no ATU behavior) study subject represents with table 23 with the contrast of the frequency of ATU (the ATU behavior is arranged) study subject that behavior is arranged.Owing to there are 14 class substance abuse variablees to be verified, have only those to draw the p value and be considered to significant less than 0.0036 (0.05/14) variable with the Bonferroni correction method.Here the variable that only shows those p＜0.01.It is an exception that alcohol dependence is only arranged.The data of showing has illustrated such fact, that is: only when comparing with the study subject that medicament dual relies on the study subject of those drug dependences or ethanol, in the study subject of alcohol dependence≤335bp allele just almost do not increase on frequency.On the contrary, only variable of drug dependence has provided peak (x ²=17.4, p=0.00003).

Regression analysis.Allele group (＜335 pairs≤335) has provided following result: r=0.25, r to the regression analysis result of drug dependence diagnosis ²=0.0625, T=4.305, p＜0.0001.

Substance abuse.Use enzyme level (Wiberg etc., 1977; Gottfries etc., 1975; Devor etc., 1994; Vonknorring etc., 1991) and the previous research of hereditary variant (Vanyukov etc., 1993) hinted the effect of MAOA gene in substance abuse.Present result is consistent with those conclusions, and is especially all the more so for drug dependence.Multivariate analysis of variance has shown remarkable related between MAOA allele and ethanol and the medicine score has many common morbidities that caused by the substance abuse of these two kinds of forms.Shown in table 23, when drug dependence and alcohol dependence are examined dividually, drug dependence and allelic related related much bigger than alcohol dependence.

Male's advantage.ADHD, Tourette syndrome, conduct disorder, disobedience obstacle, dyslexia, learning disorder, stutter, drug dependence and alcoholism, all these demonstrates male's advantage.Male's advantage part is possibly imputed hormone and factor of environmental to, and in addition, the X linked gene may also be a factor.In the tourette's syndrome group, as regression coefficient, to r ²Judgement demonstrate: to different QTV, the MAOA gene can illustrate at most any one QTV variation 2.5% or still less, this shows that the chain MAOA gene of X can not the explanation male advantage relevant with tourette's syndrome, ADHD or relevant obstacle.On the contrary, the diagnosis of drug dependence and the≤allelic existence of 335bp related r whether whether ²Value shows: reaching as high as 6.2% variation may be caused by the MAOA gene.It can play certain effect in male's advantage of drug dependence.

Specific QTV's in longer moonlet allele and the Tourette syndrome group is related little, shown in table 21, all on the trend between QTV compelling homogeneity degree is being arranged.Because this may be a kind of accidental, association at random, the inventor attempts to determine that whether these results can fully independently be able to repetition in study subject and the collator's group at another.The data of this group (substance abuse group) demonstrates: compare with observed situation in the tourette's syndrome group, longer MAOA VNTR allele, especially≤allele of 335bp, with exist between the substance abuse related more closely.This pattern of two groups is remarkably similar, and≤335bp allele has all obtained top score, and minimum allele (＜320) obtains time high score and gives, and 334-335bp allele has obtained intermediate section.It is this that to repeat allelic size and the interrelated of phenotype be consistent with moonlet allele from the probability of playing a role in the adjusting of MAO gene.

Table 20-D

MAOA VNTR polymorphism: the comparison of the different behavior scores that different allele groups is undertaken by variance analysis

Allele group behavior＜320 320-333,334≤335 F-that divided by base pair length are than p F ₂N 82 43 110 52 manic 1.43 ^*1.8 1.36 ^*1.9 1.59 ^*2.27 2.59 2.8 3.59 0.014 6.39 ¹OCD 2.18 2.8 2.18 2.4 2.80 2.8 3.31 3.2 2.16 0.093 5.89 ¹Property 0.62 ^*1.1 0.47 ^*0.9 0.66 ^*1.2 1.25 1.6 3.91 0.009 5.82 ¹Sleep 0.36 0.7 0.38 0.8 0.49 0.9 0.76 1.1 2.31 0.075 5.56 ¹Primary school 2.60 1.7 2.90 2.0 2.98 2.1 3.46 2.1 1.89 0.131 5.14 ¹Gambling 0.13 ^*0.9 0.27 0.8 0.22 0.8 0.61 1.6 2.49 0.060 4.82 ¹Stutter 0.13 0.3 0.11 0.3 0.25 0.4 0.23 0.4 2.23 0.084 4.41 ¹Study 0.52 ^*0.9 0.65 1.0 0.54 ^*0.9 1.00 1.1 3.32 0.020 4.37 ¹Inattention 6.69 5.2 7.37 4.9 7.67 4.7 8.48 5.3 1.42 0.235 4.15 ¹ADHD 19.43 14.9 20.95 13.5 21.18 13.9 24.80 15.3 1.53 0.206 3.74 ¹ADDR 4.63 4.9 4.84 4.6 5.18 4.7 6.42 5.2 1.57 0.196 3.74 ¹Get excited 5.74 4.9 6.30 4.8 6.49 4.7 7.46 5.0 1.35 0.257 3.68 ¹Do shopping 0.58 1.3 1.00 2.2 1.32 2.8 1.09 2.4 1.61 0.186 3.23 ¹

Allele group behavior＜320 320-333,334≤335 F-that the size of being represented by bp is divided are than p F ₂N 82 43 110 52MDE 3.00 2.8 3.29 3.1 3.45 3.1 3.92 3.1 1.01 0.385 2.90CD more than 2.78 2.4 2.90 2.1 2.97 2.2 3.54 2.6 1.15 0.328 2.54 moving 6.91 5.6 7.27 5.2 7.00 5.4 8.86 5.9 1.59 0.190 2.29 terrified 2.00 2.7 2.15 3.0 2.45 2.7 2.65 3.3 0.71 0.548 2.10 division samples 1.31 2.3 0.68^*1.3 1.34 2.2 1.84 2.3 2.26 0.081 1.92 generalized anxiety disorder, 0.21 0.4 0.18 0.3 0.28 0.4 0.29 0.4 0.85 0.463 1.60 somzatization, 2.13 3.0 1.58 2.4 2.17 3.0 2.90 3.7 1.16 0.325 1.34 medicines 0.36 1.2 0.45 1.6 0.40 1.3 0.75 2.0 0.81 0.489 1.28 are read 1.86 1.9 1.36 1.8 1.78 2.1 2.34 2.5 1.75 0.156 1.27ODD 3.07 3.3 3.02 2.7 3.16 3.1 3.73 3.4 0.57 0.633 1.02 and are twitched 0.51 2.3 0.81 2.9 0.31 1.8 1.11 3.2 1.41 0.241 0.45 terrified 2.91 2.0 3.09 2.1 3.18 2.2 2.98 2.2 0.27 0.845 0.20 smokings 0.07 0.3 0.11 0.3 0.05 0.2 0.77 0.3 0.53 0.662 0.10 of 2.81 3.7 3.04 3.4 2.84 3.5 3.57 4.1 0.55 0.642 0.73 alcohol¹Significance,＜0.05, the F-ratio that the F2=linear equation is analyzed ^*Significantly less than 〉=335, α=0.05 (Tukey check) (providing mean and standard error) (n=287)

The ethanol of table 21 substance abuse group and medicine score are to the multivariate analysis of variance of MAOA allele group

(N=257 name male)

Variable F-compares P

Ethanol score 3.72 .012

Medicine score 5.85 .001

Sum (Wilks) 2.99 .007

Table 22

Substance abuse group alcohol and medicine score to the variance analysis allele group N mean standard deviation F-of MAOA allele group than p alcohol score＜320 94 2.27 3.1320-333,81 1.49* 3.0334-335 56 1.94 2.7≤335 26 3.73 3.52 3.72 0.012 medicine scores＜320 94 3.59* 5.2320-333,81 1.94* 3.8334-335,56 3.34* 4.9≤335 26 6.42 6.2 5.85 0.0007

* significantly be lower than 〉=335 allele group means α=0.05 in the Tukey check.

Table 23

Matched group to the ATU study subject of no behavior in the ATU study subject that behavior is arranged

Carry≤the linear chi-square analysis of the allelic study subject number of 335bp

Contrast no ATU ATU is arranged

Behavior N % N % N % card side P only has the use of use overdose 160 6.3 71 12.7 25 28.0 11.00 .0009 barbiturates 160 6.3 61 13.1 32 25.0 10.56 .0011 of pharmacological dependence 160 6.3 58 15.5 62 27.4 17.4 .00003 intravenous (IV) drugs 160 6.3 56 14.3 27 29.6 13.65 .00022

The use of use 160 6.3 68 14.7 28 21.4 8.05 .004 opioids 160 6.3 58 15.5 38 18.4 6.88 .009 of the use 160 6.3 14 7.1 82 18.3 8.35 .004 heroin of the use 160 6.3 25 12.0 67 19.4 8.86 .003 hemps of the use 160 6.3 67 14.9 26 23.1 8.96 .0027 cocaines of use 160 6.3 19 5.3 77 19.5 9.37 .0022OSH* of DUI 160 6.3 34 8.8 62 21.0 9.92 .0016 amphetamines only has alcohol dependence 160 6.3 98 24.5 22 9.1 non-linear

* other opiates of OSH=(except that heroin or methadone), tranquilizer and somnifacient embodiment 3 d2 dopamine receptor polymorphisms are related with division sample/avoidant disorder behavior

The selection of study subject and trial.Enlist the white race volunteer.58 men and 71 woman are arranged, the mean age be respectively 40.9 ± 1.8 years old and 47.1+1.5 the year (mean+SD).All carried out the inspection of the computerization test of " the clinical polytropism investigation of rice Long Shi " second edition (MCMI-II) in advance, volunteer is dedicated 15 milliliters of blood to through venipuncture.It should be noted that: owing to consider race and ethnic group layering, blood is to gather on one's body from the Northern Europe of non-Spain system and the white people in West Europe, also has some to make an exception, because be to come from this group about collator's data the most widely.In order carry out this research, 129 psychiatric patients that are with or without the medicine sent out altogether or alcohol abuse are selected altogether comes out, and carries out gene type, and tests with the clinical polytropism investigation of rice Long Shi (MCMI-II) computerization and to assess.This group is made of following member: spirit depressing person's (22.1%), generalized anxiety disorder person (8.4%), unipolar disorder person (24.5%), two-phase obstacle person (12.9%), schizophrenic's (5.2%), attention-deficient obstacle person (21.9%) and substance abuse obstacle person (18.1%).

The clinical polytropism investigation of rice Long Shi (MCMI-II) is used to 11 known personality disorders of assessment in each study subject, comprises division sample/avoidant disorder group.This test comprises 175 projects, and most people can finish it with 20～30 minutes time.Come from personality and psychopathology The Theory Construction 22 clinical scales measure yardstick as the operability of symptom.These clinical directed scales are directly to cooperate with the diagnostic system and its syndrome classification (DSM-IV) of official.According to the DSM-IV model construction other scale, to distinguish acute clinical obstacle (Axis I) and patient's more persistent personality characteristics (Axis II).Actuarial basic valuation data, rather than standard score converting data by analysis, the calculating that is used for the scale project is with quantitative.Basis appraisal is 60 just to hint and have certain obstacle.Between 75 and 83, certain obstacle chronic or certain order of severity is arranged, just represent to exist uncontrollable pathologic obstacle and surpass or reach 84 with regard to expression.In this research, the inventor selects by using a rice Long Shi division sample/avoidant disorder group to determine seminar in study subject, reaches 84 or the percentage classification evaluated of higher basis in this special colony.The inventor classifies them according to the MCMI-II score of study subject, and according to following classification method, by basic assessment values difference the proband is divided into 4 groups: (1) score is below 60; (2) score from minimum equal 60 to the most high in 73; (3) score from minimum equal 74 to the most high in 83; (4) score is minimum equals 84 to the most high in 100.The demography data of all study subjects is shown in table 24.

Table 24

The diagnostic characteristic that is used for the psychosis colony of embodiment 3

Deagnostic category study subject percentage ratio

Dysthymia 27.1

Anxiety disorder 8.4 widely

Single-phase 24.5

Two-phase 12.9

Schizophrenia 5.2

Attention-deficient obstacle 21.9

Substance abuse obstacle 18.1

MCMI-II test scoring has obtained confirmation in different inpatient colonies, promptly the personality disorder scale of 104 patients and Minnesota leggy personality inventory (MMPI) is compared.The conservation significance level is used to guarantee the effectiveness of conclusion.The scale significant correlation of division sample, avoidance, dependence, performance desire and autophilia, passive-attack, disintegrated type and critical scale do not have dependency with corresponding MCMI-II scale, so just verified that this test it is can be used for confidently measuring unusual personality character (Shuler etc., 1994; Zimmernman etc., 1994), for example resemble division sample and elusive behavior.

30 " super " contrast study subject in this research is screened, lack behavior to get rid of some recompenses, these behaviors comprise family history, ADHD, pathological gambling and a kind of Axis II diagnosis (comprising SAB) of alcoholism, multiple substance depilatory, cigarette smoking, carbohydrate gluttony behavior, BMI＞25, substance abuse obstacle, and carry out DRD ₂A ₁And A ₂Allelic gene type.Other contrasts are from the volunteer in 3 sources: A) the Tourette patient's adoptes father and mother, fosters father and mother or stepparent; B), suffers from the experimenter of thyroid carcinoma or noninsulindependent diabetes from the endocrine clinic; And C) hospital internal personnel comprise professional, technician and service worker.142 contrasts are all screened altogether, get rid of the abuse of ADHD, ethanol, medicine and Nicotiana tabacum L..The inventor is divided into DAT to 91 people in these contrasts by gene type ₁9 and 10 allele, 51 people are divided into D β HB in addition ₁And B ₂Allele.An important caution of need considering is: in order to estimate the related of dopaminergic allele and SAB, study subject should be selected.The inventor has realized that: altogether the study subject (promptly having 18.1% people to have the substance abuse obstacle in this colony) of the RDS behavior of sending out may be obscured causing aspect the statistical procedures of data.The inventor also has realized that simultaneously: it will be very difficult getting rid of all RDS behaviors in having the patient of SAB.Therefore, inventor's complementary studies that must wait in the future just can address this problem.

Gene type.In this research, the gene type of 271 study subjects belongs to one or more dopaminergic genes altogether, and all study subject all respectively carries out gene type and reads the result with identification number of a neutrality, and does not know it is which is by typing.

DRD ₂Polymorphism.Of following article (Blum etc., 1990a:Comings etc., 1995), D2A ₁, D2A ₂Gene type uses is Southern Blot hybrid method.In addition, a great deal of sample is to use PCR ^TM(Noble etc. 1994d) carry out gene type to technology.

D β H polymorphism d ' Amato and colleague (d ' Amato etc., 1989) have reported two kinds of existence that are called as the TaqI D β H polymorphism of A and B.D β H cDNA clone AII (Lamouroux etc., 1987 have been used; Lamouroux etc. 1993), contain the insertion fragment of a 2.7kb in the EcoRI site.In order to improve labelling, carrier is produced 5 bands by BamHI and SalI enzymic digestion.A 3.5kb fragment is labeled to be used to detect the β polymorphism.After the digestion of TaqI restriction endonuclease, electrophoresis in agarose gel, the Southern method is transferred on the nylon membrane, with ³²The probe hybridization of P labelling carries out autoradiography again, determines 2.8kb (B ₁) and 1.4kb (B ₂) fragment.

DAT ₁Repeat polymorphism.Allele PCR at 3 ' UTR ^TMMethod is determined, oligonucleotide PCR ^TMCondition is by reports (1992b) such as Vandenberg.Through the product behind the pcr amplification in 8% acrylamide gel with one group of mark electrophoresis that is used to show the fragment length size.

Statistics.Use Pearson came (Person) card side and Fei Xier (Fisher) accurately to check the proportion grading that carries out the 2X2 table.Bigger table as the 2X3 table, then uses Pearson came card side and linear trend Mantel Hensel check to carry out statistical test.Difference between the mean uses student t-method of inspection to test.Polynary logistic regression is used to analyze the contribution of one or several variable, with the probability of prediction at acquisition high score aspect division sample/avoidant disorder.This logical model uses the Hosmer-Lemeshow test of goodness of fit, represents the c of recipient's operating characteristic (ROC) area under curve to add up and assessed by the r-side of SAS computer program Proc Logistic demonstration or the existence of variance.In addition, in any logistic regression analytic process, be used for the also machine processing (Dunn and Clark, 1995) as calculated of odds ratio of predictor.

The result.For division sample/avoidant disorder, utilized X 2 method to carry out data set analysis, restriction dopaminergic allele divides the possible related of sample obstacle and MCMI-II avoidant disorder with the MCMI-II that surpasses 84 fens, with the stringency of the phenotype of security reserve association.Find in the research in 50% the study subject of suffering from avoidant disorder (12/27) of suffering from division sample obstacle (11/22) and 44%, can find DRD ₂A ₁The position gene; Can find DAT in the study subject (13/18) of division sample obstacle and the 62% avoidant disorder study subject (13/21) 72% ₁480bp (VNTR 10/10 allele); Found D β HB in the study subject (14/17) of the study subject (13/16) of division sample obstacle and 82.4% avoidant disorder 81.3% ₁Allele.According to card side, with DRD ₂A ₂Allele is compared, DRD ₂A ₁There is more significant association (x between allele and division sample/avoidant disorder group's (MCMI-II score 〉=84) the patient ²=7.6, df=1, p＜0.006).DRD ₂A ₁Allelic homozygote demonstrates: it accounts for the highest percentage ratio in the study subject of SAB colony (＞84).In SAB colony, the percentage ratio that heterozygote occupies approximately is half of study subject of homozygote group.DRD is only arranged ₂A ₂The group that allele is expressed then shows: it occupies minimum percentage ratio (linear trend analysis: p＜0.005, A in SAB colony ₁/ A ₁=83%, A ₁/ A ₂=41% A ₂/ A ₂=23%).

But, with DRD ₂A ₁Allele data difference, chi-square analysis fail to represent D β HB ₁Allele and DAT ₁10/10 allele is related with division sample/avoidant disorder behavior.Utilize a plurality of variablees and the SAB score of dividing is carried out related, use the logistic regression method to test itself and DRD ₂The significance relation of allele, age and sex, DRD ₂A ₁Allele and sex all are the remarkable predictors of SAB seriousness.The inventor finds that the Hosmer-Lemeshow goodness of fit is when p=0.78.The odds ratio of DRD2A1 and SAB is 2.79 (p=0.018), with the odds ratio of sex be 3.6 (p=0.0031).In the super collator of 30 screenings (get rid of alcoholism, multiple substance depilatory, the body quality index is below 25, cigarette smoking, family history, pathological gambling, ADHD, division sample/avoidant disorder behavior), DRD ₂A ₁Allelic prevalence rate is 1/30 or 3.3%.In the collator of 91 screenings, DAT ₁10/10 allelic prevalence rate is 34/91 or 37.4%.And in 51 garbled collators, D β HB ₁Allelic prevalence rate is 27/51 or 53%.

As for DRD ₂A ₁Allele (18/37 or 48.6%), can find remarkable related when comparing with following two contrasts: the document contrast is 185/714 or 26% (x ²=9.2, df`=1, p=0.0024; OR=2.71), super contrast is 18/37 or 48.6% (x ²=16.8, df=1, p=0.00004; And SAB＞84 OR=27.5).In addition, when comparing, at DAT with the contrast of screening ₁Found remarkable related (x between 480bp (VNTR10/10 allele) and those individuals's (18/28 or 64.3%) with SAB diagnosis ²=6.3, df=1, p=0.012; OR=3.0).Be assessed as D β HB with SAB ₁That carries out between the contrast of allelic carrier (17/23 or 73.9%) and screening has more also found similar tendency (x ²=2.89, df=1, p=0.09; OR=2.52).Utilization is called the statistical technique that logistic regression is analyzed, compared the MCMI-II score be lower than 84 and the MCMI-II score be higher than the whole circumstances of 84 (seriousness score), DRD ₂A ₁Allele is 8.3% to the contribution rate of this degree of variation, and this is (the x that significance is arranged aspect statistics ²=7.5, df=1, p=0.0059).In contrast, (DAT ₁10/10 allele) contribution rate to this degree of variation is 1.6%; D β HB ₁Allele is 0.0025% to the contribution rate of this degree of variation, and this is not the significance contribution.

When sex was verified in the logistic regression analytical model as unitary variant, the sex factor was 9.9% (x to the contribution rate of total degree of variation separately ²=9.4, df=1, p=0.0022).In a logistic regression analytical model, utilize DRD2A1 allele and sex factor as predictor, prediction surpasses 84 minutes high score in the MCMI-II score of division sample/avoidant disorder, for these predictors, DRD ₂A ₁The odds ratio of gene is 2.79, and male's odds ratio is 3.55.Hosmer-Lemeshow goodness of fit p value=0.778, C statistics (ROC area under curve) equals 0.714, and they are 17.9% to the associating contribution rate of degree of variation.In addition, 67 gene types are being belonged to DRD ₂A ₁And A ₂In the allelic preliminary study, use the process come into question in the above, the inventor does not find that they do not have related with other any one of 10 kinds of personality character of using MCMI-II to test to assess.

As conclusion, the inventor has used MCMI-II to carry out relatedness research from report type computerization test, with the contribution to the unusual personality character that is called division sample/avoidant disorder group of the polymorphism that discloses 3 kinds of dopaminergic genes.Because it does not rely on the age of ethnic division, sex or study subject, so it seems that observed correlation be not because the layering factor of population.In addition, be 1-4% based on frequency in generally division sample/avoidant disorder behavior of crowd.DAT ₁With D β HB ₁(10/10 repeats allele) allele lacks the significance contribution because the sample size size can't be excluded to total degree of variation.In fact, in the sample size size only is under the situation of 44 (because the values of losing), these three kinds of gene pleiomorphisms are about 7.6% to the merging contribution rate of hereditary variation in multiple regression analysis, this with this complex behavior obstacle in relate to a plurality of genes and other the notion of factor resemble the sex is consistent.

Embodiment 4

The correlating method of Fructus Cannabis ester acceptor gene and intravenous injection Drug abuse:

Study subject.Race's factor is one and causes the factor of obscuring easily that for it being reduced to minimum, all study subject all is limited in white people's scope of 91 non-Spain systems.Matched group is made of 62 older students (SB contrast) and 52 study subjects (LL contrast).The latter is by 40% maintainer, 43% secretary and office worker and, 17% the doctor of medicine or the doctors of philosophy constitute.Totally 114 collators are counted in two combinations.

Assessment.According to independent interview, exist the people of medicine or alcohol abuse/dependence to be excluded out the LL matched group.All contrast is all assessed by the self management's questionnaire with " test of Michigan alcoholism seriousness " (revision has comprised drug dependence) 24 projects (MAST-R).All MAST-R scores the SB collator above 4 minutes all be left out.In addition, described as embodiment 4, all the ATU study subject is all assessed with the diagnosis meet program (DSM-III-R version) of clinician's management, whether has the substance depilatory obstacle with diagnosis; Addiction seriousness index the 5th edition with clinician's management is assessed a series of ethanol and drug dependence variable.

Gene type.Process by standard from whole blood is extracted DNA.The use of DNA sample is following increases as the described primer of Dawsom (1995): 5 '-GCTGCTTCTGTTAACCCTGC-3 ' (sequence numbering: No. 3), and 5-TACATCTCCGTGTGATGTTCC-3 ' (sequence numbering: No. 4).It discerns those the allele of the repetitive sequence of one group of three base of (AAT) n.With fluorescence HEX Amidite (Applied Biosystems, Inc., Foster city, California) each primer of each 0.1uM is carried out labelling, so that labelling PCR ^TMProduct.PCR as the VNTR polymorphism of description among the embodiment 2 ^TMPolymorphism analysis is such, the PCR that 2ul10 doubly dilutes ^TMProduct is uploaded in 6% polyacrylamide gel and analyzes.

The statistical analysis of allele and genotypic frequency.The frequency of different allele in contrast and in the ATU study subject checked, the RxC program of having used the George Carmody by Canadian Ottawa Carleton university to write, use the check of regression Monte Carlo to send out check significant difference (Roff, and Bentzen, 1989).The advantage of this Computerized method is: can finish the simulation of card side with the standard error of estimation, and that cell is folding with observation in a small amount (especially zero).Different genotypic frequencies is to use the chi-square analysis method to compare.

Factorial analysis.In order to solve the statistical problem of big figure variable analysis, carried out a kind of preliminary factorial analysis, restrictive condition is only to produce two factors.Factor 1 is tended to combination medicine and is relied on relevant variable, and factor 2 is tended to the relevant variable of associating alcohol dependence.

Allele group and gene type.Although the inventor has observed 9 different allele that constitute with 3～10 repetitive sequences by 1, in the contrast or ATU study subject of this research, none has carried 2 allele.9 allelic distributions demonstrate in contrast and the ATU study subject: modal allele is #4.It is present among 36.4% the collator, in the ATU study subject of 31.7% ATU study subject and 20.2% intravenous injection Drug abuse.Inventor's working hypothesis (Comings, 1996b; Comings 1996a) is: moonlet is formation (Hamada etc., 1982 by Z-DNA at first; Schroth etc., 1992) may in Gene regulation, play direct effect, and the size of action effect depends on the length of repetitive sequence.Because main allele group is 4 repetitive sequences, and 1 or 3 allele are seldom arranged, this just allows 9 allele are divided into two groups naturally: short (＜5) allele group and long (≤5) allele group.So just produced 3 genotype:＜5/＜5, heterozygote and≤5/≤5.In order to eliminate statistical complicated factor, do not verify other allele, genotype or allelic combination.

Variance analysis.In order further to eliminate the complicated factor on the statistics relevant,, carried out preliminary test to the data that are used for the inspection of 2 pairs of above-mentioned three genotype relations of factor 1 and factor by variance analysis with the big figure variable.This be used for determining afterwards test whether can propose a preliminary regression model (＜5/＜5 pairs of heterozygote +≤5/≤, or≤5/≤5 pair of heterozygote+＜5/＜5); Perhaps dominant models (＜5/＜5+ heterozygote is right≤5/≤5, or≤5/≤5+ heterozygote is right＜5/＜5; Perhaps hybrid vigor (heterozygote right≤5/≤+ 5≤/≤5)).The result is right≤and 5/≤5 pair of heterozygote+＜5/＜5 mode proposed regression model.Again, in order to eliminate the complicated factor of statistics, other any model is not all verified.When using regression model,, will be studied by the utilization variance analysis method by the little branch of factorial analysis hint if wherein any one has produced remarkable result for factor 1 and factor 2.

Chi-square analysis.Each variable with significant factors is further used 2 * 3 chi-square butters to assess.* 2 dimensions are made up of two genotype groups :≤5/≤5 repeat allelic homozygous genotype other genotype to remainder.* 3 dimensions are composed as follows: (1) contrast, (2) are that the ATU study subject of 0 (ATU feminine gender) and (3) scores on given variable in score on the given variable is the ATU study subject of 〉=1 (the ATU positive).It is for ruled it out that the negative group of ATU is necessary to be added the reason of coming in: promptly in the ATU study subject 〉=increase of 5/ 〉=5 genotypic frequency may be actually because it sends out the related of obstacle or variable altogether with certain, and is rather than relevant with that variable of being verified.Inventor's expection has 3 kinds of models:

A. significantly-linearity.If 〉=5/ 〉=5 genotype are relevant with given variable, the inventor is expected at the growth that there is a kind of so significant similar linearity in case percentage ratio aspect, that is: 〉=5/ 〉=5 group, to shining the negative case (that variable is negative influence) of ATU, progressively increasing to ATU positive case (that variable is positive role).The significance of this progressive growth uses linear chi-square butter (the Mantel-Haenzel card side of the statistical packages of SPSS, SPSS Inc., Chicago, Illinois) to check.Because 〉=5/ 〉=5 genotype may take place relatedly with material or variable more than a kind, the inventor requires: in the positive group of ATU 〉=and the percentage ratio of 5/ 〉=5 carrier is at least than in that ATU is negative is high by 20% in organizing.

B. significantly-non-linear.If carry 〉=percentage ratio of 5/ 〉=5 genotypic case, in the negative group of ATU than high in the positive group of ATU, so, 〉=5/ 〉=5 genotype just be considered to certain given material do not have related, even the p value is significant.

C. not significantly-non-linear.In addition, if the p value be＞0.05, 〉=5/ 〉=5 genotype also are considered to do not have related with certain given material.The result

The study subject feature.The mean age of ATU study subject is 39.7 years old (standard deviation is 7.0, and scope is 23～52 years old), and collator's mean age is 38.4 years old (standard deviation is 12.6 years old, and scope is 21～71 years old).(F-is than=0.76, p=0.38) not have significant difference between them.All the ATU study subject all is the man, is men and 45 (39.5%) collators are arranged, and 69 collators (60.5%) are the woman.

Gene frequency.The reason selected by the ATU study subject of intravenous administration is: clinical experience shows: its representative be those study subjects with relevant with drug dependence serious problems.When repeating check 10,000 times that carry out the Monte Carlo check, in matched group was organized ATU, whole allelic frequencies all had been carried out relatively, do not have significant difference (x between them ²=7.09, p=0.54).When collator and intravenous drug abuser compared, difference was marginal value (x ²=14.49, p=0.065).

41 people in the contrast or 36.0% are 〉=5/ 〉=5 homozygote that on the other hand, 43 people in the ATU study subject or 46.7% are 〉=5/ 〉=5 homozygote (p=NS).In 32 intravenous drug abusers, there are 20 people or 62.5% to be 〉=5/ 〉=5 homozygote (x ²=7.23, p=0.007).

Although the woman is arranged in matched group, women collator (34.8%, n=69) with male collator (37.8%, n=45) between, genotypicly do not have significant difference, x on popular 〉=5/ 〉=5 ²=0.106, p=0.74.

The factorial analysis factorial analysis is set to and only produces two factors.Factor 1 is tended to comprise and the related variable of drug dependence.These variablees comprise that the DSM with the form of service life, medicine score and the drug dependence of the intravenous injection abuse of medicine, the numeral of expenses for medicine, various medicines diagnoses related variable.Factor 2 is tended to comprise for example DUI number, used up money, the score of ethanol seriousness on ethanol, is carried out ethanol antidotal number of times, ethanol score and DSM diagnosis alcoholic dependence etc. and ethanol correlated variables.

Factor 1 and 2 The results of analysis of variance are represented with table 25.The average of factor 1 is the highest (0.198) in the homozygous study subject of 〉=5/ 〉=5 allele, all high (F-is than=2.85, p=0.060) than the study subject (0.151) of heterozygote or right＜homozygous study subject of 5/＜5 allele (0.085).This means exist one about 〉=5 repeat allelic homozygosity right 〉=regression model of the heterozygosity+homozygosity of 5 repetition allele (other).When this model is checked with variance analysis, the mean of factor 1 for those have 〉=5/ 〉=5 genotypicly significantly have ' other ' genotypic people (p=0.019) greater than those.On the contrary, no matter use 3 kinds of genotype or regression model, 2 means of factor all do not have significant difference.

These initial results be relevant with the allele of CNRI gene with drug dependence but with irrelevant identical of views of alcohol dependence.In order further to check this point, the inventor has tested regression model (table 26) for the mean of the numeral of ethanol score, medicine score and intravenous drug.Have 〉=5/ 〉=5 genotypic those people in, the mean of medicine score is than the mean obviously bigger (p=0.038) that has at those among its genotypic people.In addition, in 〉=5/ 〉=5 homozygous those people, the mean that uses medicine (#IV) by intravenous injection is than the mean obviously bigger (p=0.005) that has at those among other genotypic people.When Bonferroni correction α value was 0.5/3 or 0.016, the result was still significant.Homozygote does not have significant difference in fact comparing with ' other ' genotype aspect the mean of ethanol score on the contrary, 〉=5.In order to get rid of the probability that the age may be the covariant of hiding of an explanation results, the age has been taken as a covariant.The p value at age is 0.404, and the p value of the main effect of CNRI is 0.004, and this variation that demonstrates on the age can not be explained this result.

The CNRI gene is proposed whether and demonstrate closer related between certain specific medicine with the related inventor of making of average medicine score.4 class drug dependences are used 2 * 3 chi-square analysis methods and have carried out testing (table 27), as described in the method part.Between whole 3 groups: cocaine, amphetamine and Fructus Cannabis dependence group, 3 class drug dependences show significant increase on 〉=5/ 〉=5 genotypic frequencies.This equipotential gene relies on significantly not related with opioid.It is 0.05/4 or 0.0125 that Bonferroni proofreaies and correct the α value, and these related neither ones are significant.

In addition, alcohol dependence is included just to comparing purpose.Carry 〉=5/ 〉=5 genotypic but do not have the ATU study subject (50.7%) of alcohol dependence to compare with the study subject that alcohol dependence is arranged (29.6%), the former has higher percentage ratio.

The route of administration of cocaine, amphetamine, heroin.That table 28 shows is the result of the route of administration of these medicines.For cocaine, 〉=5/ 〉=5 genotypic popular in different study subjects, be distinguishing: in those ATU study subjects that do not use cocaine and collator is 36.2%, in the people of those suction cocaines is 32.3%, in those smoke the people who sucks cocaine is 55.6%, is 68.4% (p=0.006) in the people of those injection cocaines.Also obtaining similar result aspect the route of administration of amphetamine: carry 〉=5/ 〉=5 genotype but not use the popularity among the people of amphetamine be 35.7% at those, using among people of amphetamine with intravenous methods at those then is 65.2% (p=0.007).Have been noted that at those and use among the people of heroin through intravenous injection, exist identical high-frequency 〉=5/ 〉=5 genotype.

Regression analysis.In order to obtain the assessment percentage ratio to the three kinds of quantity score degrees of variation that are attributable to the CNRI gene, the inventor has carried out a regression analysis: wherein those have 〉=5/ 〉=5 genotypicly counted 2 fens, other genotype is counted 1 minute (table 29).The CNRI gene can not illustrate any one ethanol score in fact, and it can illustrate 3.8% (p=0.005) of 2.1% and intravenous drug score of whole medicine scores.

The result shows that CNRI gene and some different types of drugs rely on (cocaine, amphetamine, Fructus Cannabis) remarkable association is arranged, and aspect the intravenous drug abuse, it with drug dependence between have the related of strict especially form.Present result provides, and it is related that special genes and between the intravenous injection Drug abuse first are clearly described.In contrast, significantly not related between variable relevant and the gene with alcohol abuse/dependence.

A kind of potential substituting explanation to the result is: if do not consider the study subject of two groups should right and wrong Spain a kind of like this restriction of white people of system, to for the collator, the ethnic group layering factor of hiding in the ATU study subject can illustrate this result.It is to need the problem be concerned about in relatedness research always, although similar population statistical information is arranged with the study subject that the behavior of ATU drug dependence is arranged, 〉=5/ 〉=5 genotype only exist related with the drug dependence variable, but lack related with the variable of alcohol dependence, compare with ethanol, most drug all has bigger upset to dopaminergic recompense approach, interaction closely between the metabolism of dopamine and Fructus Cannabis, be consistent between these phenomenons, this unlikely describes the layering factor of ethnic group to the result.In addition, the age as complicated variable also by except.

Table 25

CNR1 genotype in ATU study subject and the contrast is to the variance analysis of factor 1 and 2

(n=205) score genotype N mean standard deviation F-is than p factor 1＜5/＜5 32-0.085 0.97 (medicine) heterozygote. and 91-0.151 0.65

≥5/≥5????83?????0.198????1.26??????2.85????0.060

Other are 123-0.134 0.75 years old

〉=5/ 〉=5 83 0.198 1.26 5.62 0.019 factor 2＜5/＜5 32-0.098 0.95 (ethanol) heterozygote. 91-0.042 1.00

≥5/≥5????83?????0.084????1.02??????0.52????0.593

Other 123-0.056 .98

≥5/≥5????83?????0.084????1.02??????0.98????0.32

Show of the variance analysis of 26CNR1 genotype to number of times, medicine score and the ethanol score of intravenous drug,

ATU study subject and contrast (n=206) score genotype N mean standard deviation F-than p ethanol score other 123 2.52 3.41

〉=5/ 〉=5 83 2.57 3.02 0.010 0.921 medicine score other 123 3.37 4.98

〉=5/ 〉=5 83 4.95 5.81 4.33 0.038# intravenous drug other 123 0.16 0.54

≥5/≥5????83?????0.48????1.05??????8.08????0.005

Table 27

CNR1 gene 〉=5/ 〉=5 homozygosity in the positive study subject of collator, ATU feminine gender and ATU

Variance analysis (n=155) the behavior contrast ATU-ATU+ of percentage ratio

N % N % N % card side p cocaine 114 36.0 72 38.9 20 70.1 5.36 0.020 amphetamines 114 36.0 58 37.9 34 58.8 4.46 0.034 hemps 114 36.0 66 39.4 26 61.5 4.41 0.035 opioids 114 36.0 82 45.8 10 50.0 3.01 0.155 only have alcohol dependence 114 36.0 71 50.7 21 28.6 0.27 0.602

Table 28

The route of administration of CNR1 genotype and medicine.8A. cocaine approach N % 〉=5/ 〉=5 card side ^*P does not use 138 ¹36.2 per nasal 31 32.3 smog 18 55.6 intravenous injections 19 68.4 7.52 0.006 total 2068B. amphetamine approach N % 〉=5/ 〉=5 card sides ^*P does not use 129 ²35.7 oral 11 9.1 per nasal/smog 43 48.8 intravenous injections 23 65.2 7.19 0.007 total 2068C. heroin approach N % 〉=5/ 〉=5 card sides ^*P does not use 177 ³37.3 per nasal 6 33.3 smog 2 50.0 intravenous injections 21 66.7 6.42 0.011 sums 206

* the linear card side of Mantel-Haenzel

¹Contain 114 contrasts, wherein 36.0% is 〉=5/ 〉=5 homozygote,

With 24 ATU study subjects, wherein 37.5% is 〉=5/ 〉=5 homozygote.

²Contain 114 contrasts, wherein 36.0% is 〉=5/ 〉=5 homozygote,

With 15 ATU study subjects, wherein 33.3% is 〉=5/ 〉=5 homozygote

³Contain 114 contrasts, wherein 36.0% is 〉=5/ 〉=5 homozygote,

With 64 ATU study subjects, wherein 40.6% is 〉=5/ 〉=5 homozygote.

Table 29CNRI genotype (〉=5/ 〉=5=2, other=1) vein of ATU study subject and contrast is annotated

Penetrate regression analysis (n=206) the score r r of medicine number of times, medicine score and ethanol score ²T P ethanol score 0.007 0.0000 00.099 0.921 medicine score 0.144 0.0208 20.081 0.038 intravenous drug score 0.195 0.0381 20.844 0.005

The hereditary variant of embodiment 5 people's obesity (OB) genes and the correlating method and the material of body quality, psychiatric disease and d2 dopamine receptor gene (DRD2)

Health promotion organization center (CHP).Study subject is made of the white people from 211 non-Spain blood lineages of health promotion organization center research project.According to the efficiency analysis to other researchs related with the behavior variable of the polymorphism of heredity, the inventor searches out the sample size of 200～225 study subjects, and their age is from 29～75 years old, 54 years old mean age, standard deviation (S.D.) 10.2 years old.98 men and 113 woman are arranged in this group.Each study subject has all been determined the age, sex, body weight, the ratio of height and waist-hip.Total body fat is decided by weighing in water.The all ages and classes that each study subject all is required to provide the age now from 16 years old to them at interval, relevant their data of the heaviest body weight.For detecting glucose, cholesterol and the insulin in the blood, extracted blood sample on an empty stomach.Sample after the numbering is used to analyze, and analyzes under the blind situation of the data sheet of health promotion organization center and carries out.

Each study subject has all been finished NEO 5-factor personality check questionnaire (Costa and McCrae, 1992) and disease is checked catalog-90 (SCL-90).study subject has also been answered a following cover problem: are you because starve just feed? gluttony? between dinner, eat a piece? take food because feel blue? take food owing to feeling pressure? have breakfast? overfeeding when dinner? to the sweet food addiction? do you like vegetable? do you like food fat or fry? possible answer is: 1: never; 2: once in a while, 3: often, 4: very frequent, 5: such was the case with.

Genetic research.The inventor uses PCR ^TMMethod, the D7S1873 that increased, D7S1875, D7S514 and D7S680 dinucleotide repetitive sequence, as being described (Green etc. 1995) in the past, they are present among the YAC that contains people OB gene.In these sequences, D7S1875 is nearest from the OB gene.The inventor is called OB1875 with this section sequence.DRD2 gene TaqAl allele is also checked with aforementioned (.1994b such as Noble) primer.

OB ₁₈₇₅Genotypic statistical analysis.A previous hypothesis is: if the allele repetitive sequence of or length shorter than D7S1875 dinucleotide and BMI or other variable are relevant, have short (perhaps long) allelic homozygous individual and will get the highest mark, those heterozygotes will get intermediary mark, and those have long (weak point) allelic homozygote and will get minimum mark.Like this, the mean of the behavior score of the study subject in the different allele groups is just compared, the method of using is from SPSS (SPSS Inc., Chicago, the Illinois) the variance analysis statistical procedure of company, wherein subcommand " multinomial " is set as 1, so that carrying out property linear relationship is checked.In the time of at least 3 groups, the Tukey analytical method is used to detect the significance individual variation between any group, wherein α=0.05.Covariance analysis (ANACOVA) is used for checking OB ₁₈₇₅Genotype is to using the influence as the SCL-90 anxiety score aspect of covariant of BMI, waist-hip ratio, age and sex.In addition, to three OB ₁₈₇₅Genotype has carried out having the chi-square analysis of two disjunction point variables.Because previous supposition is: at these 3 OB ₁₈₇₅Between the genotype, will there be a kind of progressive linear increasing or minimizing, so they have carried out checking (the Mantel-Haenszel chi-square butter in the SPSS statistical package) with linear Chi-square method.Finally, use regression analysis to determine the dependency (r) between OB and DXD2 genotype or the allele.R ²The part explanation that provides the combination of relevant gene or gene to provide to degree of variation (variance).The result

The BMI meansigma methods is 27.9, and standard deviation (S.D) is 7.2, and scope is 17.7～57.6.In preliminary study, by allele being divided into the group of a long and weak point, the allele of whole 4 kinds of polymorphisms all has been verified.Study subject is divided into by gene type: those short allelic homozygotes, long allelic homozygote and short and long allelic heterozygote.The BMI that carries out in 3 genotype groups checks that demonstrating the D7S1875 polymorphism has the highest F-ratio.Research like this, afterwards all uses this polymorphism that other variable is studied.

OB ₁₈₇₅Allele distributions.The OB of health promotion organization center study subject ₁₈₇₅The different allele of polymorphism, its length range are 199～225bp.The inventor to the existing method (Comings etc., 1996) of carrying out the dinucleotide repetitive sequence checking in psychiatric disturbance is, if there is the conatus of bimodal distribution, just at first will be divided into about equally group to allele.OB ₁₈₇₅The situation of polymorphism comes to this, and cut-point fixes on 208bp.The genotype grouping situation that produces is:＜208bp/＜208bp,＜208bp/ 〉=208bp and 〉=208bp/ 〉=208bp.

Fat variable.For comprising the men and women whole group only has significance related with body weight, progressively reduces (＜208/＜208 n=47,183.83kg, standard deviation (S.D) 45.39 between 3 genotype groups; Heterozygote n=95,177.56kb, standard deviation (S.D) 44.00; 〉=208/ 〉=208 n=39,161.29kg, standard deviation (S.D) 45.17, F-is than=5.22, p=0.023).For BMI, exist a kind of progressivity but be not significant reduction, the BMI value is from＜208/＜208 homozygous 28.92, to heterozygote 27.96 and 〉=208/ 〉=208 homozygous 26.62.On the mean of plasma insulin, fasting blood glucose, cholesterol and percent fat, the difference that different genotype causes is not remarkable.When considering each sex, for the male, tendency is identical.The BMI value is near significantly (p=0.053), and body weight different (p=0.038) significantly.Tendentiousness also is identical, and the neither one variable is significant to the woman only.

BMI according to the age.Table 30 is presented in the study subject (comprising the men and women) of all ages and classes, the comparison of being undertaken by variance analysis between the mean of BMI.Concerning the study subject in 26-30 year, the difference that different genotype causes is significant, and concerning the study subject in 16-20 year, difference almost is significant.When only the women being tested, same, only the 26-30 age bracket has shown significant result (p=0.028).When only the male being tested, any one age group is not remarkable.The study subject age is when being 26～30 years old, their BMI divided for＜25,25～34 and 〉=3 groups of 35.Between these 3 BMI groups, OB ₁₈₇₅＜208/＜208 genotypic frequencies are progressivity to be increased, and promptly from 24% to 33% again to 56% (linear card side=4.46, d.f=1, p=0.034.)。

SCL-90。The result of SCL-90 is shown in table 31.＜208/＜208 homozygous marks are obviously higher aspect anxiety, depression, psychosis, hostility, paranoid idea, obsession, symptom summation, the general overall summation of disease exponential sum.In the above-mentioned score 7, p value＜0.025.

Covariance analysis.In order to check for body weight question or fat type (waist-thigh ratio) the psychiatry variable this probability that only occupies a secondary and subordinate position, score is checked to the SCL-90 anxiety, uses OB ₁₈₇₅Genotype is as main effect, and BMI, waist-hip ratio, age and sex are as covariant (table 32).The result shows: in these 5 variablees, have only between demonstration of OB1875 genotype and the anxiety mark to have significant association.Related same result arranged also to what the SCL-90 mark of depressed, the total positive symptom and symptomatology carried out.

DRD2 and BMI age group.Carried out D ₂A ₁The variance analysis whether allele exists is so that determine whether the DRD2 gene also works in the BMI of given age.When study subject is that 31-40 is in the time of year, for carrying the allelic individual of A1, the obviously bigger (D of its average BMI value ₂A1+n=54,28.78, standard deviation (S.D)=8.56; D ₂A1-n=70,25.67, standard deviation (S.D) 5.21, F-is than=6.04, p=0.015).The same with the OB gene, concerning older age group the F-ratio tend to lower, although sample size is also less.

The OB that merges ₁₈₇₅＜208/＜208 homozygosity and D ₂A1 allele.For whether the effect of determining OB and DRD2 gene has accumulative action, by study subject is divided into OB ₁₈₇₅The people that＜208/＜208 allele isozygoty and/or carry D ₂The allelic people of A1, and then their BMI carried out checking (table 33).These are highly significant as a result.Those are OB ₁₈₇₅＜208 allelic homozygotes also/or carry the allelic people of DXD2 D2A1, be 16-20 when their age, 21-25,26-30, in 31-40 and 41-50 when year,, they have significantly higher average BMI value.When the effect of these 2 genes is merged, and a complete set of fat variable is when all being carried out inspection, and concerning waist/hip ratio (p=0.033), body weight (0.013) and insulin level (0.042), difference is significant.

Regression analysis.Univariate regression analysis allows the variance (r by OB gene or OB+DRD2 gene BM1 explanation, given age ²) percentage ratio checks (table 34).Comprise the men and women all, only for the OB gene, r and r ²Only the BMI to 16-20 year and 26-30 year age group is significant.Here, the variance of these 2 BMI of OB gene pairs provides roughly 3% contribution.Yet, when the effect of OB and DRD2 gene is merged, at 16-20,21-25,26-30 and 31-40 age group, they have reached 8.5-9.9% to the variance contribution of these BMI.When only the women being tested, only with regard to the OB gene, this association is significant (p=0.029) to 31～40 age brackets only.When the effect of the merging of OB and DRD2 gene is verified, when study subject is 16-20,21-25,26-30,31-40 and 41-50 are in the time of year, and this association is significant to study subject.Significant association is at 26～30 years old (p=0.0004) and 31～40 years old (p=0.0005) age group.These 2 genes have illustrated 22.8% of BMI variance here.

N.E.O, for pleasant, conscientious, export-oriented, nervousness and open these 5 N.E.O factors, it is significant only having nervousness to improve.These 3 kinds of OB ₁₈₇₅Genotypic score is respectively 21.16,17.82 and 14.86, F-ratio=7.29, p=0.008.Conscientious property reduction is that critical significant (32.97,34.67 and 35.65, F-is than=3.71, p=0.056).

The reason of feed.In those problems about the feed reason that are asked, 2 reasons are only arranged promptly: taking food and have breakfast owing to feeling pressure is and OB ₁₈₇₅Genotype has remarkable association.For these three genotype, the score of " taking food owing to feeling pressure " is 2.55,2.32,1.85, F-ratio=4.54, p=0.034.The score of ' having breakfast ' is 3.50,4.15,4.14.F-ratio=4.89, p=0.028.

When the BMI that uses based on the present body weight of study subject, OB ₁₈₇₅Genotypic phenotype effect does not have any evidence.Though there is a kind of tendency, homozygote promptly＜208/＜208 has the highest BMI value, and heterozygote is placed in the middle, and the homozygous value of 〉=208/ 〉=208 allele is lower, and the result is not remarkable.Difference on ratio, percent fat, plasma insulin and the blood glucose of waist-hip is not remarkable, but the significant difference on body weight (p=0.02).

When the study subject age is 16～30 years old, OB ₁₈₇₅Have significance or the related tendency of significance almost between the BMI value of＜208/＜208 homozygosity and study subject.But when they are 41～70 years old, the BMI value and the OB of study subject ₁₈₇₅Just have only few between＜208/＜208 homozygosity or do not have association.And this tendentiousness is only carrying out female gender and OB1875＜208/＜208 homozygosity also to exist when related.On the contrary, only male gender and OB ₁₈₇₅When＜208/＜208 homozygosity are carried out association, can only see non-significant tendency (table 30).Exist related between these results have hinted the OB gene and youthful BMI makes a variation.When the age be divided at 26～30 years old men and women's BMI＜25,25 to 34 and＜35 groups in, the isozygoty percentage ratio of study subject of＜208 allele has the increase of significant and progressivity, promptly from 24% to 56%.

Related more remarkable with based on the psychiatry variable of SCL-90.Between 2 kinds of behaviors and the obesity the most frequent generation related-be anxiety and depression, they and OB ₁₈₇₅＜208 allelic homozygosity have remarkable association (p=0.003-0.0005).For anxiety, the average (0.85) of＜208 allelic 36 study subjects that isozygoty be those heterozygotes (0.277) or 〉=more than the twice of 208/ 〉=208 homozygote (0.203) study subject average.OB ₁₈₇₅Exist a related reasonable dismissal to be between＜208 allele and anxiety and the depression: these allele are causeed fat, and the effect of anxiety and depression is accessory.In order to examine this point, the inventor uses BMI, waist-hip ratio, age and sex as covariant, OB ₁₈₇₅Allele has carried out covariance analysis as main effect.For anxiety, analysis result shows: with the unique variable of SCL-90 anxiety score significant correlation be OB ₁₈₇₅〉=208 allelic exist (tables 32), this just shows that the psychiatric disease of OB gene pairs has main influence, and may be to playing direct inductive effect with related depression of obesity and anxiety.

For the effect of check DRD2 gene in health promotion organization center study subject, the D that has that variance analysis was used at the various ages ₂The allelic study subject of A1 and do not carry in the allelic study subject of Al relatively their BMI.To OB ₁₈₇₅The allele carrier, the group at lower age, 31-40 year age group for example, its RMI and this equipotential gene have just in time significant association p=0.015.

In order to check 2 or more polygenic cumulative effects, the inventor is to OB ₁₈₇₅The allelic homozygote of＜208bp and/or carry D ₂The allelic study subject of A1 to the BMI group of different given age, has carried out variance analysis.For the situation that the men and women combines, OB ₁₈₇₅＜208/＜208 or D ₂There are significant association (table 34) at allelic existence of A1 and age between the high BMI value of all groups of 16～50 years old.Whole p value of 4 groups at 16～40 years old age all is＜0.005.When only checking the woman, although the number of study subject still less, the result is more remarkable.The p value scope of 16～40 years old age group is from 0.0017～0.0004.Waist-hip the ratio, body weight and the plasma insulin level that carry the study subject that any one or both in allele or the genotype carry all have significant association with this equipotential gene or genotype.

As (Comings etc., 1996f in the research formerly; Comings etc., 1996; Comings etc., 1997b), the regression coefficient r between the genotype of discussing at present and given mark determines, feasiblely can determine r ²Or by the ratio of the mark amount of variability of the gene that is being studied explanation.The result of the BMI value of these given age represents with table 34.Merged the men and women under the situation of research, at 16-20 and 26-30 year age group, the OB gene illustrated alone respectively BMI amount of variability 3.2 and 3.0%.Under the situation that only has the woman to be studied, these values are increased to 6.2 and 4.1%.When the effect of OB gene and DRD2 gene is added up, merged the men and women under the situation of research, when study subject is 16～40 years old, these genes illustrated BMI variation 8.5～9.9%.When only the woman was verified, at 16～40 years old age group, these 2 genes illustrated 19.1～22.8% (p=0.0017～0.0004) of the BMI mark amount of variability of study subject.

Table 30

OB ₁₈₇₅The linear equation analysis of the BMI value that genotype is carried out in the CHP study subject according to the BMI genotype N mean standard deviation F-at age than p masculinity and femininity 16-20 year＜208/＜208 38 24.26 3.95

Heterozygote 63 22.66 3.74

〉=208/ 〉=208 27 22.50 3.64 3.58 0.06121-25 year＜208/＜208 38 25.20 4.96

Heterozygote 65 24.22 4.56

〉=208/ 〉=208 27 23.54 4.80 2.02 0.13926-30 year＜208/＜208 38 26.83 6.44

Heterozygote 65 24.79 4.67

〉=208/ 〉=208 27 24.22 5.57 4.05 0.04631-40 year＜208/＜208 37 27.83 6.11

Heterozygote 65 26.92 6.35

〉=208/ 〉=208 26 25.79 9.14 1.31 0.27541-50 year＜208/＜208 32 28.51 6.50

Heterozygote 60 27.86 8.07

〉=208/ 〉=208 24 27.38 6.21 0.33 0.56451-60 year＜208/＜208 20 30.19 8.24

Heterozygote 43 27.52 6.61

〉=208/ 〉=208 14 28.40 9.89 0.64 0.42461-70 year＜208/＜208 14 29.35 7.32

Heterozygote 23 26.66 3.57

〉=208/ 〉=208 7 26.61 6.98 1.67 0.202 according to the BMI genotype N mean standard deviation F-at age than p women 16-20 year＜208/＜208 19 24.00 4.09

Heterozygote 35 21.78 3.72

〉=208/ 〉=208 21 21.83 3.48 3.18 0.07821-25 year＜208/＜208 19 25.30 5.73

Heterozygote 37 23.17 4.89

〉=208/ 〉=208 21 22.86 4.86 2.20 0.14026-30 year＜208/＜208 19 28.04 8.00

Heterozygote 37 23.93 5.11

〉=208/ 〉=208 21 23.60 5.87 4.99 0.02831-40 year＜208/＜208 18 28.94 7.28

Heterozygote 37 26.35 7.77

〉=208/ 〉=208 21 25.46 10.08 1.61 0.20841-50 year＜208/＜208 14 28.33 7.00

Heterozygote 34 28.28 8.84

〉=208/ 〉=208 19 27.51 6.85 0.10 0.75251-60 year＜208/＜208 9 30.01 7.90

Heterozygote 20 27.11 5.22

〉=208/ 〉=208 12 29.07 10.26 0.03 0.85961-70 year＜208/＜208 6 26.93 4.55

Heterozygote 10 25.23 4.28

〉=208/ 〉=208 6 26.69 7.65 0.006 0.939 according to the BMI genotype N mean standard deviation F-at age than p male 16-20 year＜208/＜208 19 24.51 3.89

Heterozygote 28 23.77 3.53

〉=208/ 〉=208 6 25.27 3.22 0.001 0.97321-25 year＜208/＜208 19 25.09 4.19

Heterozygote 28 25.52 3.79

〉=208/ 〉=208 6 25.90 4.07 0.24 0.626-30 year＜208/＜208 19 25.62 4.25

Heterozygote 28 25.94 3.80

〉=208/ 〉=208 6 26.39 4.01 0.18 0.67131-40 year＜208/＜208 19 26.79 4.71

Heterozygote 28 27.67 3.76

〉=208/ 〉=208 5 27.19 3.38 0.25 0.61941-50 year＜208/＜208 18 28.64 6.28

Heterozygote 26 27.31 7.07

〉=208/ 〉=208 5 26.88 3.74 0.49 0.48851-60 year＜208/＜208 11 30.38 8.89

Heterozygote 23 27.88 7.71

〉=208/ 〉=208 2 24.45 0.15 1.21 0.27861-70 year＜208/＜208 8 31.18 3.08

Heterozygote 13 27.74 0.71

≥208/≥208????1?????26.11???????1.98????????0.174

Table 31

OB ₁₈₇₅The linear equation of polymorphism and SCL-90 score is analyzed SCL-90 score genotype N mean standard deviation F-than p anxiety＜208/＜208 36 0.850 1.36

Heterozygote 65 0.277* 0.33

〉=208/ 〉=208 29 0.203* 0.29 12.72 0.0005 depression＜208/＜208 33 0.935 0.79

Heterozygote 63 0.552* 0.59

〉=208/ 〉=208 30 0.453* 0.52 9.14 0.0030 psychosis＜208/＜208 35 0.426 0.60

Heterozygote 65 0.198* 0.28

〉=208/ 〉=208 30 0.183* 0.24 6.58 0.011 hostility＜208/＜208 36 0.509 0.66

Heterozygote 66 0.255* 0.36

〉=208/ 〉=208 30 0.227* 0.38 6.36 0.013 paranoid idea＜208/＜208 37 0.527 0.56

Heterozygote 67 0.335 0.42

〉=208/ 〉=208 30 0.278 0.38 5.23 0.024 force＜208/＜208 37 0.876 0.69

Heterozygote 64 0.703 1.03

〉=208/ 〉=208 31 0.443 0.42 4.34 0.035, somatization＜208/＜208 33 0.715 0.72

Heterozygote 65 0.549 0.48

〉=208/ 〉=208 28 0.467 0.34 3.41 0.067 interpersonal sensitivity's property＜208/＜208 34 0.732 0.73

Heterozygote 66 0.569 0.84

〉=208/ 〉=208 30 0.496 0.57 1.55 0.214SCL-90 score genotype N mean standard deviation F-is than the terrified anxiety of p＜208/＜208 37 0.212 0.46

Heterozygote 66 0.076 0.18

〉=208/ 〉=208 30 0.138 0.41 1.05 0.305 score＞0＜208/＜208 29 33.72 21.48 symptoms sum heterozygote, 52 21.23* 15.50

〉=208/ 〉=208 37 20.85* 18.65 7.33 0.008 score f0＜208/＜208 29 62.51 58.86 symptoms sum heterozygote, 52 36.42* 33.15

≥208/≥208???27?????31.44*?????29.51?????8.24????0.005

* the Tukey assay is remarkable, α=0.05.

Table 32

SCL-90 anxiety variable is with OB ₁₈₇₅Polymorphism is as main effect, and BMI, waist-hip compare, year

Age and sex amount to d.f. card side mean F p covariant 1.369 4 0.342 0.518 0.677RMI 0.763 1 0.263 0.447 0.505 waist-hip than 0.046 1 0.046 0.078 0.780 age, 0.255 1 0.255 0.433 0.512 sex, 0.374 1 0.374 0.634 0.427 main effect OB as the analysis of covariance anxiety card side of covariant₁₈₇₅7.855 2 3.928 6.669 0.002 explicable 9.495 6 1.58 2.687 0.012 remaining 71.258 122 0.589 sums 81.345 128 0.636

Table 33 is OB in the CHP group ₁₈₇₅＜208/＜208 homozygosity and/or DRD2 D ₂A1 allele

The relevant BMI genotype N mean standard deviation F-of the BMI analysis of variance of age of existence-all ages and classes than p masculinity and femininity 16-20 year OB+ and/or D ₂A1+ 51 24.52 4.15

OB and D ₂A1-39 22.03 3.24 9.45 0.0028 21-25 year OB+ and/or D ₂A1+ 51 26.05 5.31

OB and D ₂A1-39 23.00 3.49 9.60 0.0026 26-30 year OB+ and/or D ₂A1+ 53 27.25 6.36

OB and D ₂A1-39 23.69 3.46 9.94 0.0022 31-40 year OB+ and/or D ₂A1+ 52 29.11 7.26

OB and D ₂A1-39 25.29 4.48 8.38 0.0048 41-50 year OB+ and/or D ₂A1+ 44 29.92 8.18

OB and D ₂A1-37 26.34 7.08 4.32 0.0408 51-60 year OB+ and/or D ₂A1+ 51 30.06 7.39

OB and D ₂A1-39 26.77 6.97 2.83 0.0984 61-70 year OB+ and/or D ₂A1+ 16 28.23 5.42

OB and D ₂A1-14 36.49 4.22 0.94 0.339 women 16-20 year OB+ and/or D ₂A1+ 25 24.50 4.42

OB and D ₂A1-24 20.98 2.74 11.07 0.0017 21-25 year OB+ and/or D ₂A1+ 25 26.32 6.32

OB and D ₂A1-24 21.61 2.76 11.24 0.0016 26-30 year OB+ and/or D ₂A1+ 27 28.53 7.91

OB and D ₂A1-24 22.19 2.12 14.45 0.0004 31-40 year OB+ and/or D ₂A1+ 26 31.09 8.98

OB and D ₂A1-24 23.66 4.06 13.97 0.0005 41-50 year OB+ and/or D ₂A1+ 20 32.05 10.19

OB and D ₂A1-23 25.90 5.73 6.08 0.018 51-60 year OB+ and/or D ₂A+ 9 31.10 7.23

OB and D ₂A1-16 27.01 5.61 2.48 0.128 61-70 year OB+ and/or D ₂A1+ 28.46 4.28

OB and D ₂A1-7 24.56 4.64 2.19 0.169

Table 34 is about OB in CHP group all ages and classes study subject ₁₈₇₅＜208/＜208 homozygosity and/or

DRD2 D ₂The allelic unitary variant regression analysis that has situation of A1

R r ²T POB1875＜208/＜208=1; Other=0: comprise masculinity and femininity 16-20 year 0.173 0.030 1.98 0.04921-25 year 0.131 0.017 1.49 0.13726-30 year 0.181 0.032 2.09 0.03831-40 year 0.108 0.012 1.22 0.22241-50 year 0.063 0.004 0.68 0.49851-60 year 0.103 0.010 0.98 0.36661-70 year 0.215 0.046 1.45 0.153OB1875＜208/＜208=1, other=0: women 16-20 year 0.204 0.041 1.77 0.07921-25 year 0.171 0.029 1.49 0.14126-30 year 0.248 0.062 2.22 0.02931-40 year 0.146 0.021 1.28 0.20541-50 year 0.039 0.001 0.32 0.75151-60 year 0.028 0.000 0.18 0.85961-70 year 0.017 0.000 0.08 0.938OB₁₈₇₅＜208/＜208 and/or D ₂A1=1, other=0: masculinity and femininity 16-20 year 0.312 0.097 3.07 0.002921-25 year 0.313 0.098 3.09 0.002626-30 year 0.315 0.099 3.15 0.002231-40 year 0.292 0.085 2.89 0.004841-50 year 0.228 0.052 2.08 0.04151-60 year 0.227 0.051 1.68 0.09861-70 year 0.181 0.032 0.97 0.339

R r ²T p OB ₁₈₇₅＜208/＜208 and/or D ₂A1=1, other=0: women BMI 16-20 0.436 0.191 3.32 0.0017 BMI 21-25 0.439 0.193 3.35 0.0016 BMI 26-30 0.477 0.228 3.80 0.0004 BMI 31-40 0.472 0.223 3.72 0.0005 BMI 41-50 0.359 0.129 2.46 0.018 BMI 51-60 0.312 0.097 1.57 0.128 BMI 61-70 0.423 0.179 1.48 0.169

Embodiment 6

Additive effect with three dopamine gene-correlations

In this research, study this three additive effects that influence the important gene of dopaminergic neuron by determining whether the behavior scoring that heredity has the experimenter of the special sign of all these three genes to tend to than those heredity to have less than three experimenter higher (relatively poor).By determine whether those do not have these signs of heredity tend to have the behavior scoring of lower (better) and detect subtracting property effect, the effect opposite with additive effect.By determining whether that those heredity have the tendency of 1 or 2 sign to have intermediary scoring and detect this two kinds of effects.This additive effect that has detected the essence-several genes of needs of polygenic inheritance produces clinical significant effect on the phenotype.Method

Experimenter experimenter is patient that TS is treated or patient's relatives.The diagnosis of TS, chronic exercise tic disorder or chronic sounding tic disorder is based on DSM-III-R standard (the sick community of Americanism, 1987).TS proband (n=225) is defined as the individuality of seeking Medicare in this clinic.Among the proband, 82% has TS and remaining 18% has chronic exercise tic disorder or chronic sounding tic disorder.In non-proband TS relatives (n=60), 54% suffers from TS, and 31% suffers from the chronic exercise tic disorder and 15% suffers from chronic sounding tic disorder.Nobody has chronic exercise or sounding twitch among the non-TS relatives (n=132).For all individual's meeting and inspections of all proband.Relatives more than 80% also individual meet with.Inquire each proband and their relatives race and ethnic background about they four grand parents.For DRD2 research, only comprise non-Spain white race people proband, relatives and have the north and the contrast of western European background.All experimenters are all above 6 years old.

The ethnic group layering.In this research, avoided the ethnic group layering.All experimenters all are limited to the non-Spain white race people of the north or western European pedigree.Determine each experimenter's ethnic background by all four grand parents.The quantity of different lineages is between 1 to 12, and most subjects has 4 to 6 different lineages.For D2A1 allele, 13 different researchs have detected 714 contrasts of getting rid of ethanol and drug dependence through screening altogether.These research in the allelic prevalence rate average out to 25.9% of D2A1, scope is 12.5% to 34.8%, all be lower than in 432 TS cases 40.7%.(also seeing the super comparative study of in this application, showing previously).When detecting average behavior scoring in D2A1 carrier and non-carrier, several averages also have significance when getting rid of contrast from analyze.Joint-detection for all three dopamine genes also is like this.Different experimenters tend to scoring height on different behavior scorings.However, when reaching " case has " three groups, " contrast does not have ", " case does not have " still have many significant results when detecting.All these results, the stratified result whether of the existence as implicit ethnic group layering but not by institute's detection variable in fact, as if its probability very little.Utilize contrast and experimenter's identical polygenes group, all three tested dopamine genes are all obtained positive findings.As if relate to the stratified probability of implicit ethnic group in these three genes at all very little.These positive findingses are not only to be that therefore accidental are arranged is significant owing to detected a plurality of behavior variablees.The inventor is total has obtained non-significant result for all behavior variablees of several other genes of being surveyed.

Contrast.Contrast (n=67) is from three sources: a) TS patient's adoptes, brings up or stepparent, b) suffer from the experimenter of thyroid carcinoma or non-insulin-dependent diabetes mellitus from an endocrinopathy clinic, and c) hospital personnel comprises professional, technical staff and service worker.Because two kinds of situations all are easy to treatment, have high cure rate and daily life is produced minimum interference, exist in the range of age widely one, and patient's basis identical with TS experimenter, so select the endocrine patient in contrast.Screen all contrasts to get rid of ADHD, ethanol, medicine and tobacco abuse.

The structuring application form.From 1987, require all patients and the first degree relative that can find is filled in one according to (DIS) 31 pages of detailed behavior application forms of (Robins etc., 1991) design of diagnostic meet flow sheet (Diagnostic InterviewSchedule).Complete application form provide elsewhere (Comings, 1990a).Except the DIS problem, also have many about the required whole DSM-III of demographic variable, ADHD diagnosis and the problem of DSMIII-R variable, and type, location, persistent period and seriousness problem (the sick community of Americanism, 1980 of twitching about mobility and sounding; The sick community of Americanism, 1987).To import a SPSS data base to these questions answer then.The various aspects that the use of this instrument and syndrome use provide (Comings, 1995c elsewhere; Comings, 1994c; Comings, 1994a; Comings, 1995a).

Blood preparation.Though TS proband or the relatives to each prescription on individual diagnosis all do not adopt whole blood sample, in the limit of consideration below, selecting is at random basically.At first, it is fully voluntary that blood preparation is provided, and many experimenters particularly year selection lightly do not draw blood.Secondly, owing to about race and the stratified consideration of ethnic group, tend to from non-Hispanic the north and western European white people, draw blood, because most of data of contrast is from this colony.The 3rd, if it is two father and mother all can provide, then preferred from these families but not the family that one of father and mother do not provide obtains blood sample.

Gene type.All patients are all carried out gene type based on a neutral ID, and under not knowing, read sheet by the typing individual instances.Carry out D according to the hybridization by the dna molecular trace described above ₂A ₁Gene type (Comings etc., 1991).Some is also according to the hereinbefore described PCR that passes through ^TMTechnology is carried out gene type (Dawson, 1986).According to the described D β H gene type that carries out in an embodiment.Repeat the polymorphism gene type according to the DAT1 that carries out described in embodiment 3.

Be total to the standard of morbidity.The structuring application form allows serial related indication detection.Before this research, determined the wherein standard of each, comprise: alcohol abuse (Comings, 1994b), drug dependence (Comings, 1994c), compulsion (Comings, 1994a), major depression outbreak (Comings, 1995c), dry mad (Comings, 1995b), somatization disorder (Comings, 1995b), panic attack (Comings, 1995b), phobia (Comings, 1995b), conduct disorder (Comings, 1995a), disobedience obstacle (ODD) (Comings, 1995a), sexual disorders (Comings, 1994c), learning disorder (Comings, 1995b) and twitch seriousness (Comings, 1995a).The summation of all 22 used ADHD variablees in application form represent in ADHD scoring, does not wherein have or never=0, once in a while=1 and frequent=2.Based on the early stage research (Knell and Comings, 1993), with those have the scoring be 21 or higher being thought of as suffer from ADHD.Because the special interest of inventor in ADHD, the inventor also utilize another set of strictness based on the standard detection of DSM-III-R diagnosis it, wherein need 8 (the sick community of Americanism, 1987) among 14 of top 22 variablees at least.

A kind of method based on application form for accuracy, practicality and the susceptiveness of symptom by the use of other people a kind of like this instrument by more all being used for same subject and structuring instrument that a kind of person of meet uses as being used for affective disorder and schizoid Kiddie flow sheet be confirmed (Karp, 1994; Grossman etc., 1994).

Be standard or scoring below for some symptoms, they write the application's fashion do not deliver or may need the clarification.Be to estimate the general performance (primary school's symptom) in school, the inventor asks following problems: " from 1 to 6 grade, your school shows a) mathematics, b) read and c) in the writing aggregate performance be lower than on average, on average or be higher than average.Possible answer is to be lower than average (2), and average (1) is higher than average (0).The result is the summation of three subjects, and difference is 6, is preferably 0.For two variation per minutes, 4 minutes or the primary school's problem positive that is considered to more.Education was disadvantageous, study is disadvantageous if they once had to be placed in, learning disorder or the class of remedying, then these individualities problem (learning disorder symptom) of being considered and having learning disorder.The experimenter is inquired in DIS the problem that has (phobia symptom) about all 16 phobias, and if they on 3 or more items, have any problem, then be considered to have the problem of phobia.For a given experimenter, the phobia scoring is the sum of the listed phobia of DIS.If individual to the DIS problem " you once had in the unafraid place of most people quail suddenly, anxiety or very uneasy a period of time or outbreak (disease of body no thanks to)? " answer be then think panic attack (panic attack symptom) some the time existed.To a given individuality, the sum of DIS panic attack symptom is represented in terrified scoring.Be to estimate reading problem (reading problem symptom), individual by inquiry " you feel to fall behind the biggest year number with you equal people in reading, if having; be what? for example; if when you at 6 grades, and you are only 4 grades level reading, you may fall behind 2 years so." those were answered 2 years or more are considered to have reading problem.The stutter (stutter symptom) be by problem " you once had the stutter problem? " estimate.For being the male individuality of extensive anxiety (extensively anxiety symptom) by scoring, two DIS problems below must answering, " you overanxiexty was once arranged or worry you live in stage of this or that? " and " if; when time that their exist when not existing, these persistence of sensation 6 months are still more of a specified duration? "If individual to 3 in the DIS problem that relates to somatization symptom or more answers be then be considered to exist the problem (somatization symptom) of somatization.To a given individuality, the sum of DIS somatization symptom is represented in the somatization symptom scoring.If any one in following: the problem of falling asleep night, night-walking, night fear, early awakening and can't fall asleep again, sleep-talking or nightmare, every day or almost all exist every day, then be considered to have sleeping problems (sleep symptom).The sum of the sleep scoring symptom of sleeping scoring above-mentioned every day of representative or almost taking place every day.If have the individuality of sexual disorders exist below in any one (Comings, 1994a): 1. frequent show-and-tell, 2. the property requirement is much higher than the ordinary people, 3. like the same sex or two kinds of sexes, 4. on the sexual behavior thing, has precocious interest, 5. as a child, draw dirty picture greatly more than other children of the same age, 6. feel always that they have been given birth to sex wrong, as opposite sex wear and be not for the All Saints' Day or the makeup party, 8-11.6 individual month or longer a period of time are by object, or child, or be subjected to cruel or execute cruel illusion and by sexual arousal, then positive by scoring.In a given individuality, the sum of the above-mentioned sexual behaviour problem of sexual behaviour scoring representative.Individuality with disruptive behaviour (division sample behavior symptom) is if they are to about 2 in the DIS problem of the symptoms of schizophrenia or more answers being then marked positive.The division sample behavior scoring of given individuality is represented the sum of DIS division sample symptom.By twitching about 8 different motion and the problem at age of existence that 9 different sounding are twitched and beginning is come that the experimenter is twitched (twitch symptom) and detected.The quantity addition of this twitch is produced a twitch scoring.So the existence scoring of multiple twitch is higher than less twitch.Above for chi-square analysis provides two fens results, for the average that compares in D2A1 allele carrier and the non-carrier provides scoring.

The case grouping.Above-mentioned information makes the experimenter be placed in two kinds of dissimilar groupings.Whether first kind be based on existence that chronic exercise and/or sounding twitch.These groups are the TS proband, have the TS proband relatives of TS or chronic twitch, do not have the TS proband relatives of chronic twitch and the contrast of not having twitch.Second kind of grouping is by a) no medicine, ethanol or tobacco abuse and do not have contrast (contrast does not have), a b of the behavioral aspect considered) the TS proband or their relatives (case has) that do not have the TS proband or their relatives (case does not have) of the behavioral aspect considered and c) have a behavioral aspect of being considered form.Therefore, for example, because contrast and experimenter have finished the structuring application form, for compulsion can not have a substance abuse and do not have compulsive contrast and do not have compulsive TS proband and relatives and having between compulsive TS proband and relatives compare.Only have 3% to be slight in polygenes group's TS proband, 74% is moderate, and 23% be severe.

Statistics.When detecting individual behavior, for example during behavior x, compare in having the TS proband of behavior x of prevalence rate to(for) a certain sign significantly increases by two possible reasons are arranged with the contrast that does not have behavior x: a) gene that is indicated may work in behavior x, perhaps b) gene that indicated since its be correlated with behavior y relevant itself and may on frequency, have increased with behavior x.In order to distinguish a) and b), the inventor requires between all 3 groups, contrast does not have (no x and do not have y most probably), case not to be had (no x and y is arranged usually) and case has (x is arranged and y is arranged usually), one of the prevalence rate existence of this sign significant, progressive, the linearity increase.SPSS (SPSS Inc., Chicago, IL) statistics software kit have been used.When between the progressive series that contrast does not have, case does not have and case has, comparing two variation per minutes, utilized SPSS Manrel-Haenzel chi-square statistics for gradual linear trend.For carrying D ₂A1 allele (D ₂A1A1 or D ₂A1A2) and not carry A1 allele (D ₂A2A2) the student t that relatively utilizes of a certain given behavior scoring average checks among the experimenter.

For each is studied the gene unanimity, the inventor has measured mutually 3 heterogeneic genotype of experimenter on the same group.This group is called the polygenes group.The experimenter is prepended among this group at its D β H of mensuration or the allelic genotype of DAT1.Polygenes group's screening criteria requires the experimenter to be necessary for non-Spain white race people, must fill in the structuring application form, must agree blood drawing, the DNA that must prepare q.s carries out the gene type of all three genes, and must be as a contrast, TS proband or a proband's relatives.Owing to detected identical experimenter, so make can be to the average of different genetic comparison behavior scorings for this.

319 experimenters are arranged in the polygenes group.In addition, a considerable amount of experimenters have been measured one of them gene but not whole three genotype.For each gene, this is called total group and different genes is varied in size.In each case, total group comprises that the polygenes group adds extra non-polygenes group case.For avoiding obliterated data or dynamics, also detect total group for each behavior.Yet,, only provide the p value at the last string of suitable table for total group for conserve space.For the experimenter with allele to be measured or sign and those individualities that do not have this equipotential gene or sign, the average of the more different behavior variablees of t check analysis.Equally, in the time of suitably, in the last string of each table, provide total group p value.

For the quantitative testing of different behavior scoring averages, the inventor has envisioned the strategy of two some opposition.First is many more based on the experimenter's quantity that detects, and the more little hypothesis of chance of II type mistake is checked a large amount of as far as possible experimenters.Strategy detects contrast, relatives and TS proband hereto.Second strategy promptly contrasts and the TS proband for only relatively more extreme, and this is based on comparing the hypothesis that mutant gene has individuality minimum and that at utmost express.To provide the result for first strategy, and if second result they provide more information than detecting bigger group, then will come into question.In suitable table, provide the gauged α value of Bonferroni, promptly divide apoplexy due to endogenous wind experimenter's polynary comparison for mutual eliminating.

Utilize linear contrast to carry out ANOVA and analyze (Dunn and Clark, 1995) determine between 4 groups, wherein there is 0 in 2,3 signs in 3,3 signs in 3 signs 1 and 3 signs, in the average of many continuous behaviors, whether has significant linear the reduction.Tukey check introduced determine whether that any in each group scoring is significantly different to each other in analyzing, α=0.05.Because the percentage ratio of the variation that three dopaminergic genes cause, utilize 1 as a token of to exist and 0 different behavior scorings are carried out multiple linear regression analysis to three genes in order to estimate simultaneously for sign does not exist.R ²Provide the ratio of the variation that causes owing to each gene, and R ²And be the contribution of all three genes.This provides every kind of special behavior, the toatl proportion of the variation of being come together to explain by all three genes.

The result

For total group, experimenter's quantity, mean age and sex distribute and are shown in the table 35 in four groups of experimenters.The mean age of estimating the TS proband may be lower than other group, and this is true.Estimate that also TS proband and the gender's ratio with relatives of TS may be higher than non-TS group, and this also is true.Yet because institute's test cdna is autosomal, sex ratio itself should not be considered a factor.For studying this, the inventor is to the existence of unlike signal or lack with having carried out chi-square analysis between sex.These are inapparent.

The Taq A1 allele of DRD2 gene." contrast do not have " be " case does not have " together between " case has " together.For polygenes group and total group, D in TS proband, the TS relatives that the TS relatives of chronic twitch, no chronic twitch are arranged and contrast four groups ₂The allelic prevalence rate of A1 is shown in the table 36.In both cases, the allelic prevalence rate of A1 exists significant progressivity to increase from contrast (23.5 and 26.9%) to TS proband (41.5 and 41.7%) by the linear chi-square analysis of Mantel-Haenszel.

Determine D ₂The result of A1 allele prevalence rate in " contrast do not have ", " case does not have " and " case has " and the p value of total group of significant correlations is shown in the table 37.The most significant dependency is with the mania shape, wherein with no mania shape case 28.7% and have 52.2% of symptom case to compare, D is carried in 21.2% the contrast of having no precedent the mania shape ₂A1 allele (p=0.00024).Other significant variable is followed successively by disobedience, property, ADHD-R, division sample, ADHD, twitch, severe depression and conduct.For total group, the most significant dependency is the same sex (p=0.0007), stutter (p=0.0008), division sample (p=0.0016) and dry mad (p=0.0017).

To D ₂A1 carrier and D ₂The T statistics of A2A2 carrier average.Because most behavior evaluation relates to successive scoring, therefore all experimenters are carried D based on them ₂A1 allele (D ₂A1A1 and D ₂A1A2) (D whether ₂A2A2) it also is useful checking the average of these scorings.These are listed among the table 38A according to the t test value that falls progressively for the result of experimenter in the polygenes group.The p value of the remarkable behavior in total group is shown in the last string.Significant variable is followed successively by ADHD, dry mad, ADHD-R, conduct, twitch, disobedience, division sample and property.For total group, three extra variablees are stuttered, are forced and somatization symptom also is significant.When the deletion contrast, the major part in these is (table 38-B) still significantly.In order to compare with being used for all three dopaminergic gene results together, contrast and TS proband significantly the results are shown among the table 38-C for the polygenes group's.The rank of conduct is the highest herein, is followed successively by dry mad, ADHD, twitch, division sample subsequently, forces and disobeys.For determining whether D ₂Allelic homozygosity (the D of A1 ₂A1/D ₂A1) provide and be higher than heterozygote (D ₂A1/D ₂A2) behavior scoring detects the average of these two groups of all behavior scorings.Each variable for except twitching scoring is lower than heterozygote for homozygous average.This significance exists only in three variablees-ethanol, primary school and reading.

The Taq B1 allele of the Taq B1 allele of D β H gene in various mental disorders.In 148 that are tested non-Spain white race people contrasts, 60.8% carries D β H B1 allele.In screening those that get rid of ethanol, medicine and tobacco abuse or dependence, 52.9% carries B1 allele (table 39).Utilizing α is 0.05, and the allelic prevalence rate of B1 exists significantly to be increased, and reaches 70.5% (p=0.012) among the 352 routine TS proband.For the seriousness that determines whether TS plays a role, the TS proband is divided into slightly (too gently need not to treat any aspect in the TS scope), moderate (some aspect needs treatment) and severe (the serious interference that some aspects in the TS scope cause them to live according to population rate, Comings and Comings, 1985).The allelic prevalence rate of B1 has increase to 72.1% of moderate to 72.7% of severe by slight 54.3%.The allelic prevalence rate of B1 is significant p=0.0071 for the moderate case.

The allelic prevalence rate of B1 is 73.1% (p=0.019) and be 73.1% (p=0.012) to 104 smokers for 78 experimenters with ADHD.The significance that do not have that the allelic prevalence rate of B1 is compared in other group in shining increases.And B1 allele is significant there not being Bonferroni under proofreading and correct only with the dependency of TS and smoking, but some worries that a kind of like this correction may increase II type mistake (Rothman, 1990) inadequately.In the hoc post analysis of B1/B1 homozygosity in the TS of Three Estate with the B1/B2 heterozygosity, the severe TS case of noticing 37.1% slight, 49% moderate and 62% be the B1/B2 heterozygote (the linear card side of Mantel-Haenszel=6.25, p=0.012).

Contrast does not have, case does not have, case has.The D β allelic prevalence rate of H TaqB1 the results are shown in the table 40 among 319 experimenters among the polygenes group.ADHD is for the most remarkable, and B1 allele prevalence rate is 47.1% for the contrast of the no ADHD of no material abuse, is 70.6% for the case of no ADHD, and is 81.9% (p=0.0001) to the case with ADHD.Other significant behavior variable is followed successively by study, primary school, ADHD-R, disobedience, twitch, dry mad, ethanol, reading, drug dependence, sleep, stutters and forces.Total group's result is similar, also is that ADHD is the most remarkable.When only detecting the male in total group, sleeping is the most remarkable (p=.00005), is ODD (p=0.002) and ADHD (p=0.005) then.

To the T statistics of B1 carrier with B2B2 carrier average.Table 41-A shows the result when the polygenes group is limited to contrast and TS proband.ADHD is positioned at the top of tabulation again and is significant p=0.020.The remarkable behavior of unique other is a primary school, p=0.029.Total group significantly the results are shown among the table 41-B.Disobedience behavior, sleep, ADHD and reading are significant at α=0.05 place.For determining whether that the allelic homozygosity of B1 (B1/B1) provides the average behavior scoring that is higher than heterozygote (B1/B2), detect for the average of these two groups of all behavior scorings.Homozygote and heterozygote all do not have marked difference for any behavior variable.

10/10 genotype of DAT1 gene.The allelic frequency of different DAT1 for whole crowd that is detected experimenter is shown in the table 42.To simplify the analysis, the variety classes to Tourette syndrome and unsociable and eccentric disease and behavior disorder compares 10/10 genotypic prevalence rate with 10/x or the genotypic prevalence rate of x/x.In 91 collators, 37.4% carries 10/10 genotype.It increases to 52.3% (p=0.015) for 241 TS proband.In slight, moderate and severe TS experimenter, there is not significance difference (p=0.031).Result for TS proband and TS relatives still has significance after proofreading and correct through Bonferroni.The detection of 10 gene frequencies provides suitable result (last two row in the table 42).

Contrast does not have, case does not have, case has.The results are shown in the table 43 for these of 319 experimenters of polygenes group.Having, the variable of high chi-square value is a somatization symptom, 10/10 genotype is carried in the contrast of 21.1% no somatization problem, the TS proband of no somatization problem or relatives are 46.8%, and TS proband or relatives with somatization problem are 60.3% (p=0.02).Other significant variable according to the order of card side size be ethanol, ADHD-R, severe depression, terrified, force, extensive anxiety and dry mad.In total group the last string that significantly the results are shown in table 43 that increases for 357 experimenters' appropriateness.Main difference is that disobedience, property, reading and ADHD are as the significantly adding of variable.

To the T statistics of 10/10 genotype with non-10/10 genotype average.When detecting total group time, those have 10/10 genotypic experimenter, and somatization and severe depression argument table reveal significantly high average (table 44-A).It is significant not having variable for all experimenters of polygenes group.For the polygenes group, have only contrast and TS proband's result in table 44-B, to provide.This is group used in the check of this additive effect of three.Xia Mian variable is significant successively herein: extensive anxiety, severe depression, ADHD-R, ADHD and ethanol.

The comparison of all three dopaminergic genes.For the results are shown in of being studied in the table 45 at all three intergenic each behavior scorings.Grouping is made up of none (group 4) institute among (group 1) of those all three signs of heredity, 2 (groups 2), 1 (group 3) in 3 in 3 and 3.Showing with 4 genomes, being distributed as altogether of remarkable linear correlation is ADHD (p=0.0002).For example, ADHD scoring for those heredity have in 3 signs 3 be 30.03, for those heredity have in 32 be 24.74, in 31 be 20.42, and in 30 be 14.07.The average of group 4 (in 3 none) significantly is lower than the average score of group 1 and 2, and organizes 3 average and significantly be lower than group 1.Secondly the most significant be scoring-1.17,1.06,0.94 and 0.46 (p=0.0002) to stuttering.Scoring for group 1 to 4 disobedience behavior is respectively 5.04,3.91,3.38 and 1.93 (p=0.023).Scoring for conduct disorder is respectively 4.08,3.05,2.87 and 1.93 (p=0.023).Other significant variable for twitches, force, dry mad, ethanol and extensive anxiety.Though other behavior is inapparent, 16 in 20 show identical gradual linear reduction the with less hereditism's load.It is similar using whole polygenes group time result, but has lower slightly significance.

For further detecting these relations in detail, the most significant behavior scoring ADHD is divided into 8 groups, represents the institute of these three genetic markers might make up (table 46).This has confirmed adding up property and subtracting property trend.Twitch scoring and also provide a similarly classification.These results are significant with the similar of those 4 gene kinds and for identical variable.

The probability that the aspect drove an of the unknown that may be contrasted in a way for testing result is only utilized experimenter (proband and the relatives) replicate analysis with TS.Though scoring scope that narrows down and the higher p value that causes thus, ADHD, ADHD-R, somatization and severe depression are significant p＜0.05, and conduct, disobedience and dry mad be slightly remarkable p＜0.07.For example, group 1 to 4 is respectively 29.9,26.1,23.0 and 22.9 (p=0.01) for the value of ADHD scoring; Scoring is 5.1,4.0,3.8 and 3.2 for conduct; And mark for disobedience is 3.8,3.3,2.9 and 2.7.Variation ratio results estimated for more remarkable behavior variable is shown in the table 47.Substantially, DRD2 gene pairs variation contribution is maximum.For the main behavior of being correlated with TS, the variation between 3.0 to 7.6% is explained by three dopamine genes.

In this research, once attempting eliminating may be because hereditism's variation of ethnic background or other similar obstacle.This relates to these points.1. all experimenters are non-Spain white race people.2. they are estimated not only to contrast screening ADHD, medicine, ethanol and tobacco abuse/dependence, and by the same structurized instrument that is used for TS proband and relatives.This makes the contrast with the behavior of studying be excluded.The huge effect of doing is like this confirmed by following true institute: for total group, the variation of gained is from 35.0% to 23.8% the scope in the D2A1 allele prevalence rate of contrast, and the variation in qualified contrast number is for different behaviors from 67 to 40.3. detect a large amount of experimenters and avoid II type mistake.Therefore, 484 experimenters are being studied in the DRD2 seat in total group, and are 319 to the polygenes group.4. based on DIS, give identical structuring psychotic symptoms to all experimenters and observe.TS is a kind of complicated pedigree obstacle (Comings, 1995d, Comings, 1990), and its allow to many different behaviors detect test D2A1 allele may be with some behavior strong correlations not relevant probability with other behaviors, rather than rely on diagnosis (TS or non-TS) in single two minutes.5.TS the behavior scoring that is introduced as more wide region to be measured of proband, the TS relatives that have and do not have TS and contrast provides chance.6. in order to eliminate the worry about inappropriate selection contrast, also analysis result under the situation that does not comprise contrast.7. because individuality may have hereditary largely load, comprise but not have a mind to get rid of to have the common proband who falls ill disease.8. collect more serious ill proband but not concentrate on the extended familys that are used for chain research type, it is ill that wherein non-proband patient is more slight usually.9. detect the effect of the two or more gene pairs phenotypes of associating, be the Taq A1 allele of DRD2 gene, the Taq B1 allele of D β H gene and 10/10 genotype of DAT1 gene in this case.For finishing this step, in the experimenter of same cluster, detect all three kinds of genes.

Dopamine D ₂The D2A1 allele of acceptor gene in the TS proband organizes other.As shown in the table 36, for total group, 41.7% TS proband carries D ₂A1 allele, and be 35.0% for the relatives with TS is 28.8% for the relatives of no TS, is 26.9% for the contrast of no material abuse obstacle.Progression is a significance, P=0.0038.Use result's (26.9%) of inventor's contrast to coexist and get rid of among 714 non-Spain collators altogether of ethanol and drug dependence/dependence 25.9% D through screening ₂A1 allele prevalence rate be as broad as long (Comings etc., 1996e).Prevalence rate (41.7%) in inventor's TS proband is 40.7% as broad as long with among 432 the TS experimenters altogether that carried out gene type by inventor and other people in fact also.Though these results are with the effect unanimity of DRD2 gene in TS, they do not determine which part in the behavior spectrum is mainly influenced.For determining this point, the inventor detects D in three groups of " contrast does not have ", " case does not have " and " case has " behavior of studying ₂The allelic prevalence rate of A1.

20 the different syndromes (table 37 and 38) that relate to impulsion, pressure, addiction, emotion, anxiety, sleep and learning behavior have been detected.(21 comprise the dual evaluation of ADHD).Same D ₂The variable of A1 allele significant correlation is property, stutter, force, divide sample, dry mad, ADHD-R, twitch, ADHD, behavior, disobedience, alcohol abuse, study and sleeping problems.All other behaviors that show same trend but are inapparent.For total group, the result of two (stutter and property) the most significant and least significant (phobia) behavior.For card side's research of polygenes group, dry mad behavior is the most significant, and rank is the highest in table.This is consistent with the behavioral study of inventor in TS proband relatives, its show the mania shape represent the expression of high form of Gts gene (Comings, 1995d).

When only detecting the TS proband, at D ₂The allelic existence of A1 is with lacking dependency between the seriousness of twitching, and the result of (1994b) such as this same Devor (1992) and Gelernter is consistent.Yet when comprising contrast and non-TS relatives, it is significant twitching scoring.In addition, when with case during according to other behavior layering, D ₂A1 allele is with many being correlated with in them.

Dopamine.The allelic prevalence rate of D β H Taq B1 (70.5%) in 352 TS proband is significantly higher than in 148 contrasts (60.8%).Experimenter's unique other group that shows the B1 allele prevalence rate of remarkable rising is the smoker, is 73.1%.

For 319 experimenters' polygenes group, be followed successively by ADHD, study, primary school, ADHD-R, disobedience, twitch, dry mad, ethanol, reading, drug dependence, sleep, stutter and force with 13 variablees of D β H B1 allele significant correlation.Yet, several in them, boundary significant those (ethanol, drug dependence, stutter, force) particularly, the allelic prevalence rate of B1 are higher than those and have the behavior in the TS of no behavior proband and relatives, show the lower frequency drives of p value quilt in contrast.Except adding somatization and severe depression and not having the stutter of significance, the result is similar for total group of 455 the bigger experimenters.

For the polygenes group, the allelic prevalence rate of B1 is in contrast from 46.9 to 56.5% the scope.When the average to polygenes group and total group's contrast and TS proband's behavior scoring all detected, ADHD, primary school, disobedience and sleep were significant.The inventor studies confirm that, the disobedience behavior always than conduct disorder with B1 allele significant correlation more.

For detecting may acting on of DAT1 gene, as in the research of DRD2 gene and D β H gene, the inventor has compared 10/10 genotypic prevalence rate to different behaviors in contrast, TS proband and TS experimenter's relatives.This demonstrate with comprise somatization, alcohol dependence, ADHD, severe depression, force, a plurality of variablees of the extensively common morbidity behavior of anxiety, dry mad, property and disobedience obstacle have remarkable related.

Though this result support shows a kind of tangible physiological effect those results with the 40bp repetition allele of DAT1 gene relevant (19904a) such as Gelernter, it is relevant with 10 allele rather than 9 allele that inventor's result also points out psychosis.This is by finding that showing in the behavior of remarkable effect 9 genotypic frequencies at each exists gradual reductions and further support.For example, for somatization, 9 allelic frequencies are reduced to 0.29 among the relatives of no somatization by 0.37 in the contrast, to the relatives with somatization 0.20, and 10 allelic frequencies increase to 0.70 and 0.77 by 0.54 between identical group.What is interesting is that as if the frequency of common allele 10 still further increase when the behavior symptom of each population exists.A possible explanation is to be normal allele on the 9 allele roots, and 10 allele are owing to it has increased by the screening frequency with the one or more relevant of listed behavior.

All three dopaminergic genes.Be adding up property of research and subtracting property effect, require the inventor from a considerable amount of serious ill relatively TS proband, their relatives and contrast acquisition DNA specimen, and need be to a group experimenter all genes being carried out typing.319 samples have been done like this.A previous hypothesis is that one of two strategies can provide more dynamics.First strategy relates to the whole polygenes group of detection, comprises uninvolved relatives substantially, comes the increase dynamics.Second strategy is the subgroup that only detects the polygenes group who is made up of contrast and TS proband.This can provide the scoring of wide region and the two bigger differentiation of seriousness.Though two kinds of technology all provide positive findings, it is more effective that the latter is proved to be.The result show for except 4 (somatization, major depression, sleep and readings) detect to some extent and all exist an experimenter of 3 from carry 3 signs to have in 3 in 2,3 to those that 0 progressive linearity reduces in 1 and 3 in the scoring of behavior.

The inventor infers that TS and ADHD are identical hereditism's obstacle (Comings and Comings, 1984 basically; Comings and Comings, 1987a; Knell and Comings, 1993; Comings and Comings, 1993).The most significant two behaviors are ADHD and twitch when these three dopaminergic genes of associating, and this proves and confirmed this hypothesis on the molecular genetics level.They have formed a hereditism such notion of behavior spectrum that is mutually related with the significant correlation support of other behavior.The detection of other gene can make phenotype tend on the less direction that is influenced by the dopaminergic gene.To the influential tryptophan 2 of 5-hydroxy tryptamine level, 3-dioxygenase gene be an example (Comings etc., 1996d).Do not find the linear additive effect evidence in addition of three genes, promptly do not have the evidence of epistasis.

When by list these three signs might make up more detailed detection with the behavior ADHD of the significant correlation of these three genes the time, these results are also consistent with subtracting property effect with adding up property.Data shows, the load of three genes can be explained a series of clinical significant scoring from the diagnosis of no ADHD to clear and definite diagnosis.Indicate that owing to still having some experimenters to have all three not having ADHD symptom and some does not have any in three signs that clearly ADHD is but arranged, so very clear other gene that also relates to.Therefore, the screening to TS and ADHD can comprise the allele variant that has been proved to be other gene that participates in these obstacles.

Be not that all behavior scorings show with each all has the dependency of this degree in eight displacements.For the proof this point, for the scoring of twitching, what same kind two was broken up the results are shown in the table 46.Though different displacements all also has one roughly and significant linear the reduction in the scoring of twitching, the D2A1 allele positive and the experimenter of D β H B1 and DAT1 10/10 genotype feminine gender shows tangible discontinuity, average is 5.0.This be one since in every group relative in a small amount experimenter's statistics unusual, perhaps DRD2 allele prompting that twitch is had the effect stronger than other gene, research that must products for further.Studies show that of DRD2 gene itself with the significance association of twitching seriousness.

Find that at present the level of platoon leader of stutter own only is lower than ADHD and is higher than twitch, support that stutter is that another of Gts gene shows this supposition.About conduct and disobey obstacle these to observe with these two behavior disorders that it has been generally acknowledged that be that the hypothesis of because socio-psychological factors (comprise bring up poor) opposes fully.Though nobody can suspect qualified important effect of bringing up, when bringing up form or environmental factors not to have defect, genetics factor may and be disobeyed in the obstacle in conduct and be played an important role.

The variation ratio that the dopamine gene is explained.Utilize multiple linear regression analysis that the calculating of correlation coefficient is made and can estimate the different behavior variance ratio examples of being explained by these three dopaminergic genes.For all three genes, its explanation from the variation of ADHD scoring 7.6% to stuttering 1.3% range of variation (table 47).Be the estimation of the relative importance that obtains three genes, add up to the r of all behavior variablees ²Value.This shows that the relative importance of three genes is that DRD2, DAT1 and the about ratio of D β H gene are respectively 3: 2: 1.This conclusion is supported by such fact: for DRD2, gene r is significant to eight behavior variablees (conduct, dry mad, division sample, primary school, twitch, OCD, ODD and phobia), for the DAT1 gene, r is significant to three variablees (depressed, extensive anxiety and alcohol abuse).Though is not significant for D β H gene r to arbitrary variable, three with peak are ADHD, ODD, conduct, reading and learning disorder.These results also for different genes and gene be combined in phenotype will how to be provided support by the understanding that plays a role in expressing (Comings, 1995a).For example, all three genes participate in ADHD and ODD approximately equably, and the participation in conduct disorder then is DRD2＞D β H＞DAT1.For study with read, order be D β H＞＞DAT＞DRD2.

Owing to be lower than 100% (Price etc., 1985) for the concordance rate of chronic twitch in identical twins, part variation (about 10 to 20%) is for factor of environmental.The Taq polymorphism that is used for DRD2 and D β H gene is not to be responsible for the sequence change that gene function changes, and only be the linkage disequilibrium of congenerous sudden change, the estimation of these variation percentage ratios may have been underestimated, and they may be higher if promptly detected function mutation itself.At last, these estimate that representative comprises all cases, contrast and proband's meansigma methods, no matter whether they have the behavior of being studied.For example, only have 41 or 14.5% in 282 cases to have the problem of stutter, and 43.7% have the conduct problem.Those variation percentage ratios of explaining of the behavior of studying may be much higher by in fact having.

These results are consistent with the polygenes of these obstacles, multifactor characteristic.Though do not have significant dependency for " contrast does not have ", " case has " and " contrast has ", but what linkage analysis is the low relatively ratio of the variation of being explained by these three genes be illustrated as is issueless, and how responsive this correlating method has to polygenes.

Table 35

The relatives 37 23 60 62 that the relatives 60 27.72 14.73 that the age of different experimenter's groups and Sex, Age group N mean age S.D.TS propositus 225 16.83 12.11 have TS do not have relatives' 132 38.07 11.95 contrasts 67 42.31 14.86 of TS to add up to the thin N male sex N women of 484 sexes N sum % male sex TS propositus 188 37 225 84 to have TS do not have relatives' 51 81 132 39 contrasts 27 40 67 40 of TS to add up to 484

Table 36

Various experimenters divide apoplexy due to endogenous wind D ₂The allelic prevalence rate classification of A1 N A1A1 A1A2 A2A2 %A1 frequency x ^2*A. polygenes group, (n=319) relatives 39 0 11 28 28.2 0.14 of TS propositus 142 9 50 83 41.5 0.24 with TS do not have the total group of relatives' 104 4 24 76 26.9 0.15 contrasts 34 17 26 23.5 0.14 6.99B. of TS, (n=484) relatives 60 2 19 39 35.0 0.19 of TS propositus 225 13 81 131 41.7 0.24 with TS do not have relatives' 132 4 34 94 28.8 0.16 contrasts 67 3 15 49 26.9 0.16 8.35* of TS based on D₂The linear card side of the Mantel-Haenzel of A1 prevalence rate

Table 37

To polygenes group, D ₂A1 allele is with the relevance score contrast case case of the various behaviors of falling ill altogether

Not having has x ^2*p ^*Total group's p ^*

N ％ N ％ N ％ 33 21.2 216 28.7 69 52.2 13.47 0.00024 0.0017 34 23.5 206 29.6 79 46.8 8.31 0.0039 0.0274 27 25.9 204 28.4 88 46.6 8.19 0.0042 0.0007ADHD-R 34 23.5 210 30.5 75 45.3 6.63 0.010 0.0085 32 25.0 194 30.4 88 44.3 5.92 0.015 0.0016ADHD 34 23.5 163 30.1 116 41.4 5.42 0.020 0.0110 34 23.5 104 26.9 181 38.7 5.27 0.022 0.0088 25 24.0 176 30.1 109 41.3 4.70 0.030 N.S. 26 23.1 161 30.4 124 39.5 3.93 0.047 0.0135 32 21.9 216 32.4 69 40.6 3.59 0.059 0.0013 18 16.7 116 31.0 169 36.7 3.01 0.082 N.S. 29 20.7 190 32.6 95 37.9 2.70 0.100 0.041 28 21.4 202 32.7 83 38.6 2.64 0.103 0.049

Not having has x ^2*p ^*Total group's p ^*

N % N % N % panic attack 28 21.4 178 32.6 100 38.0 2.58 0.107 N.S. extensive anxiety 30 23.3 187 34.2 88 34.1 0.60 0.436 N.S. of 26 15.4 226 35.4 67 32.8 0.98 0.322 N.S. primary schools of 33 21.2 241 32.8 41 39.0 2.53 0.111 0.0008 alcohol abuses, 34 23.5 262 33.6 23 43.5 2.53 0.111 0.037 neurosis, 23 17.4 182 33.0 103 36.9 2.40 0.120 N.S. drug abuses, 34 23.5 258 33.7 27 40.7 2.09 0.147 N.S. somzatizations, 19 26.3 154 32.5 78 39.7 1.78 0.182 N.S. that stutter

Contrast does not have=does not have the contrast of ethanol or medicine or tobacco abuse/rely on and do not have the behavior of studying

Case does not have=does not have the TS proband and the TS relatives of the behavior of studying

Case has=has the TS proband and the TS relatives of the behavior of studying

^*The linear card side of Mantel-Haenszel or based on the p value of Mantel-Haenszel linearity card side.

Table 38

To D ₂A1 carrier and D ₂The comparison A. polygenes group of average behavior scoring between the A2A2 carrier, contrast, TS relatives and TS proband (N=319, D ₂A1=106, D ₂A2A2=213) scoring |----D2A1-----| | the total group of-----D2A2A2-----| t p p ^*

S.D. S.D.ADHD 20.77 13.22 15.53 13.60 3.28 0.001 0.003 2.00 2.49 1.16 1.83 3.19 0.002 0.003ADHD-R 4.97 4.42 3.40 4.30 3.02 0.003 0.012 3.01 2.39 2.24 1.99 2.87 0.005 N.S. 3.05 3.74 1.92 3.00 2.70 0.008 0.042 3.11 3.22 2.22 2.79 2.44 0.016 N.S. 1.66 2.19 1.11 1.34 2.41 0.017 0.007 0.78 1.33 0.46 0.96 2.19 0.030 0.017 2.90 3.05 2.28 2.64 1.79 0.074 0.009 0.59 1.68 0.34 1.24 1.34 0.181 N.S. 2.62 3.31 2.11 2.87 1.24 0.217 0.048 0.57 0.89 0.45 0.81 1.17 0.245 N.S. 3.76 3.09 3.35 3.01 1.13 0.260 N.S. |-----D2A1-----| |----D2A2A2---| t p p^*

Average S.D. average S.D. terrified 3.17 2.20 2.94 2.07 0.92 0.360 N.S. of 0.16 0.37 0.11 0.32 1.11 0.268 0.002 neurosis, 2.61 2.82 2.28 2.74 1.00 0.319 N.S. that stutter read 1.78 1.93 1.57 1.99 0.91 0.365 N.S. extensive anxiety 0.23 0.43 0.21 0.41 0.40 0.692 N.S. of 2.75 1.93 2.57 1.95 0.75 0.457 N.S. alcohol, 0.60 2.27 0.41 1.93 0.72 0.471 N.S. of 0.47 0.79 0.39 0.79 0.77 0.443 N.S. primary school that sleeps

^*Total group: N=417, D ₂A1=153, D ₂A2A2=264B. have only total group, TS relatives and TS proband (N=417, D ₂A1=153, D ₂A2A2=264) scoring |----D2A1-----| | the total group of-----D2A2A2----| t p p ^*

Manic 2.03 2.48 1.37 1.94 2.83 0.005 conducts of average S.D. average S.D. 3.15 2.36 2.50 2.13 2.82 0.005 division samples 1.81 2.24 1.22 1.74 2.80 0.006 stutters 0.24 0.43 0.13 0.34 2.69 0.008ADHD 22.26 12.87 18.71 12.94 2.60 0.010 forces 3.24 3.11 2.48 1.69 2.51 0.013 somzatizations, 2.93 3.51 2.01 2.70 2.49 0.014 property, 0.80 1.29 0.50 1.00 2.44 0.015ADHD-R, 5.28 4.46 4.36 4.62 1.99 0.048 twitches, 3.11 3.54 2.52 3.32 1.67 0.096C. to only have polygenes group, contrast and TS propositus. Scoring |----D2A1-----| | the total group of-----D2A2A2-----| t p p^*

Average S.D. average S.D. conduct 3.65 2.47 2.64 2.10 2.80 0.006 manic 2.31 2.50 1.40 1.95 2.54 0.012ADHD 25.57 12.16 21.23 14.53 2.12 0.036 twitch 4.25 3.88 3.06 3.44 2.06 0.042 division samples 1.91 2.58 1.20 1.53 2.02 0.046 and force 3.40 3.24 2.44 2.80 2.01 0.047 to disobey 4.31 3.27 3.30 3.17 2.01 0.046

Table 39

The allelic universality obstacle of D β H Taq B N 11 12 22 %1 O.R. freq1 x in various mental disorders ^2*Contrast 148 21 69 58 60.8 0.37 through contrast 51 6 21 24 52.9 0.32TS of screening 352 71 177 104 70.5 1.54 0.45 6.29 slight 35 6 13 16 54.3 0.76 0.36 0.014 moderates, 251 58 123 70 72.1 1.67 0.48 7.24 severes, 66 7 41 18 72.7 1.72 0.42 4.82ADHD, 78 18 39 21 73.1 1.75 0.48 5.46 alcoholism, 23 3 13 7 69.6 1.47 0.41 1.77 unsociable and eccentric disease 40 6 21 13 67.5 1.34 0.41 1.95 depressed 28 68 14 50.0 0.64 0.36 0.06 drug abuses, 29 2 17 10 65.5 1.22 0.36 1.18 gamblers, 111 11 56 44 60.4 0.98 0.35 0.78 smokers, 104 29 47 28 73.1 1.75 0.51 6.18 totals 913^*At the contrast (alcohol-free, medicine or tobacco abuse/dependence) of screening and the gauged α=0.05/8=0.0065 of comparison O.R.=odd ratio Bonferroni of the B1 allele prevalence rate between obstacle of studying.

Table 40

To the polygenes group, the anosis example of D β H Taq B1 allele (11+12 genotype) the scoring no case of contrast in contrast, TS proband and relatives has x ^2*p

N ％ N ％ N ％ADHD 34 47.1 163 70.6 116 81.9 15.14 0.00010 29 48.3 190 72.6 95 81.1 10.13 0.0014 30 46.7 187 72.2 88 79.5 9.48 0.0021ADHD-R 34 47.1 210 74.3 75 78.7 8.52 0.0035 34 47.1 206 74.8 79 77.2 7.21 0.0070 34 47.1 104 75.0 181 75.7 7.17 0.0074 33 48.5 216 75.0 69 76.8 6.02 0.014 34 47.1 262 76.0 23 69.6 5.92 0.018 18 44.4 116 73.3 169 76.9 5.42 0.019 34 47.1 258 76.0 27 70.4 5.33 0.021 28 53.6 202 74.3 83 78.3 4.63 0.031 33 48.5 241 76.3 41 73.2 4.56 0.032 32 46.9 216 75.9 69 73.9 4.33 0.037 32 50.0 194 76.8 88 73.9 2.94 0.086 x^2*p

N % N % N % somzatization 19 47.4 154 76.0 78 75.6 2.62 0.105 panic attacks 28 50.0 178 76.4 100 74.0 2.43 0.119 major depressions 25 52.0 176 75.6 109 75.2 2.40 0.121 conducts 26 50.0 161 77.0 124 73.4 1.61 0.203 property 27 55.6 204 74.5 88 72.7 1.12 0.288 neurosis 23 56.5 182 76.4 103 73.8 0.70 0.401 extensive anxieties 26 53.8 226 75.2 67 70.1 0.62 0.430

The relatively scoring of average behavior scoring between table 41D β H Taq B1 carrier and B2B2 carrier |----B1-----| |-----B2B2-----| t p

Average S.D. average S.D.A. polygenes group, contrast and TS proband.(N=176, B1=124, B2B2=52) the total group of 3.26 1.96 2.55 1.88 2.22 0.029B. of ADHD 24.48 13.57 19.07 12.72 2.37 0.020 primary schools, contrast and TS proband (N=292, B1=207 B2B2=85) disobeys 4.16 3.19 3.08 3.12 2.68 0.008 sleeps, 0.63 1.03 0.36 0.70 2.60 0.010ADHD 25.17 13.73 20.86 14.66 2.31 0.023 and reads 2.14 2.04 1.57 2.09 2.11 0.037

Table 42

DAT1 40bp repeats the polymorphism genotype in different patient's groups

Prevalence rate and allelic frequency experimenter N 6/6 8/9 9/9 8,/10 9,/10 10,/10 9,/11 10,/11 11/12 %10/10 O.R. f9 f10 x ^2*P x ^2**P contrasts 91 10 12 0 40 34 130 37.4 0.36 0.61Tourette syndromes, 241 21 15 0 92 126 041 52.3 1.84 0.25 0.72 5.89 0.015 7.93 0.0048 slight 15 002058000 55.3 1.92 0.25 0.74 1.37 N.S, 1.75 0.184 moderates, 132 0150 56 66 031 50.0 1.92 0.30 0.70 3.47 0.062 4.27 0.039 severes, 94 2080 31 52 010 53.3 2.08 0.25 0.72 5.99 0.014 5.15 0.023 TS relatives, 103 0070 37 57 020 55.3 2.08 0.25 0.72 6.27 0.012 5.43 0.019 unsociable and eccentric diseases 36 0021 12 21 000 58.3 2.35 0.22 0.75 4.62 0.032 4.63 0.031 and adds up to 471

The O.R.=odd ratio

^*In the contrast and the 10/10 genotype prevalence rate card side relatively between the experimenter of studying

^{* *}In the contrast and the 10 gene frequencies card side relatively between the experimenter of studying

Bonferron; Gauged α=0.05/3=0.017.

Table 43 dopamine transporter 10/10 genotype has x with the anosis example of association (1=10/10 genotype) the scoring no case of contrast of the various behaviors of falling ill altogether ^2*p ^*Group's p ^*

N ％1 N ％1 N ％1 19 21.1 154 46.8 78 60.3 9.51 0.0020 0.0009 34 35.3 262 50.4 23 69.6 6.37 0.011 0.0027ADHD-R 34 35.3 210 49.5 75 58.7 5.02 0.024 0.004 25 24.0 176 50.0 109 55.0 5.39 0.020 0.006 28 28.6 178 50.0 100 55.0 4.53 0.033 0.010 32 31.3 216 50.5 69 56.5 4.59 0.032 0.010 26 26.9 226 50.9 67 56.7 4.94 0.026 0.011 33 33.3 216 50.5 69 56.5 4.07 0.044 0.012 34 35.3 206 50.5 79 55.7 3.32 0.068 N.S. 27 33.3 204 50.0 88 55.7 3.41 0.064 0.013 18 38.9 116 49.1 169 53.8 1.63 0.201 0.043ADHD 34 35.3 163 51.5 116 53.4 2.29 0.129 0.049 28 35.7 202 50.5 83 55.4 2.66 0.102 N.S. 33 33.3 241 52.3 41 51.2 1.96 0.164 N.S. x^2*p ^*Group's p ^*

N %1 N %1 N %1 drug abuse 34 35.3 258 51.9 27 51.9 1.99 0.158 N.S. learn 29 37.9 190 51.1 95 53.7 1.56 0.210 N.S. and twitch 34 35.3 104 55.8 181 49.7 0.49 0.482 N.S. division sample, 30 36.7 187 52.4 88 47.7 0.19 0.659 N.S. conducts, 25 34.6 161 54.0 124 49.2 0.21 0.644 N.S. neurosis, 23 21.7 182 54.9 103 46.6 0.40 0.525 N.S. of 32 37.5 194 52.6 88 51.1 0.78 0.370 N.S. primary schools

^*The linear card side of Mantel-Haenszel or based on the p value of Mantel-Haenszel linearity card side.

Table 44

The relatively scoring of average behavior scoring in contrast and the case |----10/10-----| |-----10/x x/x-----| t p

The total group of average S.D. average S.D.A., contrast, TS proband and their relatives (N=357,10/10=182,10/x, r/x=175) somatization 2.69 3.30 1.94 2.68 2.11 0.036 major depressions 3.87 3.05 3.20 2.98 2.11 0.036B. polygenes groups, have only contrast and TS proband (N=176,10/10=85,10/x, x/x=91) extensive anxiety 0.33 0.47 0.16 0.37 2.55 0.012 major depression 4.00 3.04 2.87 2.83 2.53 0.012ADHD-R 6.60 4.75 4.91 4.45 2.43 0.016ADHD 25.44 13.94 20.42 13.28 2.43 0.016 ethanol 0.68 2.46 0.09 0.70 2.11 0.037

Table 45

The comparison behavior class mean S.D. F of contrast and TS proband behavior scoring average compares p ^*ADHD 1 30.04 10.97

2?????????????24.74???????????13.53

3?????????????20.42 ^*?????????13.77

4 14.07 ^*# 13.11 14.77 0.0002 stutter 1 1.17 0.49

2?????????????1.06????????????0.55

3?????????????0.94????????????0.62

4?????????????0.46 ^*#????????^0.64????????14.61??????0.0002ADHD-R????????????1?????????????7.75????????????4.18

2 6.43 4.74 behavior class mean S.D. F compare p ^*

3???????????4.82 ^*????????4.53

4 3.53 ^*4.12 9.87 0.0020 disobey 1 5.04 3.05

2???????????3.91??????????3.20

3???????????3.38??????????3.25

4 1.93 ^*2.84 9.56 0.0023 twitch 1 4.95 4.41

2???????????3.71??????????3.30

3???????????3.28??????????3.65

4 1.40 ^*2.97 9.56 0.0023 conducts 1 4.08 2.51

2???????????3.05??????????2.35

3???????????2.87??????????2.23

4 1.93 ^*1.22 8.61 0.0038 force 1 3.37 3.51

2???????????3.02??????????3.03

3???????????2.75??????????2.91

4 1.13 1.99 5.48 0.020 manic 1 2.29 2.52

2???????????2.11??????????2.22

3???????????1.40??????????2.12

4 0.87 1.59 5.21 0.024 ethanol 1 1.21 3.45

2???????????0.35??????????1.70

3???????????0.20??????????1.07

4 0.00 0.00 4.50 0.035 extensive anxieties 1 0.33 0.48

2???????????0.29??????????0.46

3???????????0.20??????????0.40

4 0.07 0.25 4.39 0.038 behavior class mean S.D. F compare p ^*Terrified 1 3.45 2.39

2?????????????3.37??????????2.44

3?????????????2.97??????????2.15

4 2.13 1.12 3.79 0.053 division samples 1 1.91 2.88

2?????????????1.66??????????2.24

3?????????????1.26??????????1.45

4 0.78 1.42 3.41 0.067 sleeps 1 0.83 1.04

2?????????????0.64??????????0.86

3?????????????0.44??????????0.89

4 0.46 0.74 2.10 0.149 property 1 1.12 1.96

2?????????????0.62??????????1.13

3?????????????0.58??????????1.16

4 0.53 0.99 2.01 0.158 medicines 1 0.87 2.29

2?????????????0.34??????????1.32

3?????????????0.34??????????1.32

4 0.20 0.77 1.92 0.168 major depressions 1 3.83 3.07

2?????????????2.91??????????2.10

3?????????????2.94??????????2.79

4 2.80 2.93 1.88 0.173 study 1 0.87 0.99

2?????????????0.80??????????0.90

3?????????????0.78??????????1.03

4 0.47 0.74 1.67 0.197 terrified 1 2.83 2.82

2 2.67 2.81 behavior class mean S.D. F compare p ^*

3????????????2.41?????????2.79

4 2.00 1.96 1.00 0.318 primary schools 1 3.33 2.00

2????????????3.09?????????1.97

3????????????2.97?????????2.02

4 2.80 1.74 0.71 0.399 somatizations 1 3.09 3.02

2????????????2.68?????????3.74

3????????????1.69?????????2.14

4 2.69 4.02 0.43 .525 read 1 2.00 1.88

2????????????1.98?????????1.98

3????????????2.10?????????2.23

4????????????1.86?????????2.41?????????0.02????????.893

Group 1=D2A1+, D β HB1+, DAT1 10/10+n=24

2+n during group 2=3 is individual is 67

1+n=70 during group 3=3 is individual

Group 4=D2A1-, D β HB1-, DAT1 10/10-n=15 adds up to 176

^*Check in α=0.05 place 1 remarkable difference on the same group by Tukey.

# checks in α=0.05 place 2 remarkable differences on the same group by Tukey.

^ checks in α=0.05 place 3 remarkable differences on the same group by Tukey.

_ utilize ANOVA to 4 correlated F-ratios of genomic linearity

^*P value based on the F-check

Table 46

By whether DRD2, D β H and DAT1 allelic all,

The average behavior scoring of some behaviors is contrasted in some or none existence

Assortment of genes N average S.D.

ADHD scoring D2+D β H+DAT1+ 24 30.04 10.97D2+D β H+DAT1-22 25.04 12.72D2-D β H+DAT1+ 34 25.91 14.83D2+D β H-DAT1+ 11 20.54 10.75D2+D β H-DAT1-10 21.50 12.90D2-D β H+DAT1-43 19.96 13.20D2-D β H-DAT1+ 16 20.94 16.38D2-D β H-DAT1-14 14.07 12.10 twitch scoring D2+D β H+DAT1+ 24 4.95 4.40D2+D β H+DAT1-22 4.00 3.18D2-D β H+DAT1+ 34 3.91 3.59D2+D β H-DAT1+ 11 2.54 2.54D2+D β H-DAT1-10 5.00 4.96D2-D β H+DAT1-44 2.84 3.31D2-D β H-DAT1+ 16 3.43 3.52D2-D β H-DAT1-15 1.40 2.97

Table 47

Multiple linear regression analysis and by DRD2, D β H and DAT1

The ratio of the variation of explaining; Only there are contrast and TS proband behavior scoring DRD21 D β H DAT1 to add up to

R r ²R r ²R r ²% ^*Conduct 0.217# 0.047 0.114 0.013 0.030 0.0009 6.0 manic 0.197# 0.039 0.055 0.0003 0.084 0.0007 5.0 behavior scoring DRD21 D β H DAT1 adds up to

R r ²R r ²R r ²% ^* 0.169# 0.028 0.046 0.0021 0.048 0.0023 3.3 0.165# 0.026-0.025 0.0006 0.0004 0.0000 2.9 0.155# 0.024 0.109 0.0118 0.108 0.0116 4.7OCD 0.153# 0.023 0.040 0.0016 0.066 0.0043 3.0ODD 0.149# 0.022 0.136 0.0184 0.108 0.0117 5.2 0.149# 0.022-0.041 0.0016 0.016 0.0002 2.5 0.142 0.020-0.023 0.0005 0.061 0.003 2.6 0.141 0.020-0.053 0.0028 0.180# 0.032 5.9ADHD 0.125 0.016 0.166 0.0275 0.186 0.0346 7.6 0.123 0.015 0.004 0.0000 0.113 0.0128 3.0 0.104 0.011 0.047 0.0022 0.024 0.0006 1.4 0.086 0.007-0.005 0.0000 0.067 0.0045 1.3 0.080 0.006 0.035 0.0012 0.075 0.0056 1.4 0.073 0.005 0.050 0.0025 0.160# 0.025 3.4 0.070 0.005-0.023 0.005 0.183# 0.033 4.1 0.041 0.002 0.088 0.0077 0.119 0.014 2.3-0.013 0.0002 0.125 0.0156 0.040 0.016 1.7-0.063 0.004 0.127 0.0161-0.049 0.002 2.4 0.32 0.11 0.20^*Based on r from a plurality of r of all three genes ²#p＜0.05

Embodiment 7

D1 dopamine receptor genetic method in addictive behavior

Three groups that detected are Tourette syndrome (TS) group, smoking cessation group and pathological gambling group.Experimenter in all three groups is limited to non-Spain white race people.

The TS group.This group comprises the contrast of alcohol-free or drug dependence, most of TS proband (Comings, 1995b that is had a strong impact on by multiple relevant behavior disorder wherein; Comings, 1990a) and TS proband's relatives.All all meet the DSM-IV standard of TS, and all are carried out the individual per capita meet with.The contrast of TS group is by TS proband's foster parents and stepparent, the experimenter with non-mental disorder and occupation and non-professional corpsman,hospital.TS experimenter and contrast all are described in detail (Comings etc., 1996f in the elsewhere; Comings, 1995b; Comings, 1994b; Comings, 1994c; Comings, 1995a).

Behavior scoring.Each contrast and TS proband or relatives all require to fill in the application form of a DSM-III-R standard based on diagnostic meet flow sheet (Robins etc., 1981) or a series of obstacles (the sick community of Americanism, 1987).Symptom is divided into 23 different quantitative variations, estimates problem concerning study, smoking, sexual behaviour, division sample, somatization, depression, sleep disorder, extensive anxiety, stutter and the related indication number of twitch in same attention deficit hyperactivity disorder (ADHD) (2 minutes), ethanol, medicine, compulsion, learning disorder, reading problem, gambling, mania shape, phobia, panic attack, disobedience behavior, conduct disorder, the primary school.The problem that is used for these behavior scorings is described in detail (Comings, 1995a in the elsewhere; Comings, 1994a; Comings, 1994b; Comings, 1995b; Comings, 1996a; Robins etc., 1981; Comings, 1995c).Coexist identical described in the embodiment 2 of the principle that detects the behavior of falling ill altogether.

Especially relate to Nicotiana tabacum L., ethanol and drug dependence, force feed and gambling with some relevant symptoms of this research.Ethanol scoring is by being formed (Comings, 1994b for " no " of 8 problems of the MAST test of alcohol abuse or the summation that "Yes" is answered; Comings, 1990a).Medicine scoring is based on to 9 diagnostic meet flow sheet (Robins etc., 1981 about drug dependence/dependence; Comings, " no " of problem 1994c) or the answer of "Yes".The smoking variable is based on following problem, " you once enfleuraged cigarette, cigar or tobacco pipe every day above one month or more? " wherein "Yes" scoring is 1 and the "No" scoring is 0.For the scoring of shopping variable based on summation to the answer of following point: " you once bought really need buy more than you and satisfied your requirement? you are once owing to buy and exceed the thing that you can bear and be absorbed in financial difficulties? you once added on your all credit cards overall balance and greater than your every month net income? you once did shopping and filled up a kind of feeling of unreality? you once did shopping and obtained a kind of happy sensation? you are once by thing and arrearage?The answer scoring of " nothing " is 0, and the answer of " once in a while " is 1, and the answer of " often " is 2.Gambling scoring from in front as relevant nine "Yes" of " gambling scoring " described gambling seriousness or " no " problem (Comings etc., 1996e).Force the evaluation of feed be based on to problem " you thought once that yourself was one and forces into trencherman? " "Yes" or the answer of " no ".

The smoking cessation group.Form by individuality for second group to anti-smoking clinic.Described in more detail in these experimenters and they self research that independently a former DRD2 gene of control group acts in smoking (Comings etc., 1996a).Variable for the smoking evaluation is the average packet number of cigarette that enfleurage every day herein.The all types of substance abuses that comprise ethanol, Nicotiana tabacum L. and other medicines are got rid of in the screening contrast.

Pathological gambling.Form by pathological gambling person for the 3rd group from a former research that in pathological gambling, acts on about the DRD2 gene.Patient conclusive evidence and the details of estimating be described in detail in the elsewhere (Comings etc., 1996e).

Gene type.Be to detect the DRD1 gene, the inventor utilize by the A in 5 ' UTR to G change form, pass through foregoing PCR ^TMThe DdeI polymorphism that step detects (Cichon etc., 1994a).Sign for the DRD2 gene is TaqI A1/A2 polymorphism (Grandy etc., 1989).

Statistical analysis.In TS group, the ANOVA statistics program of utilizing SPSS statistics software kit (SPSS Inc., Chicago, Yin state, Illinois) compares the behavior scoring average of experimenter with different genotype.When detecting two groups when above, with Turkey analytical test significant individual variation between any each group.In the average that is expected at different scorings between different genotype, have under the situation of gradual increase, carry out linear ANOVA by in SPSS statistics software kit, subcommand multinomial=1 being set.Based on the result that ANOVA analyzes, carry out chi-square analysis come more below three not on the same group in the relevant genotypic frequency of the highest average behavior scoring.First is made up of the contrast that does not have tested behavior.Because contrast also needs to fill in one or more application forms, be possible so determine whether in contrast, to exist or lack specific behavior.For TS group, as described in the research in front (Comings, 1995b), the existence of certain behavior or lack is based on two fens breakpoints in contrast and experimenter.There is not the contrast of given behavior to be called as " contrast does not have ".Two fens identical breakpoints make TS proband and theys' relatives be divided into two groups, do not have tested specific behavior those and have those of the sort of behavior.These groups are called as " case does not have " and " case has ", and form second and the 3rd group.If existing being assumed to be between a given genotype and institute's research behavior has significant dependency, this genotypic frequency should have a gradual increase in these three groups so.The utilization of " case does not have " and " case has " group controlled the sort of genotypic frequency in TS experimenter may since with institute's inspection behavior relevant probability that increases of a certain behavior in addition.Because the inventor infers that genotypic frequency has gradual increase in these three groups, so used linear X 2 test (the Mantel-Haenszel check in SPSS statistics software kit).For determining that result's part in three groups meets linear increase, the inventor also require tested genotypic frequency in " case has " than in " case does not have " organized high at least 20%.As (Comings etc. 1996f), find that regression analysis is helpful in the variation percentage ratio of the different behavior scorings of determining to be explained by a specific gene in the dopaminergic gene studies among the TS.This provides r, and r ²The variation mark of a given quantitative trait of being explained by that gene is provided.

The correction of multiple analysis.Owing in the TS group, detected 23 different behaviors, needed so in the level of significance, adjust.Though 0.05 α value may be too free, 0.05/23 or 0.002 α value is considered to too cautious.Therefore select one intermediary 0.05/10 or 0.005 α value.Because the research of smoking cessation group that utilizes previous hypothesis is based on the result in the TS group, and to check unitary variant: the bag number that smoke every day, so utilized 0.05 α value.At last, because pathological gambling person's detection only relates to the comparison of genotypic frequency, so utilized 0.05 α value.The result

The TS group.For contrast (n=61), the gene frequency of DdeI polymorphism is 0.34 to 1 allele, and is 0.66 for 2 allele.For TS proband (n=227), allelic frequency is 0.37 to 1 allele, and is 0.63 for 2 allele.These do not have significance difference, x ²=0.43, d.f.=1, p=0.51.For contrast and TS group, the genotype of DdeI polymorphism distributes and is shown among the table 48-A.11 genotype are present in the TS group of 4.9% contrast and 17.5%, x ²=3.75, d.f.=1, p=0.053.Carry 11 or 22 genotypic in those difference of percentage ratio be 57.3% among 41.3% and the TS proband in contrast.This is significant, x ²=5.08, d.f.=1, p=0.024.

In the allelic research of TaqI A1/A2 of DRD2 gene, having observed very large-scale quantitative scoring 12 heterozygotes are shown the highest scoring in front, is medium scoring to 22 homozygotes, then is minimum scoring to 11 homozygotes.Based on these research, detect the percentage ratio of 12 heterozygotes in three subject group.These the results are shown among the table 48-B for the TS group.Among 73 collators, 19.2% is 12 heterozygotes, 35.3% is heterozygote then among 345 TS proband, x ²=7.19, d.f.=1, p=0.073.

Recently test in DRD1 and the intergenic interaction of DRD2 by the percentage that detects DRD1 DdeI 11 or 22 allele experimenter that isozygoty and DRD2 TaqI A1/A2 allele heterozygosis.Numerical value separately is 33.2% for contrasting 17.9% TS proband, x ²=5.71, d.f.=1, p=0.016.

Detect dependency (table 49) between DdeI genotype and 23 quantity trait variables by ANOVA.Though none is significant at α=0.005 place, the p value of marking for ethanol is 0.0096.In seven scorings of p value＜0.20, five same addictive behavior-alcohol abuse, smoking, force feed, gambling relevant with shopping.Those have 11 and genotypicly have the highest scoring.For example, for the alcohol abuse variable, those have 11 and genotypicly have an average score of 1.30, by comparison, those have 12 genotypic be 0.23, those have 22 genotypic be 0.42.In 23 variablees altogether, those carry 11 genotypic variablees all except learning disorder and somatization and have the highest average.

Based on these results, the inventor has detected between three groups of " contrast does not have ", " case does not have " and " case has " whether have gradual increase (table 50) for different behavior 11 genotypic frequencies then.It is significant linear the increasing of α 〉=0.005 place that three kinds of behaviors are arranged, and wherein 11 genotypic frequencies in " case has " than height at least 20% in " case does not have " organized.According to the order of significance, these are gambling, alcohol abuse and forced purchase.For example, for gambling scoring, 11 genotypic prevalence rates increase to by 4.6% of the contrast of no gambling problem no gambling problem case 15.5%, to 33.3% (p=0.00095) of the case that the gambling problem is arranged.When with α 〉=0.005, three extra variable-drug dependence, force feed all relevant with addictive behavior with smoking.

For two kinds of behavior-gambling and alcohol abuse, single argument regression analysis (table 51-A) is significant in p 〉=0.005 place.α 〉=0.05, other four variablees are for forcing feed, smoking, twitch and reading.Secondly the most significant behavior is shopping.Based on r to the DdeI polymorphism ², the DRD1 gene is contributed 3.6% of gambling scoring, 2.8% of ethanol scoring, 1.6% of 1.9% and the smoking scoring that forcing takes food marks.

Before multivariate regression analysis detection DRD1 and the potential interaction of DRD2 gene, the inventor utilizes the single argument regression analysis at first to detect the effect of DRD2 gene based on TaqI A1/A2 polymorphism.Carry DRD2 11 and 22 homozygous experimenters scoring is 1, and the scoring of 12 heterozygotes is 2.In α=〉=0.005 place, the DRD2 gene with disobedience behavior, conduct disorder, force feed, smoking, gambling and ADHD significant correlation (table 51-B).In α 〉=0.05 place, extra variable be dry mad, stutter, force and divide the sample behavior.Based on r ²Value, the allelic heterozygosity of Taq A1 are responsible for disobeying 4.2% of scoring variation, 3.8% of conduct disorder scoring variation, 4.1% of feed scoring, 2.9% of 3.3% and gambling scoring of smoking scoring.

Multivariate regression analysis shows that DRD1 and DRD2 gene all are significant (table 51-C) for gamble, force feed and smoking r value.For in these each, adding up property of result, when associating, DRD1 and DRD2 gene be responsible for these scoring variations percent 5.9 to 4.8.Ethanol scoring included has significant effect with 11 genotype that show the DRD1 gene, and the allelic effect of DRD2 A1 is inapparent in this group.

Utilize the single argument regression analysis also to detect the adding up property effect of DRD1 and DRD2 gene, wherein DRD1 DdeI 11 and DRD2 Taq A12 are genotypic is 3 by scoring, having those wherein a kind of scorings is 2, and the scoring that two kinds of genotype all do not have is 1 (table 51D).Those are followed successively by gambling, smoking, feeding,forced, disobedience, ADHD, conduct disorder at α 〉=significant variable in 0.005 place, force, dry mad and alcohol abuse.

Also utilize DRD1 DdeI 12 or 22 and DRD2 TaqI 11 or 22 (all not having); DdeI11 or DRD2 TaqI 12 (one); Or the linear ANOVA of three groups of DRD1 Dde 11 and DRD2 Taq12 (all having) detects the additive effect of DRD1 and DRD2 gene.In α 〉=0.005 place, for feeding,forced, smoking, disobedience, ADHD, conduct disorder, force, dry mad and ethanol behavior variable, its show average score from all do not have, to one, to the significant gradual increase that group is all arranged.

Also utilize the additive effect (table 53) that DRD1 Dde 11 genotype or DRD2 Taq A12 genotype or the linearity card side that has frequency that both all have is detected DRD1 and DRD2 gene.Those at α 〉=0.005 place significantly and also for the average of " case has " group than the variable of the height at least 20% of " case does not have " group be followed successively by dry mad, ethanol, force, conduct disorder, division sample, property, feeding,forced and major depression outbreak.For variable alcohol abuse, feeding,forced and dry mad result.

Owing to have marked difference between contrast and TS proband aspect the percentage ratio that carries DRD1 DdeI11 or 22 genotypic experimenters, the inventor has also detected " contrast does not have ", " case does not have " and " case has " three groups of percentage ratios that carry 11 or 22 genotype experimenters.Variable below α 〉=0.05 place is significant, and average " case has " group comparison " case does not have " group height at least 20%-gambling, alcohol abuse and primary school's problem.

The smoking cessation group.DdeI 1 allelic frequency is 0.35 in the contrast of 61 smokings, and it is 0.34 (table 48A) in 177 smokers.In contrast, 4.9% carries 11 genotype, and corresponding smoker is 17.5%, x ²=5.88, d.f.=1, p=0.015.In contrast, 39.3% carries 11 or 22 allele, the smoker corresponding to 66.1%, x ²=13.45, d.f.=1, p=0.0002.

For the DRD2 gene, 26.2% carries 12 genotype in contrast, the smoker corresponding to 42.8%, x ²=5.69, d.f.=1, p=0.017.For contrast, carry DRD1 11 or 22 genotype and DRD2 12 genotype in the time of 24.1%, the smoker corresponding to 45.5%, x ²=8.25, d.f.=1, p=0.0041.

The variable that is detected in the smoking cessation group is by being inhaled the bag number every day.Because all contrasts have all finished the behavior application form, it is included in the used same problem about smoking in the TS group, therefore gets rid of that those once inhaled medicated cigarette, cigar or tobacco pipe etc. and those contrasts with medicine or alcohol abuse are possible.These individual packages every day 0 of forming.Experimenter in the smoking cessation group be divided into inhale those every days 1 to 1 1/2 and 2 to 2 1/2 the bag.(, there is not to inhale every day the experimenter who is lower than 1 bag owing to only there is the permission of inhaling 1 bag those every days at least to participate in research.) the results are shown in the table 54 for the DRD1 gene.Carry 11 genotypic experimenter's percentage ratios and between these three groups, be increased to 16.4% to 18.0%, x by 4.9% ²=5.14, p=0.023.Carry or 11 or 22 genotypic experimenters' percentage ratio increases to 61.8% to 68.0%, x by 39.3% between these three groups ²=12.87, p=0.00033.The experimenter's of DRD2 TaqI A1/A2 polymorphism heterozygosis percentage ratio increases to 34.5% to 46.3%, x by 26.2% between these three groups ²=7.99, p=0.0047.For have simultaneously the DRD1 11 that isozygotys or 22 and the allelic experimenter's of DRD2 TaqIA1/A2 of heterozygosis percentage ratio between these three groups, increase to 34.5% to 50.4%, x by 24.1% ²=23.48, p=0.0001.

Utilize the multiple linear regression analysis also to detect the interaction (table 55) of DRD1 and DRD2 gene for smoking.This shows, DRD1 and DRD2 gene, and as being indicated by these polymorphisms, each contributes impartial.Uniting of they is responsible for wrapping every day 10.5 percent of number variable variation.

Pathological gambling.Different with the smoking cessation group with TS, the pathological gambling group does not have its oneself matched group.Therefore,, TS and smoking cessation contrast combined form total matched group for purpose relatively, and the person that uses it for the pathological gambling.For the gambler, 14.1% isozygotys for DRD1 DdeI1 allele, x ²=5.39, p=0.020, and 55.8% be that 11 or 22 genotype are isozygotied, x ²=6.75, p=0.009.For the DRD2 gene, 45.7% gambler carries TaqI12 group, x ²=1 8.61, p＜0.0001.23.3% is genotypic for carrying DRD1 11 or 22 genotype and DRD2 TaqI 12 among the gambler.

Although many researchs prompting is the interaction between dopamine D 1 and D2 receptor in many mental disorders, but still lack the research of the potential dependency of detection between hereditism's variant of hereditism's variant of DRD1 gene and the potential dependency between the behavior or DRD1 and DRD2 gene.The inventor is interested in the additive effect of the variant of DRD1 gene or DRD1 and the DRD2 gene this probability that may also play a role in these behaviors.The inventor select DRD1 DdeI polymorphism (Cichon etc., 1994a) because it is a kind of PCR-based ^TMTest, and this little effect allele is general in general crowd.Because the research of front DRD2 gene has shown the value that an above interaction of genes detects, detected the dependency between the potential additive effect of the TaqI A polymorphism of DRD1 gene genetics variant and DRD2 gene separately.For the effect with ethnic group minimizes, only studied non-Spain white race people.For the effect with occasionality minimizes, the inventor attempts by detecting three not on the same group experimenters, and wherein two matched groups with them come overlapping any discovery.

Allele and genotypic frequency.The results are shown among the table 48A of allele and genotypic frequency.For the DRD1 variant, the allelic frequency of DdeI the contrast with the experimenter in almost be identical.For three groups of TS proband, smoker and gamblers, 1 allelic frequency is respectively 0.37,0.34 and 0.35.For TS and smoker's matched group, frequency is respectively 0.34 and 0.35.Many reports in the research of mental disorder relatedness are limited to themselves in contrast and comparison with the gene frequency between the experimenter of a particular obstacle.Similar restriction will show that the DRD1 gene does not all play a role in these obstacles any.Yet, because genotypic distribution may be different, so although the similarity of gene frequency is arranged, the inventor has still detected genotypic frequency.Studies show that 11 genotypic prevalence rates significantly increase in smoker and gambler, and be that dividing value significance (p=0.053) increases in the TS proband.For all four groups, there be significant increasing in each allelic homozygosity, TS proband, smoker, gambler and totally have 0.024,0.0002,0.009 and 0.0001 p value respectively.If utilize＜0.5/4 or 0.0125 the gauged α value of Bonferroni, the back is still significant for three groups.

The allelic allele of Taq A1 of DRD2 gene and genotypic frequency are shown among the table 48B.Based on these results, the inventor has detected the percentage ratio of A1/A2 in this research.In TS proband, smoker, gambler and overall subject group, the result is respectively 35.3,42.8,45.7 and 40.0 percent.For TS, smoker and overall contrast, the result is respectively 19.2,26.2 and 22.4 percent.In all four groups, the prevalence rate of A1/A2 heterozygote is significantly higher than in the experimenter in the contrast, have 0.0073,0.017,＜0.0001 and 0.0001 p value.In four groups three, according to the gauged α of Bonferroni 〉=0.05/4 or 0.0125, the result is still remarkable.

Above the result show significance negative correlation with the allelic heterozygosity of DRD1, and with the significance positive correlation of DRD2 allele heterozygosity.Be to detect the potential interaction of DRD1 and DRD2 gene, the inventor is divided into those of (being 11 or 22 homozygotes) of the non-heterozygosity of DRD1 allele and DRD2 allele heterozygosity with case.Because two kinds of genes have all been optimized these genotype, this group is called as " all having ".Second group, be called " one ", form by those of those or DRD1 11 or 22 homozygotes or DRD2 heterozygote.The 3rd group, be called " all not having ", for DRD1 allele heterozygosis, and for DRD2 allele 11 or 22 homozygotes.These the results are shown among the table 48-C.The percentage ratio of experimenter in " all having " group is significantly higher than TS contrast (17.9), p=0.016 for TS proband (33.2); Be significantly higher than smoker contrast (24.1) for smoker (45.5), p=0.0041, and be significantly higher than overall contrast (20.8), p=0.0033 for overall experimenter (34.3).Experimenter's percentage ratio does not increase for gambler (23.3) is significant in " all having " group.

The TS group.Because TS is organized the preliminary exploratory study that has carried out DRD1 gene latent effect in behavior, and owing to studied more than one behavior, so will show these results in more detail.The inventor at first utilizes ANOVA to compare the average of 23 different behavior groups in different DRD1 genotype, and in given seven behaviors that are lower than 0.2 p value, five is addictive behavior-alcohol abuse, smoking, feeding,forced, gambling and shopping.Though the alcohol abuse variable is the most remarkable (p=0.0096), none is significant (square method) at α 〉=0.05 place.In all quantitative traits shown in the table 49 and all the other 16 variablees 14 have the highest average for 11 homozygotes.Though the average score in 22 homozygotes is usually above 12 heterozygotes, but for 11 and 22 heterozygotes, the relative size of scoring shows that the dependency that 11 allelic homozygosity show with the scoring that raises in experimenter's TS group is higher than 22 allelic homozygosity.

Based on the result of ANOVA, the inventor has detected the percentage ratio (table 50) that carries 11 genotypic experimenters between " contrast does not have ", " case does not have " and " case has " group.In α＜0.005 place, three kinds of behaviors, gambling, alcohol abuse and shopping all are significant.For gambling, this percentage ratio increases to 15.2 to 35.2 (p=0.00095) from percent 4.5.Significant six character are addictive behavior in α+＜0.05 for all.

Utilize the single argument regression analysis to obtain similar result, wherein the scoring of isozygotying of those DRD1 DdeI 1 allele is 2, and those to have 12 or 22 genotypic scorings be 1.Gambling and alcohol abuse variable are significant (table 51) at α＜0.005 place.For the single argument regression analysis of DRD2 gene, wherein A1/A2 heterozygote scoring is 2, and homozygote be 1, seven different character is significant (table 51B) in α＜0.005 place.These comprise three addictive behavior, feeding,forced, gambling and smoking.

By multivariate regression analysis, only all provide remarkable result, i.e. gambling, feeding,forced and smoking (table 51C) for three character DRD1 and DRD2 gene.Utilize the single argument regression analysis to obtain identical result, wherein those scorings in " all having " group are 3, are 2 at those of " one " group, and are 1 (showing 51D) at those of " all not having " group.Herein, the p value for gambling, smoking and feeding,forced is＜0.0001.

Be to estimate the size of scoring, the inventor has detected average (table 52) by ANOVA in " all not having ", " one " or " all having " are organized those.Herein, eight character that comprise feeding,forced, smoking and alcohol abuse are significant at α＜0.005 place.Average is the highest all the time in " all having " group.Last check is the detection (table 53) of DRD1 11 homozygotes or DRD1 A1/A2 heterozygote or experimenter's percentage ratio of all having between " contrast do not have ", " case does not have " and " case has " are organized.Alcohol abuse and feeding,forced are to be significant in α 〉=0.05 place in eight character.For alcohol abuse, percentage ratio increases to 46.9 to 70.6, p=0.00005 from 23.9 between three groups.

The result of associating with the DRD1 gene in many addiction and other behavior effect and be consistent in the additive effect of DRD1 and the DRD2 gene hereditism of place variant.Though in TS group, all provide the scoring that is higher than 12 heterozygositys, provide the strongest dependency with many character for 1 allelic homozygosity for DRD1 and 2 allelic homozygosity.

The smoking cessation group.For determining whether in another diverse subject group inventor can repeat these any one in finding, the inventor be used to the early-stage Study that in the smoker, acts on from the DRD2 gene individuality (Comings etc., 1996a).All experimenters in this group inhale at least one cigarette package every day, and once attempt to stop smoking unsuccessfully.As discussed above, when as one group, the smoker shows the remarkable increase of DRD1 DdeI 11 genotype and 11 or 22 genotype prevalence rates.Be the dependency of more detailed discussion between DRD1 gene and smoking, the inventor has detected and has inhaled bag this quantitative trait of number (table 54 and 55) every day.Inhale the contrast of 0 bag and inhale the smoker of 1-1 1/2 bag every day and inhale every day between three groups of the smokers of 2 to 2 1/2 bags in every day, percentage ratio with DRD1 11 genotypic experimenters exists percent 4.9 to 16.4 to 18.0 gradual remarkable increase, p=0.023.Opposite with the TS group, for the smoker, 11 or 22 homozygote groups provide more significant result.Therefore, the experimenter's of isozygotying percentage ratio from every day 0 bag contrast 39.3 be increased to the smoker's that inhales 1-1 1/2 bag every day 61.8, inhale to every day 2-2 1/2 bag the smoker 68.0, p=0.00033.The experimenter's of DRD2 A1/A2 allele heterozygosis percentage ratio increases to 34.5 to 46.3, p=0.047 from 26.2 percent between three groups.As the TS group, the effect of DRD1 and DRD2 gene adds up.Therefore, experimenter's percentage ratio increases to 34.5 to 50.4, p=0.0001 by 24.1 between these three groups in " all having " group.For further detecting DRD1 and the additive effect of DRD2 gene in the smoker, the inventor utilizes multivariate regression analysis to detect and wraps number variable every day.There is suitable effect in two genes, unite they then be responsible for wrapping every day number variable variation 10.5 percent.

The gambler.As discussed above, compare with overall contrast, DRD1 11 homozygotes or 11 or 22 homozygous experimenters' percentage ratio significantly increases in the gambler, and the experimenter's of DRD2A1/A2 heterozygote percentage ratio also significantly increases.Organize differently with TS and smoker, these effects adding up property not in the gambler is not because experimenter's percentage ratio increases than overall contrast in " all having " group.

Heterozygosity.The inventor shows that to the observation of the multiple behavior average score in many different subject group hereditism's variant of DRD1 and DRD2 gene may also show hybrid vigor.Effect opposes in two genes, and this point highly significant.Therefore, DRD2 gene TaqI A1/A2 heterozygote has how unusual scoring to most of variablees, and DRD1 genetic heterozygosis has how normal scoring.The inventor infers that two kinds of polymorphisms all are (Comings etc., 1991) with other sudden change linkage disequilibrium that influences DRD1 and DRD2 gene function.The mechanism of action of this tangible heterozygosis advantages/disadvantages is unknown.Opposition effect in two genes is because they have the fact of the effect of opposition to ring AMP, or simply because also be unknown by linkage disequilibrium with the fortuitous variation in their other relevant mutation types.This result shows that in addictive behavior DRD1 adds the effect of the polygenic inheritance of DRD2 gene.Though the pith of polygenic inheritance is the pivotal role of gene associating, this research also confirms the pivotal role of allele associating.

The importance of experimenter's conclusive evidence.Can be three effects that independently confirm the DRD1 gene between experimenter group, and at two importances of DRD1 and DRD2 gene additive effect among the experimenter group independently? the inventor is to supporting the effect of DRD1 gene because of the early-stage Study that is diagnosed as the experimenter that alcoholism or drug dependence prove conclusively first, and DRD1 and DRD2 gene are not the additive effect in all substance abuse at some.This is not unexpected for polygenic inheritance.Because DRD1 and DRD2 gene are responsible for a given character separately less than percent 6 variation, and a character is responsible for less than percent 11 variation, so these moderate effects can be surpassed by the difference in experimenter's conclusive evidence at an easy rate in the associating back.For example, the polytype substance abuse of unusual participation of many receptors probably comprises dopamine, 5-hydroxy tryptamine, Fructus Cannabis ester, nitric oxide, nicotine hydroxycholine, GABA and other.The syndromic diagnosis of Tourette relies on the existence that motion is twitched, and dopamine plays an important role in muscular movement is regulated.Therefore, can expect that the substance abuse of sending out altogether in TS or other addictive behavior should be than hereditism's defectives that more may relate to dopamine receptor in the subject group of proving conclusively based on the substance abuse of any kind.

In the symptom level but not the importance of diagnostic level test.(Comings and Comings 1987b) show that independent two minutes diagnostic classifications may be too wide, and cause that dynamics significantly reduces in correlational study for these and former research.For example, TS experimenter may be in that destructiveness be united twitch, stutter, ADHD, is forced to having from the individuality that has slightly gentle twitch and do not have an other problem, change the individuality of conduct, anxiety, emotion, substance abuse and learning disorder.It is relevant with stutter especially if a given gene pairs TS has effect, and stutter only exists in 20% the case, then the effect of that gene may be left in the basket in contrast and all TS proband's comparison, but may be detected in the contrast of not having stutter and stutter TS proband's comparison.This notion particular importance in this research.In TS group, relate to those of addictive behavior with the behavior variable of DRD1 gene significant correlation.This can not be one and chances on, because zooscopy also shows the effect of DRD1 gene in the addiction character, and because has repeated this discovery in three different subject group.

About this research several explanations are arranged.Because being neutral base, DRD1 DdeI polymorphism changes, so the inventor infers that it coexists and influences dopamine D ₁The DRD1 site of Rd or the sudden change linkage disequilibrium in the near zone.A common worry of association study is that the possibility of result is because the ethnic group of hiding or the probability of ethnic layering rather than gene itself.Though the inventor attempts to find it is minimized to non-Spain white race people's research and by independently repeating this in the subject group at three by being confined to, it is still a possible explanation.Another potential problem in the TS group, is the detection of 23 varying number character particularly.Come it is compensated by the α that utilizes very conservative 〉=0.05.When detecting the joint effect of DRD1 and DRD2 gene, many p values are lower than 0.05/23 or 0.0022 complete Bonferroni correction less than 0.001.In addition, some internal consistencies are provided around hiving off of addictive behavior (alcoholism, feeding,forced, gambling, shopping and smoking) significant result.Last explanation is that DRD1 11 allelic homozygosity have been emphasized in the table of TS group, and 11 or 22 homozygosity have been emphasized in the table of smoking group.Yet as shown in the table 48,11 or 22 homozygosity are significant between all three groups.The characteristics that all three groups have jointly are for the reduction of DRD112 heterozygote average score and for the increase of DRD2 A1/A2 heterozygote average score.Conclusive evidence bias discussed above may be also decision why 1 allelic homozygosity may be in a subject group more important and 1 and 2 allelic homozygosity play a role in may be in another group more important.

Table 48

Allele and genotypic frequency in all three subject group

N 11 12 22 p q %11 %11,221A.DRD1 DdeII.Tourette syndrome group contrasts 63 4 37 22 0.34 0.66 6.4 41.3TS proband 227 36 97 94 0.37 0.63 15.9 ¹57.3 ²II. the smoking cessation group contrasts 61 3 37 21 0.35 0.65 4.9 39.3 smokers 177 31 60 86 0.34 0.66 17.5 ³66.1 ⁴III. gambling group gambler 163 23 72 68 0.36 0.64 14.1 ⁵55.8 ⁶Overall contrast 124 7 74 43 0.35 0.65 5.7 40.3 overall experimenters 567 90 229 248 0.36 0.64 15.8 ⁷59.6 ⁸

¹x?sq＝3.75???p＝0.053

²x?sq＝5.08???p＝0.024

³x?sq.5.88????p＝0.015

⁴x?sq.13.45???p＝0.0002

⁵X sq.5.39 p=0.020 (between gambler and overall contrast)

⁶X sq.6.75 p=0.009 (between gambler and overall contrast)

⁷x?sq.8.81??????????p＝0.003

⁸x?sq.15.38?????????p＝0.0001

Table 48 (continuing)

N 11 12 22 p q %121B.DRD2 TaqII.Tourette syndrome groups contrast 73 3 14 56 .15 .85 19.2TS proband, 345 17 122 206 .23 .77 35.3 ¹II. the smoking cessation group contrasts 65 6 17 42 .22 .78,26.2 smokers, 194 7 83 104 .25 .75 42.8 ²III. gambling group gambler 186 3 85 98 .24 .76 45.7 ³Overall contrast 138 9 31 98 .20 .80 22.4 overall experimenter 725 27 290 408 .24 .76 40.0 ⁴

¹x?sq＝7.19????p＝0.0073

²x?sq.＝5.69???p＝0.017

³X sq.18.61 p=＜.0001 (between gambler and overall contrast)

⁴x?sq.15.26????p＝0.0001

Table 48 (continuing)

N does not all have one and all has % all to have 1C.DRD1 DdeI 11,22 to add DRD2 Taq1 12I.Tourette syndrome group to contrast 67 4 51 12 17.9TS proband 214 19 124 71 33.2 ¹II. the smoking cessation group contrasts 58 17 27 14 24.1 smokers 176 64 32 80 45.5 ²III. gambling group gambler 154 32 86 36 23.3 totally contrasts 125 21 78 26 20.8 overall experimenters 544 115 242 187 34.3 ³

¹x?sq.5.71??p＝0.016

²x?sq.8.25??p＝0.0041

³x?sq?8.63??p＝0.0033

All do not have: DRD1 12 and DRD2 11,22

One: DRD1 12 and DRD2 12, perhaps DRD1 11,22 and DRD2 11,12

All have: DRD1 11,22 and DRD2 12

Table 49

In the TS group, utilize the DRD1 result of ANOVA

|----------the allele group-------------|

11??????????????12???????????????????22

N=43 n=136 n=125 behavior M S.D. M S.D M S.D. F-ratio ^*P ethanol 1.30 3.3 0.23 ^*1.2 0.42 2.2 4.17 0.0096 smoking 0.97 0.4 0.77 ^*0.5 0.81 0.5 2.80 0.062 forces feed 1.17 0.5 0.94 0.6 0.94 0.5 2.71 0.068 to twitch 4.27 3.0 3.64 3.1 3.25 2.9 1.88 0.154 shopping 1.56 2.9 1.00 2.3 0.78 2.2 1.74 0.177 that show effect of 4.51 4.5 3.17 3.6 3.23 3.8 2.18 0.115 gambling, 0.41 1.2 0.12 0.7 0.15 0.8 2.00 0.136 major depression

^*Significantly be lower than genotype 11 by the Tukey check at α=0.05 place.

Table 50

In Tourette syndrome group for DRD1 Dde 11

The anosis example of the genotypic chi-square analysis behavior no case of contrast has card side p

N % N % N % 67 4.5 217 15.2 17 35.3 10.91 0.00095 alcohol 70 5.7 216 15.3 18 33.3 9.08 0.002 shopping, 54 7.4 182 13.7 52 26.9 7.85 0.005 medicines, 70 5.7 209 15.8 25 24.0 6.50 0.011 gluttonies, 49 6.1 185 15.1 43 23.3 5.35 0.020 smokings 70 5.7 218 16.4 15 20.0 5.02 0.024 of gambling

Table 51

In Tourette syndrome group for DRD1 DdeI 11 genotype

Single argument regression analysis behavior r r with different behavior scorings ²T behavior r r ²T31A.DRD1 DdeI (12; 22=1; 11=2) gambling 0.190 0.036 3.36 alcohol 0.168 0.028 2.97 forces feed 0.139 0.019 2.33 smoking 0.129 0.016 2.26 to twitch 0.119 0.014 2.98 and reads 0.112 0.012 1.97 shopping, 0.095 0.009 1.6731B.DRD2 TaqI (11; 22=1,12=2) opposition disobeys 0.205 0.042 3.59 conduct disorder 0.197 0.038 3.43 and forces feed 0.202 0.041 3.37 smoking 0.184 0.033 3.21 gambling 0.173 0.029 2.98ADHD 0.163 0.027 2.82 manic 0.156 0.024 2.70 stutter 0.148 0.022 2.55 to force 0.146 0.021 2.53 division sample 0.136 0.018 2.3431C.DRD1 and DRD2 gambling DRD1 0.171 0.029 3.00DRD2 0.168 0.028 2.94F 0.243 0.059 9.10 to force feed DRD1 0.124 0.015 2.8DRD2 0.201 0.040 3.38F 0.237 0.056 7.91 cigarette smoking r r²TDRD1 0.119 0.014 2.09DRD2,0.181 0.032 3.18F 0.219 0.048 7.37 alcohol DRD1,0.189 0.035 3.30DRD2 0.060 0.000 1.05F, 0.199 0.040 6.0731D.DRD1 11 and DRD2 12 all have=and 3; One=2, all do not have=1 (N=293) gambling, 0.240 0.057 4.21 smoking 0.219 0.047 3.84 forces feed 0.236 0.055 3.97 opposition to disobey 0.196 0.038 3.42ADHD, 0.176 0.031 3.05 conduct disorder 0.174 0.030 3.01 to force 0.172 0.029 2.99 manic 0.171 0.029 2.98 alcohol abuse 0.162 0.026 2.82 to twitch 0.134 0.018 2.32 division sample 0.118 0.014 2.23

Table 52

In the TS group, utilize the DRD1+DRD2 result of ANOVA (all not have=DRD1 12,22 Hes

DRD2 11,22; One=DRD1 11 or DRD2 12; All have=DRD1 11 and DRD2 12)

|--------the allele group----------| behavior does not all have one all has F-to compare p

n＝168??????????????????n＝112?????????????????n＝15

M S.D. M S.D M S.D. forces feed 0.87～0.6 1.06 0.5 1.42 0.5 15.76 0.0001 smoking 0.73～0.5 0.91 0.4 1.13 0.3 14.71 0.0002 opposition to disobey 3.05 ^*3.1 4.21 3.3 5.06 3.7 11.70 0.0007ADHD 20.31 ^*14.5 24.63 13.6 28.60 12.7 9.31 0.0025 conduct disorders 2.58 ^*2.1 3.54 2.4 4.26 3.0 9.15 0.0027 force 2.47 ^*3.0 3.40 3.2 4.27 4.1 8.93 0.0030 manic 1.43 2.0 2.06 2.4 2.80 2.7 8.89 0.0031 ethanol, 0.24 ^1.5,0.53 ^2.0 1.93 4.1 7.99 0.0050

The linear ANOVA of #

^*Check in α=0.05 place by Tukey and significantly to be lower than the one group

^ checks significantly to be lower than in α 0.05 place by Tukey all group

～check in α=0.05 place by Tukey significantly to be lower than one or group is all arranged

Table 53

In Tourette syndrome group for having

DRD1 11 or DRD2 12 genotype or all have

The chi-square analysis behavior contrast case of experimenter's percentage ratio does not have case card side p

N???????????％???????N???????????％???????N?????????％??????????squar

Manic 63 22.2 153 44.4 75 57.3 16.82 0.00004 alcohol 67 23.9 211 46.9 17 70.6 16.51 0.00005 of e force 64 23.4 155 44.5 73 57.5 15.87 0.00007 conduct disorders, 54 22.2 94 41.5 134 53.7 15.45 0.00008 division samples, 62 25.8 144 44.4 82 57.3 13.94 0.0002 property 52 26.9 158 41.1 85 56.5 11.94 0.0005 to force feed 46 23.9 181 47.0 41 58.5 10.76 0.0010MDE 46 28.3 134 44.0 94 55.3 9.12 0.0025

Only provide the experimenter that " case has " is higher than " case does not have " 20% at least

Table 54

DRD1 and DRD2 number (1%) in the smoking cessation group

Inhale bag number genotype 0 1-11/2 2-21/2 every day and add up to DRD1 11 3 (4.9) 9 (16.4) 22 (18.0) 204

12，22??58????????????46??????????????100?????????????34

Amount to 61 55 122 238 5.14 0.023 ^* DRD1 11,22 24 (39.3) 34 (61.8) 83 (68.0) 141

12??????37????????????21??????????????39??????????????97

Amount to 61 55 122 238 12.87 0.00033 ^*

Inhale bag number genotype 0 1-11/2 2-21/2 every day and add up to DRD2 12 17 (26.2) 20 (34.5) 63 (46.3) 100

11，22????48?????????????38??????????????73??????????????159

Total 65 58 136 259 7.99 0.0047 ^*DRD1 and DRD2 all do not have 17 23 41 81

One 27 13 19 59

All have 14 (24.1) 19 (34.5) 61 (50.4) 94

Add up to 58 55 121 234 23.48 0.0001 ^*

^*Linear card side d.f.=1

^*Perason card side d.f.=4

Table 55

DRD1 and DRD2 gene in the smoking cessation group

Multiple linear regression is analyzed

r???????????????r ²??????????T??????????pDRD1?DdeI?12????????????0.232????????????0.054????????30.73?????0.0002DRD2?TaqI?12????????????0.220????????????0.048????????30.54?????0.0005F???????????????????????0.325????????????0.105????????13.63?????＜0.0001

Embodiment 8

Effect with endorphine enzyme inhibitor and releasing agent treatment

In the present embodiment, the interaction that has detected d-phenylalanine (other endorphine enzyme inhibitor), Tyr-D-Arg (a kind of endorphins releasing agent) and naltrexone (narcotic antagonist) is released into the effect in the volt nuclear (Acb) of Lewis (multiple medicines hobby) and Fischer (non-multiple medicines hobby) rat for dopamine.Lewis and Fischer rat all are divided into two groups: acute and chronic.Chronic group of dosage that gives three kinds of medicines every day is: every morning 500mg/kg the Tyr-D-Arg (TDA) of d-phenylalanine (DPA), 1-5mg/kg and the naltrexone (NX) of 1-2mg/kg (Du Pont, Wilmington, DE) totally 18 days.At the 19th day, chronic treatment or acute group are begun to carry out the microdialysis sampling.The following variation of the associating of medicine: give all three kinds; Any two kinds associating; Independent each medicine.The acute dose of three kinds of medicines is with as follows: DPA=500mg/kg i.p., TDA 5mg/kg i.p., NX i.p.2mg/kg.

Microdialysis method before research seven days, all rats will be implanted microdialysis probe: A+2.0, L1.2 and V-8.0 (apart from skull) by perform the operation under the solid location is collaborative (with respect to bregma) down in sodium phenobarbital (50mg/kg) anesthesia in the Acb of left side.Probe design be concentric and with 0.5mm external diameter stainless steel tube make (Small PartsCompany, Roanoake, VA).The 2mm that each probe has 250 μ m external diameters and covers the Acb vertical range basically expose dialyzer (Spectra/Por doughnut cellulose fiber, MWCO5000, Spectrum Medical Industries, Lip river China fir base, CA).

The dialysis response rate determines by in vitro study, the relative recovery of probe will be approximately 3.9% dopamine (DA), 4.1% 3,4-dihydroxyphenyl acetic acid (DOPAC) and 3.2% 4-hydroxy-3-methoxy-.alpha.-toluic acid. (HVA).Just after implantation, will begin to continue in probe infusion and the whole research.Utilization is coupled to the dual vitreous carbon working electrode of two LC-4C detectors, and (bio-analysis system company, West Lafayette IN) carry out electrochemical measurement, all are located at corresponding to Ag/ACl reference electrode+0.7V place.Electrode is used for that DA detects and another is used for DOPAC and HVA detects.Recovering 20-24h from operation after, animal begins the dialysis sampling.Behind DA, the DOPAC of dialysis solution and HVA horizontal stable, collect the dialysis specimen.Collect the preceding specimen of injection of three 20min baselines then.Before any chronic injection, will analyze pre-dialysis solution and determine the baseline amount.After setting up, this baseline amount begins chronic injection.For acute rat group, before collection, begin medication combined administration with a predetermined interval that changes according to research.Collect more than the 2h specimen of six 20min altogether after the administration.Also will beginning from research to move observation with stereotyped behavior of the rat chronic and acute group.After all collections were finished, the histologic analysis that the brains of all rats is carried out standard was proved conclusively the position of probe.

The oneself selects also will to carry out other and relates to the medication combined research of the oneself being selected to influence between Lewis and Fischer rat and come to determine specifically that independent DPA and TDA and NX unite the influence of ethanol, cocaine, sugar juice, Fructus Cannabis and nicotine being thirsted for to reduce systemicly.Suppress GABA and transmit that uniting of active DPA and TDA actively to be suppressed at the amount aspect that remarkable increase DA enters Acb to central member lid GABA should be the strongest.The utilization of NX seems to prevent ethanol inductive glad.

Embodiment 9

The correlating method of DRD2 A1 allele and body fat percentage ratio

Obesity experimenter.The target of this research is to obtain main and subject group that needn't be made up of the morbid obesity experimenter utterly.Have 34% or the women of bigger fat content and have 28% or the male of bigger fat content be considered to morbid obesity.

Contrast.Contrast is made up of the twinborn father and mother from " Minnesota twins family study ".Since simply based on have 11 or 17 years old twins determine these experimenters with whole state, so the group that their representative is organized than university student all social economies more at random and education.Because the result of substance abuse evaluation also can't provide in the twins contrast, so the inventor classifies these as " not Shai Xuan contrast ".Owing to the experimenter in the fat sample is not screened substance abuse, so this makes that contrast is suitable with the experimenter for the existence of obesity yet.

The result.Fat group is made of 76 women and 15 male 91 experimenters.Among the women, 60 or 78.9% is morbid obesity, and remaining is 16 or 21.1% for overweight but have and be lower than 34% fatty %.Among the male, 12 or 80% is morbid obesity, and remaining is 3 or 20% for overweight but have and be lower than 28% fatty %.Do not allow independently statistical analysis owing to the ratio that is not MO experimenter is too little, be used for analyzing so merge two groups.

DRD2 D in obese subjects and contrast ₂The allelic prevalence rate of A1 is shown in table 56.Overall for masculinity and femininity, 67.0% carries D in the obese subjects ₂A1 allele, and the screening contrast is not 32.3%, x ²=32.95, p＜0.00001.In fact this value is identical (table 57) with 32.9% value of being reported in the document of nineteen ninety for 980 white people's contrasts of not screening.68.4% prevalence rate is significantly higher than contrast.For the results are shown in the table 58 of DAT1 gene.Opposite with the DRD2 gene, compare the not significant increase of 10/10 genotypic prevalence rate in obese subjects with contrast.

Table 56

The Minnesota twins contrast-does not screen substance abuse

D2A

11 12 22 Tot %12M+F N, 14 83 203 300% 0.047 0.277 0.677 1 0.323F N, 7 46 99 152% 0.046 0.303 0.651 1 0.349M N, 7 37 104 148% 0.047 0.250 0.703 1 0.297 obesity sample-% fat numbers reach and do not screen substance abuse

D2A

11???????????12??????22???????Tot????％12M+F????N?????????5????????????58??????30???????92％???????????????0.055????????0.615???0.330????1??????0.670F??????N?????????5????????????47??????24???????76％???????????????0.066????????0.618???0.316????1??????0.684M??????N?????????0????????????9???????6????????15％???????????????0.000????????0.600???0.400????1??????0.600

Table 57

Contrast 11 12 22 and add up to %1

The unknown Bolos of contrast-state etc., 1,990 4 17 41 62 33.9Grandy etc., 1,989 2 14 27 43 37.2Comings etc., 1,995 0 21 67 88 23.9Gelenter etc., 3 21 44 68 35.3Uhl, 1,992 3 27 71 101 29.7Goldman Finns, 4 38 70 112 37.5Goldman, 1,993 11 42 114 167 31.7Nother etc., 5 26 38 69 44.9Noble etc., 1,994 2 18 38 58 34.5Jonsson etc., 1,993 4 13 38 53 32.1Hedebrand etc., 1,993 4 19 38 61 37.7O ' Hara etc., 1993 (the non-user of white man) 6 39 115 160 28.1 adds up to=44 278 658 980 32.9

Table 58

Other total of DAT1

.10/10M+F???????????N??????????????160?????????????????122????????????282％???????????????????????????0.567???????????????0.433??????????1F?????????????N??????????????89??????????????????63?????????????146％???????????????????????????0.568???????????????0.432??????????1M?????????????N??????????????77??????????????????59?????????????136％???????????????????????????0.566???????????????0.434??????????1M+F???????????N??????????????44??????????????????47?????????????91％???????????????????????????0.484???????????????0.516??????????1F?????????????N??????????????38??????????????????38?????????????76％???????????????????????????0.500???????????????0.500??????????1M?????????????N??????????????6???????????????????9??????????????15％???????????????????????????0.400???????????????0.600??????????1

Embodiment 10

Be used for the allelic analysis of other polygenes of the detection of RDS

Tryptophan 2,3 dioxygenases.In multiple mental disorder, being in the news unusually of the metabolic defective of 5-hydroxy tryptamine and blood 5-hydroxy tryptamine and tryptophan levels.Tryptophan 2,3-dioxygenase (TDO2) is the rate-limiting enzyme that tryptophan is degraded to the N-formylkynurenine.The functional variety of this gene may be responsible for viewedly increasing in 5-hydroxy tryptamine and the tryptophan in various disorders or reducing.Four kinds of different polymorphisms of people TDO2 gene have been identified.Association study shows one or more significant correlations that move obstacle (ADHD) and drug dependence with Tourette syndrome (TS), attention-deficient of these polymorphisms more.Intron 6 ^{G → T}Variant is with the horizontal significant correlation of platelet 5-hydroxy tryptamine.Through the Bonferroni timing, having only the dependency with TS is significant (p=0.005).

The experimenter.The TS of the state-run medical center, city (COH) that TS proband, TS family member, ADHD proband, 2/3rds unsociable and eccentric disease proband and most contrast are hope and the patient or the patients' relatives of other clinic treatment.The diagnosis of TS, chronic exercise twitch or chronic sounding tic disorder, ADHD and unsociable and eccentric disease is based on DSM-III-R (the sick community of Americanism, 1987) standard.The TS proband is restricted to the TS individuality of seeking therapeutic treatment in this TSS-ADHD clinic.All proband and most relatives individual are met with and check by D.E.C..In the proband, 82% has TS and remaining 18% has chronic exercise tic disorder or chronic sounding tic disorder.The TS family member is TS proband's father and mother, and no matter whether they have TS.A unsociable and eccentric experimenter of/3rd is from Sagamore children's hospital, Dix Hills, NY (J.S.).Each proband and their relatives are all putd question to ethnic group and the ethnic background about they four grand parents, and only comprise that all four grand parents are non-Spain white race people's experimenter.All experimenters have signed written agreement, and research is studied administration committee (Institutional Review Board) approval.The smoker (Comings etc., 1996a) and the pathological gambling person (Comings etc., 1996b) and have the research that the individuality of alcohol dependence (Comings etc., 1994) and drug dependence (Comings etc., 1994) acts in these obstacles from the DRD2 gene.These experimenters' screening and source are described in detail in these files separately.

Schizophrenia and depressed experimenter's DNA specimen be from from " national neurological studies the storehouse " (V.A.Wadsworth hospital, isolating in experimenter's CA) the brain specimen with these diagnosis.Schizoid experimenter has chronic schizophrenia, and the long-term spirit history of being in hospital is arranged usually.Having depressed experimenter once committed suiside and the chronic depression history was arranged.Schizophrenia and depressed diagnosis are based on DSM-III or DSM-IIIR standard under an above psychiatrist's agreement.The experimenter who has alcoholism and drug dependence simultaneously has been excluded.

Contrast.COH contrast is from four sources: a) from the irrelevant grand parents of CEPH (Centre d ' Etude duPolymorphisme Humain) family; B) TS patient's adoptes, brings up or stepparent; C) has the experimenter of thyroid carcinoma or noninsulindependent diabetes from an endocrinopathy clinic; And d) comprises occupation, half occupation, technical staff and surfaceman's corpsman,hospital.This contrast widely is used for avoiding issuable potential screening problem when utilizing the source of more limiting to.Select the endocrine patient in contrast,, have high cure rate, and daily life is produced minimum interference, be present in the range of age widely, and the patient basis is identical with TS experimenter because two kinds of morbid state all are medicable.All contrast all screened ethanol, medicine and the tobacco abuse got rid of.

The PCR of intron 6 saltation zones ^TMAmplification.The PCR of amplification TDO2 target sequence ^TMReact as follows: 10mM Tris HCl, pH8.3,50mM KCl, 1.5mM MgCl ₂, 0.05%Tween20,0.05%NP-40, each 100 μ M dATP, dCTP, dTTP, dGTP and 0.1 μ M primer.Primer is No. 116 GACACTTCTGGAATTAGTGGAGG (SEQ ID NO:5) and No. 117 GAAGTTAAATCCATGTGGCTC (SEQ ID NO:6).With following material add to 20ul:0.5U AmpliTaq (Po Jin-Ai Ermo, Foster City, CA), 1 μ l (250ng) genomic DNA.Operate in a PE-9600 thermal cycler (Po Jin-Ai Ermo) or PTC-100 thermal control instrument able to programme (MJ research company below utilizing, Watertown, MA) react on: 94 ℃ of 5min, 94 ℃ of 30sec of 30 circulation then, 60 ℃ of 30sec, 72 ℃ of 1min, 72 ℃ of 5min then.For determining whether amplification takes place, the reactant mixture of getting 10 μ l in tbe buffer liquid on 1.5% agarose gel electrophoresis.

Clone and order-checking.PCR with primer 116 (SEQ ID NO:5) and 117 (SEQ ID NO:6) ^TMThe product ethanol precipitation, and be resuspended in the TE buffer (Tris HCl 10mM, EDTA 1mM).Fragment cloning is arrived among the modified Blue Script pBdT (Hoton and Graham, 1991).Contain following composition in the 20 μ l coupled reactions: 20 μ l 10 * connection buffer (Bao Lingman), every kind of PCR of 100ng ^TMProduct, pBdT and 1 μ l T4 ligase.It is hatched 18h at 11 ℃.(FosterCity CA) measures fragments sequence with terminal primer T3, T7 and No. 129 GCTGATTTTCAGACTGAGTGTG of inner primer (SEQ ID NO:7) and No. 130 CTACAAACATATTTAAACATATGTT (SEQ ID NO:8) on the automatic sequencer in an Applied Biosystems company.

Denaturing gradient gel electrophoresis.Pass through PCR ^TMIncreased DNA between intron 6 oligomers No. 116 (SEQ ID NO:5) and No. 117 (SEQ ID NO:6) produces the fragment of a 1359bp.Digest the fragment of this fragment generation 816,470 and 60bp with RasI.With its in 6.5% acrylamide gel of 20-80% degeneration in 60 ℃, 70V electrophoresis 16hr (Grey, 1992).At 10mM Tris HCl, 50mM KCl, 1.5mM MgCl ₂, 0.05%Tween20,0.05%NP-40, pH8.3 buffer in carry out PCR with the various deoxidation NTP of various primers of 0.2 μ M and 0.2mM ^TM(MA), (in 1 μ l) carries out 20 μ l PCR with the 5U enzyme for New EnglandBiolab, Beverly to utilize 10 of 2.3 μ l * reaction 1 buffer ^TMThe RsaI of product digests, and digests at 37 ℃ and differentiates segmental size yesterday.

BslI digestion.From top PCR ^TMIn the reaction, the portion of getting 10 μ l digests with the restricted enzyme BslI of 1.5U and final 1 * buffer (by New England Biolab, Beverly, MA provides), and 55 ℃ of overnight incubation.(ME) 100V carries out electrophoresis 1h in the gel for F.M.C. product, Rockland in 4% metaphor agarose in 1 * TBE (Tris-borate 100mM, EDTA 1mM) with the portion of 10 μ l in the digestion product.Gel is dyeed in ethidium bromide.Expection has the fragment of three different sizes.When pleomorphism site was G/G, DNA was produced the fragment of 673bp and 359bp by catapepsis.When pleomorphism site was A/A, the 1032bp fragment was not digested.The G/A heterozygote has three fragments, 1032bp, 673bp and 359bp.

Oligonucleotide connects test (OLA).Oligonucleotide used in the OLA to G → T variant is as follows: for G specificity oligomer OLA-G CTATTCTTATCCCTCTTTTCTTAA-(HEO) 1 (SEQ ID NO:9).For T specificity oligomer OLA-TATATTCTTATCCCTCTTTTCTTAAT-(HEO) 3 (SEQ ID NO:10).G specificity oligomer has 1 and T specificity oligomer has the six oxirane phosphamides (Grossman etc., 1994) that 5 ' end changes these two oligomer molecular weight that add to of 3U.General oligomer is FAM-TATATATTACGGTTTATTACCGT-PO4 (SEQ ID NO:11), wherein FAM be 5 ' CF 5(6)-Carboxyfluorescein phosphoramidite (Applied Biosystems company, FosterCity, CA).When the G specificity was connected by coupled reaction with general oligomer, the expection size was 50.5bp.When the T specificity was connected with general oligomer, the expection size was 56.5bp.The reactant mixture of OLA reaction is made up of following compositions: 20mM Tris HCl, pH7.6,100mM KCl, 10mM MgCl ₂, 10mM dithiothreitol, DTT, 1mM NAD, 0.1%Triton X-100,10nM oligomer.The PCR of 10U ligase (New England Biolab) and 1 μ l ^TMProduct adds in the reactant mixture of each 20 μ l.Cycling below utilizing on a PE-9600 thermal cycler is reacted: 94 ℃ of 30s of 20 circulation, 54 ℃ of 2min30s.Each 2 μ l reaction mixed and be heated to 92 ℃ of 2min with the deionized formamide of 0.5 μ l come degeneration.Used 2.5 μ l all go up in 8% acrylamide gel of sample in TBE and 8M carbamide and electrophoresis 3h.Carry out electrophoresis or dye the identification reaction product by the primer that in Applied Biosystems dna sequencing instrument, utilizes fluorescence staining by silver.The latter is made up of double staining, at first uses Stains-All, dyes with silver then.(NY) 20-30min in the solution decolours in 2% glycerol then and spends the night for Eastman Kodak, Rochester gel to be placed at 0.01% the Stains-All for preparing among 50% the Methanamide pH7.5.In the acetic acid of 10% ethanol and 0.5%, gel is washed twice then, each 30min.Then with gel at 0.1% AgNO ₃Hatch 20-30min in the solution, wash twice, place the 1.5%NaOH in 1 liter of distilled water, the 0.01%NaBH of prepared fresh then with deionized water ₄, 4ml 37% formalin in.In 10-20min, show band, use 0.75% Na then ₂CO ₃Cessation reaction, and fixing in 5% acetic acid.

By the evaluation of DpnII digestion to G → T variant.The sequence that is right after G → T 3 ' is GATA.GATC is the recognition site of DpnII restricted enzyme.The oligomer of design 3 ' 23bp is with the ATC sequence that is right after G → T variant 3 ' following (oligomer underscore and for the following stroke two-wire in two g sites of variant): the 5 '-TCATTAATCCTCTGGGTATTGTAAATGTGGATTTAGGTTAATGTATTATATATAAT GCCAAATAATGGCAGATAAGAATAGGGAGAAAAAGAATTA-3 ' (SEQ ID NO:12) that matches, 5 '-ATTAATCCTCTGGGTATTGT-3 ' (SEQ ID NO:13), 5 '-TAGTCTTATCCCTCTT TTTCTTA-3 ' (SEQ ID NO:14).This mismatching on the 3rd site seldom damaged it as PCR ^TMThe effect of primer.Select 5 ' primer that the product of one 92bp is provided.As G → when the T variant is A, only there is the fragment of a 92bp to occur.

PCR ^TMThe condition of reaction is as follows: the various primers of 0.1 μ M, the various dNTP of 0.2mM, 50mM KCl, 10mM Tris HCl, 1.5Mm MgCl ₂, 0.001% (w/v) gelatin, the AmpliTaq of per 100 μ l 2.5U ^Archaeal dna polymerase (Po Jin-Ai Ermo, Foster City, CA), the 80ng genomic DNA.PCR ^TMCirculation is 94 ℃ of 4min, 94 ℃ of 30sec of 30 circulation then, 52 ℃ of 90sec, 72 ℃ of 120sec, 72 ℃ of 5min then.The condition of DpnII digestion is the PCR of 10 μ l ^TMProduct, the 10U μ l of 0.05 μ l ^-1DpnII; 1.5 μ l's: 1M NaCl, 0.5M Bis HCl, 0.1M MgCl ₂, 10mM dithiothreitol, DTT, pH7.9; 3.5 μ l H ₂O spends the night in 37 ℃.With product electrophoresis in 4% metaphor agarose.

The multiple amplification of CCCCT.Be used to detect the reverse complemental that the multiple oligomer of intron 5 CCCCT is No. 166 5 '-CTCTTACAATAGAAGAAACCATTT-3 ' (SEQ ID NO:15) and No. 167 5 '-TCTCCTCTCTTTCCCTTCCC-3 ' (SEQ ID NO:16).Amplification condition is 95 ℃ of 5min, 95 ℃ of 1min of 30 circulation then, 50 ℃ of 1min, 72 ℃ of 1min, 72 ℃ of 5min subsequently then.Final concentration in reactant mixture is 50mM KCl, 1.5mM MgCl ₂, 10mM Tris HCl, dATP, the dCTP of pH8.3,0.1 μ M primer, each 200 μ M, dTTP, the dGTP of each 100 μ M and the template of 7-denitrogenation-dGTP and 0.5 μ l.

Exon 7 A → C (Asn → His) the discriminating of polymorphism that suddenlys change.The PCR of A → C variant is used to increase ^TMPrimer is 5 '-GCATGGCTGGAAGAACTCC-3 ' (SEQ ID NO:17) 5 ' primer and 5 '-TCTTCCAGGCCTCTGGTCATAT-3 ' (SEQ ID NO:18), 3 ' primer.The 89bp product that produces is digested to 67 and the fragment of 22bp in C variant site by NdeI.

Association study.Used method is the comparison proband and various allelic prevalence rates between contrast that have nothing to do in the same person kind of groups.Bonferroni correction α is 0.05/10 or 0.005 in table 59,60 and 61.In the time in several years, finish these research.Owing to identified new polymorphism,, then do not had the experimenter and further detected if a hundreds of experimenter's of beginning research prompting variant same behavior phenotype is not a significant correlation.As a result, the quantity for different experimenters that variant is studied is different.

Table 59

The n of the group as a result GG GA AA %A OR CI x of intron 6G → ATDO2 polymorphism ²P contrasts depressed 16 14 11 12.6 3.88 0.7-21.9, the 2.70 NS pharmacological dependences of unsociable and eccentric disease 65 61 22 6.2 1.78 0.5-6.8 of 141 136 41 3.5ADHD, 113 110 30 2.7 0.74 0.2-3.2,0.16 NS alcohol dependence, 65 65 00 0.0 0.00---2.36 NS 0.72 NS 71 69 20 2.8 0.79 0.2-4.2 0.78 NS pathologic 166 158 80 4.8 1.38 0.4-4.3 0.30 NS gambling schizophrenia 41 41 00 0.0 0.00---1.45 NS smokers 93 92 10 1.1 0.30 0.03-2.6 1.37 NSTS 299 268 29 2 10.4 3.15 1.1-7.8 5.93 0.015TS carrier 151 135 14 2 10.6 3.22 1.1.9.0 5.43 0.020

OR=odd ratio GA, AA/GG

The confidence interval of CI=OR

Appropriate location uses the Fisher Precision Test

Table 60

The up of the group as a result n GG GT TT %T OR CI x of intron 6G → T TDO2 polymorphism ²P contrasts depressed 19 16 30 15.8 1.00 0.2-3.6,0.00 NS of 197 166 30 1 15.7ADHD 108 81 25 2 25.0 1.78 1.0-3.2,3.89 0.048 alcohol dependence 65 61 31 6.1 0.35 0.1-1.0,3.88 0.049 unsociable and eccentric disease 65 58 52 10.8 0.65 0.3-1.5 0.97 NS^*Pharmacological dependence 73 52 19 2 28.7 2.16 1.1-4.1 5.81 0.016 pathologic 165 127 33 5 23.0 1.60 0.9-2.7 3.10 0.078 gambling schizophrenia 43 33 10 0 23.3 1.62 0.7-3.6 1.41 NS smokers 108 88 18 2 18.6 1.22 0.6-2.2 0.388 NSTS 320 263 52 5 17.7 1.16 0.7-1.9 0.37 NSTS carrier 134 119 14 1 11.1 0.67 0.3-1.3 1.37 NS

OR=odd ratio GT, TT/GG

The confidence interval of CI=OR

^*The Fisher Precision Test

Table 61

The n of the group as a result G/G A of intron 6G → T TDO2 polymorphism or T A+T %A or T OR CI x ²P contrasts depressed 15 10 50 33.3 2.43 0.8-7.8,2.36 NS of 135 112 23 0 17.0ADHD 96 68 27 1 29.1 2.01 1.1-3.7,4.80 0.028 alcohol dependence 64 60 40 6.3 0.32 0.1-1.0,4.31 0.038 unsociable and eccentric disease 64 53 11 0 17.2 1.01 0.4-2.2 0.01 NS^*Pharmacological dependence 70 47 23 0 32.9 2.38 1.2-4.7 6.63 0.010 pathologic 165 123 38 4 25.4 1.66 0.9-2.9 3.10 0.078 gambling schizophrenia 34 26 80 23.5 1.50 0.6-3.7 0.76 NS smoker 84 69 15 0 17.9 1.06 0.5-2.1 0.24 NSTS 271 190 80 1 29.9 2.08 1.2-3.5 7.81 0.005TS carrier 98 74 21 3 24.5 1.58 0.8-3.0 1.96 NS

OR=odd ratio A or T, A and T/GG GG

The confidence interval of CI=OR

^*The Fisher Precision Test

Nonrandom allelic association.Owing to do not carry out family's research,, alleviated classical linkage disequilibrium analytical method (Lewontin and Kojima, 1960 successively so can not determine phase specificity haplotype frequency; Lewontin, 1964).Estimate the degree of nonrandom allelic association between four different polymorphisms being studied by the cross tabulating in the experimenter, wherein same individuality is carried out two or more tests.All experimenters in all diagnostic classifications include among these are analyzed.

Analysis by specific behavior.(Comings etc., 1996c), the inventor finds can provide suitable information by different behavior symptom groups' analysis in the research that d2 dopamine receptor gene DRD2, D β H and DAT1 gene act in TS.This is to detect specific allelic prevalence rate among the contrast of the behavior of studying not having and TS proband and the relatives with the behavior of studying.

5-hydroxy tryptamine and tryptophan levels.The technology that is used for platelet 5-hydroxy tryptamine level, 5-hydroxy tryptamine/platelet ratio and the horizontal analysis of blood plasma 5-hydroxy tryptamine provide in the elsewhere (Comings, 1996b).The result

Contrast.Irrelevant CEPH grand parents forms 60% contrast.For any one that determines whether inventor's self-controlled group provides different results, not on the same group between the frequency of many state properties.There is not the significance difference by chi-square analysis.

Intron 6 DGGE polymorphisms.On polyacrylamide gel electrophoresis, judge polymorphism.The results are shown in the table 62 of original intron 6 DGGE polymorphisms.91 contrasts 12.1% in go out to represent the band of the lower frequency of allele 2.It significantly increases to 27.5% in 40 TS patients, to 50%, and reaches 37.5% in 8 depressed experimenters that once committed suiside in 10 drug dependences.Yet these numerals are all less, and inventor's intention is identified related variant before the research that develops into a large amount of experimenters.Two polymorphisms have been identified in order-checking research, G → T variant and another G → A variant, different two base pairs (Comings etc., 1995).The inventor at first utilizes to connect to test and detects these variants respectively, utilizes modified primer and restricted enzyme to develop a more simple steps (square method) then.

Table 62

The PRELIMINARY RESULTS group n 11 12 22 %2 OR CI x of the DGGE polymorphism of TDO2 intron 6 ²P contrasts 91 80 11 0 12.1 depressed 8530 37.5 4.36 0.91-20.8 3.91 0.083 ^* Drug dependence 10 541 50.0 7.23 1.8-29.2 9.71 0.008 ^*Pathologic 26 16 73 38.5 4.55 1.6-12.3 9.55 0.007 ^* Gambling TS 40 29 92 27.5 2.76 1.1-7.0 4.72 0.031

OR=odd ratio 12,22/11

The credibility interval of CI=OR

^*The two tail Precision Test of Fisher

G → A variant.The results are shown in the table 59 of intron 6 G → A variant.In 141 contrasts, have only 3.5% to carry A allele.In 10 groups that detected, be that A allele increases to 10.4% among 299 TS experimenters (odd ratio 3.15) in the unique remarkable result in α=0.05 place, and in 151 TS first degree relatives 10.6%.These are all not remarkable in gauged p value 0.05/10 of Bonferroni or 0.005 place.

G → T variant.The results are shown in the table 60 of G → T variant.In 197 contrasts, 15.7% carries T allele.It increases to 25.0% (p=0.048) in 108 experimenters with ADHD, at 73 experimenters to 28.7% (p=0.016) with medicine or many substance depilatories and 6.1% (p=0.049) in 65 experimenters with alcohol dependence.These are all not remarkable at α=0.005 place.

G → A or G → T variant.The inventor has detected the prevalence rate (table 61) of G → A or G → T polymorphism existence.In the contrast that 135 two kinds of detections have all been carried out, 17% carries any allele.It increases to 29.1% (p=0.028) for 96 experimenters with ADHD, to 70 experimenters to 32.9% (p=0.01) with drug dependence, and to 271 experimenters to 29.9% (p=0.005) with TS.For 64 experimenters with alcohol dependence, it reduces to 6.3% (p=0.03).The result who only has TS at α=0.005 place is significant.

A → the C of exon 7,748 Asn → His variant.The inventor may can provide information to be filled with unbounded confidence to Asn → His variant, because it is a kind of exon variant, and relates to important aminoacid in the haemachrome combination.C or His allele be present in 48 contrasts 6.3% in, and this value for experimenter with ADHD, unsociable and eccentric disease, schizophrenia and TS by a little variation, but none is remarkable.

The CCCCT polymorphism.For all tested experimenters, at intron 53 ' the allelic base pair length in terminal GCCCT repeat region place and frequency be as follows:

Gene frequency

Size

210bp?????????0.018?????????240bp????????????0.838

215bp?????????0.053?????????245bp????????????0.018

220bp?????????0.023?????????260bp????????????0.005

230bp?????????0.045

So far, 240 allele and 240/240 genotype are modal.For the purpose of analyzing, compare with 240/240 genotypic frequency with non-240/ non-240 genotypic frequency 240/ non-240.For 125 contrasts, 31.2% carries 240/ non-240 or non-240/ non-240 genotype.For the prevalence rate of other group much at one, in 22.1% to 30.5% scope, and none is significantly different with contrast.

Nonrandom allelic association.Table 63 is presented at the result of nonrandom allele dependency estimated value between four TDO2 variants of being studied.Though 1245 experimenters have altogether been measured G → A and G → T variant, only there are 10 experimenters to be heterozygosis for A and T allele, one is TT homozygote and A heterozygote, shows that two kinds of sudden changes are on same chromosome.Each variant allele is rare relatively, one of 18.0% T allele and the two of 24.1%.These results show that the sudden change (2bp distance) of these two vicinities is present on the different chromosomes, and observed distribution is as broad as long with the accidental associating of independent event.Among 312 experimenters that tested G → T and A → C variant (table 63-B), both are heterozygosis neither one.For total group, have only 7.4% carrying C allele (AC heterozygosis in the AA homozygote) on the phase homologous chromosomes.Among 621 experimenters that tested G → A and CCCCT polymorphism (table 63-D), have only six for A and non-240bp CCCCT allele heterozygosis, and one non-240/ non-240 homozygotes are the GT heterozygote.These distributions do not have difference with occasionality, show that they are on complete isolating chromosome.

At last, among the experimenter that 696 have been tested G → T and CCCCT polymorphism, (table 63-E), 80 (47.6%) are the GT heterozygote in 169 240/ non-240 heterozygotes, 19 (70.3%) are GT or TT in non-240/ non-240 homozygotes, and 10 (100%) is non-240/ non-240 homozygotes in the TT homozygote.This relation is highly significant (x ²=421.07, p＜0.00001), show the nonrandom allelic association that between non-240 CCCCT variants and T allele, has height.The result shows that at least 50% non-240 allele occur on the T allele chromosome, and at least 78% T allele occurs on the non-240 allele chromosomes.

Table 63

Nonrandom equipotential between four different TDO2 variants

Between the estimation of gene-correlation (df=4) 63A. intron 6 T → A and T → G

G GT TT adds up to x ²PG 944 194 19 1157GA 70 10 1 81AA 7007 2.74 0.60 totals 1,021 204 20 124563B. introns 6 G → T and exon 7 A → C (Asn → His)

G GT TT adds up to x ²PA 227 60 6 293AC 18 00 18CC 1001 5.43 0.24 add up to 246 60 6 31263C. intron 6G → A and exon 7 A → C (between Asn → His)

G GA AA adds up to x ²PA 254 16 0 270AC 20 11 22CC 2002 9.72 0.045 add up between 276 17 1 29463D. introns, 6 G → A and intron 5 CCCCT polymorphisms

G GA AA adds up to x ²P240/240 426 17 5 448240/x 144 60 150x/x 21 10 23 1.97 0.74 add up between 591 25 5 62163E. introns, 6 G → T and intron 5 CCCCT polymorphisms

G GA TT adds up to x ²P240/240 473 28 0 501240/x 88 80 0 168x/x 89 10 27 421.07＜0.00001 add up to 569 117 10 696

The comparison of single behavior.When for the average of A or the more different behavior scorings of T allele, the result who has only polyphobia disease to exist is significant, is 2.89 (p=0.043) for those average score of " all not having ".When the contrast that is not having phobia, the TS proband who does not have phobia and relatives and have the TS proband of phobia and relatives between when the prevalence rate of each variant compared, have the linear x2 (3.45) of topnotch for the result of phobia, but not significantly (p=0.06).Every kind prevalence rate increases to 24.15% to 34.1% from 18.8% between these three groups herein.None is with the non-240bp allele significant correlation of CCCCT polymorphism in each behavior.

TDO2 polymorphism and 5-hydroxy tryptamine level.From before research can obtain platelet 5-hydroxy tryptamine and blood tryptophan levels data (Comings, 1990b).Because some experimenters participate among two researchs, so determine that in having the allelic experimenter of different TDO2 whether platelet 5-hydroxy tryptamine or blood tryptophan levels exist any difference is possible.These the results are shown in the table 64.Analyze or by the Mann-Whitney non parametric tests, for G → A variant, compare with G/G experimenter by ANOVA, 5-hydroxy tryptamine among G/A and the A/A/platelet ratio does not have significance and changes.The distribution of 5-hydroxy tryptamine/platelet rate value has GG for those or GT, TT are genotypic.

Though numerical range is similar, distributing is different from normal distribution.For the GG that carries G → T polymorphism genotypic those, many values concentrate in the scope of 0.5-1.25.On the contrary, carry T allelic those seldom in this scope.Also has one 10.9 super inclined to one side value for T allele group.When removing should be super inclined to one side value, for carry T allelic those, average 5-hydroxy tryptamine/platelet ratio significantly higher (p=0.003).When this value of this worthwhile eliminating, the p value reduces to 0.081.Yet with more suitably Mann-Whitney non parametric tests, T allele group still has significantly high average.Repeat polymorphism for CCCCT, as expected, carry non-240 allelic those and have higher 5-hydroxy tryptamine/platelet ratio.Yet difference is not remarkable.Tryptophan levels is all not remarkable to any polymorphism.

Embodiment 11

The adding up property of hereditism's association

The PS1 gene pleiomorphism.Diallele sign during senilism element-1 (PS-1) polymorphism is positioned on No. 14 chromosomes, has proved that it participates in Alzheimer.In view of the smoker develops into the report that the risk of Alzheimer reduces, the inventor has determined at a series of 132 that its ethanol and medicated cigarette are used among the non-Spain white race people experimenter of pattern and has detected the PS-1 gene.The inventor finds that the homozygote (genotype 1/1 and 2/2) of this gene is significantly more common for smoker (p＜0.05), and has the scoring (p＜0.01) of remarkable rising in (MAST) at " Michigan alcoholism filler test ".

From whole blood, extract experimenter's genomic DNA by the method for standard.Use PCR ^TMMethod (Saiki etc., the 1988) target DNA that increases utilizes each primer (5 ' CACCCATTTACAAGTTTAGC3 ' (SEQ ID NO:19) and 5 ' CACTGATTACTAATTCAGGATC3 ' (SEQ ID NO:20)) of 0.1 μ M in different reactions.Reaction is carried out second step 94 ℃ of degeneration 30 then at first 94 ℃ of degeneration 5 ' ", 50 ℃ of annealing 30 " and 72 ℃ of extensions 30 ".Second step was repeated 34 circulations, and extend step at last at 72 ℃ 5 '.Restricted enzyme BamHI with 2.5U spends the night 37 ℃ of digestion with the amplified production of 199bp.

The product that is digested is run 2h in 150V on 10% PAGE, and use ethidium bromide staining.Determine genotype based on two segmental restriction enzyme site A-C that produce 181bp and 18bp.

The ADRA2C dinucleotide repeats polymorphism.The ADRA2C dinucleotide repeats polymorphism on chromosome 4p, crosses over the 179-193 base pair.The people's gene group data base registration number of this gene is M94915.The inventor has detected this hereditism's sign and has found that (≤181bp) existence is with more serious drug dependence pattern (cocaine, amphetamine and heroin use low base pair allele), but less relevant with severe alcoholism in a series of 53 non-Spain white race personage matter misusers.The inventor has made up the homozygosity that dinucleotide repeats allelic genotype: genotype 1=≤181bp in the following manner; Genotype 2=heterozygosity (allele 1≤181bp, the homozygosity of allele 2 〉=183bp) and genotype 3=〉=183bp.Genotype 1 uses year number (p＜0.05) relevant with spending in the high money amount (p＜0.01) on the medicine, high amphetamine use year number (p＜0.05) and high heroin.On the contrary, high base equity position gene (genotype 3) uses many (p＜0.01), high ethanol to use year number (p＜0.05) with ethanol in the past 30 days and alcoholism (rather than medicine) is appointed as to seek the first cause (p＜0.05) for the treatment of relevant.

The people's gene group registration number of this polymorphism is GDB:196352 and sequence is M94915.From whole blood, extract experimenter's genomic DNA by the method for standard.Use PCR ^TMMethod (Saiki etc., 1988) the amplification target DNA utilizes each fluorescently-labeled primer (5 ' AGTGGGCAGGGCGGGGCAGGT3 ' (SEQ ID NO:21) and 5 ' CGCTGCCTCCCTTCCACCTGTTG3 ' (SEQ ID NO:22)) of 0.1 μ M in different reactions.Reaction is carried out second step 94 ℃ of degeneration 30 then at first 94 ℃ of degeneration 5 ' ", 57 ℃ of annealing 30 " and 72 ℃ of extensions 30 ".In a circulation, second step was repeated 29 times, and program finishes 72 ℃ 5 ' extension step with one.For gel analysis, each reaction is by the amplification PCR of 0.5 μ l dilution ^TMProduct (15 μ l are in 75 μ l deionized waters), 2.5 μ l contain the mixture composition of the blue color doxtran of the ROX500 standard+0.25 μ l dyestuff of 2.0 μ l deionized formamides+0.25 μ l, and 92 ℃ of degeneration 2 '.The sample of degeneration is splined on AppliedBiosystems 373 DAN sequenator (GeneScan ^TM) 6%PAGE in, and under 1500 volts and 300W, will run glue 5h.Treatment gel, and utilize internal standard (ROX500) analysis.Discern two peaks based on color and segmental size by gene type instrument (genotyper) (1.1 version).The value of being distributed is the mark that its allele size accuracy of performance is judged.Go out the complete information of each specimen from each gel file printout, and edited data is analyzed.

The inventor has detected the multiple polymorphism of dinucleotide in a series of 64 substance abuse persons, 3 ' terminal CA of PENK gene repeats.The inventor finds that the experimenter that high base equity position gene (〉=80 base pairs) isozygotys has significantly more serious alcoholism and drug dependence.High base equity position gene uses the high frequency of a year number (p＜0.05), high Drug therapy number of times (p＜0.05) and methadone maintenenace to use (p＜0.05) relevant with high drunk year number (p＜0.05), high medicine drug addiction treatment number of times (p＜0.01), high heroin.

Embodiment 12

Determine using PHENCAL ^TMThe polymorphic allele that treatment is responsive

Go out pharmaceutical intervention is suppressed the most responsive individuality of specific RDS behavior for determining which kind of RDS gene, particularly which kind of polymorphism are diagnosable, to detect specific polymorphism in the RDS gene to potential candidate, carry out various psychometrys and identify the RDS behavior, and/or the compositions of utilizing in this to be announced is carried out pharmacological treatment.Can the special RDS gene of statistical evaluation or the existence of gene pleiomorphism with the existence of the RDS behavior of identifying by psychometry and/or with the dependency that utilizes the change of treatment in the RDS behavior of the compositions of announcing in this.The RDS gene pleiomorphism that showing the RDS behavior that is suppressed by neurotrophic compositions together has remarkable (for example p＜0.05) dependency of statistics is identified as diagnosis more to utilize neurotrophic compositions successfully to treat individual sign.In table 4, provided the overall prescription of RDS therapeutic combination composition, and considered that listed one or more compositions will be effective in the people of the one or more RDS behaviors described in treatment shows this.In addition, table 6-16 contains preferred special formulation in the treatment of the specific RDS behavior described in this.And the simplified diagram that how to influence by analeptic (cocaine etc.), opiate and recompense that calmness-somnifacient is induced for specific composition sees Table 17-19.Below being described in for the instantiation of the Therapeutic Method of the associating genetic diagnosis of RDS behavior:

Carbohydrate gluttony or obesity.About hereditism's contact for the treatment of with carbohydrate gluttony and success, the inventor is just measuring many proband's genotype at present, and provides experimenter's every day 6 PHENCAL ^TMThe capsule of prescription, and be determined at the poundage that stage of 90 days loses, carbohydrate gluttony scoring, and the increase again of after ensuing 6-8 month, determining body weight.This will contrast placebo group and carry out.To determine by research whether maximum improvement takes place under the range gene type.For the research of obesity, the inventor will determine following gene and may other and their polymorphisms separately, already provided in this with using PHENCAL ^TMRelevant having: D1, D2, D4, DAT1, D β H, COMT, MAOA (x), TDO2, APO-D, chromosome-2 sign, UCP-2, CNR1, GABAB3, HTR2A, HTR1A and nNOSla (seeing Table 5) are treated in success.Concrete operations: to PHENCAL ^TMThe clinical response different with the variant that contains the nicotinic acid chromic salts according to genotype

The research participant.The body fat that participant in this research will have 34% body fat and have 28% for the male between the 130-180% of ideal body weight or for the women for morbid obesity person and body weight.The participant will be at 18-65 between the age in year.If having hypertension, diabetes or other weak property disease, the patient will be excluded.

Research design.This clinical experiment will be an experimenter's a double blinding placebo-controlled study, compare them with the placebo that matches (by the Weider nutrition group, salt lake city, Utah manufacturing) and be given 6 PHENCAL every day ^TMCapsule (see Table 4 for overall prescription, and special formulation is seen Table 7).This will be a 14wk research.The last fortnight comprises deep work-up, comprises that screening of blood and buccal swab DNA collect and the platform scale body weight detects.In case the experimenter enters this research and has signed the written agreement of a standard, then will utilize Collins Helium dilution pulmonary functional unit (Warren E.Collins company by local lung laboratory independently, Braintree Mass.) measures remaining lung volume.Test (alternative method) under water with utilizing the Whitmore volumetric type meter (Whitmore Eenterprises, San Texas) of weighing height correlation and having a testing reliability again (Ward etc., 1978) of 0.96 to 0.99 with hydraulic pressure.A kind ofly calibrate to 1/10 pound the commercial platform platform scale of Deteco (Web City Missouri) obtains the platform scale body weight for 8850 types, Deteco Scale company using.In addition, the inventor will utilize dual energy X line absorptionmetry (DE﹠amp; A) survey bone density.

During finishing 2 weeks of all these tests, will offer the experimenter, and select the PHENCAL that contains that encodes in advance at random except the test result of their genotype (they will maintain secrecy until prepare research is analyzed) ^TMOr in the bottle of its placebo that matches one.The experimenter will be required to take every day two of about 1 hour clothes of 6 capsules-before breakfast, lunch and dinner are had a meal.Maker will be as the manager, and will encode every batch before distribution.Among the investigative technique person of researcher, dispensing product or the experimenter nobody is known which numbers corresponding PHENCAL ^TMOr the placebo that matches.With other some researchs particularly single with or those differences of finishing of associating d-Fenfluramine or phentermine, diet or exercise information and experimenter are not provided as long as they strictly according to prescribe and take supplement and then will be required to carry out any activity that they wish at test period.(in the experimenter of a specific quantity, the inventor may add the Kaats by Dr.Gilbert, San Antonio, " the suitableeest health diet plan " of Texas development).The experimenter will select another bottle supplement or placebo weekly to the center, and will measure the platform scale body weight.When the summary of test phase (from taking the extra 12wk that capsule begins), the experimenter will finish a last health and form test.All data are carried out Computer Processing and will analyze by the computerization research system at the University of Texas health science center of Texas Houston-San-Antonio.When analyzing, the manager will be required publication number.Will be by the product operating position PHENCAL that calculating consumes of themselves report ^TMActual quantity with placebo.

In this research, it will be to accept placebo or PHENCAL that main results is weighed ^TMThe experimenter between different health form the average comparison that changes.To estimate PHENCAL ^TMAnd Without Chrome Salt and contain the PHENCAL that pyridine carboxylic acid chromic salts, nicotinic acid chromic salts or both have ^TMTo in the group that divides according to genotype, form variation by average health.As described above, as the method for being discussed earlier, analyze more intergenic haplotypes of all detections with the regression analysis of standard polygenes and (see OBKIT about the gene type details by the Comings utilization ^TM).

Anti-smoking.Utilize Nicorest ^TMPrescription is measured genotype according to inventor's basic plan to many heavy smokers.At 6 Nicorest every day ^TMBehind the therapeutic scheme of capsule to 90 day, will be according to comfortable until smoking cessation; Withdrawal symptom (comprising depression); Recurrence comes weighing result.Corresponding gene above all is estimated this data.According to the work before (1996) such as Comings, will adopt complete smoking history (to see NICOKIT about the gene type details ^TM).

Concrete grammar: Alcotrol ^TMAnd Cocotrol ^TMAs genotypic function.Research will comprise will be given the outpatient of anti-ethanol and anti-cocaine prescription, and will be according to early stage detoxifcation withdrawal symptom with by not getting back in the therapeutic scheme or knownly getting back to the relapse rate that medicament selection measures and come weighing result (seeing Table 5).

The experimenter.This research in, will utilize Minnesota Multiphasic Personality Inventory (Minnesata Multiphasic Personality Inventory) (MMPI) and the hemanalysis (CBC/SMACO24) that carries out through agreement all patients are estimated.Further blood testing is not taked in end in research.Be grouped into: ethanol-Alcotrol ^TMEthanol-placebo; Analeptic misuser's (cocaine)-Cocotrol ^TMAnaleptic misuser-placebo.

Research design.For the dependency evaluation, will and enter BAL as possible covariant test age, body weight, sex, race.In inpatient treatment place, doctor, nurse and experimenter and collection of data person all which experimenter nobody is provided about and accepts Alcotrol ^TMOr Cocotrol ^TMOr the information of the placebo that matches.Dosage is each prescription two capsule every day (seeing Table 8 and 9), and the branch agent gave one hour ante cibum.Test event

Skin conductivity level (SCL).The electrical characteristics of skin have been widely used in the evaluation of emotional response.This technology has confirmed that a kind of detection as stress level among the patient (for example, anxiety or angry degree) is very reliable.So, this is an indirect detection of stress level among the patient.SCL, the inverse of dermal resistance impedance (GSR), the absolute skin conductivity level (Edelberg, 1972) that monitoring is measured by microhm.There is dependency between the anxiety reaction that causes skin electric conductivity to increase in the location with by sympathetic activation.Therefore, the reduction of electric conductance is with the reduction relevant (Luthe, 1969) of oneself's awakening.Detect for carrying out these, Autogen 3000 is connected on three fingers in centre of dominant hand of each patient, and obtains a reading.Detect so that the patient not can be appreciated that when he is with detected in the unscheduled time for each selected patient who studies.

Clinical detection.Health scoring (giving up evaluation [CIWA-A] unanimity with the clinical research of ethanol) and BESS scoring are to be used for each patient's subjectivity to detect during staying inpatient therapeutic community in whole these 28 days.(Blum etc., 1989) were all described in health scoring and BESS scoring.In therapeutic process, will observe the change of these factors every day by clinical guidance person, doctor and clinical care director, and their observation will be discussed and coordinate to reaching an agreement.Each patient all is assigned with one 5 baseline scores value when entering.To in the improvement scope of 6-10 and in the scope that falls back of 0-4, indicate one every day and improve or fall back, and 5 refer to getting the hang of no change.

Gene type.As SUDKIT in the application ^TMDescribed in, get permission to enter each clinical patient and will be given the detection of DNA-buccal swab.All genotype will be maintained secrecy to all personnel who comprises medical treatment guidance person.When preparing analysis of data, genotype is only offered the computational resource system of University of Texas of Texas-Houston/San-Antonio for statistical purpose.Gene to be measured comprises: D1, D2, D4, DAT1 (10/10,9/9), TDO2, nNOS1 α, CNR1, GABAB3, HT2R, MAOA (X), COMT.

The statistical analysis of data.Will be with placebo to comparing independent ethanol and analeptic misuser group and associating group analysis Alcotrol ^TMAnd Cocotrol ^TMThe effect of treatment.The most important thing is, the inventor will estimate to stress therapeutic response, Clinical detection comprises that health scoring and Bess mark, as genotypic function.Concrete operations: HyperGen in the ADHD proband ^TMAs genotypic function

The research participant.The inventor utilizes 6 HyperGen ADHD child every day of ADHD scale, TOVA test and the diagnosis of CCPT outpatient clinic in the research of a double blinding placebo ^TMThe capsule of prescription.Testing evaluation that the inventor is relatively more preceding then and the back is tested and result are with the dependency of the range gene of being studied.This research is included in the ADHD experimenter in the outpatient clinic of two places (Texas and Tennessee).By a large amount of psychological tests experimenter that comprised TOVA and ADHD scale (Cull and Blum, 1997) and Wisconsin card sorting testing and diagnosing.The participant has the negative history of the psychosis situation eliminating of AxisII diagnosis.All experimenters all do not write a prescription when test, and each experimenter all is required to sign a written agreement and agreement is taken capsule according to method.

Research design.The experimenter carries out twice complete test (comprising TOVA, Cull/BlumADHD scale and performance task (face as follows)), and it forms the test model of a test-again.Repeat once (back test) after 90 days then testing (Pretesting) on the 0th day at first.Between twice, the experimenter is required to take HyperGen with 6 capsular dosage every day (respectively obeying 21 hour about ante cibum in breakfast, lunch and dinner) ^TMPerhaps a kind of placebo that matches.HyperGen ^TMComponent be shown in Table 12.This is a double blinding placebo-controlled study, and manufacturer holds information encoded in advance, and it offers the statistician when research finishes.Nobody knows the information about the bottle of coding among staff or the experimenter.

The performance task.Use two performance models to bring out electrophysiologic response:

Space orientation.This is a response time task (SOT), (Posner etc., 1988) wherein cause the prompting be presented in the left side or the visual field, right side.Relatively when the response time that provides or do not provide when causing prompting.Comparison during by reaction between part, it makes can estimate the individual ability that smoothly distracts attention between the visual field.In this research, task is to have structurizedly, so it allow to estimate the more basic stage of attention process, its attention operation with deep layer more is relevant, and trends towards load heavier (Defrance etc., 1993) on the more automatic stage of noting.

The emergency sustained performance.Emergency sustained performance task (CCPT) is the variant of a classical theme, and it incorporates the factor of selectivity and persistence attention into.Analyze this task and represent the more controlled stages of attention process with the quality of guaranteeing to show.A letter once appears at the center of screen.Basically, require his dominant hand of body and function a specific lexicographic order to be reacted by the left button of clicking the mouse (right hand crowd): for example, if catch up with last one " T " by another " T " immediately.First " T " in a pair of " T " is as a warning prompt, and second is that " T " is target.All other letters are considered to chaff interference, and it should be ignored by the experimenter.The basic calculating of data is according to former work (Defrance etc., 1997).It should be noted that prefrontal cortex seemingly participates in sustained attention power and makes great efforts the heavyliest, it be an important factor and to the unusual sensitivity (Sostek etc., 1980) of stimulant substance about the total performance on this task.

The record scheme.(Defrance etc., 1996) as described, reference electrode is connected ear-lobe (A1-A2), effectively writes down site record EEG by 28.Each stimulate to occur begins to cause 800msec.Therefrom constitute the EEG sampling of ERP: sample comprise the stimulation of 100msec in each signal time before, it is used for baseline calibration.

Analysis.Fundamental analysis is with former work (Defrance etc., 1997) and comprise the evaluation of three parameters substantially, and wherein each may be according to the efficient of individual attention process and difference.These parameters are: the incubation period in the ERP composition, amplitude, symmetry (spatial distribution) (Defrance etc., 1996).

This work may be used in HyperGen ^TMThe attention process change with having shown (as previously discussed and the HYPERKIT described in this application with the relevant allele of ADHD ^TMIn indicated) association.At least the expection experimenter that carries the polymorphism of the VNTR10/10 repetition of DRD2A1 allele, D β H B1 allele, DAT1 and TDO2, MAOA (X) and HTR1A gene will show the strongest reaction.

Uprising power and attack.Owing to have been found that, so estimate the use of attacking prescription with being logical as these behaviors described in table 14 and 15 at the dependency between " pathologic violence " and dopaminergic polymorphism (being DRD2 and DAT1 allele) and with the dependency between division sample and elusive behavior.Dosage will be 6 capsules every day, and the standard placebo compares.Result's measurement will be the reduction of non-motivation outburst number of times in various proband (being teenager and convict) and the cardinal principle calmness that hostility reduces.With end value in 6 months time frame with top described genotype (seeing TEMPKIT) association.In this research, the inventor will utilize with table 14 or 15 and fill a prescription accordingly.Finish among the youth of 40 imprisonment that this research will be in the youth center of Larned Kansas.

Embodiment 13 is used for a kind of multiple the adding up property corresponding technology (MAA) of polygenes obstacle gene identification: right

In ADHD with the result of the obstacle of falling ill altogether

ADHD is a kind of modal obstacle of being treated by most of pediatric psychology man and psychiatrist, is present in percent 2 according to estimates in 8 school age population.Having clearly, evidence shows that ADHD is a kind of organ, nerve and/or hereditism's obstacle.Compare with contrast, the father of ADHD or HAC has the frequency of higher ADHD, alcoholism and antisocial personality disorder (ASPD), and mother has higher ADHD, alcoholism and " hysteria " or hysterical personality's frequency (Morrison and Stewart, 1971; Cantwell, 1972).ADHD child's compatriot has frequency (Augest and Stewart, 1972 of higher ADHD; Pauls etc., 1983; Weiner etc., 1977), if particularly among the father and mother one have ADHD or if the child is identical twins (Hechman, 1994).At the ratio of these behaviors between compatriot and half sib in from 2.0 to 5.2 scope, with hereditism's etiology unanimity of ADHD.Compare with normal children, child with ASP has remarkable increase (Cadoret etc., 1978) among the father and mother on ADHD, the angry and frequency of flying into a rage.

Compare with contrast relatives' 8%, the more relatives of ADHD proband have ADHD.For severe depression obstacle, oppositional defiant disorder (ODD), conduct disorder, antisocial personality disorder, alcohol dependence, medicine or alcohol dependence, comprise that the anxiety disorder of generalized anxiety disorder also has the risk (Biederman etc., 1990) that significantly increases.ADHD may have the identical susceptibility (Biederman etc. of family with the dysthymic disorder, 1990), ADHD with conduct disorder may be the different hypotype (Farone etc. of of ADHD, 1993a), and ADHD and anxiety disorder (Hodge etc., 1986) and ADHD and learning disorder (LD) (Farone etc., 1993b) independent delivery in family.

Observed the etiology that the metabolic defective of dopamine participates in ADHD for a long time.That the damage of the dopaminergic neuron of limbic system (ventral tegmental area) causes in Rodents is many moving, overresponse, to stress Low Response and a series of other obstacle (Lemoal etc., 1976; Lemoal etc., 1991).The chemical depletion of back frontal lobe dopaminergic neuron of just being born produces animal model (Shaywitz etc., 1976 of a kind of ADHD to the stimulant reaction; Shaywitz etc., 1991).In child, detect low-level HVA (Cohen etc., 1979k with Tourette obstacle (TD); Butler etc., 1979k).In the child of ADHD, reported low CSF HVA (Shaywitz etc., 1979k), yet, also reported the positive correlation between the scoring of the how moving and conduct disorder of CSF HVA and ADHD.The brain video studies show that defectiveness in the striatum that is rich in dopamine in ADHD (Comings etc., 1991k; Noble etc., 1994k), and in ADHD and TD the hypofunction (Zametkin etc., 1990) of frontal lobe.Research has shown the hyperkinetic syndrome in the knock out mice that lacks dopamine transporter or DRD3 gene.The dopaminergic agonist has been successfully used in the treatment of ADHD.

Total hypothesis of inventor is that ADHD is a kind of owing to influence the polygenes obstacle of additive effect of the gene of dopamine, norepinephrine, 5-hydroxy tryptamine, GABA and other neurotransmitter.Some related special genes are dopamine gene (DRD1, DRD2, DRD3 and DRD4 acceptor gene, dopamine and dopamine transporter); Norepinephrine (NE) and epinephrine (EPI) gene (ADRA2A and ADRA2C receptor, PNMT, norepinephrine transporter, MAOA, COMT); 5-hydroxy tryptamine gene (TDO2, serotonin transporter, 5-hydroxytryptamine receptor gene); GABA gene (GABA acceptor gene GABRB3 and other) and also unidentified other gene.

The inventor has detected three dopaminergic gene DRD2, DBH and DAT (Comings etc., 1997b), three adrenergic gene DBH, ADR2C and ADR2C (Comings etc., 1998a), and androgen receptor gene AR, add DRD2 and serotonin transporter gene, (Comings etc. are 1998b) for the adding up property effect of ADHD and other obstacle of falling ill altogether for HTT.These studies have shown that two principles: the first, when the experimenter being divided into the group that the variant existence is arranged in 0,1,2 or 3 that represents these genes, on the size of QTV, there is a gradual increase, and its dependency of comparing any individual gene is more remarkable.The second, these studies show that this technology can be used for identifying the gene that plays a role by increase QTV scoring in phenotype, and QTV marks and bring into play gene minimum or insignificant effect in phenotype because their reduce.

The inventor has enlarged these principles and has detected 29 heterogeneic adding up property effects of total in ADHD.It is multiple adding up property association (MAA) technology that the inventor has named this.The inventor has checked several hypothesis about the MAA technology.Because it is very big that different QTV may change in scope of marking and size, in order to carry out the comparison of a plurality of gene size effects on different Q TV, the inventor has utilized the percentage comparison result of the variance r2 that returns dependency r and explained to analyze and draw.Be below some hypothesis and introduce these research in method.

The MAA technology that is used for ADHD and other behavioral study is based on this hypothesis, it thinks that most of psychiatric disorders are the allelic result of many variations (Blum etc., 1990 that influence the gene of dopamine, 5-hydroxy tryptamine, norepinephrine, GABA, NMDA and other neurotransmitteies from biparental inheritance; Comings etc., 1996a; Comings etc., 1998a).The inventor selects the candidate gene that detects to be based on this hypothesis in this report.Selected with the irrelevant gene of neurotransmitter based on show the research that they play a role in the past in the behavior phenotype.A principle of being added for screening is with being the relevant useful polymorphism of available candidate gene.

The second, twins study shows most of polygenes obstacles, comprises ADHD and behavior disorder, and percent 50 is hereditary (Sherman etc., 1997 to 90; Slutske etc., 1997).Because each gene only is responsible for percent 1 to 2 variation, must relate at least 20 to 40 genes so simple mathematical shows.Quantity can be littler in a given individuality.The 3rd, for avoiding annular factorial, show that independently having effectiveness for the scoring of each gene genotype in the group by experimenter's that inventor or other people studied be important.The gene that the inventor detected is listed in the table 65." confirmation " row refer to that those are at one of the experimenter used confirmed gene of scoring in the group independently.The 4th, with a value in 0,1 or 2 each gene is marked.For example, if relate to a diallele polymorphism and independently studies show that 1 allelic dominance, then scoring is 11 or 12=2 and 22=0.If independently studies show that genotype is codominance, then scoring is 11=2,12=1 and 22=0.Similar three scoring technologies are used for the repetition polymorphism.This is convenient to the test and feasible can the scoring to each gene with one 0 to 2 similar yardstick of the additive effect of each gene.The 5th, because the polygenes obstacle is common, itself must be very common so the fact that many genes are related to shows related hereditism's variant.Because this, the inventor wishes to utilize polymorphism and genotype to hive off and makes that scoring is 1 or 2 experimenter's percentage ratio maximization.These the results are shown in %1 in the table 65, under 2 row.Attention is in most of cases, and the summation of 1+2 percentage ratio is greater than percent 25, and usually greater than percent 50.

Table 65

The gene of MAA technology for detection

Gene pleiomorphism gene scoring %1,2 confirm

012 dopamine gene 1.DRD1 RFLP DdeI 12,22 11-, the 12nd, (Comings etc., 1997b)

(Cichon etc., 1994) 2.DRD2 RFLP TaqI A (Grandy etc., 11,22 12-, the 36th, (Blum etc., 1995; Comings and

1989) MacMurray, 1998) 3.DRD3 RFLP McsI (Lannfelt etc., 1992) 12 11,12-, the 47th, (Crocq etc., 1992; Comings

Deng., 1993; Asherson etc., 1996) 4.DRD4 SS repetition (Lichter etc., 1993) other any 6-8-, the 52nd, (Comings etc., 1997a) 5.DRD5 DN repetition (Ravindranathan etc. ²151/ ²151 ³151/ ³The 151 25, the 61st, (not delivering)

1994) 6.DAT1 SS repetition (Vandenbergh etc., 1992) x/x the 10,/10 42, the 51st, (Cook etc., 1995)

Be (Blum etc., 1997) 5-hydroxy tryptamine gene 7.HTT promoter insertion/disappearance (Collier SS LL SL 27, the 47th, (Kauck etc., 1997; Lesch etc., 1996)

Deng., 1996) 8.HTRIA MR (Bolos etc., 1993) 127bp other-, the 24th, (not delivering) 9.HTR1Db RFLP HincII 12,22 11-, the 59th, (Lappalainen etc., 1995)

Gene pleiomorphism gene scoring %1,2 confirm

01 210.HTR2A RFLP Mspl (Arranz etc., 1995; 11,22 12-, the 51st, (not delivering)

Williams etc.; 1996) 11.HTR2C RELP HinfI (Lappalainen etc., other M1, F11-, the 79th, (not delivering)

1994) X12.TDO2 RFLP BslI (Comings, 1995) GG GA, AA-, the 5th, (Comings etc., 1996) noradrenaline plain gene 13.DBH RFLP TaqI (Wu etc., 1997) 22 11,12-, the 79th, (Wei etc., 1997) (do not deliver) 14.ADRA2A RFLP MspI (Lario etc., 1997) 11 12 22 38, the 7th, and (not delivering) 15.ADRA2C DN (Riess etc., 1992) heterozygosis homo ^*-, the 27th, (not delivering) 16.NT RFLP MnlI ^{* *}(St ber etc., 22 11 12 45,43

1995) catecholamine metabolism gene 17 .MAOA SS repeat X (Hinds etc., 1992) other ³320 ^{* * *}-, the 65th, (Gade etc., 1998) 18.COMT RFLP NlaIII (Lachman etc., 22 12 11 51, the 74th, (Lachman etc.,, 1996) (not delivering)

1996) GAGA gene 19.GABRAA3 DN (Hicks etc., 1992) X other ³168#-, the 67th, (not delivering) gene pleiomorphism gene scoring %1,2 confirm

01 220.GABRAB3 DN (Mutriangura etc., 1992) het＜188/＜188 39, the 14th, (Gade etc., 1996) (not delivering)

³188/ ³188 Fructus Cannabis ester acceptor gene 21.CNR1 SS repeat (Dawson, 1995) ²5/ ²5 ²5/ other other 43,10th, (Comings etc., 1997c; Johnson

Deng., 1997) nicotine cholinergic 22.CHRNA4 SS repeats (Weiland and x/x x/9 the 9/9 32,7th, (not delivering)

Steinlein, 1996) nmda receptor gene 23.NMDAR1 RFLP HpalI ^*(Rupp etc., 1997) the 11 22 12 10, the 44th, (not delivering) enkephalin gene 24.PENK SS repetition (Konig etc., 1996) other ²178/ ²178-, the 64th, (Comings etc., 1997d) androgen receptor gene 25.AR GGC (Sleddens etc., 1993; Other ²16###-, the 58th, (Coetzee and Ross, 1994; Lrvine

Sleddens etc., 1992) etc., 1995) the gamma interferon gene

Gene pleiomorphism gene scoring %1,2 confirm

01 226.INFG RFLP NlaIII (Wu and Comings, the 11 12 22 50, the 27th, (Comings etc., 1998c)

1996) 27.CD8A SS repeat (Polymeropoulos etc., 142/142 other-, the 73rd, (not delivering)

1991) senilism element-128.PS-1 RFLP BamHI (Scott etc., 1996) 12 11,22-, the 54th, (not delivering) CRF gene 29.CRF RFLP XmnI (GDB) the 11 12 22 8, the 2nd, the explanation of (not delivering) table 65

X=X-is chain

* homo 154=＜154bp homozygosity and ³The 154bp homozygosity; The hetero=heterozygote

* homo 183=＜183bp homozygosity and ³The 183bp homozygosity; The hetoro=heterozygote

***A1970G

* * * 2=male ³320bp.The women is any ³320

Among the #2=male ³168, among the women ³168/ ³168,0=other

##2=a-c/a-c or d-g/d-g homozygote, the 0=heterozygote

###2=male ²16 repeat and the women ²16/ ²16,0=other

The MR-mononucleotide repeats;

The DN dinucleotide repeats;

SS is short, and sequence repeats;

The single base pair polymorphism (restriction fragment length polymorphism) of RFLP-

Do not deliver-from the inventor breadboard one independently in the subject group this gene and polymorphism to the observation of not delivering of the effect of one or more other phenotypes.

The GDB-genome database

The 6th, if hereditism's variant related in the polygenes obstacle is common, they must be fundamentally with those differences that cause the single-gene obstacle, it is the change that they must only cause the gene expression moderate, because if the change that they cause gene expression severe then they may cause the single-gene obstacle (Comings, 1998b).Observe based on these, the inventor has shown the length variable by formation Z-DNA, and this important effect of performance in polygenic inheritance of common dinucleotide repetition polymorphism (Comings, 1998b).These repeat is a large amount of, and allele is general, and formed Z-DNA is to the effect of an appropriateness of gene expression performance.Significant phenotypic effect can be only comes together to produce by being added in the effect of the variant at many difference in functionality related genes place.For this study its influence for inventor when exist especially to utilizing short placed in-line repetition polymorphism interested, and any single base pair polymorphism is possible same or the multiple allele of connecting of the weak point of a plurality of and this gene-correlation is linkage disequilibrium.Because the latter, the inventor supposes any single nucleotide polymorphism, even it is irrelevant with exon or promoter, may provide valuable information for the MAA technology.The 7th, for detecting the additive effect of two or more genes influence a given neurotransmitter or function group, a fictitious variable that is called polygenes (PG) is set up in the simple addition of marking based on each individual gene.Detection can be identified the subgroup that a certain amount of scoring is had the gene of additive effect to the effect of variance percentage is feasible various associating of gene in a given functional group.Then this group gene is used for forming a trans polygenes scoring (TPG) that between the difference in functionality group, detects additive effect of gene.PG or TPG scoring are high more, and the quantity of the phenotype correlation gene variant that an individuality has is many more.

The 8th, as (Comings etc., 1996b in the former research; Comings etc., 1998a; Comings etc., 1998b), linear regression analysis is used for detecting the gene scoring of additivity.This makes can calculate r ²Or detected gene contributes to the percentage ratio of the variance of character, as long as F estimates the size of effect, and p is the significance of effect.Gene is only explained variance true of character about percent 1 and is not meant that this effect is unessential.According to the quantity of the factor, it can provide predictability (Rosenthal and Rubin, 1982 up to percent 10 (r) of a phenotypic effect; Ozer, 1985).The 9th, because the various actions scoring has different magnitude range, so with cumulative r ²Value but not behavior scoring is used to provide the accurate comparison of the effect of 29 genes between different phenotypes.

The tenth, the attention deficit that the inventor has selected at first to detect based on DSM-IV (diagnosis of APA,American Psychiatric Association and statistics handbook IV, 1994) standard moves (ADHD) scoring more.The inventor has utilized by the average that relatively has ADHD scoring in the genotypic individuality of distinct group and has utilized the method for gamut scoring, rather than emphasizes a diagnosis of two minutes.The 11, there are two potential methods to be used for regression analysis: single argument and multivariate.For univariate analysis, the scoring phase Calais of each gene is produced an one scoring, and determine the dependency between this scoring and QTV.This is the expansion of 0,1,2,3 methods of marking.The advantage of this method is to have only that the r value just increases when one or more gene pairs QTV have additive effect.When adding had the gene of subtracting property effect, r reduced.This is to final r and r ²The determining of value guarded.Yet, because all gene scorings are merged into an one variable, so no matter the quantity degree of freedom of institute's gene mentation is generally 1.This is non-conservative about the p value.

For second method, in according to the multivariate regression match, estimate each individual gene scoring also with the dependency of analyzing QTV.Because along with the adding degree of freedom of each gene increases, so it is guarded for the p value.Yet, because additivity and subtracting property gene all have contribution to r value, thus though this method for gene on its effect be additivity or subtracting property and all r and the r of increase ²Value is non-conservative.

The inventor since two former thereby selected univariate method.The first, because r and p value all reduce when adding the gene that QTV not have to contribute, so it is more conservative.The second, because gene is not considered the quantity of the gene that detects by a single degree of freedom representative, so it has the effect that detects a hundreds of gene simultaneously and the potentiality of not losing a power.

The 12, have the hypothesis of identical gene for detecting most of pathogenic obstacles with ADHD, the inventor has also detected with 29 genes of a group to the additive effect in the quantitative scoring of oppositional defiant disorder (ODD), behavior disorder (CD), tic, learning disorder (LD) and other QTV.For greater than 14 years old experimenter, the inventor has also detected the QTV to alcohol abuse/dependence, drug dependence/dependence and smoking.The 13, the detection of these pathogenic obstacles also makes the inventor can detect this hypothesis (Comings etc., 1996b; Comings etc. 1996a), think except shared gene, and different pathogenic obstacles utilizes some genes and to the combination of the unique gene of particular phenotype.The 14, for exploring for the dependency between an individual gene and a given phenotype, whether that gene has on the additive effect and has in all senses the significance level of p＜0.05, and the inventor has detected about whether they have the r of 145 gene-phenotypic correlations of an additivity or subtracting property effect in the phenotype scoring ²And the relation between the p value.At last, an initial operation that utilizes all candidate genes identify those have additive effect after, each additive gene is added in the new TPG scoring gradually, and analyzes dependency with the QTV in the problem.This provides and can be accredited as the overall r that the contributive gene of QTV in the problem is obtained by only utilizing those ²An estimation.

The advantage of an additivity scoring.The most unique aspect of many polygenic inheritances is excavated out in the utilization of one additive gene scoring---and the effect of heterogeneic additive effect, heterogeneic subtracting property effect, hybrid vigor and epistasis and when many different genes may be contributed same phenotype in any given individual or individual group only has a subgroup of those genes may have this fact.The most important thing is that this method can compensate the existence of distinct group gene in Different Individual.Therefore, many next correlational studyes are not that whole follow-up studies repeats by some.This is because such fact, and promptly this gene only has a group of contribution and gene may participate in a subject group to the variance of a little percentage ratio and the different subject group of another group participation one.An equal effectively conclusion is that different gene groups can produce identical phenotype in different groups of individuals, and be not one of hint or other result be incorrect.Because the total group's of related gene effect is weighed in the additivity scoring, so the MAA technology may be more repeatably between different subject group.In order to provide a specific example, though DRD2 (TaqIA1 allele) (Comings etc., 1996b; Blum etc., 1998), DRD4 (48bp7 repetition) (Lahost etc., 1995), DBH (TaqI B1 allele) (Comings etc., 1996b) and variant (Comings etc., the 1996b of DAT1 (10 are repeated allele) gene; Cook etc., 1995; Gill etc., 1997; Waldmaqn etc., 1996) all participate in the etiology of ADHD, but each may be in some subject group but not other in have a significant effect.Yet, if for the physiological effect of each gene is that similar (change in the dopamine metabolism) should prove consistent and be significantly higher than contrast in all ADHD experimenters' group for all additivitys of four scorings so, show that it is hereditism's defective general in the dopamine metabolism rather than any individual gene important on the ADHD etiology.Adopt identical principle for the additive gene between different neurotransmitter groups.A final dynamics of additivity scoring is the evaluation that a maximum information group can be optimized for a given phenotype by the various combination that utilizes gene.With the smallest effect difference of individual gene, such group who optimized can provide important prediction and diagnostic value.

Suppose.First is assumed to be, because the polygenes obstacle relates to the additive effect of a plurality of genes, so the MAA technology will be in the evaluation of related gene than bigger strength is provided in the detection of next gene, and have only the adding of those genes just can make r with an additive effect ²Maximize with the p value.For checking this, the inventor has detected the effect of 29 genes in 336 white people's individualities that the contrast of having been weighed ADHD seriousness by 274 Tourette syndromes and 62 is formed.Second is assumed to be, if detect two independently samples, the inventor infers because different samples may utilize different gene groups, so many and be not whole in a sample, may in test sample book again, be additivity for the gene of additivity, many and be not whole may be in subtracting property in the test sample book again in a sample for the gene of subtracting property, some genes in a sample may be additivity and in test sample book again subtracting property, and for the additive effect of two a plurality of genes of sample may be significantly greater than the effect of any individual gene.Inventor r when also detection of desired has a less N ²Fluctuation may be bigger.For checking this, 336 experimenters are divided into two independently groups of 168 experimenters at random, in each group, have identical TS and contrast experimenter's quantity and identical masculinity and femininity quantity.Dependency between PG that has detected at ADHD for each group and TPG scoring.The 3rd is assumed to be, and when detecting different phenotypes, the inventor supposes that pathogenic obstacle should more shared ADHD genes, and the combination of some genes and gene hole is unique for pathogenic obstacle.For checking this, the inventor has detected 29 gene pairs different Q TV---the additivity of oppositional defiant disorder, behavior disorder, tic, study and other obstacles and subtracting property effect.

Test subject.Seminar is made up of 336 irrelevant, non-Spain white race people experimenters.In these, 274 diagnosis with a Tourette syndrome (TS), and 62 for the contrast.TS experimenter comes leisure to wish the Tourette syndrome clinic of city medical centre.All all meet the DSM-IV standard of TS, and all are met with by the inventor individual.Though these mainly are identical experimenter (Comings, the 1995a that detects in the former research; Comings, 1990), but when experimenter's used in the research formerly DNA specimen is eliminated, added some new TS proband.The inventor is divided into those with inventor's TS experimenter in the past and has slightly (1 grade, chronic tic is too light and need not treat), (2 grades of moderates, enough serious and need treatment) and (3 grades of severes, have unusual effect on aspect some of their life) (Comings and Comings, 1987b).In TS experimenter, 30% is 3 grades, and 62% is 2 grades and 8% be 1 grade.TS is that similar obstacle and the major part of coming the TS experimenter of clinic have pathogenic ADHD (Comings and Comings, 1987b with ADHD; Comings and Comings, 1984).Among the TS experimenter, 54% meets the DSM-IV standard of ADHD.Contrast and have and do not have the TS experimenter's of ADHD existence make this group be particularly suitable for detecting as a successive variable at the dependency between iso-allele and ADHD not.TS experimenter's age average out to 18.0 years old (S.D.13.2).Though great majority are bigger child and teenager, 29% is 21 years old or bigger.The mean age of contrast is 46.3 years old (S.D.15.38).(Comings etc., 1996b were all described in TS experimenter and contrast in the elsewhere; Comings, 1995a; Comings, 1994b; Comings, 1994a; Comings, 1995b).

Each contrast and TS proband all are required to fill in one and meet with program (Robins etc. based on diagnosing, 1981), DSM-III-R (diagnosis of mental disorder and statistics handbook, 1987) and DSM-IV (APA,American Psychiatric Association diagnosis and statistics handbook IV, 1994) for the application form of the standard of a series of obstacles.Whether the problem inquiry (scoring=0) never or seldom occur child and adolescence DSM-IV ADHD symptom, occurs (scoring=1) once in a while or has (scoring=2) always.Oppositional defiant disorder (ODD) and behavior disorder (CD) are estimated also the total based on the quantity of these obstacles DSM-IV standard.Be the existence that evaluation has the problem of learning disorder, three problems of inquiry experimenter.1。Are were you once placed in an education and obstructed (EH), learns to be obstructed (LH) or special class of learning disorder (LD)? 2.Did you once go up recreation class? 3.Did were you once apprised of you and have learning disorder? each problem score for not=0 or=1 and addition form the LD scoring.Place above-mentioned special class any one all to need the comprehensive assessment of one or more educational psychologists in California, and be evaluated as more backward more than 2 years or 2 years than its companion.(Comings is 1994b) based on the summary of the DSM-IV symptom of alcohol abuse/dependence for ethanol scoring.The summary that the scoring of twitching exists based on a series of mobilitys and sound tic (Comings, 1995a).The specific problem that is used for behavior scoring has had detailed description (Comings etc., 1996b in the elsewhere; Comings etc., 1998a; Comings etc., 1995a; Comings, 1990; Comings, 1994a; Comings, 1995b; Robins etc., 1981; Comings, 1995c; Comings, 1994c).Each experimenter has all been checked application form to guarantee their accuracy.Use confirmation (Gadow and Sprafkin, 1994 of having passed through more a kind of instrument based on its accuracy of application form, usability and the sensitivity of symptom evaluation methodology by other people; Grayson and Carlson, 1991).

Even for detect the probability that the allele group who marks at random may show a significant effect when only utilizing additive gene, close the scoring that random number is specified each gene 0 and 2 according to the circulation in an algorithm, those the identical final gene frequency that it produces as is reported in table 65.

The inventor utilizes dstatt10.exe journey (http; //odin.mdacc.tmc.edu/anonftp/) is provided at the accurate p value of the big F value that obtains in the linear regression analysis.Utilizing linear chi-square analysis to check is not having and those have the significance difference of gene dosage between the experimenter of ADHD.Inc (Chicago, SPSS statistics software IL) have been utilized from SPSS.

The result.The type, one that table 65 is listed name, the polymorphism of 29 genes for the reference of polymorphism it comprise measure genotypic technology, how to genotype scoring, scoring for 1 or 2 experimenter's percentage ratio, whether independently proved conclusively the method for genotype scoring in the subject group and when its reference be provided one.Be null hypothesis #1, calculate r, the r of gradual PG and TPG gene scoring corresponding A DHD variable by linear regression analysis ², F and p value (table 66).In present result, the inventor is divided into group based on the function of neurotransmitter or their influence with gene.

Table 66

Individual gene, PG and TPG scoring are to the regression analysis result (N=336) of ADHD QTV

R r ²F p dopamine gene DRD1 .064 .0041 1.38 .240DRD2 .091 .0084 2.83 .093DRD3 .016 .0003 0.08 .772DRD4 .007 .0000 0.02 .901DRD5 .047 .0022 0.74 .388DAT1 .090 .0081 2.73 .099PG:D1+D2 .111 .0125 4.23 .040PG:D1+D2+D3 .095 .0092 3.08 .080PG:D1+D2+D3+D4 .084 .0071 2.39 .122PG:D1+D2+D3+D4+D5 .096 .0093 3.13 .078PG:D1+D2+D3+D4+D5+DAT .117 .0139 4.70 .031PG:D1+D2+D5+DAT (D) .144 .0208 7.10 .0081

R r ²F p5-hydroxytryptamine gene HTT .103 .0106 3.57 .059HTR1A .071 .0050 1.68 .20HTR1D β .030 .0009 0.31 .57H7R2A .040 .0016 0.53 .46H7R2C .015 .0002 0.08 .78TDO2 .050 .0025 0.73 .39PG:HTT+1A .120 .0143 4.84 .028PG:HTT+1A+1D β .117 .0137 4.65 .032PG:HTT+1A+1D β+2A .115 .0145 4.92 .027PG:HTT+1A+1D β+2A+2C .118 .0139 4.71 .031PG:HTT+1A+1D β+2A+2C+TO .125 .0158 5.38 .021PG:HTT+1A++2A+TO (S) .126 .0167 5.68 .018TPG:D+S .192 .0370 2.83 .0004

R r ²F p noradrenaline plain gene DBH .087 .0077 2.57 .105ADRA2A .110 .0122 4.11 .043ADRA2C .154 .0238 8.14 .0046NET .081 .0066 2.23 .136PG:DBH+2A .130 .0170 5.79 .017PG:DBH+2A+2C .188 .0355 12.30 .00052PG:DBH+2A+2C+NET (N) .205 .0422 14.70 .00015PG:2A+2C .185 .0344 11.89 .00064TPG:D+N .249 .0624 22.23 .000036TPG:S+N .231 .0535 18.87 .00025TPG:D+S+N (DSN) .272 .0741 26.73 .0000040 catecholamine degrading genes MAOA .156 .0244 8.35 .0041COMT .099 .0098 3.32 .069

R r ²F pPG:MAOA+COMT (C) .182 .0333 11.51 .00077TPG:D+C .215 .0465 16.29 .00067TPG:S+C .213 .0453 15.85 .00084TPG:N+C .272 .0741 26.71 .0000041TPG:DSN+C .317 .1005 37.31 .000000028GABA acceptor GABRA3 .090 .0082 2.77 .097GABRB3 .081 .0066 2.21 .14PG:A3+B3 (G) .119 .0143 4.85 .028TPG:D+G .178 .0318 10.97 .0010TPG:D+N+G .262 .0686 24.62 .000011TPG:DSN+G .281 .0792 28.14 .0000021TPG:DSN+C+G .329 .1087 40.72 .000000005

R r ²F p hemp ester acceptor gene CNR1 .041 .0017 0.57 .45TPG:D+CN .149 .0223 7.62 .0061TPG:DSN+C+G+CN .333 .1109 41.64 .000000003 nicotine courages can acceptor gene CNRA4 (NC) .119 .0143 4.87 .028TPG:D+NC .179 .0320 11.06 .0010TPG:DSN+C+G+CN+NC .342 .1171 44.31 .000000001NMDA acceptor gene NMDAR1 .076 .0058 1.96 .16TPG:D+NM .163 .0267 9.17 .0027TPG:DSN+C+G+CN+NC+NM .350 .1226 46.67 .000000000

R r ²F p enkephalins PENK .034 .0011 0.39 .53TPG:PENK+D .137 .0187 6.37 .012TPG:DSN+C+G+CN+NC+NM+PE .351 .1234 47.03 .000000000 (NT) androgen receptor gene AR .106 .0111 3.77 .053TPG:D+AR .168 .0284 9.77 .0019TPG:D+N+AR .271 .0731 26.36 .0000048TPG:DSN+AR .287 .0829 30.19 .00000078TPG:NT+AR .364 .1327 51.13 .000000000 immunogene CD8A .026 .0007 0.23 .63INFG .031 .0009 0.32 .57

R r ²F pPG:CD+ING (I) .039 .0016 0.52 .47TPG:NT+AR+I .354 .1258 48.08 .000000000 early ageing elements-1PSI .071 .0051 1.71 .191TPG:NT+AR+PS .348 .1212 46.06 .000000000 corticotropin releasing factor gene C RF .071 .0050 1.67 .196TPG:NT+AR+CF .369 .1362 52.67 .000000000

The dopamine gene.All five dopamine receptor genes and dopamine transporter gene (DAT1) have been detected.Wherein, DRD2 (Comings etc., 1996b; Comings etc., 1991), DRD4 (Lahoste etc., 1996; Swanson etc., 1998) and DAT1 (Comings etc., 1996b; Gill etc., 1997) gene has shown and has participated in ADHD.DRD1, DRD2, DRD5 and DAT1 gene pairs ADHD scoring have the strongest effect, its r ²Value is in from 0.0022 to 0.0088 scope.Divide other p value in from 0.388 to 0.093 scope.Though the DRD4 gene does not play a role in ADHD in this research, but be the document before the conclusive evidence, the inventor marks to this gene and emphasizes 6 to 8 long multiple allele, although inventor oneself studies show that these 2 allele may also be important (Comings etc., 1997a).

PG scoring shows that DRD1 and DRD2 gene are additivitys in their effect, make their r separately ²The summation (0.0041+0.0084=0.0125) of value is identical with observed PG scoring (0.0125) for D1+D2.The inventor claims that these are additive gene.On the contrary, when the scoring of DRD3 gene being joined in those D1+D2 PG scorings, the r that is obtained ²(0.0092) is lower than for DRD1 and adds the DRD2 gene.Therefore, about the ADHD phenotype, the inventor claims that these DRD3 are subtracting a property gene.Owing to add DRD4 gene r ²Continue to reduce (0.0071), subtracting property gene when it also is considered to.Because r after adding DRD5 and DAT1 gene ²Value increases once more, and these are considered to additive gene.When including in the PG scoring with additivity and subtracting property gene 0.0138 compared, when in the PG scoring, only comprising additive gene, and total r ²Be 0.0208.

Based on these results, the inventor has formulated this principle, increases r if promptly add a gene ², increase F and reduce p, it plays a kind of additivity effect in phenotype, however if it reverses these values, then it has subtracting a property effect to phenotype.In the remainder of these researchs, the p value that the inventor selects itself but not an arbitrary decision is as for the standard that comprises this gene in PG or TPG scoring.Therefore, the inventor selects DRD1, DRD2, DRD5 and DAT1 additivity dopaminergic gene (D) group as the TPG scoring.These four dopaminergic genes are responsible for 2.08 percent (p+=+0.0081) of ADHD scoring variance together.

The 5-hydroxy tryptamine gene.R for six 5-hydroxy tryptamine genes ²Value from for 5-hydroxy tryptamine transhipment (HTT) 0.0106 in 0.0002 scope for HTR2C.When the HTR1A gene is joined the HTT gene, r ²Value increases to 0.0143 from 0.0106.When adding HTR1D β, r ²Value reduces, r behind the adding HTR2A gene ²Value increases, r when adding the HTR2C gene ²R when value reduction and adding TDO2 gene ²Value increases.The suitableeest group of additivity 5-hydroxy tryptamine gene when this shows HTT, HTR1A, HTR2A and TDO2 gene possibility.The r of these genes ²Be 0.0161.Yet, owing to r for the HTR2A gene ²Value (0.0016) is minimum in four, so the inventor has also determined the r for HTT, HTR1A and TDO2 ²Value.Because its higher (0.0167) is so place the HTR2A gene outside the additivity group of 5-hydroxy tryptamine energy gene.Get rid of and have time low r ²The gene of value does not further increase additivity r ²These four genes are responsible for 1.67% (p+=0.018) of ADHD scoring variance together.When adding additivity dopaminergic and 5-hydroxy tryptamine energy gene, they are responsible for 3.7% (p+=0.0004) of ADHD scoring variance.

The noradrenaline plain gene.R for four noradrenaline plain genes ²Value is in 0.0066 to 0.0122 scope.In the case, all four genes are at total r ²Has a gradual additive effect on the value.Their are responsible for 4.22% (p+=0.00015) of ADHD scoring variance together, show that the noradrenaline plain gene bring into play than dopamine or 5-hydroxy tryptamine or dopamine and 5-hydroxy tryptamine to unite more important role in ADHD.In fact, two responsible variances (3.44%) of almost uniting the ADHD scoring of as much of norepinephrine energy alpha-2 receptor gene itself with dopamine and 5-hydroxy tryptamine gene.These adrenergic genes concrete in ADHD act on the elsewhere provide (Comings etc., 1998a).When the associating dopaminergic added the effect (DSN TPG scoring) that 5-hydroxy tryptamine can add the norepinephrine energy gene, they were responsible for mark 7.41% (p+=0.0000040) of variance of ADHD.

The catecholamine degrading genes.The r2 value of two catechol amine degradation gene pairs ADHD scoring from for the MAOA gene 0.0244 in 0.0098 scope for the COMT gene.When adding (C), they are responsible for 3.33% (p+=0.00077) of ADHD scoring variance.When joining the DSN scoring, they are responsible for 10.05% (p+=2.8 * 10 of ADHD scoring variance ^-8).

The GABA acceptor gene.Two r that the GABA acceptor gene is marked to ADHD ²Value from for the GABRA3 gene 0.0082 in 0.0066 scope for the GABRB3 gene.R for two associatings ²Value is 0.0143 (p+=0.028).When with DSN and C scoring associating, they are responsible for 10.87% (p+=5.9 * 10 of ADHD scoring variance ^-9).

Other neurotransmitter gene.R for four extra neurotransmitter genes ²The value for for Fructus Cannabis ester acceptor gene CNR1 0.0017, for nicotine cholinergic α 4 gene C HNRA4 0.0143, for nmda receptor gene NMDAR1 0.0058, for 0.0011 of endorphins protogene PENK.When in the gene that each is added to the front, the percentage ratio of ADHD scoring variance increases to 11.09%, 11.78%, 12.26% and 12.34% gradually.For whole neurotransmitter related gene (NT) group, p+=3.4 * 10 ^-10

Androgen receptor gene.Because all behaviors that the inventor detects are more common in the male, so the inventor has also detected the AR gene.Provide in the elsewhere the concrete effect of this gene in ADHD, ODD and CD (Comings etc., 1998b).The r of AR gene pairs ADHD scoring ²Value is 0.0111.When in the gene that is added to the front, they are responsible for 13.27% (p+=5.5 * 10 of ADHD scoring variance ^-11).

Immunogene.Because to the interest of immune factor latent effect in ADHD (Warren etc., 1995) and TS (Swedo etc., 1998), the inventor has detected two immunogenes, gamma interferon and CD8A gene recently.Both are proved to be subtracting property gene and are not included among the additivity group.

Other gene.Two other genes have been detected, senilism element-1 (PS1) and corticotropin releasing factor gene (CRF).Though for both r ²Being worth similar (0.0051 and 0.0050), is additivity but the CRF gene is only arranged.When all additive genes were united, they were responsible for 13.62% (p+=2.8 * 10 of ADHD scoring variance ^-11).

The ADHD scoring.Fig. 4 shows the variant additive gene of the accelerating effect to the ADHD scoring.It shows a kind of from those only being had 1.0 trend to 25.0 the gradual increase that those is carried 15 variant genes of 4 or 5 variant genes.P value for the linear X 2 test of a gradual increase in the ADHD scoring is＜10 ^-8Fig. 5 shows for the r that adds 29 genes that institute tests to some extent gradually ²Value.Next-door neighbour's genetic marker left side gets the prompting that the slope of curve provides the Relative Contribution of that gene pairs ADHD scoring.When slope increased, this gene pairs ADHD scoring had contribution.The angle of slope is big more, contributes big more.When slope reduced, gene was a subtracting property, and compared with other gene, in the ADHD scoring performance less or do not play a role, although the scoring of initial gene is 1 or 2 a kind of bigger phenotypic effect of genotype performance usually to scoring.Determined that DRD1, DRD2, DRD5 and DAT1 dopamine gene pairs ADHD scoring have contribution DRD3 and DRD4 gene then not to have.In the 5-hydroxy tryptamine energy gene, HTT, HTR1A and TDO2 gene pairs ADHD scoring have contribution HTR2A and HTR2C gene then not to have.Four norepinephrine energy genes are all bigger than the gene pairs ADHD scoring contribution of any other neurotransmitter group.When comprising all 29 adding up property and subtracting property gene, final r ²Value is 0.113, p+=2.7 * 10 ^-9

If the inventor has checked the gene of same cluster based on being appointed as allele at random, and only utilizes the gene that produces positive correlation with the ADHD scoring, the r that is obtained so ²It may be significant this probability.The results are shown in the bottom of Fig. 5.Square by sky is expressed r ²Gradual the adding up property effect of value, and express the adding up property effect of only utilizing positive correlation by the square that contains x.The common final r that utilizes adding up property and subtracting property gene ²Be 0.0001.Only use the final r of adding up property random gene ²Be 0.0004.Both are all not remarkable.In addition, though the alternative r at PENK gene place at random ²Up to 0.008, but it reduces to 0.0004 during adding up property gene (CD8A) at random when adding last.

Other potentially obscures effect for inspectability, and the inventor has detected masculinity and femininity respectively.Two groups all show r ²The remarkable rising of value, it is removed by the reduction of the experimenter's of smallest number in every group dynamics basically.

Therefore, the check of supposing #1 shows that the adding up property effect of 29 genes produces the r that is significantly higher than any individual gene ²With the p value, and when only utilizing adding up property gene r ²Be maximized with the p value.The assay of supposing #2 is shown among Fig. 6.It shows that viewed result conforms to the result of hypothesis.14 genes are adding up property in two groups, 2 are subtracting property in two groups, and 13 is different in two groups.Substantially, the strongest gene of adding up property in total group, as DRD5, DAT1, HTT, HTR1A, all four adrenergic genes (DBH, ADRA2C, ADRA2C, NT), MAOA, COMT and AR gene is the gene that is adding up property in two groups, and in total group the strongest gene of subtracting property, DRD3, DRD4 are for being subtracting property in two groups.The most visibly different gene is CNR1, CNRA4, NMDAR1 and PENK gene in two groups.For example, the NMDAR1 gene is suitable adding up property in 2 in group, subtracting property then in the group 1.Yet, though in two groups the less r of N ²Gradual increase, it is to two groups final p value＜+5.0 * 10 ^-4When including only adding up property gene, both are all remarkable, p＜10 ^-6When in total group, only utilizing those to two groups of genes that are adding up property, r ²=0.108, F=40.82, p+=5.6 * 10 ^-9When in total group, utilizing those in two groups all to be or only in one group, being the gene of adding up property, r ²=0.11, F=43.71, p+=1.5 * 10 ^-9These results and then be used for show the common difficulty in repetition when detecting individual gene.On the contrary, the MAA technology produces strong result under a series of conditions.Fig. 4, Fig. 5 and Fig. 6 show and send out the check that disease is enjoyed identical gene with ADHD and utilized this hypothesis of some special genes #3 altogether.

ODD and CD.Detected the r of 29 genes in ODD and CD scoring ²Effect on the value.When adding up property and subtracting property gene are all comprised, for the r of the maximum of ODD scoring ²Be 0.0775, p+=3.1 * 10 ^-6, and for CD scoring be 0.0225, p=0.0059.Equally, be easy to identify the contributive gene of marking by observation to ODD and CD.When only utilizing adding up property gene, for the maximum r of ODD scoring ²Be 0.10, p+=3.2 * 10 ^-8, and for CD scoring be 0.075, p+=+3.7 * 10 ^-6For the most important gene of ODD is DRD1, DRD2, DRD3, DAT1, HTT, HTR1A, HTR2A, HTR2C, DBH, ADRA2A, ADRA2C, MAOA, GABRA3, GABRB3, CNR1, CHRNA4, NMDAR1, PENK, AR and CD8A.To the most important gene of CD is DRD1, DRD2, HTT, HTR1A, HTR2A, HTR2C, DBH, ADR2C, CHRNA4, AR and CD8A.Therefore, 20 in 29 genes play a role in ADHD, and 20 genes play a role in ODD, and 10 genes play a role in CD.The ODD gene is not only similar but also different with the ADHD gene, and equal identical with in the ADHD scoring of all CD genes.

LD and twitch.Detect 29 gene pairs LD and twitched scoring r ²The effect of value.When adding up property and subtracting property gene are all comprised, for the maximum r of LD scoring ²Be 0.011, p=0.054, and be 0.014 for what twitch scoring, p+=0.029.When only comprising adding up property gene, for the maximum r of LD scoring ²Be 0.043, p=0.0005, and be 0.061 for what twitch scoring, p=0.00005.Equally, observing the gene that shows participation twitch scoring is DRD1, DRD5, HTR1A, HTR1D β, HTR2C, TDO2, DBH, ADR2C, COMT, GABRA3, CNR1 and CHRNA4.The gene that participates in the LD scoring is DRD1, HTR2C, TDO2, DBH, ADRA2A, ADR2C, MAOA, CNR1 and CNRA4.Therefore, 12 genes participate in twitching, and 9 participation in learning obstacles.

Ethanol or drug dependence/dependence and smoking.Detected test result at 164 29 genes of marking for alcohol abuse/dependence among the experimenter more than 14 years old.When N hour, r ²There is bigger fluctuation in value.Therefore, when only being detected as man-hour, be easy to distinguish adding up property gene and subtracting property gene.For alcohol abuse/dependence QTV, utilize the final r of two groups of genes ²Be 0.049, p+=0.0044.Yet, when only utilizing adding up property gene, r ²Value is 0.14, p=+7.5 * 10 ^-6As discussed below, many in the past showing in these genes or the neurotransmitter system plays a role in alcoholism.

Those of the same alcohol abuse of result/dependence of drug dependence/dependence QTV (except the smoking) are similar.Yet, because drug dependence is more rare than alcohol abuse/dependence, so the amplitude of r2 value level is lower.Main difference is that HTR1D β and HTR2A, ADRA2C gene are subtracting property in drug dependence/dependence, but adding up property in alcohol abuse/dependence, and HTRC and NT gene adding up property in drug dependence, but subtracting property in alcohol abuse/dependence, INFG alcohol abuse be subtracting property but be neutral in drug dependence.Smoking is more common, and has detected these results., QTV scoring is smoking once more than one month=1 herein, never inhaled=0.In this research of experimenter, the DRD2 gene plays an important role, and it oneself produces 0.055 r ²Value.Other adding up property gene is NMDAR1, PENK, AR and INFG.Final r ²=0.10, p=0.00038.

Other QTV.The inventor has also detected often fall ill altogether several other QTV of behavior with ADHD.Total r ²Value, p value and main adding up property gene are as follows: manic-r ²+=0.0721, p+=5.8 * 10-6, DRD1, DRD2, DRD3, DAT1, HTR1D β, HTR2C, TDO2, ADR2AC, GABRB3, CNR1, CHRNA4; Division sample-r ²=0.058, p=8.3 * 10 ^-5, DRD2, DRD3, HTR1D β, HTR1A, HTR2A, HTR2C, DBH, CNR1, CHRNA4; OCD-0.46,1.0 * 10 ^-4, DRD1, DRD2, HTR1D β, GABRB3, CNR1, NMDAR1; Generalized anxiety disorder-r ²=0.032, p=0.001, DRD1, DAT1, HTR1D β, CHRNA4, PENK; And severe depression-r ²=0.0271, p=0.0025, DRD1, DAT1, HTT, HTR1D β, TDO2, CNR1, CHRNA4, NMDAR1.

The quantity of related gene.For detecting in the relative distribution that has or do not have variant gene quantity among the experimenter of specific two minutes phenotypes, the inventor has drawn the quantity (Fig. 7) of adding up property variant gene between the experimenter of the DSM-IV standard that does not meet ADHD and those DSM-IV standards that meet ADHD.The par of variant is approximately 10 in those of no ADHD symptom, and is approximately 12 in having those of ADHD diagnosis.Those do not have the ADHD symptom in, be distributed in from 3 to 14 the scope, and in having those of ADHD diagnosis, be distributed in from 7 to 18 the scope.

Adding up property effect and p value.By this research, the inventor has drawn 145 r of 5 quantitative scorings of all 29 gene pairss (ADHD, ODD, CD, LD and twitch) ²With p value result, and identified to research phenotype to be those of those and subtracting property of adding up property.This demonstration when detecting gene one at a time, one be made as arbitrarily＜0.05 p value and gene be adding up property and also to what phenotype had contribution or a subtracting property phenotype is not had the few of dependency of judgement of contribution.When single detection, adding up property of major part gene has p value and the many p values that has between 0.10 to 0.40 greater than 0.05.Though the p value greater than 0.45 is accredited as subtracting property gene widely, the p value between 0.12 and 0.45 in the scope more then is that some of adding up property then are not.

Discuss.Worry about the difficulty of identifying the gene that participates in the polygenes obstacle is often proposed.In this research, the inventor attempts to change the single principal character of polygenes obstacle, many heterogeneic adding up property effects into by the adding up property effect that detects a plurality of candidate genes advantage.Owing to utilized the adding up property and the subtracting property effect of a plurality of association studies, so the inventor is called the multiple corresponding technology that adds up.When utilizing linkage technology to detect compound obstacle, show that the significance that must utilize strict especially level avoids false positive (Lander and Krugyak, 1996).Because this suggestion, the inventor has provided total length but not the p value of reduction in table 66 and Fig. 5 and Fig. 6.This shows the MAA technology of utilizing, and 29 genes and 344 experimenters only can obtain up to 10 ^-11The p value.

Because this is a kind of new technology, so the used statistical method of scrutiny is rational.It is ready to accept several potential criticisms significantly.If first is for only to study in a subject group, mark when maximizing its effect on given phenotype when setting gene, those genes scorings are added to together may produce more and more higher r simply ²Value.Because this, the inventor is the scoring of careful check gene in the independently subject group by other people research in inventor or the document.As shown in the table 65, all genes all satisfy this standard.For some genes, a variable scoring is only arranged.For example, for gene with two polymorphic alleles, when 1 allelic frequency therefore low and when seldom having 11 genotype, 22=0 and 11 or the scoring of 12=2 be unique.

If second potential criticism is if detected enough genes and only utilized adding up property gene, it is possible obtaining significant result based at random basis so.For checking this probability, the inventor has set up another set of scoring, wherein to scoring of each " virtual " gene random assortment of each individuality, be provided with its frequency with in observed result, found those match.The effect that adds positive and negative findings gradually is shown in (open squares) among Fig. 5.Till during to the 29th gene, positive and negative findings cancels each other out and final r ²Be 0.001.The more important thing is, when only utilizing the positively related random gene of generation, substitute r ²Value increases to the maximum for PENK gene 0.006.Yet, add latter two positive random gene after, r ²Value is returned and is reduced to 0.004, p＞0.25.This subtracting property effect may be because even the effect that CD8A and CRF mark at random is male, but r ²Value is in 0.0000 to 0.0003 scope, and subtracting property of generation effect.Though only utilize multiple repetition of those identical process can finally produce a significant result undoubtedly with positive r, but the important point is that final p value is apparently higher than (10 when utilizing the candidate gene that the same phenotypic effect of independent confirmation is relevant in other research ^-5To 10 ^-11) random gene.

Even the 3rd potential criticism is obviously than desired strong by marking at random, utilizes the single argument recurrence and only comprise that adding up property gene still raises the p value that is obtained.Quite reasonably criticize hereto two kinds of methods are arranged.First is that the goldstandard of all gene studiess is detections of repetition or a plurality of separate groups.Inventor suggestion is that adding up property group should be included in all genes that are indicated as adding up property in any research with reasonable N for a rational standard of MAA technology.For example, in the testing research of inventor's oneself test-again, selected adding up property gene group may be included in any one group and to be those of additivity.Because some in these may subtracting property in some researchs, so this can help to be avoided r ²With artificial the raising of p value.When having done like this, final r ²Still bigger, 0.11, and also highly significant, p＜2.0 * 10 ^-9

The reason of subtracting property effect.The subtracting property effect of a moderate may be because the responsible variance percentage of institute's gene of studying is relatively little.For example, scoring has very little effect if 11 genotype of gene A are to ADHD, and another gene may be responsible for the non-A11 experimenter that the ADHD scoring is higher than A11 experimenter.As a result, non-A11 experimenter may still have significant symptom, causes the subtracting property effect of gene.When detecting many different phenotypes, the Another reason of subtracting property effect may occur.For example, 11 genotype of gene may be with the ADHD significant correlation, and 22 genotype may be relevant with different phenotypes such as depression.As a result, when scoring was 11 genotype=2, this may cause the adding up property effect of ADHD scoring but depression scores is subtracting property effect.

Meaning to compound obstacle genetics research.The inventor believes that these researchs not only have many influences to psychiatric genetrics but also to the hereditism of generalized compound polygenes obstacle.1. compare the dynamics that detects a plurality of genes with individual gene.These researchs provide some understandings to the repeated problem of the association study of a gene of one-time detection.Most likely because a plurality of gene participates in, it is crucial not having individual gene, and each gene is only contributed the sub-fraction of variance for it.Therefore, when one next when detecting gene, gene will in a research, have significance in another then not.Can not think that this is the source (Moldin, 1997) of endless setback, perhaps even think that these study none is correct, this can regard as expected result in the hereditism of compound obstacle (Comings, 1998a).Variance percentage and the p value (except alcohol abuse/dependence) of utilizing the association study result of gene and behavioral trait for 145 have been reported herein.If this demonstration utilizes the working standard for the significance of individual gene p＜0.05, the major part of adding up property gene will be excluded.This also proves why so be difficult to reach an agreement for two laboratorys when utilizing the dividing value of p＜0.05.If produce 0.0008 to 0.025 r ²The association of value is the in fact independently research from the different experiments chamber, so only has 6 or 20% can be considered to positive and remaining 24 meeting is considered to negative ' non-repetition '.Yet by the MAA technology, all these associations all have been utilized.

Because the single notable attribute of polygenic inheritance is the participation of a plurality of genes, the inventor advises utilizing this feature to identify gene (Comings etc., the 1996b that participates in compound obstacle; Comings, 1996a; Comings, 1998a; Comings, 1998b).Originally studies confirm that this notion and shown that dynamics in polygenic inheritance is from the detection of a plurality of adding up property of gene effects.The inventor advises also determining that the appropriate criteria whether a gene plays a role is that it has additivity or subtracting property effect for relevant quantitative scoring in given obstacle.This can differentiate by listing as being easy in the result as shown in table 66 or Fig. 5 and Fig. 6.2. as the association study of the polygenic inheritance best approach.Current, the most popular method that is used for identifying gene in the compound obstacle is by utilizing the chain full genome screening of lod scoring, ill compatriot to technology or haplotype relative risk technology.Though this has powerful internal logic, repeat normally difficulty and also the said gene having found in ADHD or TS, to play a role based on adding up property genetic method based on these researchs (Pakstis etc., 1991) inventor in have and manyly be excluded.Though the ability (Risch and Merikangas, 1996 that utilize the irrelevant contrast and the proband's that significantly gets involved association study to have the little effect feature that detects polygenic inheritance; Comings, 1998a), but they are also worried in the problem of non-repeatability.The MAA technology that detects the adding up property effect of a plurality of genes is utilized the ability of association study and has been avoided the problem of non-repeatability effectively.This be because its with one to accumulating r ²The low strict parameter of cumulative effect substitutes the significance standard of individual gene at p＜0.05 place.This method is also more practical, owing to utilize identical tool detection, the evaluation of single irrelevant proband and irrelevant contrast is more much easier than the right evaluation of compatriot of getting involved.In the effort of the gene that identify to participate in many different obstacles, only set up by get involved with the dna library of unburdened compatriot to forming.By comprising single proband and a considerable amount of irrelevant contrast with the screening of same test instrument, the ability of the speed of case conclusive evidence and the discovery gene in these storehouses all can obviously improve.Its efficient may be maximized by allowing different storehouses that the identical contrast of screening that provides that discloses is provided.As for CEPH (centre detude du polymorphism human) (Dausset etc., 1990) sample, its can by require one preset time section disclose all gene type results later on and further strengthen so that can put the result of a plurality of researcheres in order.

The result provides an extra reason to be interpreted as what association study than stronger based on the linkage technology of family.Even its demonstration is when utilizing this two optimal separation group times, the sum of variant gene only slight different (10 pairs 12 averages).This prompting is when comparing in family, with with the right lod scoring of the compatriot that in fact or relatively gets involved with including and do not get involved, compatriot to, haplotype relative risk and TDT analysis (Spielman and Ewens, what 1996) taken place is the same, difference even will be littler.When problem is once only detected a gene when complicated, need to detect the experimenter's of a myriad of related gene, even at critical lod scoring or p value place.For example, even very a large amount of family and born of the same parents right is arranged at present, but the major part of the chain research of behavior disorder does not identify specific gene.Result among this coexist Fig. 5 and Fig. 6 is opposite, wherein utilize less than 350 experimenters and identified 20 specific gene and 15 specific genes temporarily, and the data that produces makes and can estimate the relative importance of each gene for each phenotype for alcohol abuse/dependence for ADHD.3. the ethnic group lamination problem of Yin Zanging.About association study one of the consideration of normal proposition be the stratified potential problems of hiding of ethnic group.Owing to making this misgivings, following reason MAA technology minimizes.Though the ethnic group layering of hiding when detecting individual gene may play a role, it is impossible that the frequency of all genes will change on identical direction.Along with the increase of detection gene dosage, the stratified potential effect of hiding ethnic group reduces.4. detect the problem of many genes.Usually another opposing views that occur in gene searching and association study are if a people pays close attention to abundant gene, and some will be significant for simple occasionality so.When gene of one-time detection and end point were＜0.05 p value, this was effective especially.Yet the multiple corresponding technology that adds up is the p value that does not rely on individual gene, and inquiry is with adding up property or the mode of the subtracting property r to phenotype at the next gene of the background of other gene ²Contribution is arranged.By emphasizing this feature of polygenes obstacle, this technology inherently and also constitutionally comprise the detection of a large amount of candidate genes.This is at the total r to twitch, LD and alcohol abuse/dependence scoring ²Can see in the fluctuation up and down of value.This shows can add gene up to a hundred, and final r ²Value may constantly fluctuate.Yet, the gene that detects many more, the adding up property gene dosage of being identified is many more, final r when utilizing the gene of this subgroup ²Big more.Therefore, the method for next gene that dynamics is all lost when detecting each extra gene is opposite, and the multiple corresponding technology of adding up property is detected and increase along with increasing gene.5. send out the relevant gene group of sexual disorders utilization altogether.Even ADHD is primary phenotype, but many ADHD genes are for the cumulative effect of representative of correlation table type such as oppositional defiant disorder, conduct disorder, alcohol abuse/dependence, drug dependence/dependence, smoking, OCD, manic, division sample behavior and other RDS behavior.6. send out sexual disorders altogether and utilize different gene groups.Except utilizing similar gene group, sending out sexual disorders altogether may have different phenotypes owing to they utilize different gene groups.Observe the prompting latter for two.The first, even when only utilizing adding up property gene, final r ²Value is lower than scoring to ADHD for ODD, CD, LD, twitch and other scoring.If inventor hypothesis is similar with to ADHD for each total hereditism contributions of these phenotypes, the genes that so necessarily relate to beyond those that this place detects make r ²Value reaches suitable level.This shows that except the gene of sharing, these are sent out sexual disorders altogether and also utilize unique gene.The second, comprise the accumulation r of adding up property and subtracting property gene ²The slope of value line shows that sending out sexual disorders altogether utilizes different gene groups or different genotype.For example, the MAOA gene is for adding up property of ADHD scoring, but to alcohol abuse/dependences subtracting property of marking.The importance of this prompting MAOA gene pairs alcohol abuse/dependence scoring comparison ADHD scoring is little.7. different phenotypes can be utilized different genotype.An alternative method utilizing the different genes group is that different phenotypes can be utilized different genotype.Reuse MAOA gene be not that its importance in alcohol abuse/dependence scoring is little, but it may utilize different genotype as an example.As another example, depressed and attack usually with low CNS 5-hydroxy tryptamine level be associated (Brown etc., 1982; Coccaro etc., 1989) and same CNS 5-hydroxy tryptamine that increases of compulsion or receptor sensitivity relevant (Insel etc., 1985).Therefore, if given genotype with depression/attack positive correlation, and is not wonderful with the compulsion negative correlation.These observations propose whether the gene scoring can be to the problem of each phenotype individuation.The inventor suspect its should, but this need confirm each such scoring in the experimenter of separate group.8. utilize different polymorphisms.In this research, the inventor only utilizes polymorphism of each gene.The utilization of other polymorphism or several polymorphism are unified into haplotype can increase r potentially ²Value.Therefore, viewed herein total r ²Value may in fact significantly be underestimated the real contribution of these each phenotypes of gene pairs.9. the comparison effect of different individual genes in different phenotypes.One of aspect that the MAA technology is the strongest is that it can differentiate heterogeneic relative importance.This may be because all genes are to detect in identical subject group.An example is the effect of AR gene.The next-door neighbour is for the r in the AR gene left side of ADHD (Fig. 5), ODD and CD scoring ²The slope of a curve increase shows that this gene is in all relative importances in the disease in three.On the contrary, the AR gene is for twitching and subtracting property of LD scoring, and not effect is neutral and mark for alcohol abuse/dependence.10. the comparison effect of distinct group gene in different phenotypes.Another aspect of MAA technology is that it can identify the important function of the gene group of the specific neurotransmitter of influence for different phenotypes.Therefore, the group of four epinephrine genes significant effect of performance in the ADHD scoring.They are responsible for the population variance r based on adding up property gene together ²42% of scoring.Compare, 5-hydroxy tryptamine can gene be responsible for only total 9.5%.This and many research (Halperin etc., 1997 that show norepinephrine metabolic deficiency in ADHD strongly; Pliszka etc., 1996; Arnsten etc., 1996) unanimity.Compare, seen opposite tendency for conduct disorder.Herein, 5-hydroxy tryptamine can the responsible population variance r based on adding up property gene of gene ²30%, and the adrenergic gene only is responsible for 13%.This is with many research (Brown etc., 1982 that show maincenter 5-hydroxy tryptamine metabolic deficiency in antisocial behavior; Lidberg etc., 1985; Coccaro etc., 1997) unanimity.11. the multistage property of association study and the influence that research is announced for individual gene.The result of MAA technology shows that association study should carry out in a plurality of stages.In the phase I, need carry out the early-stage Study of next gene and determine which kind of polymorphism may be useful and how they are encoded.Yet the important discovery that inventor's suggestion is studied for single-gene is how the genotype of the polymorphism of given gene should be marked.According to size, even have＞0.05 p value, this information also may be valuable.Since most of produce＞0.005 r2 or＞single adding up property of association study of 0.07 r, so when N is enough, this may be the standard of announcement, rather than be standard with p＜0.05.

Second stage should be to utilize the use of the MAA technology of all rational candidate genes.These results are represented by the line of adding up property and subtracting property gene in Fig. 5, and represent non-selectivity ground to comprise all genes.Three phases should be only to utilize the given phenotype of adding up property gene pairs to produce one ' new model '.This is by the line representative of adding up property gene in Fig. 5.The last stage should be independently to repeat to study, and it is identified new adding up property gene and continues test and test ' new model ' based on adding up property of accumulation gene group that is obtained again.12. the gene of alcoholism, drug dependence and smoking.Though it is genetic strongly that geminus and adoptive studies clearly illustrate that alcoholism, drug dependence and the smoking of at least some forms, also not significantly effectively to the evaluation of related specific gene based on the method for next gene.In fact, in the Taq A1 allele of the DRD2 gene of being reported and the association between the alcoholism (Blum etc., repetition 1990b) and can not multiplely unite (Noble, 1993; Blum etc. 1995b) have become one of contributor of suspecting the association study ability.Utilize the result of MAA technology to provide to alcohol abuse/dependence scoring to many understandings about crapulent molecular genetics problem.The first, DRD2 gene left side r ²Slope of a curve shows that the DRD2 gene utilizes Taq A1/A2 polymorphism to play a role really in this group experimenter in the ethanol scoring.Yet it only is responsible for total r ²10% of scoring, and because total r ²Scoring only is 0.14, and this gene only is responsible for 1.4% of alcohol abuse/dependence scoring population variance.This is quite typical and is a so difficult main cause of repetition of utilizing next genetic method why in adding up property gene.The second, dopaminergic, 5-hydroxy tryptamine energy and GABA can all play an important role in the ethanol scoring by gene group.The 3rd, by the final r of highly significant ²Value (p+=7.5 * 10 ^-6) confirmed the dynamics of MAA technology.Known to the inventor, for a suitable N, it is considerably beyond any next genetic method results reported once of utilizing.The 4th, though the focus of research is on ADHD rather than alcoholism, but be consistent (Loney etc., 1981 by the ADHD gene shared (DRD1, DRD2, DAT1, HTT, HTR1A, DBH, ADRA2C, GABRB3, CNR1, NMDA and CRF) with the ADHD of alcoholism high frequency and at the alcoholism of ADHD high frequency in a large number with alcoholism; Cantwell, 1972; Alterman etc., 1983; Tarter, 1988; Biederman etc., 1995).

Result for smoking shows that the potent of DRD2 gene should.Yet, because these experimenters have itself Tourette syndrome (Comings etc., 1991) with the DRD2 gene-correlation, so may be enhanced and surpass it for only being smoker's the due dependency of experimenter with the dependency of smoking.Although the gene for ADHD is primary focus, result's demonstration of marking for alcohol abuse/dependence has been indicated as crapulent candidate gene in the literature to before many in the ADHD genes identified.13. a plurality of phenotypes of research in single subject group.A common strategy in the genetics research of behavior be collect have a certain ' pure ' obstacle, less or do not send out the individuality of sexually transmitted disease (STD) disease altogether, and two fens ground with experimenter's scoring for having or not having those of that diagnosis.This research since two former thereby may lose dynamics.The first, the individuality with many diseases of falling ill altogether more may have higher hereditism and bear integral level, thereby increases the dynamics of identified gene.The second, the behavior of screening a wide region makes and can detect many different obstacles in identical subject group.This is proved in this research.Because TS is with many diseases of falling ill altogether relevant (Comings and Comings, 1987a; Comings, 1995a; Miller etc., 1996), so detection specificity ground is possible with the relevant gene of many obstacles in a subject group.The result shows that evaluation is still possible to the assortment of genes of the uniqueness of different phenotypes.This shows the development that utilizes structuring to meet to carry out multiple DSM-IV diagnosis and allow multiple quantitative trait in single subject group with high sickness rate altogether, may be a kind of method of gene of a series of obstacles of the more effective discovery of DNA data base than many each the corresponding particular obstacle of accumulation.14. risks and assumptions and diagnosis and predictability.About one of common hypothesis of polygenic inheritance be related gene only as risks and assumptions, and opposite with the individual gene obstacle, it will be impossible utilizing the hereditism to test in diagnostic mode.Yet along with the increase of the variance percentage of being explained by the adding up property effect of gene, r also increases, thereby increases in predictability.Show by utilizing this gene group that for mark final 0.37 r value of ADHD the predictability ratio of ADHD scoring exceeds when not having genetics information and reaches 37%.Along with more polygenic adding, can obtain to double the r value of this amplitude.Along with the appearance of DNA chip technology, the quantity of the hereditism's test that must carry out no longer is a problem.As shown in FIG. 7, though show very big overlappingly for the distribution curve that does not have ADHD symptom experimenter and satisfy related gene quantity between those of DSM-IV standard of ADHD, the difference that exists is a highly significant.Adding up property gene dosage doubled or three times may finally produce most of non-overlapped curve.The inventor considers that the diagnosis dynamics will increase along with the gene that is added.15. repeat.Just because to repeat be goldstandard in chain and association study, so it also should be the multiple standard of MAA technology.Should expect that ratio between composition, contrast and the experimenter of experimenter's quantity, proband's seriousness, the scope of quantitatively scoring and the variation of other factors all change final r ²Value, and do not rely on the effect of specific gene itself.Except these factors, the inventor estimates that multiple level can change.The highest level is that wherein identical gene group is adding up property or subtracting property in different subject group, and slope of a curve is similar.Because it is one of feature of polygenes obstacle that different genes produces the ability of similar phenotypic effect, so the inventor estimate this level to repeat be unlikelyly also can't estimate, and different research relates to demonstration the Different Slope of different gene group and curve.This is confirmed (Fig. 6) by the sample check of dividing equally.

Another and more real repetition standard are such, the major part of one of them group and be not whole adding up property gene for repeating groups also adding up property, and r wherein ²Value and significance level little by little increase along with comprising more gene, but slope of a curve may be different for different genes.On this level, will expect that also the relative importance of gene group is repeated.Therefore, this research, it shows that the adrenergic gene is prior relatively in ADHD, and 5-hydroxy tryptamine can gene be prior relatively in conduct disorder, should be able to be repeated.This is proved in the sample check that divides equally, and wherein the importance of norepinephrine energy gene in ADHD is repeated in two samples.If quickened the evaluation of gene related in the polygenes obstacle, then the multiple terminal level of MAA technology will be taken place.16. technical problem.Though covered the most of important aspect of the multiple corresponding technology of adding up property, also had several aspects to be worth emphasizing.Inventor's sensation is because the several reasons quantitative trait is preferred.It needs just with the identical character evaluation contrast of experimenter.This is important because in inventor's experience, and contrast and experimenter may have the scoring of a wide region.In addition, when detecting FR quantitative character, the dynamics of analysis is bigger.For the dynamics of maximum, the quantity of contrast should be near experimenter's quantity, and the quantity that perhaps has the experimenter of low scoring should be near the quantity of high scoring.

Though final r and r ²Value is not rely on order, but for clearly identifying subtracting property gene, needs to change order that the adding gene enters so that those have at first entering of the strongest positive correlation.If initial gene is not at r or r ²Produce suitable increase in the value, identify that so subtracting property gene will be difficult.

17. treatment and pharmacology's influence.The MAA technology is differentiated related gene and the ability of weighing their relative importance based on it, and its last aspect is the ability of its potential guidance treatment and pharmacological intervention.For example, four adrenergic genes are responsible for the ADHD final r that marks ²40% observation of value shows and acts on adrenergic α ₂The medicine of receptor may be valuable in the treatment of ADHD.Adrenergic α is in fact supported in clinical research ₂Receptor stimulating agent clonidine and the guanifacine effectiveness (Hunt etc., 1985) in the ADHD treatment.

Embodiment 14

Chromium picolinate and nicotinic acid chromium diet supplement therapy obesity

In this research, chromium picolinate is primarily interested, but thinks that the preliminary data of the nicotinic acid chromium that (in conjunction with physical exercise) tested under optimum condition also will be valuable.This research that replenishes the effect of young obese women about chromic salts has two purposes.The first, determine whether that independent chromium picolinate replenishes change body weight and composition, sugar tolerance and plasma lipid, and whether these effects can enlarge with physical exercise.The second, provide for nicotinic acid chromium and replenish data in conjunction with the effectiveness of physical exercise.

Method.The experimenter is the fat women who is used to sitting of 43 health.The definition of various statistical circles points is used for obesity, but for the purpose of this investigation, obesity is defined as being higher than the body fat percentage ratio of recommendation, for young woman, is recommended as the body fat (Blackburn etc., 1994) of 20-25%.The inventor finds that the part of inventor's colony only is slight obesity.Before acceptance, application form is used for determining experimenter's health status and activity pattern.Unmanned proof has any health problem among the experimenter, also not to the treatment of this situation.Age is in 18 to 35 years old scope, average 24.4 ± 0.70 years old.Initial body weight is in 50.8 to 96.1kg scope, average 71.3 ± 1.9kg.Weigh the body fat percentage ratio measured in from 25.0% to 45.0% scope by hydraulic pressure, average out to 33.0% ± 0.91.Initial VO2max value is in 1.53 to 3.30L/min scope, average out to 2.38 ± 0.13L/min.Before signature written consent file, notify each experimenter with participating in relevant potential risk and interests.Research is through the administration of research activities committee approval of University of Texas.

Experimental design.The experimenter is assigned in four treatment groups at random: chromium picolinate replenishes no physical exercise (CP), physical exercise and placebo (E/P), physical exercise and chromium picolinate and replenishes (E/CP) and physical exercise and nicotinic acid chromium additional (E/CN).Treatment stage was 9 weeks.Do not control the influence in menstrual cycle stage.Though carried out Pretesting and back test in the different phase of 28 days menstrual cycle of supposition, its influence (if there is) in all research participants be at random and also so should not influence data.

(San Francisco CA) prepares, and transports to researcher with the bottle of each coding of 100 by U.S. Shaklee company for chromic salts supplement and placebo tablet.Give the experimenter 16 weekly and contain chromium picolinate (200 μ g), nicotinic acid chromium (200 μ g) or placebo (invalid components).Give experimenter's verbal assistance and test phase every morning and evening take 1, those that accept the chromic salts supplement are formed the dosage of 400 μ g every day in whole treatment and back.Require the experimenter to return the tablet of not taking weekly.Weigh compliance by the tablet that calculating is given back.Compliance is no problem.

Physical exercise is by being made up of the cross-training program of plurality of units.First unit is the walking aerobicise, and this is the aerobic dancing of a class, utilizes weak beat music, 12 to 24 inches high stools and directors, and the latter instructs the participant to design to be used to provide a series of actions of the aerobic exercise of whole body.These classes of 1 hour participated in twice by the experimenter of each exercise in one week, and by titular director's religion.Allow the experimenter to select the height of themselves stool.Along with progress of research, the director increases exercise intensity gradually.

Second unit is by bike.Temper a week by bike and carry out twice, at the 75%-80% maximum heart rate (at initial VO _2maxDetermine in the test) the target heart carried out 30 minutes at the place down.Utilized general aerobic bicycle (general Gym equipment company, Cedar Rapids, Iowa).When each part begins, target heart is enrolled the program of ergometer, and in exercise, monitor, keep target heart with the computer regulated resistance on the instrument.

The 3rd unit is endurance training.Utilize Powercise Fitness system (TruTrac treats Products Co., Ltd, Temecula, California) to carry out endurance training, biweekly.Utilize five independently machines, wherein set endurance by a deceleration of electrons device.This is " two positive " system that excites the concentric contraction of agonist/antagonist muscle group.Five used machines are tempered all main muscle groups of body up and down.Can not be lifted again up to load by lift the weight (recruitment of 10lb.) that repeats to increase the weight of gradually at every turn at first, determine each experimenter's peak power.In case peak power is determined, exercise weight just is made as 50% peak power automatically, instructs the experimenter to carry out 15 multiple exercises of three covers on each machine.The experimenter follows rhythm and pace of moving things lamp, and it allows each to shrink about 1.5 seconds with one heart.The time of having a rest of inner room is 25 to 28 seconds.In the not strict control of the time of machinery compartment.The encouragement experimenter increases the weight of being lifted but still finishes 15 repetitions of three covers in whole research, and writes down these increases.Also instruct the experimenter to carry out 20 " abdominal crunches " (abdominal crunch) of three covers.

Supervise all by at least one researcher and temper part.Heart rate and endurance training parameter (finishing of weight of being lifted and repetition and cover) during record is turned out for work, taken exercise by bike.Require the experimenter in research process, not change their diet.

Experimental implementation.In the week before the treatment beginning, two independently occasion utilize and one have ± Health-o-Meter platform scale (the Continental ScaleCorporation of 200g sensitivity, Bridgeview, IL) weighing experimenter's body weight, and by hydraulic pressure weigh (Behnke etc., 1974) experimenter is carried out the analysis that health is formed.The Burker method of utilize revising at the gas of using the aspirated volume measured and exhalation on the treadmill according to the description inventor's laboratory before (Yaspelkis etc., 1993) mensuration VO _2maxExtracted blood samples on an empty stomach at two days that separate, and carry out oral glucose tolerance test (OGTT) according to described subsequently.

During 9 all treatment stages, give nutritional supplementation and physical exercise according to what describe in detail above.In the 9th week of treatment, repeat to detect before all tests.No matter before the treatment or treatment back is all tested with identical order.Replenish and the lasting whole test after-stage of physical exercise.All experimenters that participate in physical exercise a few days ago participate in meeting of a walking aerobicise back OGTT's, and have a rest in a day before test then, and this makes between exercise the last time and OGTT about 40h.Collection of specimens and analysis.On an empty stomach behind the 12hr, it comprises no caffeine and nicotine, obtains to be used for the blood plasma of hormone and substrate analysis at 7 in the morning and 9.Blood (10ml) is collected in venipuncture by antecubital vein.As one of blood sample before the treatment, in the mensuration of carrying out on the same day for the glucose and the insulin response of oral glucose load.The experimenter takes in beverage (Tru-Glu 100 orange juices/carbonating that contain 100 gram glucoses of room temperature, 10oz.FischerScientific, Pittsburgh, PA), and before absorption and take in back No. 21 venous duct (BaxterHealthcare from antecubital vein in the time of 15,30,60,90,120 and 180 minutes, Deerfield IL) obtains blood sample (3ml).

All blood samples are all used ethylenediaminetetraacetic acid (EDTA) (Sigma chemical reagents corporation, St.Louis, MO) anticoagulant of 250ml.(Sigma diagnosis, St.Louis MO) will be used for glycosylated hemoglobin from the whole blood (200ml) of basic sample and measure by affinity resin post, colour comparatour, terminal point step.With remaining sample centrifugal 15 minutes at 1,000 * g; Shift out blood plasma and frozen then-20 ℃ of analyses that are used for subsequently.(ICN Biomedicals company, Costa Mesa CA) measure concentration of insulin by radioimmunoassay, RIA.By enzymatic analysis come analysed for plasma sample glucose, triglyceride and T-CHOL (Sigma Diagnostics, St.Louis, MO).LDL-C and VLDL-C post precipitation enzymatic assays HDL-C (Sigma Diagnostics, St.Louis, MO).Utilize following equation calculate LDL-C:LDL-C=(T-CHOL)-(HDL-C)-(triglyceride/5) (Sigma Diagnostics, St.Louis, MO).Each is analyzed equal testing standard product and confirms concordance.The equal double test of all samples, each experimenter's of sequential testing specimen.

Statistical analysis.Front and back value to each variable between all treatment groups is carried out multivariate analysis of variance.Before handling, repetitive operation measures the value of measuring with the back, and the time point in OGTT.If observe a significant F value (p＜0.05), analyze further to determine where these changes occur in.A predetermined significance (p＜0.05) is with standard for further testing.According to the character of data, repetition Post-hoc checks and weighs variance analysis or Fisher PSLD (p＜0.05).These low rigorous degree are used for preventing because the chance that sample size is little and therefore statistics designs the limited II of the generation type of dynamics mistake.Utilization is used for statistics software kit (SPSS) software version 6.0 of the social sciences of Macintosh and analyzes.

The result.The experimenter who participates in physical exercise finishes the appointment training session of 90.0% (± 2.2).There is not difference on the compliance between the treatment group.In process by bike heart rate always HRmax 75 and 80% between.Heart rate is very high in walking aerobicise process, in the scope of per minute 175 to 190 times (HRmax 85 to 95%).VO2max all changes less than remarkable in any treatment group.

After the treatment, body weight has significant increase in the CP group, and body weight has significant reduction in the E/CN group.Not significant change the in body fat percentage ratio, fat mass or fat-free quality.No matter fat-free quality is significantly higher than at all in other group in the E/P group before treatment or after the treatment.It should be noted that the higher initial body weight of non-significance can be almost completely owing to bigger fat-free quality in the E/P group.

There is not marked difference on pro-and basic plasma glucose in back or the insulin level.Glycosylated hemoglobin does not change after treatment yet.Before treating during the treatment in 9 weeks by 3 hours each treatment groups of oral glucose tolerance testing and drawing and back glucose tolerance and insulin response curve.The experimenter accepts chromic salts and replenishes (CP), physical exercise and placebo (E/P), chromium picolinate (E/CP) or physical exercise and nicotinic acid chromium (E/CN).

In accepting the experimenter that chromic salts replenishes (CP), physical exercise and placebo (E/P), physical exercise and chromium picolinate (E/CP) or physical exercise and nicotinic acid chromium (E/CN) before the treatment stage in 9 weeks and after from the glucose tolerance curve of oral glucose tolerance test in 3 hours or comparison shows that do not have notable therapeutic effect at the glucose curve lower area for any group.Similarly, CP (p=0.433), E/P (p=0.087) or E/CP (p=0.110) treatment back not significant improvement the in the insulin response curve.Yet the insulin response that E/CN treatment back was located after the oral glucose carrying capacity in 60,90 and 120 minutes has significant decline, and it causes the remarkable decline (p=0.041) in overall insulin response curve.Also find insulin response curve zone down significantly reduction after exercise of E/CN treatment.Before treatment and between the specimen of back, do not find marked difference for triglyceride, T-CHOL, LDL-C and HDL-C.

Discuss.Inventor's research has compared that chromium picolinate replenishes, physical exercise or both all have the effect in the young obese women.Because external work prompting nicotinic acid chromium may be also to be effectively in changing these risks and assumptions, has also collected the data of replenishing the effect of associating physical exercise about nicotinic acid chromium.

Treatment back body weight significantly increases in the CP group.This is an important discovery, because chromium picolinate is advocated usually as a kind of slimming auxiliary.Inventor's result shows does not have physical exercise, and chromium picolinate replenishes not only causing in losing weight and may be invalid but also may cause weight increase.Weight increase (Kaats etc., 1991 are not seen in the additional research of chromic salts in the past; Page etc., 1991; Riales, 1979) may be, owing to do not keep on a diet and activity pattern and experimenter's different genotype pattern (being DRD2 A1 and the allelic carrier of DRD2 A2).And inventor's experimenter group (young obese women) was not studied in the past; The difference of this possible explanation in discovery.Similarly, the amount of the chromic salts that in this research, is given (400 μ g/d) be before to the twice of the amount of women's research.Might under this concentration, have a kind of effect that helps weight increase by chromic salts, and it may have depression effect under lower concentration.

Do not see the significant change of body weight in E/P or E/CP group, showing through the physical exercise in 9 weeks before having confirmed can not seen the research (Stefanick, 1993) that loses weight usually.Yet there be significant the reduction in body weight in the E/CN group.Known to the inventor, this is to studies show that it may be a kind of effective slimming mode that nicotinic acid chromium replenishes the associating physical exercise for the first time.

The treatment back is not changing in body fat, fat mass or the fat-free quality relatively significantly.As body weight, this research and before result of study between difference may be since the study population, do not keep on a diet and the amount of activity pattern and/or the chromic salts that given in difference.

Temper the not significant increase of back VO2max level.This may be because test lacks specificity: the max test of before on treadmill, carrying out and back, and take exercise by forming with aerobicise by bike.In addition, the aerobism change may be covered or remove to the endurance events of degree of physical exercise.

Basic glucose level does not significantly change in any treatment group.This result research (Abraham etc., 1992 that other chromic salts among the euglycemia crowd replenishes that coexist; Wilson etc., 1995) and physical exercise (Wallberg-Henriksson, 1992) unanimity.When comparing with E/P group, and though plasma glucose levels to the oral glucose carrying capacity be reflected in the CP group before the treatment and after all obviously higher.The average prompting of group glucose under fat-free quality and curve is interregional a reverse relation, shows that bigger fat-free quality allows the processing faster of glucose absolute quantity.Yet individual data shows dependency poor (r=-0.26) between these two factors.

Glucose response to the oral glucose carrying capacity is not observed therapeutic effect.As basic glucose level, the experimenter shows normal glucose level at first after the oral glucose carrying capacity.Glycosylated hemoglobin, the indication of one 6 all backs plasma glucose levels does not change after treatment.Basal insulin level, in normal range, the treatment back is not significant the change in any group at first.

Insulin response treatment back to the oral glucose carrying capacity significantly reduces in the E/CN group.Shown improving in insulin response (Djordjevic etc., 1995) with the hyperglycemia experimenter in the past.In other physical exercise group, do not observe the remarkable change of insulin response; This is unexpected, can reduce (Heath etc., 1983 because put down in writing in the past by physical exercise; Mikines etc., 1989; Sharma, 1992; Wallberg-Henriksson, 1992).Yet, have been found that existence very of short duration (Heath etc., 1983 of this reduction; Mikines etc., 1989), therefore, the inventor can't detect the insulin response that the glucose aggressive reaction is reduced in these experimenters, may be because the quick decline of insulin improvement effect.What the reason that no matter lacks in E/P and E/CP group the physical exercise effect of insulin action is, it may be useful that the result points out the associating of physical exercise and nicotinic acid chromium really.

Do not change through treating basic plasma lipid.What is interesting is and find that experimenter that minority has a unusual initial lipid levels treats the back and moves towards normalization, though these variations are not remarkable.

In a word, high-caliber chromium picolinate replenishes and does not have the while physical exercise to cause significant weight increase in the fat women of youth, and physical exercise associating nicotinic acid chromium replenishes and causes several potential useful changes, comprises losing weight significantly and the insulin response of oral glucose carrying capacity being reduced.Inventor's introduction be high chromium picolinate magnitude of recruitment (400 μ g/d) in the fat women of youth for the side effect that lost weight, may to provide some protections to prevent among CAD and the NIDDM in the adjustment by risks and assumptions than simple physical exercise more useful and physical exercise associating nicotinic acid chromium replenishes.

Chromic salts replenish the multiple risks and assumptions may influence coronary artery disease and non-insulin diabetes, comprise body weight and composition, basic blood plasma hormone and substrate level and to the reaction of oral glucose carrying capacity.Research has detected in the obese women of youth chromic salts in every days 400 during μ g, be with or without the effect (Grant etc. of physical exercise to these risks and assumptions, medical science and science motion and exercise (Med.and Sci.Sports and Exercise), 29:992-998,1997).Replenishing of chromium picolinate (CP) causes significant weight increase among this crowd, and physical exercise associating nicotinic acid chromium (CN) additional causes losing weight significantly and the insulin response of oral glucose carrying capacity is reduced.The inventor is summarized as replenishing among the young fat women for the side effect that lost weight of high-caliber chromium picolinate.In addition, inventor's results suggest physical exercise associating nicotinic acid chromium is additional may be more useful for the adjustment of some CAD and NIDDM risks and assumptions than simple physical exercise.

After the treatment, body weight has significant increase in the CP group, and body weight has significant reduction (P=0.05 at least) in the CN group of taking exercise.Not significant change the in body fat ratio, fat mass or fat-free quality.Though before the treatment still after, fat-free quality all is significantly higher than in all other organized in the placebo group of taking exercise.In addition, different with the CP group, the CN treatment of exercise causes locating insulin response at 60,90 and 120 minutes and reduces significantly.After the oral glucose carrying capacity, it causes reducing significantly (P=0.041) in overall insulin response curve.

Data from this research shows that high-caliber chromium picolinate is additional and does not have physical exercise simultaneously to cause significant weight increase in the fat women of youth, and physical exercise associating nicotinic acid chromium replenishes and causes several potential useful changes, comprises losing weight significantly and the insulin response of oral glucose carrying capacity being reduced.This data point out high chromium picolinate magnitude of recruitment (400 μ g d-1) in the fat women of youth for the side effect that lost weight, and physical exercise associating nicotinic acid chromium replenish may be more useful for losing weight than simple physical exercise.The gene frequency of OB/ gene and the associating of DRD2 gene is responsible for reaching 22% of obesity variance in the obese women of youth, and this genotype may influence the effect of chromic salts on the losing weight of this specific crowd, influence the effect (people such as Comings, 1997) that chromic salts loses weight to the mankind.Can expect, resemble fat those of relating to of describing in supplement composition, the especially table 6 of the treatment RDS associated disorders that contains chromic salts such as nicotinic acid chromium and/or chromium picolinate described in table 4, will helpful effect in keeping losing weight.

Embodiment 15 induces the health group as the allelic function chromium picolinate of TaqI d2 dopamine receptor A2

The variation that becomes

Method.The inventor utilizes Standard PC RTM technology (people such as Blum, 1997) that 100 experimenters' d2 dopamine receptor (DRD2) and dopamine transporter gene (DAT1) carried out gene type in this research.Assessment experimenter's body weight grade also utilizes densitometry to assess the ratio of body fat.According to the method that people such as Kaats proposed in 1998, the experimenter is divided into placebo group and chromium picolinate (CrP) group (400mg every day).

The result.In document contrast, 3.3% (1/30) in 30 super contrasts of 26% (185/714) and fine assessment among 714 experimenters have TaqI DRD2 A1 allele.Chi-square analysis has illustrated that these two matched groups have significant difference (p＜0.006).

DRD2 A1 allele occurs in the obese subjects of 67.4% (58/86) body fat 〉=28%; Occur in the obese males of the body fat 61.5% (8/13) 〉=28% and among the fat women of 68.5% (50/73) body fat 〉=28%.DRD2 A1 allele occurs in the obese subjects of 65.3% (49/75) body fat 〉=34%; Occur in the obese males of the body fat 62.5% (5/8) 〉=34% and among the fat women of 65.7% (44/67) body fat 〉=34%.DAT1 10/10 allele appear at 91 the contrast experimenters (34/91) 37.4% in, in the obese subjects of 47.7% (41/86) body fat 〉=28%, in the obese males of the body fat 38.5% (5/13) 〉=28% and among the fat women of 49.3% (36/73) body fat 〉=28%.DAT1 10/10 allele appears in the obese subjects of 46.7% (35/75) body fat 〉=34%, in the obese males of the body fat 37.5% (3/8) 〉=34% and among the fat women of 47.8% (32/67) body fat 〉=34%.The chi-square analysis explanation is when comparing with document or super contrast, relevant with the morbid obesity significance at TaqIDRD2 A1 allele.Table 67 shows, for the masculinity and femininity of body fat≤28% when comparing with the contrast in the document (card side=62.6, df=1, p=＜.0001) and for super contrast (card side=36.6, df=1, p=＜.0001) have found being correlated with of significance.Found masculinity and femininity for body fat≤34% when comparing with the contrast in the document (card side=50.6, df=1, p=＜.0001) and for super contrast (significance of p=＜.0001) is relevant for card side=33.0, df=1.Male for body fat≤28% compares (card side=8.31, df=1, p=＜.004) and for super contrast (card side=18.6, df=1, the effect of p=＜.0001) with the contrast in the document; And compare (card side=57.34, df=1, p=＜.0001) and with the contrast in the document for super contrast (card side=36.11, df=1, the effect of p=＜.0001) for the women of body fat≤28%.Male for body fat≤34% compares (card side=5.46, df=1, p=＜.02) and for super contrast (card side=16.6, df=1, the effect of p=＜.0001) with the contrast in the document; And compare (card side=46.73, df=1, p=＜.0001) and with the contrast in the document for super contrast (card side=32.38, df=1, the effect of p=＜.0001) for the women of body fat≤34%.On the contrary, for any allele combination that comprises 10/10 genotypic DAT1 gene, in this morbid obesity crowd, do not find relevant.Masculinity and femininity for body fat≤28% is compared (card side=2.27, df=1, p=＜.132) with the contrast in the document; Compare with the contrast in the document with the male for body fat≤28% (card side=.02, df=1, p=＜.89); Compare with the contrast in the document with the women for body fat≤28% (card side=2.73, df=1, p=＜.098).Masculinity and femininity for body fat≤34% is compared (card side=1.75, df=1, p=＜.185) with the contrast in the document; Compare with the contrast in the document with the male for body fat≤34% (card side=.01, df=1, p=＜.96); Compare with the contrast in the document with the women for body fat≤34% (card side=2.02, df=1, p=＜.16).

Table 67 dopamine transporter gene and body fat body fat ratio DAT ₁(%) card side ^b

10/10 p value, the total crowd in (contrast)＜28 41 0.132, (47.7), (N=86)＜28 5 0.89 the male sex, (38.5), (N=13)＜28 36 0.098 women, (49.3), (N=73)＜34 35 0.185 total crowd, (46.7), (N=75)＜34 3 0.96 the male sex, (37.5), (N=8)＜34 32 0.16 women, (47.8), (N=67) the a=bracket represents percentage b=34/91=37.4%

Table 68 d2 dopamine receptor gene and body fat body fat ratio DRD ₂(%) card side ^bCard side ^b

10/10 p value p value

(document contrast) the total crowds in (super contrast)＜28 58 0.0001 0.0001 (67.7) (N=86)＜28 8 0.004 0.0001 the male sex (61.5) (N=13)＜28 50 0.0001 0.0001 women (68.5) (N=73)＜34 49 0.0001 0.0001 total crowd (65.3) (N=75)＜34 5 0.02 0.0001 the male sex (62.5) (N=8)＜34 44 0.0001 0.0001 women (65.7) (N=67) the a=bracket represent percentage b=1/30=3.3%

Utilize a kind ofly to be called the more super contrast of statistical technique and all body fats that logistic regression analyzes and to surpass 34% case, DRD2 A1 allele causes 45.9% variation, is significant (card side=43.47, df=1, p=＜.0001) statistically.On the contrary, when when document contrast is compared, DAT1 (10/10 allele) causes 3% variance and is inapparent contributor to this variance of this study population statistically.

According to the CrP data, sample is divided into two independently groups; There are A1/A1 or A1/A2 allelic and have only A2/A2 allelic.Each group all separately check in placebo and Therapeutic Method difference for the various different measuring projects of body weight change.These measurement projects comprise that calculating fat weight variation percentage ratio, the variation of fat weight, the variation of body weight, the variation of non-fat mass, fat weight change percentage ratio, health is formed the body weight change that exponential sum is shown with the kilogram numerical table.With the independent group t method of inspection statistical test mean difference of placebo group and treatment group.By on p=0.05, setting up their significance,statisticals relatively time of α criterion evaluation.Anyly be lower than 0.05 p value and all be construed on the mean of placebo group and treatment group, have significant difference on the statistics.

It is more responsive to the CrP effect than DRD2 A1 allele carrier that the T check analysis has disclosed DRD2 A2 allele carrier.To the variation (p＜0.011) of the variation (p＜0.032) of fat weight, body weight, weight change percentage ratio (p＜0.035) and in the assessment of the body weight change (p＜0.012) of kilogram all is significant, and those have any parameter of DRD2 A1 allele experimenter not find that significance changes.

These presentation of results dopaminergic system, especially D are discussed ₂The density of receptor, lose weight and the body fat variation aspect cause the CrP treatment effect of marked difference.As the A1 that considers surfeit and DRD2 gene during allelic the relation, this presents even bigger significance people such as (, 1995) Blum.Inventor's viewpoint, the positive metabolic effect of the positive reaction maintenance CrP of A2 DRD2 carrier, but by contrast, A1 DRD2 carrier since carbohydrate gluttony increase covered CrP lose weight and the body fat variation aspect effect.These data further specify, and the treatment of CrP or other chromic salts and amino acid precursor combines, even also can cause reducing addiction and lose weight significantly people such as (, 1997) Blum in A1 DRD2 carrier.Inventor's suggestion in addition, the melange effect of taking health composition aspect of existing observed CrP may be solved by before treating the expection patient being carried out gene type.The reason that nicotinic acid chromium or chromium picolinate have observed effect is that chromic salts produces primary metabolizing parameters and aminoacid influence time dopamine shortage, thereby reduces the addiction to sweet food.

Embodiment 16

Aminoacid and herb ingredients are for the enhancing of focusing one's attention on: at attention deficit hyperactivity disorder

In potential effectiveness

The inventor observes dopamine D ₂Dependency between receptor gene polymorphism and human Electroencephalo are unusual.The weighted linear trend analysis shows in this relation, compares with common morbid substance abuse obstacle (SUD) with DRD2 A2 genotype, and when DRD2 A1 allele existed, event related potential (EPs) had the effect [p＜0.0001] of obvious variation.The amplitude of waking related potential (ERPs) P300 ripple up descends relevant with ethanol and drug dependence for a long time with incubation period.The inventor finds that also the preclinical obvious prolongation of P300 is relevant with three kinds of risk factor: (1) parents SUD, (2) chemistry relies on (relying on as cocaine) and (3) carbohydrate gluttony.The inventor also finds, compares with the A2 form (A2/A2) of gene, and the DRD2 A1 allele (A1/A1) of two copies is obviously relevant with the preclinical prolongation of P300 ripple.In addition, the P300 amplitude is relevant with the family history of alcoholism and SUD, and S.Hill (Pittsburg university) finds that the P300 amplitude is also obviously relevant with DRD2 A1 allele.These presentation of results, DRD2 A1 allele participate in comprising focus one's attention on and the non-behavior Pathophysiology of the brain function phenotype of potential RDS corelation behaviour (as SUD and ADD/ADHD) in effect.Must be noted that P300 is well proved unusually, but it is also special to drug dependence like that not as what once thinking.They also are common in Combination ADD, schizophrenia, delirium, obesity and other psychiatric disorders.

It is unusual to improve excitation potential to have proposed many therapeutic methods and spectrum analysis, as cholinergic agent, cholinesterase inhibitor, dopaminergic and serotonin activator, analeptic, trace element, diet (low concise carbohydrate), the cranium side stimulates, biofeedback and other special and non-special methods.

Because the dopaminergic function may be by reducing D ₂Receptor and to influence Electroencephalo unusual, the treatment measure may be that development strengthens brain D ₂The technology of function of receptors.Have TaqI A1 and TaqIB1 and other allelic dopamine D ₂The acceptor gene expression product significantly reduces, and causes the dopaminergic defective.From the gene repair aspect, do not have the known technology of this receptor defects of permanent reparation at present, yet the specific part among the present invention provides and has overcome the D that this heredity causes ₂B _MaxThe probability that reduces.Observe, comprise that the agonist of bromocryptine and the nor-apomorphine of N-n-pro-pyl causes D ₂Dopamine receptor raises people such as (, 1994) Fitz.In addition, behind agonist treatment, the variation of mRNA seems to illustrate the increase of receptor.On the contrary, to studies show that of a kind of inhibitor of protein cycloheximide, the increase of protein synthesis (rise of gene expression), and not decline of protein degradation, this is because the rise that the dopaminergic agonist causes.Utilize this logic, the present invention's suggestion is united utilization and is improved to promote D via the synapse dopamine release that enkephalinase inhibition (D-phenylalanine) causes ₂The occupying for a long time and shown in the HEK-293 cell of transfection, may make D of receptor ₂Receptor propagation or rise.This has further formed basis of the present invention.

Electrophysiology and neurotransmitter function.The brain electrical acti network for location that comprises QEEG and cortex excitation potential, the experimenter who has disclosed numerous species exists meticulous neurological to change people such as (, 1987) Porjesz, comprises schizophrenic (people such as Braverman, 1990; People such as Christian, 1994; People such as Blum, 1995), criminal (people such as Lovinger, 1995; People such as Seiden, 1995), depressed (Hudson, 1995), Alzheimer disease (Scourfield, 1996; Lawford, 1995), AIDS people such as (, 1995) Meiswanger, ADD/ADHD and they are to the reaction of medicine (people such as Kokkevi, 1995; People such as Nunes, 1995; People such as Yoshida, 1984), and SUD (people such as Gilman, 1990; People such as Morrow, 1992; People such as Brown, 1994; People such as Comings, 1996; People such as Neshinge, 1991).As everyone knows, medicine can induce the neurotransmitter in the marginal texture of deep to lack (being positioned at temporal lobe) (people such as Yoshita, 1984; People such as Gilman, 1990), cause the kitchen range electrophysiological abnormality.It may be to induce individual material to use the important symbol or the ingredient of desire that those topographies change.The exciting phenomenon that limbic system has been described may be a factor (people such as Ballenger, 1980) of SUD experimenter's addiction and withdrawal.In addition, SUD sample premorbid depression may just make the proband tend to Alzheimer encephalopathy, the ADD/ADHD that sends out altogether subsequently and else comes from the temporal lobe mental sickness at premorbid.The unusual hypometabolism with PET scanning of the temporal lobe of cerebral localization is relevant, it to have between the outbreak that metabolism fails temporal lobe epilepsy show effect similar people such as (, 1993) Adams equally.Reported that DRD2 A1 allele and frontal lobe PET scan detected LG metabolism relevant (people such as Noble, 1997).This shows the brain frontal lobe dopamine D of metabolism decline and the malfunction that causes ₂Receptor is crossed low relevant.Many articles show that SUD has promoted to induce abnormal evoked potential and unusual agitated reaction (people such as Goldstein, 1994) or the electric physiology unstability of spectrum analysis.In addition, cocaine and alcohol induced agitated reaction or electric physiology unstability, and it is unusual probably to discharge the rectification excitation potential via dopamine on acute basis.Even using at material, the substance abuse person in the recovery interrupts that low P300 is still arranged later.The P300 activity has only part to recover, and it also represents the inherited characteristic prior to substance abuse, cause to alcoholic in observed similar cocaine or heroin abuse people such as (, 1990) Gilman.

Inventor's reasoning, substance depilatory have significantly increased the weight of potential premorbid state, and point out the interaction of heredity-environment strongly.Cause material automedication or behavior act (being cocaine, ethanol, nicotine, cigarette, property or the like) that volt nuclear dopamine discharges, especially medicine is alleviated these electric physiologic derangements usually, but sorry has caused increasing the weight of of cerebral disturbance.

The additive effect of different dopamine genes.The inventor to gene type up to the nearest research summary of three dopaminergic gene DRD2, D β H of the ADHD/ADD+RDS in the 4th generation family and DAT1 gene in this.Comprehensive these results, provide support to following notion in the molecular genetics level: ADHD, TD and other obstacle are with the heredity of polygenes mode, be the part of associated disorders spectrum (RDS), cause, cause, cause, cause by the gene that disturbs dopamine system (and other) by the gene of additive effect by allele common among the crowd by the gene of sharing.

To how moving obstacle (ADHD) proband of attention-deficient with until the generation association study of a plurality of kinsfolk's dopamine genes in four generations.Dopamine D ₂(RDS) perhaps impulsion-the addiction of heterogeneous pass-compulsion is relevant with " recompense deficiency symptoms " for receptor gene polymorphism; VNTR 10/10 genotype of dopamine transporter gene (DAT1) is relevant with Tourette obstacle (TD) with ADHD; The B1 allele of dopamine-gene (D β H) is also relevant with the inferior character of many RDS behaviors with TD.

The inventor to 51 come from multiple influence family nearly the experimenter in four generations carried out gene type.DNA is according to PCR-based ^TMMethod wipe away paper from the oral cavity and extract people such as (, 1997) Blum.Two initial proband are diagnosed as by many reference instruments carefully has ADHD.Subsequently, other kinsfolk also to ADHD relevant RDS behavior with other diagnose.All experimenters have been carried out the gene type of three dopaminergic genes (DRD2, DAT1 and D β H).In all experimenters, 80 (40/50) percent carries DRD2TaqA1.When with " super " contrast (1/30 or 3.3% carries DRD2 A1 allele) when comparing, observing odd ratio is 116[95% confidence interval 13.6-2,575] significant correlation (card side=41.1, df=1, p=0.00000001, Yates correction).The inventor proposes these data for the importance of highly garbled contrast is described.When utilizing document contrast (185/714 or 26% DRD2 A1 allele carrier) comparing data of 714 non-alcoholics and non-drug dependence, the inventor obtained a similar but weak slightly discovery.Found that odd ratio is 11.4[95% confidence interval 5.38-24.93] remarkable association (p=0.00000001, Yates revises for card side=63.2, df=1).In 91 garbled contrasts, DAT1 10/10 allelic prevalence rate is in 34/91 or 37.4%, 51 garbled contrast, and the allelic prevalence rate of D β H B1 is 27/51 or 53%.When comparing with garbled contrast, between two kinsfolks' (30/50 or 60%) in ADHD source DAT1 (VNTR 10/10 allele), find that odd ratio is 2.64[95% confidence interval 1.31-5.38] significant correlation (card side=7.51, df=1, p=0.0061).By contrast,, find that odd ratio is 0.63[95% confidence interval 0.28-1.4 when D β H B1 carrier (32/50 or 64%) when comparing with garbled contrast] non-significance (card side=1.27, df=1, p=0.259).The inventor believes, compares with the 40%-50% that finds in the inferior character of single impulsion-addiction-compulsion usually, and DRD2 A1 is allelic to be owing to the inferior character of a plurality of behaviors that is included under the RDS at high proportion.In one family, DRD2 A1 allele appears at 100% be diagnosed as among the experimenter with ADHD.

When data being carried out completely linkage analysis, utilize at least one the RDS behavior that appears among the kinsfolk as the co-variation amount.It should be noted that when the number of RDS behavior increased, the allelic appearance of DRD2 A1 also increased in the experimenter.First glances up, and the DRD2 A1 allele relevant with two other dopaminergic gene predicts in the sample of check at present that at least ADHD and RDS behavior are more useful.At present data of handling and other result will find that biogenetics to impulsion-addiction-compulsion (RDS) and important inferior character ADHD influences according to these and propose and discuss.

Cognition, electrophysiology and neurotransmitter function.An importance of the present invention is the concentrated influence that is subjected to the neurotransmitter function change of attention, especially in the brain position (central authorities-edge) of being responsible for " recompense " and other corelation behaviour.Via heredity or environmental factors such as medicine, property with stress may influence attention in the malfunction in " recompense level chain " and concentrate.Though many neurotransmitter approach are final to be participated in focusing one's attention on, remembering and cognitive, preferred at least four main paties substantially among the present invention: 5-hydroxy tryptamine energy, opium energy, GABA can and dopaminergic.To support the positive correlation that dopaminergic system and attention are concentrated about the brief overview of cognitive and neurotransmitter document.This dependency has proposed such conception of species: the chemical compound that activates dopaminergic system and interaction that promotes agonist on dopamine receptor or release nature dopamine will strengthen the individual and focus one's attention on.

Dopamine D ₂Receptor gene polymorphism is relevant unusually with the human brain electrophysiology.This is to dopamine D known to the inventor ₂Receptor gene polymorphism and the unusual relevant research first that evidence is provided of human brain electrophysiology.In this, the weighted linear trend analysis shows: compare with common morbid substance abuse obstacle (SUD) with DRD2 A2 genotype, and when having DRD2 A1 allele, the effect (p＜0.0001) that has event related potential (EPs) significantly to worsen.The interval check explanation of Duncan ' s is compared with DRD A2 contrast, has or does not have the allelic SUD of DRD2 A1 and obviously worsened EPs.In addition, with respect to " super contrast " (p＜0.0000033) of inventor and a large amount of document contrasts (p＜0.0021), the inventor has observed serious substance abuse obstacle (SUD) and the allelic significant correlation of DRD2 A1.The amplitude of waking related potential (ERP) P300 ripple up reduces relevant with ethanol and drug dependence for a long time with incubation period.In this research, the inventor finds the preclinical obvious prolongation of P300 and three risk factor relevant (1) parents SUD, and (2) chemistry relies on (relying on as cocaine) and (3) carbohydrate gluttony (p＜0.03).In this crowd, the inventor also finds P300 amplitude decline relevant with the SUD family history with alcoholism (p＜0.049), but not relevant with DRD2 A1 allele.These presentation of results DRD2 A1 allele relate to brain function and potential addiction the tendency non-behavior Pathophysiology phenotype in effect.

The purpose of this research is to determine whether dopamine D ₂Acceptor gene (DRD2) TaqI A1 allele is relevant unusually with the outpatient's of private clinic the Electroencephalo that has or do not have substance abuse obstacle (SUD).Present inventor's laboratory is found D ₂The A1 allele of dopamine receptor gene and crapulent strong correlation (people such as Blum, 1990), several subsequently seminar fail to repeat this observation (people such as Gelernter, 1997).The inventor has proposed two possible reasons: the first, and insufficient screening of ethanol, medicine and tobacco abuse and other corelation behaviour in the contrast, the second, according to the missampling (people such as Blum, 1996) of the chronicity and the disease seriousness feature of alcoholic.Yet, the review of document has been disclosed the DRD2 gene not only and between the alcoholism, and with comprise many substance abuses, smoking, attention-deficient how moving obstacle (ADHD), carbohydrate gluttony, Tourette obstacle, pathological gambling, posttraumatic stress disorder one group of impulsion-addiction-obsessive-compulsive disorder and be called as many positives between " recompense deficiency symptoms " division sample/elusive behavior (RDS) related people such as (, 1996) Blum.Existing people proposes the variation (alcoholism is only single performance) (people such as Hill and Neiswanger, 1997) in the DRD2 allelic variation representative very common recessive character relevant with the dopaminergic activity.In addition, from contrast and patient group eliminating " other disease " be operated in San Antonio, Los Angeles, Duarte and Pittsburgh finish (people such as Hill, 1997; People such as Blum, 1996).Inventor's argument is not find that reason related in chain or the family may be to suitable understanding not exclusively for analyzing phenotype.

Therefore, in order to reduce false results, the inventor determines to utilize and is known as the unusual non-behavior pathophysiology phenotype of Electroencephalo as the sign of subsequently RDR2 TaqI A1 allelic association being studied.This method (people such as Noble, 1994 are also advised in the research to people RDR2 TaqI A1 allelic association in addition; People such as Blum, 1994).Up to now, comprise in SUD, ADHD, conduct disorder (CD), pathologic violent behavior, Alzheimer and other disruptive behaviour obstacles at many behavior disorders, unusually all there are related (the Task Force of APA,American Psychiatric Association, 1991) with spectrum and excitation potential.

Because the inventor has found that significant excitation potential is unusual in SUD and the obesity, the assessment Priceton of inventor's decision systems, unusual brain electrical acti and the directly related probability of DRD2 A1 allele among the patient of New Jersey neuropsychiatry and medical science clinic.Positive correlation can provide important and appropriate clinical information from hereditism's diagnosis cerebral disturbance to premorbid.

Relation between DRD2 allelic variation, SUD and the spectrum analysis is checked.In having the allelic typical normal subjects of RDR2 TaqI A2/A2, determined the remarkable probability graph (SPM) of vision arousal reaction (VER).Basis of calculation difference maximum (0.34) and minima (1.00) be as SPM, inventor's matched group and standardized BEAM ^TM(brain electrical acti mapping) contrast does not have significant difference.The characteristic VER mind map that has the allelic non-SUD experimenter of RDR2 TaqIA1/A2 has excessive negative electricity to 2.92 SD of slight volume temporo, is visualized as brilliant white indigo plant.The right volume temporo that shows by weak white blue area unusually in the fluctuation of being in a bad mood, cardiopalmus, anxiety and anxiety, to be with or without among the individual people of SUD be typical.The distinctive VER BEAM of genotypic patient SUD of RDR2 TaqI A1/A2 has a left side and excessive negative electricity to 6.13 SD of right volume temporo, is visualized as brilliant white light.

The inventor also finds the preclinical significant prolongation of P300 and three risk factor relevant (a) parents SUD, and (b) chemistry relies on (being that cocaine relies on) and (c) carbohydrate gluttony (p＜0.03).The inventor also finds P300 amplitude decline relevant with the SUD family history with alcoholism (p＜0.049), but not relevant with DRD2 A1 allele.

The most important thing is that weighted linear trend shows that the event related potential when having DRD2 A1 allele has remarkable degradation effects (p＜0.0001) when comparing with the DRD2 A2 genotype and the SUD that falls ill altogether.The interval check explanation of Duncan is compared with DRD A2 contrast, has or does not have the allelic SUD of DRD2 A1 and significantly worsened EPs.

It should be noted that in this research 52% serious SUD experimenter (N=29) carries RDR2 TaqI A1 allele.When (getting rid of alcoholism, SUD, cigarette smoking with inventor " super contrast ", ADD/ADHD, carbohydrate gluttony, pathological gambling, division sample/avoidance personality obstacle behavior, violent behavior, and alcoholism, SUD, positive family history with obesity) when comparing, prevalence rate percentage ratio has significant difference, and the allelic prevalence rate of DRD2 A1 is 3.3% (1/30) [card side=17.47, df=1, p＜0.0000033].In addition, in the non-Spain white man contrast of 714 non-alcoholics, non-SUD (except Nicotiana tabacum L.), 25.9% carries DRD2 A1 allele.When comparing, found very strong dependency (card side=9.44, df=1, p＜0.0021) with non-Spain white man SUD proband.

Known to the inventor, this is to observe the remarkable related research first between unusually of VER increasing among DRD2 A1 allele and the patient who participates in the neuropsychiatry treatment and audition arousal reaction (AER) Electroencephalo.This hereditism's evidence is studied the relation that further specifies between dopaminergic gene and the analeptic abuse natural inclination with other of lacking of proper care about the electrophysiology that is easy to psychoanaleptics abuse/dependence individual.Existing report SUD increases the weight of mind map parameter (people such as Braverman, 1996; People such as Blum, 1995; People such as Lovinger, 1995; People such as Seiden, 1995; Hudson, J., 1995) and when abstinence, in most of case, occur learning the continuing of damage (people such as Scourfield, 1996 by some drug-induced Electroencephalos; People such as Lawford, 1995; People such as Neiswanger, 1995).Inventor's reasoning psychosis proband sends out interaction (people such as Kokkevi, 1995 that SUD significantly increases the weight of premorbid sneak condition and points out gene-environment consumingly altogether; People such as Nunes, 1995).The self-medication of illicit drug for example resembles cocaine, is used for alleviating the imbalance of these electrophysiology, unfortunate has caused increasing the weight of of disordered brain function, especially at two temporal lobes of long-term repeated use person's brain.In addition, the P300 amplitude is relevant (people such as Yoshita, 1984 with preclinical decline and alcoholism and drug dependence; People such as Gilman, 1990).This is irrelevant with the acute effect of ethanol and medicine, because found evidence (people such as Morrow, 1992 in the son of the alcoholic that does not use ethanol; People such as Brown, 1994).This shows that existence relates to individual gene or a plurality of gene and relevant with the excessive risk of SUD of decline of P300 wave amplitude and prolongation of latency.

On the whole, the inventor believes these observationses the effect in cerebration (being recompense) has important neuro pathology's meaning to further understanding dopamine.Chain DRD2 polymorphism is (people such as Pohjalainen, 1996) with the function association of dopamine receptor.In 33 Finland man volunteers, compare the TaqI A1 allele of DRD2 gene and the B of calibration with A2/A2 experimenter _MaxThe significant reduction is correlated with on the statistics.K _dBetween group is inapparent.This shows that when A1/A2 experimenter's acceptor quantity descended, the function of receptor seemed not change, and shows the regulating action of the polymorphism of 3 ' mediation in receptor is synthetic.In addition, DRD2 knock out mice D ₂Receptor B _MaxSignificantly reduce and K _dWithout any changing people such as (, 1989) Grandy.Therefore modulability gene element may with TaqI A polymorphism linkage disequilibrium.Therefore proof is learned unusual special molecular polymorphism such as the DRD2 A1 allele of identifying by Electroencephalo, may have deep clinical value.In logic, seem that from these researchs the polymorphism of dopamine system is related to special Electroencephalo and learns dysfunction (being VER and AER), it seems that them mediated Deviant Behavior (the inferior character of RDS).

Continue these associations of research, the inventor will enough provide better preventive measure, especially in the excessive risk group.In addition, more special targeted therapy mode finally will come from these research, according to inventor's viewpoint, and their appreciable impact behavior of men pathology.

The experimenter has utilized 294 experimenters in the present invention altogether.All experimenters are right and wrong Spain white man, and they agree in signature; The approval of PathResearch Foundation Institutional Review Board has been passed through in this research.The patient who is used for studying is from participating in about 5000 visit random chooses that PATH outpatient private clinical practice surpasses the 800 routine patients in 1 year.

The experimenter of all SUD group is through identifying that clinically having fully (DSM IV) early stage SUD alleviates people such as (, 1994) Brown.Inventor's 173 routine demographies are sorted in the table 69 and describe.Sex and psychiatric diagnosis do not have significant difference in all groups.Mean age to all groups is assessed, and does not have significant change (p＜0.00001).For this research, sex selection comprises 53.1% male and 46.9% women, finds age differences and not discovery in sex in the psychosis diagnosis.

Electrophysiology and methods of genotyping. for definite SUD phenotype (cocaine abuse, DSMIV code name No.305.60; Cocaine relies on, DSM IV code name No.304.20; Alcohol abuse; DSM IV code name No.305.00; Alcohol dependence, DSM IV code name No.303.90) choice criteria and assessment tool, the inventor utilizes inventor's same procedure of report people such as (, 1996) Braverman in the past.(DRD2TaqI A1 and A2) carried out gene type to experimenter DRD2 allelic variation, with people consistent (1996) such as Comings.Nicolette BEAM ^TMBe used for assessment: overall abnormalities of brain, overall spectral singularity, excitation potential (EP, AER, VER) and P300.Detailed description to this unusual method of assessment Electroencephalo is seen Braverman and Blum, 1996.The inventor has also comprised a P300 matched group that does not have gene type for the purpose of the comparison that matches in this research.The P300 matched group comprises 15 male's trial volunteers, and they do not have medicine, ethanol and food addiction, do not have mental sickness yet.

Statistical analysis.In order to carry out statistical analysis, all mind map data are divided into unusual and normal.Especially EEG is divided into normal and unusual two classes, and spectrum analysis is by standardization BEAM ^TM2.5 SD of contrast are divided into two classes, wherein independently repeat defective (in same site) behind the spike at three.P300 voltage is divided into two classes people such as (, 1991) Neshinge with the normal voltage 10dv that establishes, and P300 is divided into two classes with 350ms incubation period.This value is based on that 300ms and the estimation of matched group mean age get, and this is by Lexicor, Inc., and Boulder, the standard that Colorado formulates, and P300 is annual after 18 years old increases about 1.25ms.(people such as Braverman, 1996) were delivered in the description fully of statistical methodology in the past.In addition, the inventor uses the paired average of Duncan ' s interval check carrying out relatively.The alpha levels of significance is set to 0.05.

Kantroll ^TMThe enhancing that healthy people's attention is concentrated: cocaine succedaneum with enkephalinase rejection characteristic.This is about taking in special ispol Kantroll every day ^TMThe first to cognitive event related potential (ERPs) effect in human body is reported.In the normal young volunteer that grows up, cognitive ERPs produces by the reaction to two computerized visual attention work, space orientation project (SOT) and emergency sustained performance project (CCPT), each experimenter checks respectively in the front and back of aminoacid absorption in 28-30 days and does own control.Taking in Kantroll ^TMAfter, the ERPs of the significant statistically amplitude of P300 composition all seen to(for) two projects strengthens.This research observed variation in normal control illustrates strongly, at amino acid additive Kantroll ^TMAfter the absorption, the active enhancing of nervous physiology may be the behavior of recompense deficiency symptoms (as ADD/ADHD, the substance abuse obstacle, the carbohydrate gluttony, nicotine abuse, or the like) add the basis of quick-recovery.

Neurobiology one of the most attractive discovery is, a large amount of neurotransmitteies are (as dopamine, norepinephrine, 5-hydroxy tryptamine, melatonin and glycine) (they play an important role in cerebration and mood regulation) can be circulated in its precursor aminophenol nutrient level appreciable impact (as, Wurtman, 1983).Precursor aminoacid separately is L-tyrosine (or L-phenylalanine), L-tryptophan and L-threonine.All these neurotransmitter synthesis systems have two conclusive features.The first, their synthetic source aminoacid is nine essential amino acids: histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine (tyrosine is synthetic by phenylalanine).The second, be undersaturated at the enzyme of the first step utilization of route of synthesis.The result of these two features is that these amino acid whose food intakes can promote the synthetic of special neurotransmitter.Because many modulability feedback mechanisms, this might not be a linear relationship; But shown that it is modulability approach (Hernandez-Rodriques and Chagoya, 1986 of a highly significant; Wurtman and Fernstrom, 1976; People such as Wurtman, 1981).

The complexity of animal midbrain chemical transformation is measured, and comprises microdialysis, has proved variation people such as (, 1988) Hernandez of introducing back neurotransmitter output at precursor aminoacid.In animal, aminoacid is after general and central nervous system directly import, and the recompense behavior changes and obtained explanation people such as (, 1972) Blum.When L-aminoacid was the precursor of neurotransmitter and neuromodulator, their racemoid, D-aminoacid also had biologic activity.Especially D-phenylalanine and D-leucine have reduced decomposition (people such as Blum, 1987 of the important instrumentality opioid peptides of emotion and behavior; People such as Carenzie, 1980; People such as Della Bella, 1979; People such as Ehrenpreis, 1979).

The neurotransmitter activity has formed the neuro chemistry basis of behavior, and their disorder may be main to many psychosiss and behavior disorder.They also are focus (Koob and Bloom, 1988 of quite a lot of comment to the contribution of assuetude disturbance always; Wise and Bozart, 1985; People such as Blum, 1990; Blum and Kozlowski, 1990; Amit and Brown, 1982).Especially dopamine, 5-hydroxy tryptamine, norepinephrine, γ-An Jidingsuan (GABA), glutamine and opioid peptides are considered to play a decisive role in assuetude disturbance, particularly about ethanol, heroin and cocaine abuse (people such as Blum, 1977; People such as Geller, 1972).These are observed and have produced such viewpoint thereupon: the selectivity nutrient is taken in can be influenced human emotion and therefore influence behavior.

Though used nutritional programs (being Williams, 1959) several times in the past, clear and definite quantification proof has been subjected to tangible restriction.Yet nearest clinical data has illustrated the obvious effect that amino acid precursor and enkephalin inhibitor recover ethanol, cocaine and food addiction (people such as Blum, 1988; People such as Halikas, 1989).Amino acid additive Kantroll has been checked in a relevant research (people such as Blum, 1988) ^TMUse in 30 days cocaine addiction inpatient treatment project.Unique difference of experimental group and matched group is the use of this additive.Two important test and appraisal projects have been used: before finishing, leave this project or disobey medical science suggestion (AMA) and medicine hunger.The test and appraisal that medicine hunger is that the relevant illusion of cocaine clearly, body reaction, request medication, the relevant opposing reaction of medicine, project are obedient to, AMA is left in excitement and violence or threat.The amino acid additive group is all good significantly than matched group aspect two kinds of test and appraisal.The AMA ratio has reduced by nine times and the medicine hunger that measures and has also lowered significantly.In addition, the staff has reported the obvious reduction of exciting, export-oriented attention and addiction.Yet assessment is complicated, often is qualitatively, and relies on many input factors (people such as Brown, 1990).

The present invention attempts to remedy clinical experience and to Kantroll ^TMThe basis understanding of neurobiology importance between gap.Here, this method assessment and Kantroll ^TMThe quantity neuro physiology that treatment is relevant changes.Electrophysiology partly focuses on the P300 composition of Cognitive Event Related Potential (ERP), is caused by two kinds of visual attention projects.Compare with more traditional EEG analysis, the advantage of this electrophysiological method is: owing to be fixed in the performance project, important special system is activated to attention processing.These attention probes have produced a series of ERPs compositions, and each is a stage of representative information processing all.Yet ERP analyzes and focuses on cognitive ERPs, especially P300 composition here.Show that recently it is to have very much predictability that quantity ERP changes for many clinical obstacles; As, attention disorders (people such as Buschbaum, 1973; People such as Halliday, 1976; People such as Prichep, 1976), schizophrenia (Braverman, 1990), depressed (people such as DeFrance, 1995a; People such as Vasile, 1992), dementia (people such as Duffy, 1984) and substance abuse obstacle (people such as Braverman, 1990; And Braverman and Blum, 1996).

The emphasis of this preliminary report is the attention course of processing, part because attention/or the change of concentrating one's energy all prior to and substance abuse (Begleiter and Projesz, 1988 of emergency sustained; People such as Whipple, 1988; Parsons, 1990), and because many to be considered to the important mediator substrate of attention be Kantroll ^TMThe target of control.The another one purpose of this research is to understand whether the aminoacid as precursors of neurotransmitters or instrumentality influences the central nervous system.This research has been understood at normal subjects's life-time service Kantroll then ^TMThe time electrophysiology and the dependency of performance, particularly change indicated by the P300 composition of cognitive ERP.

The experimenter.This research comprises 20 normal trial volunteers; Psychiatrist by committology does not have psychology, neurological and psychiatric disorders by DSM IV criterion evaluation.All experimenters consent by signature and every all their participation in this research has been obtained compensation.Every experimenter has implemented twice check, tests pattern as check-reinspection, therefore as own control.Initial check carries out repeating then (back check) after 28-30 days the 0th day (preceding check).The experimenter takes six Kantroll every day ^TMCapsule 28-30 days altogether.Form and see Table 6.Because low two experimenters' of quality of preceding check and back inspection record data are not included.

Performance work.Two performance parameters are used as the behavior probe of electrophysiologic studies.First probe is space orientation.This is based on the project (people such as Posner, 1988) in response time, and wherein priming cues appears at the different piece in the visual field.As or when not having priming crues the response time for right and the left visual field compare.By the comparison in response time, can assess the ability of individual smooth conversion attention.Indication is, concentrates on the cross at monitor screen center and works as ^(*)When appearing at right frame by right mouse button, when ^(*)When appearing at left frame by left mouse button.According to two frames of brightness transition.To note in the response time, ERPs constitutes four classifications: (1) for the reinforcement of right visual field, does not strengthen for right visual field (2), and does not strengthen for the left visual field for reinforcement and (4) in the left visual field (3).The appearance form is at random, but four kinds of conditions are had impartial probability.Strengthen frame before target occurs 500msec by highlight.The accurate scoring and the response time of record left and right view field, and A ' separately (standard signal detected parameters) value.This pattern has been demonstrated to the example of location, attention flowability and the Cognitive Processing speed that stimulates.

Second probe is the emergency sustained performance.This project is the variant of traditional method (people such as Rosvold, 1956).Require individuality to react like this, specific lexicographic order occurs and press left mouse button: as, T ' another appears ' in ' T ' afterwards.The project of this version has constant 0.8 second stimulation intermittence (ISI) and comprises 500 experiments.The probability of non-' T ' is set at 50%, and the probability of advance notice property ' T ' is set at 30%, and the probability of target ' T ' is set at 20%.Average ERPs constitutes according to one of three conditions: interfering signal (except any letter the ' T '), advance notice property signal (first of a centering ' T '), and purpose (second ' T ' of a centering).Primary performance test and appraisal are index of inconsistency people such as (, 1989) Ringholz, and it is the conforming tolerance of performance.It should be noted that prefrontal cortex be it seems has farthest participated in persistence attention and effort, cause that good performance is arranged in such work (people such as Cohen, 1987; People such as Corbetta, 1991), and these projects are highstrung to upper.

The record scheme.Reference electrode effectively writes down site record EEG at the ear-lobe that links (A1-A2) from 28.Carry out montage based on international 10-20 system, supplemantary electrode place preceding temporo (FTC1, FTC2), middle temporo (CP1, CP2), the temporo wall (TCP1, TCP2) and wall occipitalia (PO1, PO2) district.Electrode impedance maintains and is less than 5KOhm.Supplemantary electrode is used for detecting the outer artificial signal of eye.Eyes vertically move and nictation (being VEOG) via directly placing left eye frame electrode up and down to come record.Eye level moves (being HEOG) and monitors with a pair of supplemantary electrode that is positioned at the tail of the eye.

EEG and EOG resemble instrument (0.1-40Hz, 6dB/ sound interval low pass filter, 36dB/ sound interval high pass filter) with 32 passage neuroscience mind maps and amplify.(NeuroScan, took a sample 2.56 seconds down in SCAN EP/EEG Collection and analysis system by control Inc.) by 16bit analogue-to-digital converters (TECMAR labmaster DMA) for thick EEG.In order to set up event related potential (ERPs), average waveform from more than the 800msec interval 256 set up.Sampling is carried out stimulating preceding baseline to set up before 100msec appears in stimulation, as the part of antidote.The single sweep data are carried out the baseline rectification by deducting from mean.The scanning times of record changes according to the behavior probe.Write down 200 scannings altogether in the space orientation, write down 500 scannings in the emergency sustained performance test.Each sections of behavior model.Checked three parameters, each may change according to individual attention work efficiency.These parameters are: the incubation period of ERPs composition, amplitude and symmetry (spatial distribution).

Data analysis.At first, 28 electrode site have been created T inspection statistics figure.This is a version of people's such as Duffy (1981) statistics probability graph.Though be exploratory, this method has produced an easy visual figure really, has illustrated that electrode site should further analyze.Because P300 is a target component, Pz is selected as the site of statistical analysis, because from this figure, this is the focus place of this composition in these visual item.Then, peak incubation period and peak-to-peak amplitude (in the interval of 275-325msec) have been assessed.The second stage of statistical analysis has been used paired T testing model (Statgraphics, 1988), comparison base and treatment condition.The data of performance project are analyzed with same model.

Be used for explanation, provided the electrode arrangement diagram that places 28scalp record site for CCPT project ERPs.Comprised that also being used for artificial signs corrects and vertical eye outer electrograph of getting rid of (VEOG) and outer electrograph (HEOG) record of level eye.

With the result who at first discusses from SOT.And this concrete model has produced many interesting compositions (people such as DeFrance, 1993), is included in the N2 negativity that produces in the interval of 100-200msec.The N2 composition is a kind of processing negativity, is a commitment (people such as DeFrance, 1993) of placing reaction.Find that this composition strengthens after treatment, but the variation of amplitude does not surpass the significance threshold value [F (1,17)=2.30, p=0.0259] that the inventor sets up in advance.Yet, find that the summit positivity (being the P300 composition) in the test condition of back is significantly higher than a left side [F (1,17)=8.531, p=0.0095] and right [F (1,17)=16.31, p=0.009] intensified condition on amplitude.Therefore, when being positioned a left side and right visual field, use Kantroll ^TMGroup mean P 300 compositions significantly strengthen.

Topography has been checked bottom line and the treatment condition of strengthening (promptly ' primed ') condition about the left visual field.The pattern of right visual field is seen the corresponding of zone with a left side, therefore no longer provides here.In the interval of 100-200msec, the N2 negativity appears at right temporo wall district.And, Kantroll ^TMTo the effect of this composition near significance,statistical, but bigger variation is relevant with the P300 composition and find out easily by comparison 300-400msec interval.In fact, the topography feature of P300 composition is still identical, but peak-to-peak amplitude is passed through Kantroll ^TMTreatment is enhanced.About the data of performance, the association response time that left and right view field stimulates is accelerated [F (1,17)=8.62, p=0.001] with 232 ± 0.03msec to 238 ± 0.03msec after treatment.In a word, the P300 composition there is significant effect, the N2 composition is had the enhancing in boundary line.Also can see remarkable effect to the P300 composition relevant with vigilant project CCPT.

The heterogeneity of the cognitive ERPs relevant and the many similarities that have that produce by other lasting performance projects, feature with emergency sustained performance project (CCPT) be significant P300 composition (be people such as DeFrance, 1995b; People such as Hillyard, 1973).Set up three groups of waveforms and be used for analysis-interfering signal, advance notice property signal and target-but have only target waveform to discuss.Remarkable [F (1,16)=7.422, the p=0.015] of the P300 amplitude of relatively finding of preceding check and back test condition strengthens.In order to determine this effect, compared the Pz average criterion waveform of bottom line and treatment condition.The big forward potential that reaches peak value during about 300msec is typical P300 composition.The P300 waveform is those that take during to second ' T ' of a centering (as indication) reaction as the experimenter.Measured Kantroll ^TMThe topography of the goal condition of preceding (bottom line) back (treatment) in the 300-400msec interval, and the topography feature of P300 composition is still identical, but amplitude presents enhancing when comparing preceding check with the back test condition.That is to say, under bottom line and treatment condition, the P300 composition has occupied the rear center site and equally has been symmetric with SOT.Yet the amplitude of P300 composition has strengthened under the treatment condition.Have local solution to dig to learn difference after about four week though treat, the behavior the main performance variable-index of inconsistency of probe do not have the significance difference.Possible reason is that these normal subjectses are near the optimal representation of their accuracy.Yet, performance difference also may appear in clinical crowd.

Kantroll ^TMBe designed the potential treatment as cocaine abuse, because one of performance of recognizing cocaine abuse is to have changed attention processing people such as (, 1993) Robledo.In addition, human attention processing often is related (that is, people such as Hillyard, 1973) with the P300 composition of cognitive ERP.In this, be necessary to recall the neurological of P300 composition, because it provides the environment of important discovery.As everyone knows, P300 is one of cognitive ERP compositions of several endogenouss, and its incubation period, morphology and spatial distribution height depend on psychology environment people such as (, 1965) Sutton of implantable stimulation.Therefore, because the effect that the P300 composition is inferred in attention and memory, it is the theme of many researchs.The evidence of human deep layer record shows that frontal lobe (as, amygdaloidal complex) and middle temporal lobe (as, hippocampal formation) and frontal lobe structure may participate in the adjusting of P300 composition, and may in fact cause its demonstration (Halgren and Smith, 1987 in the temporo district; People such as Wood, 1980).Yet because the temporal lobe excision art is not eliminated the P300 composition, the deep temporo structure that therefore former researcher is be sure of is that unique product survivor's of P300 composition saying is unlikely (Johnson and Fedio, 1986; People such as Smith, 1985).But, the deep of temporal lobe seems that the P300 composition is had clearly regulating action (Halgren and Smith, 1987), and prefrontal area (people such as Simon, 1977) is also like this.

Because the characteristic of any cognitive ERP is to be fixed in very much to bring out the sexual behaviour model, remember that therefore it is important that the performance probe that uses in this research has the lasting composition with the work of selective attention power.Therefore, the special component of ERP (as, P300) should be relevant with two aspects of attention.What is interesting is, nearest PET research (people such as Corbetta, 1991) illustrated that frame volume cortex height participates in the selectivity aspect of attention, and the persistence aspect seems it more may is the scope (people such as Cohen at prefrontal cortex middle part, 1987), right hemispherical portion has played more important role (people such as Pardo, 1991; People such as Wilkins, 1987).Because the activity in temporo district before frame volume cortex is regulated via uncinate fascicle, it should not be astonishing that so amygdaloidal damage also influences selective attention power.Illustrating that amygdaloidal damage has damaged selective attention power really, may be owing to can not distribute sufficient emotion value (LeDoux, 1993) to stimulating.Also known attention performance is subjected to the characteristics and the distinctive influence of the information of importing into.Have that important evidence explanation Hippocampus plays in the comprehensive process of selective attention power be exactly this effect (as, people such as Salzmann, 1993; White, 1993).Therefore, frame volume-amygdaloidal complex-hippocampal formation axle seems in the adjusting of selective attention power it is important, and wherein amygdaloidal complex may the partition characteristic value be given the stimulation complex, and hippocampal formation is the comparative characteristic value between stimulating.In this framework, P300 works-the essential basis of subsequently emotion, memory and cognitive process as the side-product of the pattern that do not rely on of the selective attention power course of processing.Therefore these performance probes have challenged the function in volume-temporo axle path.These forebrain zones participate in the reaction of brain to cocaine just.

Illustrated that also attention processing relies on the adjusting of biogenic amine (people such as Stanzione, 1990; People such as Scatton, 1982).Take in owing to the required precursor of synthetic these amine relies on diet, diet is supplemented with and may changes effectively biogenic amine storage in the brain.This caused being devoted to the brain chemistry nutrition improvement different clinical protocol with treat special obstacle (Blum, 1989c).The clearly improvement that this method has been utilized Normocellular control mechanism and can have been caused psychology prospect, behavior performance and prevent to recur.This nutrition improvement relies on the amino acid precursor of the crucial neurotransmitter of utilization and vitamin and to the synthetic important mineral of these neurotransmitteies.It should be noted that all these chemical substances (mediator, vitamin and mineral) found not only in ethanol and drug abuser's body defective and also through the convalescent period of being everlasting continue defective (Blum, 1991a).

Therefore this research is proof, and nutritional supplementation can strengthen the nervous physiology function of normal control, and this may have important results (people such as Blum, 1988 for the utilization of aminoacid addition thing in cocaine recovers; Trachtenberg and Blum, 1988).How affected attention and memory function that later plan will be studied the cocaine abuse person in recovering are.Because important different neurotransmitteies rely on the precursor of source of nutrition in attention, might increase precursor so that attention deficit is reduced to minimumly by selectivity, and/or add quick-recovery.Be each (people such as Blum, 1990 in these neurotransmitteies of provable because drug dependence and/or genetic abnormality and functional change with the meaning of recompense cascade model; People such as Noble, 1991a; People such as Noble, 1991b; People such as Blum, 1991a; People such as Blum, 1991b; People such as Blum, 1992; People such as Noble, 1992; People such as Blum, 1994a; People such as Blum, 1994b; People such as Blum, 1995b; People such as Blum, 1995a; People such as Noble, 1995), all should be conditioned to promote that improving cerebration therefore also improves sensation, emotion and behavior potentially.This common disease is at first by people such as Blum (1995a; 1996a) called after " recompense deficiency symptoms " is a kind of malfunction of " recompense cascade ".Inventor's presentation of results can be introduced technology and enkephalinase rejection characteristic to small part by aminoacid and finish treatment to " recompense deficiency symptoms ".

Table 69

Cocaine abuse person classification

The identical more male's personality LI of sex men and women of more heredity in the more multi-ring border of reason of A type Type B abuse problem and high impulsion, sensation seeking

Sensation seeking,

Young factor early stage conduct disorder is seldom avoided in high injury

Risk factor substance abuse early in initial evening at age is more not serious, the ictal multiple medicines thing of more more long-term and serious seriousness pathopsychology lower severity, high seriousness

More susceptible perception is more antisocial

After the research of several years, researcher can be identified the factor that alcoholic is categorized as A type and Type B.The nearest research explanation that NIDA subsidizes, substantially, same a plurality of standards also are effective the cocaine abuse person being divided time-like.The result can prove when explaining the different reason of abuse and be useful when designing special prevention and Therapeutic Method.

Embodiment 17

The MAA technology is used for other polymorphism character-cholesterol and low-density lipoprotein white level.

In the research that the inventor acts in cardiovascular disease about different genes, the inventor has identified four genes relevant with cholesterol and lipoprotein metabolism.They are serotonin transporter (HTT), ocytocin receptor (OXYR), dopamine D RD2 receptor (DRD2) and senilism plain gene (PS1) gene.Polymorphism separately is to insert disappearance (people such as Collier in the promoter of HTT gene, 1996), the dinucleotide polymorphism of OXYR gene (people such as Mechelini, 1995), promoter insertion/the deletion polymorphism of DRD2 gene (people such as Arinami, 1997), and the RFLP of PS1 gene (people such as Higuchi, 1996).Based on experimenter's grouping research, it is as follows to mark: HTT gene-O=SS, LL, 2=SL; OXYR gene 278/278=0,278/267=1,276/276=2; DRD2 11=0,12=1,22=2; And PS1 gene 22=0,12=1,11=2.

Fig. 8 has illustrated this four genes effect in cholesterol and LDL metabolism of utilization MAA technology assessment.

These presentation of results MAA technology can be generalized to any polygenes obstacle or character.These four genes have disclosed 16.2 and 11.5% of cholesterol and the variation of LDL level respectively.The p value of cholesterol is 0.0002, and LDL's is 0.002.

Embodiment 18

Dopamine gene, violence and division sample/elusive behavior. about " pathologic violence ".The inventor has carried out gene type to DRD2 and DAT1 genetic mutation to the age at 11 teenager of 12-19, and they have participated in San Marcos, and the resident of Texas treats project.Diagnosis having that violent behavior is attacked in impulsion and selected come out to these experimenters according to one hour structuring talk.Selected each experimenter who is used to study have by the Nicolett that authorizes the psychiatrist to explain ( ^TM) the unusual BEAM of 2.5 SD measured.6 have D2A1 allele (56%) among 11 experimenters, and 11 are carried DAT1 (VNTR10 allele).When the D2A1 allele among the experimenter and " super contrast " (1/30 or 3.3%) when comparing, observed significant dependency (X2=14.9, df=1, p＜0.0001).When with document when comparing, also found DAT1 10 allelic significant correlations (34/91 or 37.4%, X2=7.6, df=1, p＜0.006), but do not found 10/10 allelic dependency (p=0.093).

The invention describes related between different dopamine genes and the SAB.For the SAB data set, the inventor has carried out gene type to three dopamine genes (DRD2, DAT1, D β B) and 72 screening contrasts of participating in Princeton 109 experimenters of outpatient clinic altogether.According to X 2 test, the inventor finds, compares the patient's significant correlation (X2=7.6 that has SAB (scoring＞84) that D2A1 allele and Millon Clinical Multi-Axial Inventory computerization check are identified with D2A2 allele, df=1, p=0.006).When comparing with super contrast, find D2A1 allele carrier in the avoidance experimenter of 11/22 or 50% division sample and 12/27 or 44% significant correlation (X2=16.75, df=1, p=0.000044).And utilize this method, X 2 test is analyzed and can not be disclosed the related of D β HB1 allele and DAT110/10 allele and SAB, when compared with the control, in having experimenter's (18/28 or 68%) of SAB, diagnosis found the remarkable association (X2=6.3 between DAT1 480bp VNTR 10/10 allele, df=1, p=0.012).Contrasting (rather than super contrast) found D β HB1 allele (17/23 or 76%) when comparing similar trend (X2=2.9, df=1, p＜0.09) with screening.The linear trend analysis has illustrated that the allelic frequency of D2A1 increases (A1/A1=83% with SAB seriousness; A1/A2=41%; And A2/A2=23%; P=0.0.005).Utilize multiple variable association, D2A1 allele and property all are the predictors of SAB seriousness.The inventor find the allelic odd ratio of D2A1 be 2.79 (p=0.0186) and sex be 3.6 (p=0.007).The Hosmer-Lemeshow goodness of fit p=0.778 and to the variation unite the contribution be 17.9%.To SAB, seem that the DRD2 gene is more important than DAT1 gene.

Embodiment 19

The example of the special analysis of genetic polymorphism

Be the heterogeneic concrete detection method that is used to diagnose RDS corelation behaviour and other polygenic character susceptibilitys of advising among the present invention below.Identical in many these concrete analyses and the document and illustrates this analysis application for polymorphism in the MAA technology.It will be recognized by those skilled in the art and can be enough these analytical methods that detect special polymorphic allele be modified, and remove description herein other genes and analytical method can in the MAA technology, use.

DRD1. the method utilization of checking the DRD1 gene is included in the Ddel polymorphism that A changes to G among the 5 ' UTR, by the PCR that is described by people such as Cichon (1994) ^TMMethod is handled.

DRD2. the detection of this gene comprises the DNA that obtains the experimenter, by Taq I restricted enzyme to above-mentioned experimenter DNA enzyme action, separate the dna fragmentation that produces, the allelic fragment hybridization of the recombinant phage-hD2G1 of above-mentioned separated DNA fragment and labelling (ATCC#61354 and 61355) or its specific bond people d2 dopamine receptor 6.6KbA1, and determine the above-mentioned allelic appearance of people D2 receptor A1.Especially the fragment of recombinant phage-hD2G1 (ATCC#61354 and 61355) can be the BamHI fragment of about 1.7kb length.The alternative method that detects comprises PCR ^TMTechnology (people such as Noble, 1994).

DRD4. extract DNA and utilize the VENT polymerase and high denaturation temperature (98 ℃ 1 minute), renaturation and extension join together to carry out PCR in 5 minutes at 70 ℃ ^TMAmplification people such as (, 1993) Sommer.The primer that uses is people such as (, 1993) Nanko, 5 '-AGG TGG CACGTC GCG CCA AGG TGC A-3 ' (SEQ ID NO:23) and D4=42:5 '-TCT GCGGTG GAG TCT GGG GTC GGA G-3 ' (SEQ ID NO:24).

The detection that DAT1.DAT1 repeats polymorphism comprises the VNTR gene type.Extract genomic DNA and pass through PCR ^TMAmplification DAT1 40-bp VNTR.The allele of 3 ' UTR utilizes people such as Vandenbergh, the oligomerization primer and the PCR of 1992a report ^TMCondition is passed through PCR ^TMIdentify.PCR ^TMAfter the amplification, product has under the situation of molecular weight marker electrophoresis in the polyacrylamide gel 8%.

People (1989) such as D β H. D ' Amato have reported the appearance of two Taq D β H polymorphisms of called after A and B.The D β H cDNA clone AII of utilization people such as (, 1987) Lamouroux contains the insertion of EcoR I site 2.7Kb.In order to improve labelling, carrier becomes five sections with BamHI with the SalI enzyme action.Be to detect the B polymorphism, labelling the fragment of 3.5kb.With TaqI restricted enzyme, agarose gel electrophoresis, Southern be transferred to nylon membrane, with the probe hybridization of 32P labelling, autoradiography has shown the fragment of 2.8Kb (B1) and 1.4Kb (B2).

MAOA. utilize MAOA VNTR polymorphism (people such as Hinds, 1992).This complicated polymorphism contains the little satellite of GT, and the latter is directly adjacent to the new 23-bp VNTR of incomplete double motif, and allele all has difference in dinucleotide repeat number and VNTR repetition.DNA extracts with the method for standard, passes through PCR then ^TMAmplification, each bar primer is all used fluorescence HEX or FAMAmidite (applying biological system, Foster City, CA) labelling, primer [＜320; 320-333; 334; 〉=335].With the PCR of two μ l through 10 times of dilutions ^TMProduct adds in the ROX500 standard of the deionized formamide of 2.5 μ l and 0.5 μ l, and 92 ℃ of degeneration 2 minutes, goes up sample then to 6% polyacrylamide gel of applying biological system 373 dna sequencing instrument.Gel electrophoresis 5 hours under 1100 volts and constant 30W.Laser scanning gel and utilize inner ROX 500 standard analysiss then.The peak is discerned based on the color fragment of base pair length by gene type instrument (Genotyper, version 1.1) [applying biological system].

PCR ^TMIntron 6 saltation zones of amplification tryptophan 2,3 dioxygenases.The PCR of amplified target sequence ^TMReact as follows: 10mM Tris HCL, pH 8.3,50mM KCl, 1.5mMMgCl ₂, 0.05%Tween 20,0.05%NP-40, the dATP of 100 μ M, dCTP, dTTP, dGTP, 0.1 μ M primer.Primer is: #116 GACACTTCTGGAATTAGTGGAGG (SEQ ID NO:25), and #117 GAAGTTAAATCCATG TGGCTC (SEQ ID NO:26).In 20 μ L, add following material: 0.5U AmpliTAq (Po Jin-Ai Ermo, Foster City, CA), 1 μ l (250ng) genomic DNA.Be reflected at PE-9600 thermal cycler (Po Jin-Ai Ermo) or PTC-100 sequencing thermal control instrument (MJResearch, Inc., Wastertown, MA) carry out on, utilize following condition: 94 ℃ 5 minutes, then 30 the circulation 94 ℃ 30 seconds, 60 ℃ 30 seconds, 72 ℃ 1 minute, then 72 ℃ 5 minutes.In order to detect whether amplification has taken place, the reactant mixture of 10 μ l electrophoresis on 1.5% the agarose gel in tbe buffer liquid.

The above-mentioned product of every part 10 μ l with the restricted enzyme bsl I of 1.5 units and final concentration be 1 * buffer (by New England BioLabs, Beverly, MA provides) enzyme action and overnight incubation in 55 ℃.The enzyme action product of 10 μ l 100VV and 1 * TBE in the 4%metaphor agarose (the F.M.C. product, Rockland, ME) in electrophoresis 1 hour.The gel ethidium bromide staining.Can obtain the fragment of three kinds of length.When pleomorphism site was G/G, the DNA complete degestion was the fragment of 673bp and 359bp.When pleomorphism site was A/A, the fragment of 1032bp was not cut open.The G/A heterozygote has three kinds of fragments, 1032bp, 673bp and 359bp.

The sequence that is right after G-＞T sudden change 3 ' is GATA.GATC is the recognition site of DpnII restricted enzyme.The oligomer of following design 3 ' 23bp is right after ATC sequence (few primer has two g sites of underscore and sudden change that double underline is arranged) 5 ' the TCATTAATCCTCTGGGTATTGTAAATGTGGATTTAGGTTAATATATTATATATAAT GCCAAATAATGGCATAGATAAGGAATAGGGAGAAAAAGGGAATTA-3 ' (SEQ ID NO:27) of G-＞T sudden change 3 ' with cooperation

TAGTCTTATATCCCTCTTTTTCTTA(SEQ?ID?NO：28)

Seldom damage it as PCR at the unpaired of the 3rd position ^TMThe effectiveness of primer.

Select 5 ' primer so that the product of 29 base pairs to be provided.When G-＞when the T sudden change was G, the GATC site was cut open the fragment of generation 22 and 70bp.When G-＞T sudden change was A, the fragment of 92bp only appearred.PCR ^TMThe condition of reaction is as follows: the various primers of 1 μ M, the various dNTP of 0.2mM, 50mM KCl, 10mM Tris HCL, 1.5mM MgCl ₂, the gelatin of 0.001% (w/v), 2.5U/100 μ L AmpliTAq (R) archaeal dna polymerase, 80ng genomic DNA.PCR ^TMCirculation be 94 ℃ 4 minutes: 94 ℃ 30 seconds, 52 ℃ 90 seconds, 72 ℃ of 30 circulation 120 seconds, then 72 ℃ 5 minutes.The condition of DpnII enzyme action is the PCR of 10 μ L ^TMProduct, the DpnII of 0.05 μ L10U/ μ L; 1.5 μ L:1M NaCl, 0.5M Bis HCL, 0.1M MgCl ₂, 10mM dithiothreitol, DTT, pH 7.9:3.5 μ L H ₂O, 37 ℃ are spent the night.Product is electrophoresis in 4% Metaphor agarose.

HTR1A repeats polymorphism.The method that is used to detect this complexity polymorphism is by people such as Bolos, and 1993 describe.For labelling PCR ^TMProduct, every kind of fluorescent dye primer of 0.1 μ M are used for reaction.Fluorescent dye be FAM amidite (the applying biological system, Foater City, CA).Two μ l are through the PCR of 10 times of dilutions ^TMProduct joins ROX 500 standards of deionized formamide and the 0.5 μ l labelling of 2.5 μ l, 92 ℃ of degeneration 2 minutes, and last sample is to the polyacrylamide gel of applying biological system 373 dna sequencing instrument 6%.Gel electrophoresis 5 hours under 1100 volts and constant 30W situation utilizes the standard of inner ROX 500 labellings to carry out laser scanning and analysis.Each swimming lane all contains internal standard can carry out point-device length and determine.The peak is analyzed by gene type instrument (1.1 versions, applying biological system) and is determined size by base pair length.If the peak of COMPUTER DETECTION to two a similar height, short peak always place a allele row, the peak places h allele row.Have one peak if computer is inferred, the experimenter then is an a allele homozygote.

The HTR2A gene.Utilization is by people such as Williams, and 1996 single base pair polymorphisms of describing are assessed HTR2A.The method of utilizing the applicating biological system DNA sequenator as mentioned above.

The OB gene.Contain that the multiple appearance of dinucleotide is by people such as Green in the YAC contig of people Ob gene, 1995 describe: D7S1873, D7S1875, D7S514 and D7S780.D7S1875 and OB gene are the closest.Comings is called OB1875.

Fructus Cannabis ester acceptor gene.The DNA sample utilizes as Dawson 1995 described following primer amplifications, 5 '-GCTGCTTCTGTTAACCCTGC-3 ' (SEQ ID NO:29) and 5 '-TACATCTCCGTGTGATGTTCC-3 ' (SEQ ID NO:30).Identified that so a kind of (AAT) n triplet repeats allele.With standard method labelling PCR ^TMProduct is also identified product polymorphism (seeing above-mentioned).

The GABRB3 gene.The DNA sample utilizes as people such as Mutirangura, 1992 described following primer amplifications.CA chain: 5 '-CTCTTGTTCCTGTTGCTTTCAATACAC-3 ' (SEQ IDNO:31) and GT chain: 5 '-CACTGTGCTAGTTTAGA TTCAGCTC-3 ' (SEQ IDNO:32).It has identified multiple 11 allele of (CA) n that length changes between 181-202bp.With standard method labelling PCR ^TMProduct is also identified product polymorphism (seeing above-mentioned)

Neuron nitric oxide synthase gene.Nitric oxide synthase gene recent findings relevant with the mice aggressive behavior (people such as Nelson, 1995).Utilize the method for people such as Hall (1994).The inventor has checked that the dinucleotide of neural nitrogen synthase gene (nNosla) repeats the dependency of polymorphism.The method of utilizing the applicating biological system DNA sequenator as mentioned above.

The COMT gene.For the analysis of COMT gene, single base pair polymorphism is by people such as Lachman, and 1996 describe.This polymorphism has shown relevant with the active varying level of COMT (seeing people such as Daniels, 1995).

Apolipoprotein-D gene.Extract DNA and use restricted enzyme TaqI enzyme action.Behind agarose gel electrophoresis, be transferred on the nylon membrane (Hybrid N, Amersham, UK) and utilize the APO-D probe hybridization of standard method and 32P-labelling.Two allele, 2.2 and 2.7kb identified.

People's the 2nd chromosome.Identified microsatellite polymorphism D2S1788, it is positioned 2p21 (from the about 74cM of the end of shortwall) (people such as Comuzzie, 1997).Be used for PCR from the DNA of lymphocyte preparation ^TM, the fluorescent dye primer of use, it contains the 169 height polymorphic micro-satellite signs that separate about 20cM from MapParis 6aLinkage Screening Set (Research Genetics).

The UCP-2 gene.Being positioned people's the 11st chromosome and mice the 7th chromosomal this gene has been identified with obesity and hyperinsulinemia chain.The forward primer of this gene is that 5 '-CATCTCCTGGGACGTAGC-3 ' (SEQ ID NO:33) and reverse primer are 5 '-AGAGAAGGGAAGGAGGGAAG-3 ' (SEQ ID NO:34).GenBank number of people UCP2 coded sequence is U76367.

Embodiment 20

The example of MAA technology in genetic polymorphism

This embodiment has proposed the polygenic character of many other than psychotic obstacles and has illustrated that the MAA technology can be generalized to all polygenes obstacles and all polygenic characters.Hereinafter lecture the enough a series of examples of the different polygenes obstacles of what use is made of MAA technical research.Osteoarthritis

The 1st step.Polygenes obstacle or character that evaluation will be studied.Generalization osteoarthritis (GOA) is modal in the joint disease.It is maimed main cause and classifies a kind of of three kinds of health problems as in developed country.GOA surpasses 45 years old and causes pain and afunction up to 70% above in 60 years old the masculinity and femininity 10% to 15%.The etiology of GOA is complicated, comprises the E﹠H factor.The research of same sex female twin is estimated adding up property gene has caused 54% GOA people such as (, 1996) Kaprio.Single-shot GOA is as the heredity of polygenes obstacle.

The 2nd step.Set up the yardstick of assessment polygenes obstacle seriousness.The seriousness of OA is (Kellgen and Lawrence, 1957) that utilize the Kellgen scoring to identify on the basis of X ray.

The 3rd step.The candidate gene that evaluation will be checked.OA very might be because the appearance of the threshold meristic variation gene that works in cartilage synthesizes and degrades.The potential candidate gene of this notion and part is listed in the table 70.The candidate gene of OA is being regulated the effect that cartilage is synthetic and degrade based on them.

The 4th step.Identify one or more polymorphisms with each gene-correlation.Be the part polymorphism tabulation that can be used in the MAA technology of OA below.

People such as table 70 gene chromosomal polymorphism type list of references collagen gene COL2A1 12q12 VNTR-1 Wu, people such as 1990COL2A1 12q12 VNTR-2 Priestley, people such as 1990COL2A1 12q12 Mae II RFLP Loughlin, people such as 1995COL9A1 6q12 DN-1 Warmen, people such as 1993COL9A1 6q12 DN-2 Warmen, people such as 1993 aggrecans (chondroitin sulfate proteoglycan I) AGC1 15q26 RFLP Finkelstein, people (Weber and May such as 1989 insulin like growth factor and receptor IGF1 15q22 DN-1 Weber, 1989) people such as IGF1 15q22 DN-2 Polymeropoulos, people such as 1991IGF1R 15q25 TN Meloni, 1992IGF1R 15q25 inserts people such as Poduslo, people such as 1991IGF2 11p15 DN Rainer, people such as 1993IGF2R 6q25 TN Ogawa, 1993 transforming growth factor TGFB1 19q13.2 Leu-＞people such as Pro RFLP Cambien, people such as 1996TGFB2 1q41 DN Westson, 1991 interleukin-11 IL1A 2q13 TN Zulini and Hobbs, 1990IL1B 2q13 C-＞people such as T RFLP DiGiovine, people such as 1992IL1R 2q12 DN GDBIL1RN 2q14 VNTR GDB metalloproteases MMP9 20p11.2 DN St Jean, MMP tissue inhibiting TIMP1 Xp11.3 RFLP-1 Allred and the Wright of 1995MMP1,1991TIMP1 Xp11.3 RFLP-2 Allred and Wright, 1991 vitamin D3 12q Uitterman RFLP etc., 1997

The variable series connection of VNTR=repeats.

DN=dinucleotide repeats.

TN=trinucleotide repeats.

The RFLP=restriction fragment length polymorphism.

The 6th step.Set up virtual polygenes or PG variable.The scoring addition of different candidate genes obtains the PG scoring.

The 7th step.Carry out the single argument regression analysis of PG to QT or DV.Use any statistical procedure the PG with first OA (oa) candidate gene (cg) (PG+PG oacg1) scoring is carried out the single argument regression analysis, carry out (PG+oacg2) after adding second candidate gene then.Continue this process and add (PG+oacgn) up to " n " individual OA candidate gene, n is the gene dosage that adds.

The 8th step.The result is mapped.As shown in Figure 4 the adding up property or the subtracting property gene of ADHD scoring are drawn to the result like that then.

The 9th step.Only utilize this process of adding up property gene redundancy.Only adding up property gene has carried out the such mapping of adding up property gene of as shown in Figure 5 ADHD being marked.Cholesterol levels

The 1st step.Polygenes obstacle or character that evaluation will be studied.Years of researches have shown related between blood cholesterol levels rising and coronary artery disease and apoplexy.Because inherited genetic factors and diet work to cholesterol levels, identify that the gene that relates to may be for identifying that risk new drug individual and development cholesterol reducing level is important.

The 2nd step.Set up the yardstick of estimating polygenes obstacle seriousness.The blood cholesterol levels of the sample seriousness yardstick that will offer the best on an empty stomach.

The 3rd step.The candidate gene that evaluation will be checked.Many genes of participating in the cholesterol route of synthesis directly work in cholesterol metabolism by inference.For the ability of MAA technology is described, the inventor assesses the effect of cholesterol approach several genes in addition with it, to identify their may act in cholesterol levels is regulated.The inventor has identified four genes relevant with cholesterol and lipoprotein metabolism.They are serotonin transporter (HTT), ocytocin receptor (OXYR), dopamine D RD2 receptor (DRD2) and senilism element (presenilin) gene (PS1).

The 4th step.Identify one or more polymorphisms with each gene-correlation.Polymorphism separately is that the promoter of HTT gene is inserted disappearance (people such as Collier, 1996), the dinucleotide polymorphism of OXYR gene (people such as Machelini, 1995), promoter insertion/the deletion polymorphism of DRD2 gene (people such as Arinami, 1997) and the RFLP of PS1 gene (people such as Higuchi, 1996).

The 5th step.Specify scoring to genotype.Based on experimenter's grouping research, it is as follows to mark: HTT gene-O=SS, LL, 2=SL; OXYR gene 278/278=0,278/267=1,276/276=2; DRD2 11=0,12=1,22=2; And PS1 gene 22=0,12=1,11=2.

The 6th step.Set up virtual polygenes or PG variable.The scoring addition of different candidate genes obtains the PG scoring.

The 7th step.Carry out the single argument regression analysis of PG to QT or DV.Use any statistic equation, the PG with first cholesterol (c) candidate gene (PG+ccg1) scoring is carried out the single argument regression analysis, add second candidate gene (PG+ccg2) then and carry out again.Continue this process and add (PG+ccgn) up to all cholesterol candidate genes.

The 8th step.The result is mapped.As shown in Figure 5 ADHD is marked then and draw like that.Fig. 8 illustrates that the MAA technology is in these four application that gene acts on of assessment in cholesterol and lipoprotein metabolism.

The 9th step.Only utilize this process of adding up property gene redundancy.Only adding up property gene has carried out as shown in the figure to such the adding up property gene mapping of ADHD scoring.In this embodiment, all four genes adding up property all.Life-span

The 1st step.Polygenes obstacle or character that evaluation will be studied.When the individual quantity that surpasses 65 years old among increase of the mean age of mankind's survival and the crowd increased, aging had become more significant theme.Evaluation can predict that many genes of individual life length have value in the individual man-hour that evaluation has early stage mortality risk.At the treatment of specific key gene effect, aspect quality that improves life and the prolongation life suitable benefit can arranged.

The 2nd step.Set up the yardstick of estimating polygenes obstacle seriousness.The seriousness yardstick of nature is the age.Yet, in this embodiment, to mark high more (older age), genotype is good more.

The 3rd step.The candidate gene that evaluation will be checked.Though the animal model and the cholesterol metabolism of the known participation aging of some gene such as superoxide dismutase (a kind of free radical scavenger), MAA technology have the potentiality of identifying the new gene of not guessing before many that life-span is worked.Because cholesterol levels is relevant with the death that cardiovascular disease causes, the inventor selects to identify two genes that cholesterol levels is played a major role.The inventor has also comprised the APOE gene.

The 4th step.Identify one or more polymorphisms with each gene-correlation.These genes are marked in following mode.The different genotype frequency of candidate gene compares by the chi-square analysis between all ages and classes subject group.If special genes type frequency is in progressivity increase between age group these years, this genotype marks=2.The genotype that remains unchanged is according to the scoring of the relative frequency in all ages and classes experimenter=0 or 1.

The 5th step.Specify scoring to genotype.The scoring addition of different candidate genes obtains the PG scoring.Each gene joins in the polygenes scoring gradually.

The 6th step.Set up simulation polygenes or PG variation.The scoring addition of different candidate genes obtains the PG scoring.

The 7th step.Carry out the single argument regression analysis of PG to QT or DV.Use any statistical procedure, the PG that has first life-span (1) candidate gene (PG+lcg1) is carried out the single argument regression analysis, add second candidate gene (PG+lcg2) then and carry out again.Continue this process and add (PG+lcgn) up to all life-span candidate genes.

The 8th step.The result is mapped.As shown in Figure 5 ADHD is marked then and draw like that.When the data of gene comparative lifetime are mapped, when three genes of MAA technical appraisement combine with 208 experimenters' research, obtain p=1.5 as a result * 10 of highly significant ^-7

The 9th step.Only utilize this process of adding up property gene redundancy.Only adding up property gene has carried out as shown in Figure 5 to such the adding up property gene mapping of ADHD scoring.

These only are three examples how the MAA technology can be generalized to any polygenes obstacle or character, utilize obstacle or the relevant human gene's research of character any and research.When genome plan when coming decade approaches to finish, expection will identify the polymorphism of human each gene, so that each polygenes obstacle or character are checked comprehensively.Inventor's expection, the MAA technology also will be applied to the polygenic character of other animal, plant, and the application of this technology in other biological species comprises in the present invention.

Embodiment 21

The example of anti-addiction compositions and right after the quick antidotismus therapeutica of opiate stone addiction

TREXAN ^Compliance strengthens

Preface.In the past ten years, utilize anesthetis naltrexone (Trexan ^, Dupont Delaware) has caused the interest of many therapeutic communities of the U.S., Canada and worldwide many countries to methadone or the quick new method of heroin addiction antidotal.Even withdraw from the stone opiate addiction rate today also near percent 90.The basic conception of this tachyiatria is to provide pure narcotic antagonist with the inductive euphoriant effect of blocking-up opiate to patient.Utilize this method, most patients is no longer deferred to and recidivism rate surpasses 99% (S.Hall, San Antonio Methadone Clinic).Inventor's argument thinks that the main cause of no longer deferring to is because as narcotic antagonist (Trexan ^) blocked opiate or ethanol inductive when glad (O ' people such as Malley, 1992; People such as Volpicelli, 1992), medicine does not almost have effect to the addiction behavior.Because the inventor finds aminoacid treatment and reduced the addictive behavior that causes glad thing to many, whether they determine to check TREXAN ^Combine to prolong to have used with aminoacid treatment and cause glad object height and reach the stone addict in 30 years Trexan ^Compliance.

Method.This research is finished at San Antonio Methadone Clinic.The standard that enters this research is to utilize DSM III that heroin/opiate is relied on and be diagnosed as stone addiction masculinity and femininity patient.For carrying out Pre-Evaluation, each patient at first injects 0.4-0.8mg Narcan and has assessed and their withdrawal (if too serious, they just can not enter this research).If they have passed through first test, oral dose is Dupont ' the s Trexan of 12.5mg then ^And after one or one and a half hours their withdrawal symptom of assessment.If patient is by this test, they take the Trexan of 50mg every day ^In this research, 12 patients have been assessed altogether.In this research, patient is except accepting the narcotic antagonist according to above-mentioned, also accept following aminoacid every day: DL-phenylalanine (2,700mg), L-tryptophan (450mg), L-tyrosine (300mg), L-glutamine (150mg), chromium (pyridine carboxylic acid salt-200mcg), and pyridoxal 5-phosphate (30mg).Calculating is not recurred or self-report refusal is taken Trexan ^Or with the natural law of aminoacid associating agent.Assess each patient (contact failure to a certain degree) every day via phone or individual the contact.

The result.To continuing to take Trexan ^Significantly enhancing aspect of compliance, the result is theatrical.The no aminoacid treatment that San Antonio Methadone Clinic calculates was accepted average natural law that hundreds of patients of this quick antidotismus therapeutica defer near 37 days.It is shocking that 12 experimenters in this research have accepted Trexan ^With not recurrence of amino acid therapy, perhaps average 262 days of conjoint therapy (p is p＜0.05 at least) is adopted in report.

Conclusion.The result shows that the adding of this anti-addiction preparation has significantly reduced the addiction to opiate, therefore assist those stone opiates addict to prevent aspect the recurrence to be important-especially with narcotic antagonist Trexan ^The associating use.

Embodiment 22

The D2 dopamine receptor gene is as the determiner of recompense deficit syndrome

Summary.Dopaminergic system, especially d2 dopamine receptor have deep related with the recompense mechanism of brain.D ₂The malfunction of dopamine receptor causes unusual material pursuits behavior (ethanol, medicine, Nicotiana tabacum L. and food) and other corelation behaviour (pathological gambling, Tourette syndrome, and attention deficit hyperactivity disorder).The inventor infers, D ₂The variation of dopamine receptor gene is " recompense deficiency symptoms " important common hereditary factor.

The inventor has used Bays (Rosner, 1986) theorem to assess DRD as mathematical method ₂Gene A ₁The predictive value of allele in impulsion-addiction-obsessive-compulsive disorder.

Bays ' s theorem can cause the probability of another incident (disease) medically being widely used in prediction particular event (defective), for example, has DRD ₂Gene A ₁Allele will cause that unusual medicine and ethanol seeks behavior (table 72).

When the assessment screening test, sensitivity is that check is at the male probability of individual man-hour of suffering from institute's study of disease; Specificity is a probability of checking the individual man-hour feminine gender of not suffering from this disease.For Bays ' s theorem, the inventor uses down and establishes an equation:

Table 71

The summary of d2 dopamine receptor genovariation and substance abuse/dependence

% prevalence rate substance abuse allele misuser contrasts P value＜list of references alcoholism DRD ₂A ₁People such as 69 20 0.001 Blum, 1990i alcoholism DRD ₂A ₁30 19 NS Nakajima, 1989i alcoholism DRD ₂B ₁People such as 17 13 NS Blum, 1993i (lower severity) alcoholism DRD ₂C ₁People such as 57 33 0.002 Suarez, the serious alcoholism DRD of 1994i (lower severity) ₂A ₁People such as 47 17 0.001 Blum, the serious alcoholism DRD of 1991i ₂B ₁Serious alcoholism DRD ₂ ^In6-Ex7People such as 39 16 0.02 Zhang, 1994i

Haplotype 1 cocaine relies on DRD ₂A ₁People such as 51 18 0.0001 Noble, the 1993i cocaine relies on DRD ₂B ₁People such as 39 13 0.01 Noble, many substance abuses of 1993i DRD ₂A ₁People such as 44 28 0.25 Comings, many substance abuses of 1994i DRD ₂B ₁People such as 33 20 0.001 Smith, 1992i*C ₁Allele is represented only about the homozygote genotype.Alcoholic (47/82); Contrast (29/87): (X ²=9.8, df=1, p=0.002)

Table 72

Dopamine D ₂Acceptor gene relies on (seriously) substance abuse more than 12.3 12.8 chemistry dependence 28.3 overfeeding (seriously) 18.6 feeding behaviour 35.0ADHD 16.0 smokings 41.5 pathological gambling 4.6Tourette as predictor risk behavior prediction value (%) alcoholism (seriously) 14.3 cocaines of compulsive disorder, and syndrome 5.5 is impulsion-habituation-compulsive behavior 74.4 totally

The hypothesis of supporting data is people such as Blum, explains in 1995

In order to calculate specificity, ethanol, medicine and tobacco abuse (table 71) are screened in the contrast that inventor's utilization has fully been identified in some sample.Without any former research for contrast with strict exclusion standard (people such as Blum, 1995a), owing to alcoholism is not and DRD ₂The phenotype that gene pleiomorphism is really relevant, this effort are essential (people such as Blum, 1995b; People such as Neiswagner, 1995; People such as Comings, 1991).In addition, in order to calculate the sensitivity of gene type, the inventor has adopted the proband to be determined the data (table 72) of chronic or serious disease.

The positive predictive value (PV+) of check is when checking when being used to contain the colony of unsoundness and disease individuality, real male positive findings percentage rate people such as (, 1975) Galen.Taq IA ₁Genotypic PV+ is 0.744 or 74%; That is to say that positive predictive value is high; But PV-has only 0.548 or 54.8%.When the individuality relevant with impulsion-addiction-compulsion got rid of from matched group, the inventor estimated to have better negative predictive value.These obstacles patient's blended data has shown and DRD ₂The strong positive of genovariation be correlated with (Yates card side=63.38, df=1, p＜10 ^-7).

Embodiment 23

Attention deficit hyperactivity disorder symptom evaluation and test valency scale (ADHD SAS)

Background: recent document (past 15 years) shows that the diagnostic criteria of attention deficit hyperactivity disorder (ADD) and many moving obstacles often changes.Essence to ADD in the research of children ' s spirit pathology still has arguement (McGee etc., 1989).

Though people such as father and mother, teacher, teenager, pediatrician, family doctor, shrink, the consultant of school understand and observed attention deficit hyperactivity disorder and moving obstacle symptom how on one's body teenager, in the North America, we feel difficulty when describing these phenomenons.Much studies show that in the document clinician in decision many moving and attention deficit hyperactivity disorder as conditions for diagnostics whether physical presence, the conditions for diagnostics that whether coexists, also have any problem when still independently diagnosing entity.

Follow many moving attention deficit hyperactivity disorders (ADHD) and do not follow many moving attention deficit hyperactivity disorders (ADD) to distribute very wide.In the U.S. 3,000,000 to 4,000,000 child patients and numerous adult patients are arranged.

ADHD patient is bearing overweight load, that is to say that their stimulation to being subjected to, especially light, sound and tactual stimulation have very high reactivity.They are also very violent to normal irritant reaction in the environment, so that " noise " that can not filtering environmental disturb, concentrate on the work in face of theirs.They are to concentrating on a problem or a task is had any problem.Because absent minded, they can forget appointment, forget and pay the bill, miss the time limit, also often run into the law difficulty because of not noticing when problem occurs.Often hurriedly, they have any problem to being fixed in a target or purpose.

Suffer from patient's life of ADHD and tend to inorganization, the room of child patient is disorderly and unsystematic, and the desk of adult patients is confusing, and daily routines also are unordered in a jumble.No matter at what age, they have any problem for making a plan, and the trouble that runs into when acting out one's plan in order is bigger.Owing to can not focus one's attention on, ADHD patient can get into trouble when finishing the work that they have begun.They can not finish the work, and can not realize a plan.Often pile uncompleted sewing product, carpenter's product in loft and the basement, repairing article such as product and notebook; Often at randomly on the desk putting letter, outline and the project plan of not write.

The ADHD intelligence of patient is no problem.Many ill people also have very high intelligence, but their achievement does not reach the shown degree of IQ usually, because they can not concentrate or lasting a certain interest.The thing followed is that family, friend, teacher and colleague lose patience to them and expects that they fail.

ADHD patient is difficult to adapt to variation, and their life has been full of confusion, even very little variation has taken place in daily life, they also can feel to upset.Father and mother go to travel, come in the school new teacher, family to move a new city and house pet in the dust, and these bring a crisis all can for ADHD patient.

The patient who tormented by ADHD lives under the pressure of height, and they can not bear failure, and are easy on the trigger when meeting with setback.Their indignation breaks out usually, as falls door, screams and get into a temper etc.In the child, often cause and fight, adult then in a great rage, lose work and and friend become estranged.Afterwards, they can apologize, but injury has caused.

Because ADHD patient often suffers setbacks, they become and lose patience, and hatred is waited in line and any delay that they are gone mad.They wish to carry out hurry up What for---once tourism, kinema, a class, once discussion---, finish as early as possible.

The impatient ADHD of making patient easily gets excited, and the child can not consider the consequences and blindly acts, and the adult shows as and steps on the gas, and uses electrician's instrument careless, inserts in the object that is just leading to electricity and does not consider the consequences, and consequently often makes themselves or other people injured.

ADHD patient's time sense and direction feeling are poor.They have to stop the thinking which be left hand, which is the right hand; They are to acting according to instruction, see map and concluding that thing such as time feels to have any problem.

How moving many ADHD patients show as.Show as ceaselessly child or baby and to move, to twist and upset etc., show as uneasiness, be sick of easily,, often be among the activity when being required to show as resistance when arranging to act the adult.

Another difference of ADHD patient is their brain wave waveform difference.Their β ripple---brain wave relevant with attention---is lower, and θ ripple---with lax relevant brain wave---is higher, and this waveform is with sleepy relevant with daydreaming.Thereby ADHD patient is difficult to bear predictability relevant with the β ripple and the activity of dealing with problems, this no wonder.They like the sort of and allow that they are in the state of θ ripple, and the very little activity (Lubar etc., 1991) of environmental stimuli.

If these symptoms are connected together, can appear such width of cloth figure in one's mind: a people is bearing over-drastic load, attempt with one too bright, too noisy, rub very much and change the too fast world and adapt.

The cause of early stage supposition ADHD mainly concentrates on following aspect: marriage maladjustment in the family, father and mother's raising is improper, mental illness and excessive drinking or drug dependence etc.Relevant behavior comprises conduct disorder and antisocial personality, and it is relevant with genetic matter utilization obstacle (SUD) that these behaviors are afterwards shown.Nearest research has begun to show that these behavior disorders (ADHD) are relevant with special gene unconventionality.

What is the cause of ADHD or its essence? it is by the unbalance compulsive disorder that causes of neurotransmitter, and is initial by inherited genetic factors.It is in the Childhood performance just to be arranged, and is extended to adult.Its influence can be alleviated by treatment and consulting.The Basic of Biology of this disease is determined (Biederman etc., 1992) by many researcheres

With unusual simple simon says, directly reason is the filtration system that ADHD patient is bearing a defective, and in other words, their brain network can not stop irrelevant stimulation.These people can notice each sound, each object and contact each time, and they are in inorganization, the insufferable confusion.Irrelevant stimulation obtains causing the attention that they are identical with the stimulation of those necessity of working or being correlated with other people.

On darker level, ADHD is that problem has appearred in patient's brain cell or the contact between the neuron, perhaps also comprises the neurotransmitter that carries information between nerve.These brains courier is under-supply.If suppress to import into courier's shortage of stimulation, too many signal just can pass through and confusion reigned.

See that on darker level problem appears on the gene of formulating neurotransmitter manufacturing blueprint.There is a genetic flaw---DRD at least in ADHD patient ₂Gene, this makes neuron obstacle occur to replying of dopamine, dopamine is with the emotion of happiness and regulates the relevant neurotransmitter of attention.Other dopaminergic gene is pointed out in other the research about gene unconventionality, as DRD ₄Acceptor gene, dopamine gene and dopamine transporter gene, (Cook etc. 1995 all can to become the reason of ADHD; Waldman etc., 1996)

The hereditism detects because the hereditary complexity of the ADD and the ADHD cause of disease, the inventor considers that the Genetic Detection method of this detection polymorphism can present detected person's dna structure by utilizing multiple dna to detect the existence that (Multi-Plex DNA BasedReward Deficiency Gene Test) assesses ADD and ADHD for the recompense dcc gene on basis.Which gene multiple dna if exist, is for the recompense dcc gene on basis detects to point out whether there is polymorphism, and they are homozygote or heterozygote.Because the order of severity and the above mentioned allele of ADHD damage and symptom are related, so multiple dna can be predicted the seriousness of ADHD symptom and a people's behavior for the recompense dcc gene detection method on basis.

Multiple dna is that treatment is very valuable instrument to specialty for the recompense dcc gene detection method on basis.Though its shortcoming also can't corrected now, the disclosed here chemical compound as anti-dependence producing drug is proved clinically and is produced effect by the serious hope that stimulates (output of brain dopamine and the activity level of dopamine receptor site) to cause to suppressing; And they have demonstrated aspect following effect: increase the ability that the people focuses one's attention on faster, promote the focus conversion, increase " in work " behavior and increase the scope that attention is concentrated.

Though the inspection of recompense deficiency symptoms behavior (ADHD is a kind of disease of hereditary basis, falls into this syndromic scope) has very high reliability and effectiveness to RDS, the distinctiveness diagnosis is challenging.Multiple dna will provide the very data and the distinctiveness diagnosis widely of conclusive evidence for the recompense dcc gene detection on basis; Yet inventor and DNA identification experiment chamber, northern University of Texas health science center and University of Tennessee have proposed complete (finishing quite fast) research profile at present.This research will be chain research, rather than association study.The inventor believe the result of this research at present will make the inventor can: make definite difference diagnosis, reduce the false positive (with present psychological test and interview diagnosis of technique time all have many false positives) of ADHD diagnosis, the raising True Positive Rate.Reduce patient and family members' resistance, reduce mistaken diagnosis (mistakenly to the anxiety of ADHD etc.) and therefore change the situation of leaving methylphenidate mistakenly, improve therapeutic scheme and correct difference diagnosis ADD and ADHD.

Multiple dna has been made for the recompense dcc gene detection kit on basis, is used for detecting dopaminergic hereditism defective.This detection has 99.9% accuracy and right and wrong invasive, need utilize a buccal swab, and doing this detection does not need special training, does not need blood sampling (other DNA detection needs).Swab is with postal delivery or anxious poorly deliver to specified DNA laboratory and detect.At this special DNA detection, there is exclusive contract inventor and DNA identification experiment chamber, northern University of Texas health science center, and the result can obtain (assessment of " state " also needs 24 to 48 hours if desired) within 72 or 96 hours.

At second edition " mental sickness diagnostic and statistical manual " (Diagnostic and Statistical Manual of Mental Disorders) that American Psychiatric Association publishes (DSM-II) in (1968), the catalogue of diagnosis is " child's how moving reaction ", in the third edition of this book of publishing in 1980, this part is divided into two parts, is mixed with many moving attention deficit hyperactivity disorders (ADD-H) and is not mixed with many moving attention deficit hyperactivity disorders (ADD).

The revised edition of the third edition " mental sickness diagnostic and statistical manual " 1989 is published, and in this version, American Psychiatric Association merges this two parts catalogue again, the diagnosis catalogue be " attention deficit---move obstacle " more (ADHD).

In the 4th edition " mental sickness diagnostic and statistical manual " publishing in 1994, three diagnostic programs have been listed: " attention deficit/how moving obstacle combination type "; " attention deficit/how moving obstacle Inattention advantage type "; " attention deficit/how moving obstacle moves-impulsion advantage type more ".

In " goodness of fit ", great difference is arranged between " being mixed with many moving attention deficit hyperactivity disorders " of the third edition and " attention deficit hyperactivity disorder " of third edition revised edition.Diagnostic criteria constantly changes in the different version of " mental sickness diagnostic and statistical manual " or the revised edition, as if lack significant concordance in each diagnostic criteria, this has shown that the suitability that constitutes the evaluation index of diagnostic criteria in any version or revised edition is limited.This in other diagnostic means too, " child diagnostic meet program " (Costello etc. for example, 1983) and " child and teenager diagnosis meet " (Herjanic and Campbell, 1997), based on the third edition " mental sickness diagnostic and statistical manual ", abundant (Newcorn etc., 1989) inadequately in new diagnosis system, the selection of main body considered.

Attention deficit hyperactivity disorder is by the children disease of diagnosing the most widely.See that from the teeth outwards the diagnosis of this situation describes that to have experienced so many variations be stranger, yet the symptom of this disease not in institute if having time or (Brown, 1986) that all occur under the environment.Only there is 20 percent the juvenile patient of ADHD symptom (Sleator and Karowe, 1969) when department of pediatrics is checked, to occur.How moving many children are owing to consulted shrink is reported as, and becomes quiet when accepting psychological assessment in testing environment and cooperation (Tobissen and Karowe, 1969).If the unique pattern of an attention or cognitive dependency is arranged, it can distinguish ADD with other diseases, and the ADD state will further be proved conclusively (McGee etc., 1989) as a kind of disease so.

Because the transmutability of the symptom of performance, ADHD SAS designs at father and mother, teacher and other answers of being familiar with the people of children's behavior manner.Thereby these contact the most frequent the tightst people with the child be the behavior that the child was assessed and reported in the position that is in a best.The report of document shows the direct observation data height correlation of teacher to child and teenager's the same clinical evaluation of ranking, neuro physiology assessment and classroom.

ADHD SAS grows up for an assessment of ADHD symptom level fast is provided, and a child on one's body through the checking of a period of time.This estimates scale is not the conclusive diagnostic tool that is used for diagnosing child's fever or acrokinesia.Make one accurately, the diagnosis of clear and definite ADHD, comprise degree, performance and boundary, need the consultation of doctors of the profound level that the doctor of several occupational areas participates in, comprise fields such as developmental pediatrics, department of pediatrics neurological, Division 12. Clinical Psychology, neuropsychology, language, specific education and occupational therapy.This scale neither be used for substituting profound work up, and this work up needs the professional doctor's in above-mentioned each field joint efforts.So this scale is not the evaluation of psychology, nerve, spirit, neural psychology and pediatrics etc.

The professional of many frequent participation children and teenager work needs the standard that can carry out fast, mark and explain, and it can show existence and the level thereof of ADHD.Because the sickness rate data of having reported are arranged, the part of this needs is obvious.The sickness rate of this state of an illness from less than one of percentage of all school age populations to 20 percent (August and Garfinkel, 1989).By " mental sickness diagnostic and statistical manual " the 4th edition, the popular estimation of ADHD accounts for 3% to 5% of school age population, and teenager and adult's data is limited.

In many cases, father and mother and teacher have noticed the distinctive behavior of ADHD, and this is interpreted as laziness, stubbornness, shortage interest, angry and immature etc.Sometimes the personnel of teachers marquis, referring physician, main health doctor and other health and education sector when explaining ADHD child's behavior, think a kind of undisciplined or come from the performance of part father and mother's sense of discipline difference.Because the ADHD child continues consistent reveal any symptoms, so the symptom that father and mother describe is interpreted as being derived from the father and mother of the excessive parental responsibility heart, rather than child's dysfunction (because the child shows very quietly in doctor's office, behavior is also very proper).When father and mother or teacher by these hypothesis or explain that when acting, they have worsened the reaction of child to potential problems.The important value of ADHD SAS is its convenient economic practicality, makes consultant, teacher, family doctor, pediatrician, neurosurgeon and other professional doctor, easily determines children's attention defective and/or many moving levels fast.Thereby professional doctor can confirm or refuse father and mother or hypothesis or the explanation relevant with children's behavior that other people make.

ADHD SAS is a scale that comprises 43 projects, and it can be answered by teacher or father and mother, scoring in 15 minutes, analysis and explanation.This scale is an objective scoring instrument, and the people that it allows father and mother or other be familiar with children's behavior answers the performance and the frequency of children's behavior, attitude or emotion.This scale requires to answer a question objectively on 4 Likert scales.The range of choice on scale is to " most of or whole time " from " not having or little time ".

ADHD SAS is the examination design of an efficient low cost, and it is carried out easily and marks, and under professional's supervision, can be utilized by housebroken technical staff and Additional Specialty personnel.Yet ADHD SAS can not replace the complete skilled clinical evaluation to the child, because this form of estimating design is the form that father and mother report, it is subjected to conscious or unconsciously to twist especially easily.Because this reason and other special restrictions that will be described in detail below, ADHD SAS should only be used as the prompting that attention-deficient is transported symptom more, and it should not be used for instructing individually formulates therapeutic scheme or treat route.This examination design is as the replenishing of clinical skilled judgement, rather than substitutes them.

The core attention deficit that is mixed with many moving attention deficit hyperactivity disorders and is not mixed with many moving attention deficit hyperactivity disorder patients has difference (Lahey, etc., 1985).synthetically refer to attention time of concentration, easily when forgetfulness, the difficulty of acting in accordance with regulations and aspect such as immature, teachers think that comparing the two with matched group has shown same attention-deficient.Yet, in their research, more not being mixed with many moving attention deficit hyperactivity disorder and matched groups, that ADHD is described to is carefree, inwholwe-hearted, impulsion, it is that just answer a question and blowzy not think.Consider their discovery (DeLamater and Lahey, 1983; Cahey 1994), can depict the picture that a pair has two groups of different children populations of attention problem.

The formation of ADHD SAS is suitable for estimating the advantage type (August and Garfinkel, 1989) of ADHD, also can estimate the aggregate level of ADHD disease symptoms.The result of ADHD SAS reflects the existence and the degree of overall attention-deficient-hyperkinetic syndrome shape (ADHD, associative form), attention deficit symptom and hyperkinetic syndrome shape.

When this scale detected the ADHD symptom, how the degree that the result also can point out these symptoms was very light, slight, moderate, seriously or extremely serious.

ADHD SAS is used to allow father and mother describe child and teen-age behavior more than 4 years old.This scale is designed in internist's the office, the meeting with or father and mother-teacher-consultant's places such as meeting of psychotherapy doctor and father and mother.This screening method is all valuable in a lot of environment.

Yet uncooperative father and mother, be hostile to, can not link up, easily twist or to such an extent as to his or her thought is too chaotic answers in the time of can not correctly reflecting the experiencing of father and mother, ADHD SAS can not use.In addition, the impaired people of the people that the language competence is lower owing to lack the senior class reading level, people, neural psychology that ESL and level is limited, moderate have any problem to finishing scale to the people of height hyponoia.

ADHD SYS can easily be carried out by housebroken Additional Specialty personnel and technician and mark.Yet ADHD SYS uses and the ultimate liability of explanation should be born by the people with senior clinical experience and technology.Before carrying out ADHD SAS, user is theoretical principle, the method for structure, the characteristics of psychological test and the particular restriction that describes in detail in this this handbook of familiar with understanding scale comprehensively.User also should be prepared to make about the clinical judgment of effectiveness as a result of the scale in certain environment in addition, in this environment, its use need replenish test material, comprises medical condition about the child, in the behavior of school and and behavioural information during the getting along of classmate.

In order to assist in ensuring that suitable use, ADHD SAS user also should be familiar with and observe the use standard of being stipulated the detection of (1994) by American Psychiatric Association, perhaps reviews the basic reader of detection and evaluation areas.Shortage before using this scale, should be read the pertinent literature in this field about the people of the clinical training of youthful evaluation of attention deficit hyperactivity disorder and case disposal.Several pieces of goodish articles about attention deficit hyperactivity disorder (for example, Rourke, 1985 are arranged; Rourke, 1989; Rourke etc., 1983; Rourke etc., 1986).

In clinical still research, use ADHD SAS and all should observe occupation and the moral code (1981) that APA proposes.The same with any assessment process that concentrates on the child, ADHD SAS can not use before one of father and mother that do not obtain the child agree.In addition, user should also be noted that the verity that guarantees the result and their use is defined in the professional and " need know ".Should concentrate on the content of the properties of attention deficit hyperactivity disorder when exchanging with child or its head of a family, and should not concentrate on or report the analysis of answering with regard to special item with regard to the scale result.Whenever explain scale as a result the time all should help father and mother to understand and be described in detail the result, also to consider understanding and his or she the current emotional status of individual to attention deficit hyperactivity disorder.

Identify that attention deficit hyperactivity disorder is the task of a complexity, need clinical sensitivity and complete knowledge about the clinical of neuropsychology and learning disability and research.ADHD SAS can not independent utility, other diagnostic method, for example pediatrics evaluation, the evaluation of department of pediatrics neurological, the evaluation of brain visualization, computer assisted E.E.G. or BEAM, neuropsychology evaluation, PASCAL evaluation PASCAL, psychiatry evaluation etc. should be used for replenishing, proving conclusively and study testing result.ADHD SAS also has many special restrictions, should remember these when explaining testing result.At first, the purpose of this scale is not covered especially, thereby when the individual finished scale, scoring can be suffered to have a mind to or twist unintentionally.The second, scale assessment to be she or he the child that reports of father and mother putting the assessment of behavior sometime.

Attention deficit-how the symptom of moving obstacle is not to occur constantly, neither all occur in all environment, thereby ADHD SYS may not predict the of short duration variation (Brown, 1986) of ADHD symptom level exactly.

It is a pen or pencil and scale list that the needed storewide of ADHD SAS is implemented in the execution of ADHD SAS.The scale list is filled in jointly by child's father or mother or by two people, and its requires the information determined, comprises the factor of the socio-demographic of selection.Include 43 test items in the front and back of this list, and with the explanation of these projects of answer and the place of fill out answers.

After definite demographic information was inserted the top of ADHD SAS scale list fully, the tester should do following explanation: some statements are arranged here, and it can help me to understand your child's behavior better.I want to allow you read each statement, point out each statement the time total amount which be real, and in suitable column, section out with " X " symbol.For example, think of this problem, " it is fidgety to be that trick is touched? " which, your child is fit to, and is " not having or little time ", " some time ", " quite a long time ", still " most of the time "?

When the tester does these explanations loudly, be also pointed out that this project and four each that answer in the column.After child's father and mother had answered, the tester marked " X " at suitable place, space and says: " ask you to finish remaining project now, if problem is arranged, must allow me know.”

Sometimes the people who does test may not understand a speech or a notion.If father and mother have any problem to understanding certain special project, the tester should make neutral explanation as much as possible.For example, if father and mother do not understand " being on tenterhooks " this speech in " be trick is touched fidgety " the words, the tester should say: " this is to ask the child that you think you has how long be difficult to keep the trick peace and quiet.”

It is very important developing and keep the emotion harmony with the people who finishes scale.From psychologic viewpoint, setting up enough harmonious relationships is necessary to reduce answering the total amount of deliberately twisting, and especially refusal at need.Attitude when any comment that the answerer did all may be filled in scale to assessment is helpful, no matter its useful statement of whether his or she child being taken action and explaining.

What the scoring ADHD SAS of ADHD SAS need mark and explain only is the scale list.To notice that at first each project all has item number in ADHD SAS scale list for ADHD SAS scoring, and be to detect attention deficit or many moving codes with indication.For example, H01 detects first of moving more; D04 is the 4th of detection attention deficit; Also have and indicate " * " number before the code of about 20% project.The scoring model of these projects is equilibrated, and purpose is to prevent that the answerer from entering a kind of scoring model.Except that the project that has " * ", all projects are marked as follows: " not having or little time " got 1 fen, and " some time " got 2 fens, and " considerable time " got 3 fens, and " most of or whole time " got 4 fens.Those project scores that have " * " in contrast, promptly " do not have or little time " 4 minutes, " some time " 3 minutes, " considerable time " 2 minutes, " most of or whole time " 1 minute.Next step is the score addition of the project of " D " with code, and the total points of subscale of decision attention deficit hyperactivity disorder is inserted these original scores the blank space after " the original total points of attention deficit hyperactivity disorder scale " then.

With code is the score addition of the project of " H ", and the total points of the many moving obstacle subscales of decision are inserted these original scores the blank space after " how the moving original total points of obstacle scale " then.Under abridged table ADHD symptom level, mark and draw the score of your record.After marking and drawing initial data, can automatically original score be converted to the conversion score, make you can directly see attention deficit hyperactivity disorder, the level of moving obstacle and attention deficit hyperactivity disorder how.Can note you to the impression of particular patients ' or family or to the guidance of its action at comment and proposal part.

Attention deficit assessment scale that the principle of ADHD SAS and theoretical basis have been published or existing is only quoted the diagnostic criteria on the latest edition " mental sickness diagnostic and statistical manual " of being published by American Psychiatric Association.Yet this method applicability is limited, because it is free restriction.Should be noted that not only significant variation has taken place the explanation along with the past initial symptom of time, does and in fact problem is arranged here: this obstacle exist? be if exist, to exist as just attention deficit? does perhaps attention deficit interrelate with move more sometimes? it should be noted in the top review of literature that present understanding situation is constantly to change.The inventor believes the effectiveness of this table, and believes that this obstacle is general, extensive distribute and somewhat popular teen-age situation.The method that the diagnosis ADHD of a first-selection is arranged here.

In in the past 30 years with regard to attention deficit-how the various symptoms and the importance thereof of moving obstacle have been done considerable discussion.Some people with him to the diagnosis of the moving obstacle of attention deficit-how with understand " the mental sickness diagnostic and statistical manual " published with American Psychiatric Association and connect, because the continuous variation of notion, they are affected more or less easily, in the constructor that detects especially like this.According to (Newcorn etc., 1989), the suitability of the test and appraisal scale that constitutes for the diagnostic criteria of any version that adapts to " mental sickness diagnostic and statistical manual " or revised edition is limited.About test and appraisal design based on the diagnostic criteria of " the mental sickness diagnostic and statistical manual " of latest edition, (Newcorn etc., 1989) emphasize " diagnostic tool of structure; ' to child's diagnosis meet program ' (DIDC) (Costello; 1983) and ' child and teen-age diagnosis are met ' (DICA) (Herjanic and Campbell; 1977) for example; be based on the standard of the third edition " mental sickness diagnostic and statistical manual ", can not in new diagnostic system, fully take into account the selection of main body.”

The project alternatives of ADHD SAS and effectiveness consider that the project of ADHD SAS is to get the principle of technical literature narration during the decade from the past two.After the review technical literature, all principles that is supported and approves of and theoretical construct-their explain or describe attention deficit or the many moving bases-behavior that is converted into is stated.Whether for example, distractibility is the main factor that a quilt is extensively mentioned, thereby just has relevant for the project of distractibility, as " diverting one's attention easily ", " finishing the work that has begun ".

The dependency of ADHD SAS scale with other scale determined in the standardization of ADHD SAS, as if perhaps use teacher's or psychologist or the internist's or father and mother's judgement as a comparable standard, these traditional standardized methods there is no need.The examination criteria that the inventor does not find is used for detecting the effectiveness of ADHD SAS.The method that scale makes up is to be based upon on the effectiveness of scoring, defines because the operation of the structure that proposes in the technical literature in two more than ten years is in the past weighed in this scoring.

ADHD SAS is used for detecting the constituent element that has been used to define attention disorders.Scoring is to be used for detecting the symptomatology total amount that proved by individuality and by the relation between the total amount of the upper limit of scale proof.

Estimate individual according to following yardstick.If 40% of the attention deficit symptom total amount of not enough this a scale detection of people so just is interpreted as " not showing the symptom that attention deficit hyperactivity disorder is arranged "; If its value so just is interpreted as " show extremely slight attention deficit symptom is arranged " between 41% to 51%; If its value so just is interpreted as " show slight attention deficit symptom is arranged " if its value between 65% to 77%, so just is interpreted as " show moderate attention deficit symptom is arranged " between 52% to 64%; If its value so just is interpreted as " show severe attention deficit symptom is arranged " between 78% to 90%; If its value so just is interpreted as " show utmost point severe attention deficit symptom is arranged " between 91% to 100%.

Can also estimate individual according to following yardstick.If 40% of the how moving obstacle symptom total amount of not enough this a scale detection of people so just is interpreted as " do not show many moving obstacle symptoms are arranged "; If its value so just is interpreted as " show extremely slight how moving obstacle symptom is arranged " between 41% to 51%; Its value between 65% to 77%, so just is interpreted as " show has moderate to move the obstacle symptom more " if its value between 52% to 64%, so just is interpreted as " show slight how moving obstacle symptom is arranged "; If its value between 78% to 90%, so just is interpreted as " show has severe to move the obstacle symptom more "; If its value between 91% to 100%, so just is interpreted as " show has utmost point severe to move the obstacle symptom more ".

Final individual by following yardstick evaluation.If 40% of the moving obstacle symptom total amount of the attention deficit that not enough this scale of people detects-how so just is interpreted as " not showing has attention deficit-how moving obstacle symptom "; If its value so just is interpreted as " showing has extremely slight attention deficit-how moving obstacle symptom " between 41% to 51%; Its value between 65% to 77%, so just is interpreted as " showing has the moderate attention deficit-move obstacle symptom " more if its value between 52% to 64%, so just is interpreted as " showing has slight attention deficit-how moving obstacle symptom "; If its value so just is interpreted as " showing has the severe attention deficit-how moving obstacle symptom " between 78% to 90%; If its value between 91% to 100%, so just is interpreted as " showing has utmost point severe attention deficit-how the symptom of moving obstacle ".

The explanation and the clinical practice of attention deficit-how moving obstacle symptom test and appraisal scale

ADHD SAS result's explanation

Case research

AD.HD symptom test and appraisal scale John G.Cull.Ph.D. and Kenneth Blum.Ph.D.
					Children's name: sex: age: grade: father and mother's name: school address: phone is finished scale answerer's name:
Please in the string on right side, draw check the number (√) and show or be considered to show the degree of every kind of behavior to show you	Do not have or little time	A period of time	A large amount of time	Big portion or All Time
					1. be on tenterhooks, H01 is touched in trick
2. D01 easily diverts one's attention
					3. twisting H02 when being seated
4. finish the thing * D02 that has begun
					5. the H03 that has any problem that is seated of Chi Xuing

ADHD symptom test and appraisal scale (continuing)	Do not have or little time	A period of time	A large amount of time	Big portion or All Time
					6. by the instruction D03 that has any problem that does things
7. at the medium H04 that has any problem in turn of recreation
					8. then finish a movable * D04 before another activity
9. wait in line the H05 that has any problem
					10. before having asked, problem robbing answer H06
11. quietly play the H07 that has any problem
					12. adapt to new situation * D05 easily
13. as if can not stop too much talk H08
					14. bother or interrupt other people (interrupt in the talk, take part in others' recreation) H09
15. seem unlike listening or it seems and do not accept said D06
					16. write down important thing (book, pencil, char, school appoint etc.) * D07
17. do not decide the H10 that just takes action

ADHD symptom test and appraisal scale (continuing)	Do not have or little time	A period of time	A large amount of time	Big portion or All Time
					18. finish char or need directly supervision D08 when appointing
Read (comprising reading of smi request) D09 19. escape with amusement
					20. only feel setback * D10 because of difficulty
21. very easily excited H11
					22. obscure detail D 11
23. say unsuitable H12
					24. at home with school in arrange very good * D12
25. can not act D13 on schedule
					26. escape writing D14
27. painful and (not being) D15 that does things lentamente and work because mental retardation
					28. ask repeatedly or needs guidance or suggestion D16 repeatedly
29. not obedient D17
					The activity * H13 30. sit silent
31. whether (or once) damages H14 such as toy
					32. too much in bed action H15

ADHD symptom test and appraisal scale (continuing)	Do not have or little time	A period of time	A large amount of time	Big portion or All Time
					33. run or climb to H16 on some objects too much
34. the rapidly violent D18 of emotion changes
					35. make unsuitable hot-tempered sound or comment H17
The very easily angry H18 36. become
					37. to adapting to the new situation D19 that has any problem
When 38. 2 activities that you like are carried out simultaneously, the very proper * H19 of behavior
					39. can not well regulate in the place of making a lot of noise, have much to do, tend to " madness " H20
40. make great efforts very much, but the still careless H21 of work
					41. do not have quiet gentle environment, D20 can not focus one's attention on
42. do not have well-regulated environment, D21 can not concentrate one's energy
					43. because too active, can not with the nurse H22 that stays in

44. when visit friend or relatives, (once) is too active, so that can not quiet H23
							The original score of notice shortage obstacle scale of filling in the air of please not wanting below amounts to: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ how the original score of moving obstacle scale amounts to: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ the original score total of ADHD symptom measurement Yuan: _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _
??	The wide figure of ADHD symptom test and appraisal scale
															ADHD symptom level
	The lightest	Slightly	Moderate	Severe	Extremely
							Attention deficit	30++++++38	39++++++50	51++++++65	66++++++76	77++++++84
Many moving obstacles	34++++++42	43++++++56	57++++++71	72++++++83	84++++++92
							Overall ADHD symptom	63++++++80	81++++++106	107++++++136	137++++++159	160++++++176

Comment and suggestion

The self-appraisal scale is please filled in before taking POLYTROL
		Name: age: DOB: sex:
Address: phone: fax
		Email: date:
Physical trait: height: body weight: blood pressure :/resting heart rate
		Marital status: married _ _, divorce _ _, live alone as a widow _ _, separation _ _, how long pointed out:
Education (filling out top level): senior middle school's educational background _ _, institute _ _, commerce or technical school _ _, university degree _ _, graduate degree _ _, doctorate _ _
		In the form below, please in descriptions that all are fit to, make the mark form from " 1 " (expression does not have or is inapplicable) to " 5 " (problem of representation is quite serious)
Be fit to or be fit to your behavior now	Be fit to or be suitable for your kinsfolk's behavior now
		?????1?????2????3????4????5	???????????????????????????1?????2?????3?????4?????5
Excessive drinking _ _ _ _ _ _ _ _ _ _	Excessive drinking _ _ _ _ _ _ _ _ _ _

Crack/cocaine addiction _ _ _ _ _	Crack/cocaine addiction _ _ _ _ _
						The carbohydrate gluttony _ _ _ _ _	The carbohydrate gluttony _ _ _ _ _
Nicotine application or abuse _ _ _ _ _	Nicotine application or abuse _ _ _ _ _
						How moving _ _ _ _ _	How moving _ _ _ _ _
Sexual activity too much _ _ _ _ _	Sexual activity too much _ _ _ _ _
						The pathologic violence _ _ _ _ _	The pathologic violence _ _ _ _ _
Pathological gambling _ _ _ _ _	Pathological gambling _ _ _ _ _
						The Tourette obstacle _ _ _ _ _	The Tourette obstacle _ _ _ _ _
Autism _ _ _ _ _	Autism _ _ _ _ _
						I select to buy this product because I: excessive drinking or drinking problem _ _, crack/cocaine addiction _ _, the carbohydrate gluttony _ _, smoking history _ _, how moving _ _, the height stress _ _, sexual activity too much _ _, the pathologic violence _ _, pathological gambling _ _, the Tourette obstacle _ _
Read over each, be fit to your making a mark in the air then	Never	The less time	Some times	Considerable time	All Time almost
						I thirst for a kind of material or activity or behavior
I change my emotion or loosen with resembling things such as food, wine
						For regulating pressure and problem, I pretend not have any problem; I ignore problem

I abuse coffee, aspirin, medicine and wait and attempt to make me to tackle (something) better
						I have a kind of sensible attitude, and I complain and criticize
Emotion	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						The skill of here director's thing
My recovery capability (ability of from pressure and trouble, breaking away from)
						I control the serious hope or the demand of other people or situation
My spontaneity (not by the ability of " supervision " or anti-punctual action)
						My steady and calm ability of doing things
The ability of my calm action when the pressure and the state of emergency
						I under the pressure or the state of emergency to other people patient ability
I am to noise on every side and chaotic tolerance
						I am to the tolerance of glistening light and troubled waters on every side
My emotion is felt bad number of times
						The ability that I all can concentrate one's energy in all types of environment

Physiology	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						The ability that I fall asleep
The ability that I am sleeping whole evening
						The ability that I have a good sleep and sink
I dream the number of times of happy dream
						I remember the ability that I have a dream
Clear-headed and energetic number of times when I awaken
						My daylong energy level
My energy level when finishing in one day
						My sexuality
My libido
						The level that I am uncontrollably angry
The level that my calmness is loosened
						My impulsion
The number of times of my headache
						The number of times that feels like jelly in my myalgia, pain, misery, joint

Total background level of my pain
						The stability of my appetite (approximately identical appetite is arranged everyday)
The number of times of my nerve stomachache
						My the tendency that accidents happened
I feel uncomfortable amount
						Spirit	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
The sensation of my inanition
						I have lost the sensation of meaning of life
I suspect that I control oneself and the sensation of the meaning of my thing of being done
						I feel that I resemble martyr's (perceptual image domestic animal product) number of times
I find that I control oneself and wish or seek a number of times that the solution (method) of " magic power " is arranged
						I have a few " stiff " and merciless number of times to other people
I lose the sensation of direction
						Need proof myself
I am cynical (be sick of, pessimistic, suspect)

I am (cold, shortage is concerned about) indifferently
						Memory	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
My impermanent memory
						I recall rapidly (remembering the ability on a word, name, incident, date etc.)
The ability that I concentrate one's energy and learn
						I remember the ability of the thing that I read or hear
I am the ability of study easily
						I am to the interest of reading
I am to the operation of studying us or the interest of work

R.D.S. grade form John G.Cull, Ph.D. with Kenneth Blum, Ph.D. (adult's form)
	Name: date of birth: address: sex:
Marital status: married _ _, divorce _ _, live alone as a widow _ _, separation _ _, how long pointed out:
	Education (filling out top level): senior middle school's educational background _ _, institute _ _, commerce or technical school _ _, university degree _ _, graduate degree _ _,

Please point out to be fit to draw a circle " 1 "=be not suitable on all numerals that is described in expression intensity of you and your kinsfolk, " 2 "=slight intensity, " 3 "=moderate strength, " 4 "=serious intensity, " 5 "=extremely strong
		Be suitable for your behavior now or	Be suitable for your mother's behavior now or
Drink or indulge in excessive drinking 12345	Drink or indulge in excessive drinking 12345
		Autism 12345	Autism 12345
Carbohydrate gluttony 12345	Carbohydrate gluttony 12345
		Controlled have an aggressive criminal behavior 12345	Controlled have an aggressive criminal behavior 12345
Long-term lower back portion pain 12345	Long-term lower back portion pain 12345
		Use or abuse cocaine 12345	Use or abuse cocaine 12345
Because of aggressive criminal behavior is determined a crime 12345	Because of aggressive criminal behavior is determined a crime 12345
		The agitator 12345 of domestic violence	The agitator 12345 of domestic violence
The victim 12345 of domestic violence	The victim 12345 of domestic violence
		Many moving 12345	Many moving 12345
Suffer from abuse 12345 mentally	Suffer from abuse 12345 mentally
		Use or abuse nicotine 12345	Use or abuse nicotine 12345
Pathological gambling 12345	Pathological gambling 12345

Pathologic violence 12345	Pathologic violence 12345
		Suffer from abuse 12345 on the health	Suffer from abuse 12345 on the health
Stress 12345 after the wound	Stress 12345 after the wound
		Premenstrual syndrome 12345	Premenstrual syndrome 12345
Sexual activity too much 12345	Sexual activity too much 12345
		Property is subjected to cruel 12345	Property is subjected to cruel 12345
Tourette obstacle 12345	Tourette obstacle 12345

Be suitable for or be applicable to your father's behavior	Be suitable for or be applicable to compatriot's behavior
		Drink or indulge in excessive drinking 12345	Drink or indulge in excessive drinking 12345
Autism 12345	Autism 12345
		Carbohydrate gluttony 12345	Carbohydrate gluttony 12345
Controlled have an aggressive criminal behavior 12345	Controlled have an aggressive criminal behavior 12345
		Long-term lower back portion pain 12345	Long-term lower back portion pain 12345
Use or abuse cocaine 12345	Use or abuse cocaine 12345
		Because of aggressive criminal behavior is determined a crime 12345	Because of aggressive criminal behavior is determined a crime 12345

The agitator 12345 of domestic violence	The agitator 12345 of domestic violence
		The victim 12345 of domestic violence	The victim 12345 of domestic violence
Many moving 12345	Many moving 12345
		Suffer from abuse 12345 mentally	Suffer from abuse 12345 mentally
Use or abuse nicotine 12345	Use or abuse nicotine 12345
		Pathological gambling 12345	Pathological gambling 12345
Pathologic violence 12345	Pathologic violence 12345
		Suffer from abuse 12345 on the health	Suffer from abuse 12345 on the health
Stress 12345 after the wound	Stress 12345 after the wound
		Premenstrual syndrome 12345	Premenstrual syndrome 12345
Sexual activity too much 12345	Sexual activity too much 12345
		Property is subjected to cruel 12345	Property is subjected to cruel 12345
Property sadist 12345	Property sadist 12345
		Tourette obstacle 12345	Tourette obstacle 12345
I thirst for a kind of material or activity or behavior
		I change my emotion or loosen with resembling things such as food, wine
For regulating pressure and problem, I pretend not have any problem;

I abuse coffee, aspirin, medicine and wait and attempt to make me to tackle (something) better
						I have a kind of sensible attitude, and I complain and criticize
Emotion	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						The skill of here director's thing
My recovery capability (ability of from pressure and trouble, breaking away from)
						I control the serious hope or the demand of other people or situation
My spontaneity (not by the ability of " supervision " or anti-punctual action)
						My steady and calm ability of doing things
The ability of my calm action when the pressure and the state of emergency
						I under the pressure or the state of emergency to other people patient ability
I am to noise on every side and chaotic tolerance
						I am to the tolerance of glistening light and troubled waters on every side
My emotion is felt bad number of times
						The ability that I all can concentrate one's energy in all types of environment
The ability that I fall asleep

The ability that I all sleep whole evening
		The ability that I have a good sleep and sink
I dream the number of times of happy dream
		I remember the ability that I have a dream
Clear-headed and energetic number of times when I awaken
		My daylong energy level
My energy level when one day finishes
		My sexuality
My libido
		The level that I am uncontrollably angry
The level that my calmness is loosened
		My impulsion
The number of times of my headache
		The number of times that feels like jelly in my myalgia, pain, misery, joint
Total background level of my pain
		The stability of my appetite (approximately identical appetite is arranged everyday)
The number of times of my nerve stomachache

My the tendency that accidents happened
							I feel uncomfortable amount
Spirit	Far below on average	Be lower than average	On average		Be higher than average	Far above on average
							The sensation of my inanition
I have lost the sensation of meaning of life
							I suspect that I control oneself and the sensation of the meaning of my thing of being done
I feel that I resemble martyr's (perceptual image domestic animal product) number of times
							I find that I control oneself and wish or seek a number of times that the solution (method) of " magic power " is arranged
I have a few " stiff " and merciless number of times to other people
							I lose the sensation of direction
I will prove the demand that I control oneself
							The attitude of I cynical (bored, pessimistic, suspection)
The attitude of I cold and detached (indifferent, shortage is concerned about)
							Memory	Far below on average	Be lower than average	On average	Be higher than average		Far above on average

My impermanent memory
		I recall rapidly (remembering the ability on a word, name, incident, date etc.)
The ability that I concentrate one's energy and learn
		I remember the ability of the thing that I read or hear
I am the ability of study easily
		I am to the interest of reading
I am to the operation of studying us or the interest of work

If the statement of the statement below the PLEASE READ CAREFULLY is correct, it is always all incorrect if symbol is pointed out its correct time degree with " T ", marks under " do not have or seldom time " " T " number

6 months in the past	Do not have or time seldom	Some times	Considerable time	Major part or All Time
					1. I once noted commiting excesses negligent mistake less than details or I in work or other activity	Adhd Inattention advantage type
2. I have any problem to continue to keep attention to concentrate in task	Adhd Inattention advantage type
					3. when directly speaking with me, I have any problem in understanding	Adhd Inattention advantage type
4. I can not act by instruction, and can not finish odd job in the family and the responsibility in the work	Adhd Inattention advantage type
					5. I have any problem to organizing affairs and activity	Adhd Inattention advantage type
6. I escape, dislikes or exempt from the task that strong going need to make lasting mental work	Adhd Inattention advantage type
					I leave about to task or movable sine qua non (as, key, glasses, file, pencil, book or instrument)	Adhd Inattention advantage type
8. I am bothered by the stimulation in the external world easily	Adhd Inattention advantage type
					9. I easily forget things in daily routines	Adhd Inattention advantage type
10. in session the time, my trick is touched to be on tenterhooks or to be sitting on the chair and is twisted	Adhd moves the advantage type more
					11. I left meeting or other occasion that need be seated always	Adhd moves the advantage type more
12. I had uneasy sensation	Adhd moves the advantage type more

13. I reach calmness and sedately play or be engaged in extra-professional activity and have any problem	Adhd moves the advantage type more
		14. other people feel that I seem always very busy or seem to have motor driving me when doing things	Adhd moves the advantage type more
15. at my hyperphasia of some occasion	Adhd moves the advantage type more
		16. before problem had been asked, I was once robbing and was answering	Adhd impulsion advantage type
17. I was once to having waited in line difficulty	Adhd impulsion advantage type
		18. before other people finished his/her word, I was robbing and am answering	Adhd impulsion advantage type
The mobility twitched 19. I had significantly (a kind of suddenly, fast, repeatedly, arythmia, mechanical mobility's activity or sounding)	The Tourette obstacle
		20. I had the sounding twitch	The Tourette obstacle
21. once made me very painful or be subjected to very big injury in social, work or other important occasion mobility or sounding twitch	The Tourette obstacle
		22. that I take advantage of is weak, threaten or coerce other people	Conduct disorder-attack humans and animals
23. I start to fight	Conduct disorder-attack humans and animals
		24. I use the weapon that may cause serious harm to other people personal safety	Conduct disorder-attack humans and animals
25. when running into the victim, I occupy its property (as plundering, stealing wallet, extortion, armed robbery etc.)	Conduct disorder-attack humans and animals

26. I am once to people's cruelty (in behavior)	Conduct disorder-attack humans and animals
		27. I am once to animal cruelty (in behavior)	Conduct disorder-attack humans and animals
28. I force other people to carry out sexual activity	Conduct disorder-attack humans and animals
		29. with the purpose that damages, I deliberately am engaged in and set on fire	Conduct disorder-destruction property
30. my other people property of wilful damage	Conduct disorder-destruction property
		31. I rush in other people house, building or the automobile	Conduct disorder-cheat or commit theft
32. in order to obtain article or benefit, or in order to play truant, I lie (for example, I " cheat " other people)	Conduct disorder-cheat or commit theft
		33. when not running into the victim, I steal some valuable things (as, pretend to be client to duck into the store and commit theft)	Conduct disorder-cheat or commit theft
34. I fly into a rage	Oppositional defiant disorder
		35. I am with other adult's arguements	Oppositional defiant disorder
36. I initiatively revolt or refuse to observe other people requirement or rule	Oppositional defiant disorder
		37. I deliberately invite the people tired	Oppositional defiant disorder
38. I be the mistake of controlling oneself or the improper censure of behavior other people	Oppositional defiant disorder
		39. I am easily enraged by other people or get angry	Oppositional defiant disorder
40. I get angry and animosity	Oppositional defiant disorder

41. I have malice or the heart of revenging	Oppositional defiant disorder
		42. I can not resist the impulsion of picking a quarrel	Intermittent explosive disorder
43. the degree of picking a quarrel that I show has exceeded the thing that causes that I pick a quarrel	Intermittent explosive disorder
		I am to some words or thing anger 44. I only pick a quarrel	Intermittent explosive disorder
45. I thirst for or like intimate relation, comprise the part as family	Split disease sample/avoidance personality obstacle
		46. I selected independent activity	Split disease sample/avoidance personality obstacle
47. I have no stomach for a bit to carrying out sexual activity with another people	Split disease sample/avoidance personality obstacle
		48. almost there is not activity can make my happiness	Split disease sample/avoidance personality obstacle
49. outside the my family front yard, I lack the close friend	Split disease sample/avoidance personality obstacle
		50. I to others praise and criticize unconcerned	Split disease sample/avoidance personality obstacle
51. I show emotion grimness, unsocial or insensibility	Split disease sample/avoidance personality obstacle
		52. I can not feel guilty	Split disease sample/avoidance personality obstacle
53. I escape the work that a large amount of human communications is arranged	Split disease sample/avoidance personality obstacle
		54. I am unwilling to associate, and remove nonego and determine that I am very favorite people	Split disease sample/avoidance personality obstacle
55. I suppress intimate contacts, because I am afraid of to be humiliated or ridicule	Split disease sample/avoidance personality obstacle
		56. I worry that the social life is criticized or is rejected	Split disease sample/avoidance personality obstacle
57. I am restrained in the occasion of new human communication, because my sensation can not be dealt with	Split disease sample/avoidance personality obstacle

58. I think that I am incompetent socially, and am not attractive or not as other people on personality	Split disease sample/avoidance personality obstacle
		59. I am reluctant to take chances or am engaged in any new activity, because they may be embarrassing	Split disease sample/avoidance personality obstacle
60. I always the gluttony carbohydrate (as, donuts, Tian Mianquan, batter, noodles, bread, sugar etc.)
		61. I feel very bad so that the gluttony again in order to feel quite a lot of to my gluttony
62. what I ate when my gluttony wants outeat than my plan
		63. I want to give up or control my gluttony always
64. I attempt to give up or control my gluttony, but unsuccessful
		65. I consider by spended time, obtain or prepare the carbohydrate that I will gluttony
66. I am poverty-stricken for the amount that I eat in social activity or workplace
		67. I escape important social activity, work or recreation because of my gluttony
Although 68. I know what this body ﹠ mind to me means, I also continue gluttony

69. my gluttony or too many 1 * every month of eating or still less; _ _ 2 * every month; _ _ 1 * weekly; _ _ 3 * weekly; _ _ 5 * weekly _ _ 7 * weekly
		70. in case I begin gluttony, even if I do not mind no matter where dangerous (when driving) yet
Although 71. my friends, family members, doctor and other professionals warned me not want gluttony, I still continue gluttony
		72. I indulge in the gluttony carbohydrate
73. in order to control, to reduce or giving up gluttony I did unsuccessful effort
		Although 74. because the influence of gluttony has caused that the problem of society and inter personal contact is (as quarrelling with the consequence of the just excessive gluttony of spouse, health status shows variation, fights) my still gluttony
75. feel that in order to reach ideal I need increase the consumption of carbohydrate
		76. when I attempted to reduce or stop gluttony, I can't peace and quiet or irritability
77. I once came the escape problem with gluttony or mood that alleviate to constrain (as helpless, evil, anxiety, sensation such as oppressive)
		78. in order to conceal my gluttony, I said lie to family members and other people
60b. I drink wantonly (as medicated beer, whiskey, gin, Little water., wine etc.)

Feel very bad so that drink again 61b. I drink to me in order to feel quite a lot of
		62b. when I drink, I drink than I plan many
63b. I think alleviating alcohol addiction or control my capacity for liquor always
		64b. I attempt alleviating alcohol addiction or control I capacity for liquor but unsuccessful
65b. I think deeply by spended time, obtain or mix the wine that I will drink
		66b. I am poverty-stricken for the amount that I drink in society or workplace
67b. I escape important social activity, work or recreation because of my drinking
		Although 68b. I know this mind ﹠ body how it feels to me, I also continue to drink
69b. I bent one's elbow 1 * every month or still less; _ _ 2 * every month; _ _ 1 * weekly; _ _ 3 * weekly; _ _ 5 * weekly; _ _ 7 * weekly
		In case 70b. I begin to drink, even if I do not mind no matter where dangerous (when driving) yet
Although 71b. my friends, kinsfolk, doctor and other professionals warned me not drink, I still continue to drink
		72b. indulging in, I drink
73b. for control, minimizing or alleviating alcohol addiction I did unsuccessful effort

Although 74b. because the influence of wine has caused that the problem of society and inter personal contact is (as arguing with regard to the consequence of excessive drinking with the spouse, health status show variation, fight) I still continue to drink
		75b. feel that in order to reach ideal I need increase the amount that I drink
76b. when I attempted to reduce or stop to drink, I can't peace and quiet or irritability
		77b. I am once with the mood of drinking the escape problem or alleviate to constrain (as helpless, evil, anxiety, sensation such as oppressive)
78b. drink in order to conceal me, I lie to kinsfolk and other people
		60c. I suck cocaine (as inhaling cocaine, taking out crack etc.)
I feel very bad 61c. take drugs about me so that in order to make sensation better, I take drugs again (cocaine)
		62c. when I took drugs, the amount of planning than me was many
63c. I once wanted to give up or control drug abuse
		64c. I make great efforts to give up or control drug abuse unsuccessfully
65c. I think deeply by spended time, obtain or the drugs that I will inhale are installed
		66c. I am poverty-stricken because of the amount of the drugs that I inhale in social activity or workplace
67b. I escape important social activity, work or recreation because take drugs

Although 68b. I know this mind ﹠ body how it feels to me, I also continue to take drugs
		69c. I take drugs is 1 * every month or still less; _ _ 2 * every month; _ _ 1 * weekly; _ _ 3 * weekly; _ _ 5 * weekly; _ _ 7 * weekly
In case 70c. I begin to take drugs, even if I do not mind no matter where dangerous (when driving) yet
		Although 71c. my friends, kinsfolk, doctor and other professionals warned me not take drugs, I still continue to take drugs
72c. I indulge in drug abuse
		73c. in order to control, to reduce or quitting drug abuse that I did unsuccessful effort
Although 74b. because the influence of taking drugs has caused that the problem of society and inter personal contact is (as arguing with regard to the consequence of excessively taking drugs with the spouse, health status show variation, fight) I still continue to take drugs
		75c. feel that in order to reach ideal I need increase the amount that I take drugs
76c. when I attempted to reduce or stop to take drugs, I can't peace and quiet or irritability
		77c. I was once take drugs the escape problem or to alleviate the mood constrained (as helpless, evil, anxiety, sensation such as oppressive)
78c. take drugs in order to conceal me, I lie to kinsfolk and other people

60d. I exceedingly gamble
		61d. feel very bad about what I gambled, so that in order to make sensation better, I am gambling again
When 62d. I gamble, my gambling than I plan more
		63d. I want to give up or control gambling always
64d. I make great efforts to give up or control gambling unsuccessfully
		65d. I think deeply by spended time, arrange, and set up the gambling that I will participate in
66d. I am poverty-stricken because of the amount of my gambling in social activity or workplace
		67d. I escape important social activity, work or recreation because of gambling
Although 68d. I know this mind ﹠ body how it feels to me, I also continue gambling
		69d. I gambled 1 * every month or still less; _ _ 2 * every month; _ _ 1 * weekly; _ _ 3 * weekly; _ _ 5 * weekly; _ _ 7 * weekly
In case 70d. I begin the gambling, even if no matter where dangerous I do not mind yet
		Although 71d. my friends, kinsfolk, doctor and other professionals warned me not gamble, I still continue gambling
72d. I indulge in gambling

73d. in order to control, to reduce or giving up gambling that I did unsuccessful effort
		Although 74d. because the influence of gambling has caused that the problem of society and inter personal contact is (as arguing with regard to the consequence of excessively gambling with the spouse, health status show variation, fight) I still continue gambling
75d. in order to reach ideal sensation, I need increase the amount of my gambling
		76d. when I attempted to reduce or stop to gamble, I can't peace and quiet or irritability
77d. I come the escape problem with gambling or alleviate the mood constrained (as helpless, evil, anxiety, sensation such as oppressive)
		78d. in order to conceal my gambling, I lie to kinsfolk and other people
60e. I excessively indulge my sexual life
		61e. feel very bad about my sexual behaviour, so that in order to make sensation better, I carry out sexual activity again
62e.
		63e. I want to reduce or control my sexual behaviour always
64d. I attempt to reduce or control my sexual activity unsuccessfully
		65e. I think deeply by spended time, arrange, set up my property opportunity
66e. I am poverty-stricken because of my sexual behaviour in society or workplace

67e. I escape important social activity, work or recreation because of my sexual behaviour
		Although 68e. I know this mind ﹠ body how it feels to me, I also continue my sexual activity
69e. I was engaged in sexual activity 2 * every month or still less; _ _ 4 * every month; _ _ 2 * weekly; _ _ 6 * weekly; _ _ 10 * weekly; _ _ more
		70e. in case I begin disposable opportunity, even if I do not mind no matter where dangerous (when driving) yet
Although 71e. my friends, kinsfolk, doctor and other professionals warned my this thing, I still continue my sexual activity
		72e. I indulge in sexual activity
73e. in order to control, reduce or stop my sexual activity, I did unsuccessful effort
		Although 74e. because the influence of sexual activity has caused that the problem of society and inter personal contact is (as arguing with regard to the sexual behaviour consequence with the spouse, health status show variation, fight) I still continue my sexual activity
75e. in order to reach ideal sensation, I need increase the amount of my property opportunity

76e. when I attempted to reduce or stop my sexual behaviour, I can't peace and quiet or irritability
		77e. I once came the escape problem with sexual intercourse or mood that alleviate to constrain (as helpless, evil, anxiety, sensation such as oppressive)
78e. in order to conceal my sexual behaviour, I once lied to kinsfolk and other people
		79. I had the memory of the misery of some things, comprised image, idea or sensation	Posttraumatic stress disorder
80. I had the dream of the misery of some things	Posttraumatic stress disorder
		81. as if some traumatic event of my sensation recur (comprise and review experience, illusion, hallucination and some pulsating Hui Ying, comprise those generations when clear-headed and when clear-headed)	Posttraumatic stress disorder
82. be exposed to symbol or during with the similar hint of some traumatic event, I have intensive mental pressure	Posttraumatic stress disorder
		83. be exposed to the place that has symbol or similar traumatic event to hint, I feel to have the reaction on the health	Posttraumatic stress disorder
84. I make great efforts to avoid and relevant idea, sensation or the talk of offending incident in the past	Posttraumatic stress disorder
		85 I make great efforts to avoid to cause my activity, place or people to the memory of unhappy incident of past	Posttraumatic stress disorder

86. I have any problem to sleeping or lasting sleep	Posttraumatic stress disorder
		87. it is fidgety or angry that I show	Posttraumatic stress disorder
88. I have any problem to concentrating one's energy	Posttraumatic stress disorder
		89. I am " watching out for " always	Posttraumatic stress disorder
90. I have a kind of reaction (I am " jittery ") of being taken aback turgidly	Posttraumatic stress disorder
		91. I am quiet and peaceful always	Posttraumatic stress disorder
92. the sensation of my unsoundness	Posttraumatic stress disorder
		93. I am sleeping easily	Posttraumatic stress disorder
94. I can sleep whole evening	Posttraumatic stress disorder
		95. I feel other people affability	Posttraumatic stress disorder
96. I have felt delicately	Posttraumatic stress disorder
		97. I feel that I am friendly to controlling oneself	Posttraumatic stress disorder
98. I like me to control oneself	Posttraumatic stress disorder
		99. I feel that I can control me and control oneself	Posttraumatic stress disorder
100. I feel that I can control my environment	Posttraumatic stress disorder

Note:

RDS test and appraisal scale impulsion/addiction/compulsion
					I. note lacking obstacle
					If the statement of the statement below the PLEASE READ CAREFULLY is correct, mark and indicate the time with " T " symbol, as incorrect, do not mark with " T " symbol down " having or little time "	Do not have or time seldom	Some times	Often or considerable time	Major part or All Time
Inattention advantage type
					1. can not notice details or in study and work or other activities, make negligent mistake
2. when working or carry out other activity, have any problem to keeping attention to concentrate
					3. others and you when directly talking, seem not listen
4. can not act by instruction and maybe can not finish the responsibility of study, family's odd jobs or working space
					5. organization task and activity are had any problem
6. escape, dislike or be reluctant to be engaged in and pay the work that continues energy
					7. lose work or movable necessary things (as the appointing of toy, school, pencil, book or instrument)

8. bothered by the stimulation in the external world easily
					9. in daily routines, easily forget things
Many moving advantage types
					Trick touch be on tenterhooks or when being seated health disorderly turn round
2. leave classroom or other occasion that need continue to be seated
					3. exceedingly run or climb (, may be limited to quietly subjective sensation) back and forth at teenager in unsuitable occasion
4. have any problem to undisturbedly playing or be engaged in extra-professional activity
					5. always in " very busy " or when action, are as " launched machine driving "
6. over-drastic speech
					Impulsion advantage type
1. robbing before problem has been asked and answering
					2. reciprocity Hou Cixu has any problem
3. interrupt or get involved other people activity (as talk or the recreation that gets involved others)
					The pII.Toureete obstacle

Though 1. not necessarily at the same time, often take place repeatedly sometimes mobility and the twitch of one or many sounding (twitch be a kind of suddenly, fast, repeatedly, non-rhythmicity and stiff motor activity or sounding)
					2. took place in one day repeatedly to twitch (normally a burst of), almost every day or surpass that discontinuity takes place in the year, not have the time of twitch to be no more than 3 months
3. in society, work or other important events, these troubles cause tangible misery or injury
					III. conduct disorder
Attack the human or animal
					1. a little less than taking advantage of, threaten or coerce other people
2. start to fight
					3. once used the weapon that can cause serious actual bodily harm to other people
4. committed theft when running into the victim (as robbing, steal wallet, extortion, armed robbery)
					5. to people's cruelty (on health)
6. to animal cruelty (on health)
					7. force other people to carry out sexual activity
Vandalize

8. with the purpose that causes serious breaking-up, deliberately be engaged in and set on fire
					9. deliberately destroy others' property
Deception or theft
					10. swarm into other people family, building or automobile
11. in order to obtain property or benefit or to escape one's duty and lie (as cheating others)
					12. the no small thing of stealing value when not running into the victim (as pretend to be client to duck into the store and commit theft, but do not force open the door, forge)
The serious regulation of disobeying
					13. although father and mother oppose that beginning is away from home evening before 13 years old
14. when living in the procuratorial family of father and mother or father and mother, have run away from home 2 whole evenings (perhaps once a very long time does not go home) at least
					15.13 began before year to play truant
IV. oppositional defiant disorder
					1. fly into a rage
2. disagree with the adult
					3. have a mind to antagonism or refusal in accordance with the requirement or the rule of being grown up

4. deliberately enrage other people
					5. to his or her mistake or uncomfortable behavior, criticize other people
6. easily enraged by others
					7. angry and indignant
8. malice or cherish hatred
					V. intermittent explosive disorder
1. the thing of the impulsion of picking a quarrel intermittently takes place to resist, and consequently causes severe attack behavior and vandalize
					2. during getting excited, the degree of picking a quarrel of performance is well beyond the ratio of the living social mentality of any burst stresser
3. the incident of picking a quarrel can not be explained preferably and not be because the direct physiological effect of a kind of material or body situation with other mental disorder
					VI. divide sample/avoidance personality obstacle
1. both be unwilling also to dislike intimacy, comprised a part as one family
					2. it is movable separately to select
3. be not interested in to carrying out sexual activity with other people

4. seldom there is activity can bring happiness
					5. except the closeest relatives, lack close friend or close friend
6. praise or the criticism to others seems unconcerned
					7. show the emotion indifference, unsocial or do not have an emotion
8. lack and regret, to injured, abused, when having been stolen thing by others' sensation it doesn't matter or think rational
					9. escape relates to the work activities that needs a lot of human communications, because fear to criticize, oppose or refusal
10. be unwilling with people's dealing, unless determine to be liked
					11. show the restriction intimacy, humiliated or make fun of because fear
12. be laden with anxiety because of criticizing or refusing in social contexts
					13. can not deal with because of sensation, be restrained in the occasion of new human communication
14. think and on personality, do not have the incapability socially of controlling oneself captivation or be not so good as other people
					15. it is be reluctant to take chances or be engaged in any new activity, poverty-stricken because they may prove
VII. material is supervised and is used obstacle

1. tolerance appears, promptly in order to reach the intoxicated effect or the effect of serious hope, need obviously to increase the consumption of material, when perhaps continuing to use the material with quantity, effect significantly reduces
					2. the symptom that withdrawal syndrome is arranged because in order to alleviate or escape its symptom, is used certain material or identical material, and withdrawal symptom has been described
3. when using material, than the amount of planning Duo or the time longer
					4. wish always or did the use that certain material was given up or controlled in unsuccessful effort
Obtain certain material, take it or the activity that from its influence, recovers etc. on spended time
					6. because the cause that certain material uses, abandon or reduce and participate in important society or recreation
7., still continue to use certain material although knowing has physiology lasting or repeatedly and psychological problems
					The material that meets foregoing description is what types of materials (all materials that meet are all sectioned out) is ethanol _ _ amphetamine (or amphetamine sample material) _ _ carbohydrate _ _ crack/cocaine _ _ heroin _ _ Fructus Cannabis _ _ nicotine _ _ other stimulus object _ _
Substance abuse (behavior not necessarily meets the standard of substance depilatory)

1. recurrent material uses and causes in work, school or family can not be fulfiled main obligation, and (poor as repeatedly absence or work performance, this uses relevant with material; Use relevant cutting classes with material, temporarily drop out of school or discharged from by school; Carelessness child and housework)
					2. the adventurous place of health is being used repeatedly material (as when being used damage, drive or move machine) by material
3. the material that has related to legal issue use repeatedly (as, because of the behavior that does not conform to rules of something qualitative correlation is caught)
					4. although the problem of continuing of having that effect by certain material causes or worsen or society repeatedly or human communication is (as the consequence quarrel of spouse with regard to excessive application something; The significant change of health status; Fight), but still continue to use certain material
The material that meets foregoing description is what types of materials (all materials that meet are all sectioned out) is ethanol _ _ amphetamine (or amphetamine sample material) _ _ carbohydrate _ _ crack/cocaine _ _ heroin _ _ Fructus Cannabis _ _ nicotine _ _ other stimulus object _ _
					VIII. pathological gambling
1. indulge in gambling (as indulge in the experience of experiencing gambling last time, stop or plan is taken a risk next time or the method for money to gamble is raised in thinking)
					2. in order to reach desirable stimulation, need increase stake
3. there was repeatedly unsuccessful effort to attempt control, reduce or give up gambling

4. when attempting to give up gambling, sitting is word or easily enraged not
					5. will gamble as a kind of method of escape problem or the oppressive mood of alleviation (as helpless sensation, guilty-feeling, anxiety, oppressive)
6. in order to reach desirable stimulation, need increase stake
					7. after in gambling, losing money in gamble, when come next time with more (he loses " recovering ")
8. for the category of concealment related to gambling activities, family members, doctor or other people are lied
					9. did illegal behavior as forging, swindle, steal or corruption for money to gamble
Since gambling make important relation, work, education or job opportunity become impaired or lose
					11. rely on others to provide money to alleviate the economic situation of the bad luck that causes because of gambling
IX. posttraumatic stress disorder
					If this people once went through once traumatic accident, his experience, witness or met some incidents in this accident, comprise genuine death, or be subjected to death threats or seriously injured or control oneself or others' health integrity is on the hazard, this people's reaction comprises in panic fear, helpless or terrified and following one
1. unexpected repeatedly remembers about the painful memories of that part thing, comprises image, idea or sensation
					2. the dream about the misery of that part thing often appears

3. feel traumatic incident recur (comprise and review experience, illusion, hallucination and some pulsating Hui Ying, comprise those when clear-headed and unclear awake event)
					4. when directly or indirectly being exposed to symbol or resembling in a certain respect the hint of traumatic event, intensive mental anguish is arranged
5. when directly or indirectly touching symbol or resembling in a certain respect the hint of traumatic event, physiological reaction is arranged
					Continue to escape the stimulation relevant and general reactive numb (before wound, not existing), point out as the statement of following three (or more) with traumatic event
1. make great efforts to avoid thought, sensation or the talk relevant with wound
					2. make great efforts to avoid activity, place or the people that can cause about the memory of wound
3. can not remember the importance of traumatic event
					4. important active interest or participation number of times are obviously reduced
5. sensation is become estranged other people
					6. the scope sensation of love (as can not have) of restriction emotion
7. feel to have an uncertain future (as not expecting the work that has, marriage, child or ordinary life)
					The lasting symptom that awakening improves (not occurring before wound) resembles two (or more) of following narration
1. sleeping or maintenance is slept difficult

2. irritability or get into a temper
					3. concentrate one's energy to have any problem
The height insomnia
					5. undue startle response
Suggestion and note

The RDS scale of testing and assessing
		I. note lacking obstacle Inattention advantage type
Total points (the highest six)
		The branch (6 * 2.5) that diagnosis needs	????15
RDS level: 15-20 moderate 21-24 severe
		Many moving advantage types
Total points
		The branch (6 * 2.5) that diagnosis needs	????15
RDS level: 15-20 moderate 21-24 severe
		Impulsion advantage type
Total points
		The branch (3 * 2.5) that diagnosis needs	????15
RDS level: 7-9 moderate 10-12 severe
		II Tourette obstacle
Total points
		The branch (3 * 2.5) that diagnosis needs	????15
RDS level: 7-9 moderate 10-12 severe
		The III conduct disorder
Total points
		The branch (3 * 2.5) that diagnosis needs	????15
RDS level: 7-9 moderate 10-12 severe
		The IV oppositional defiant disorder
Total points
		The branch (4 * 2.5) that diagnosis needs	????15
RDS level: 10-13 moderate 14-16 severe
		The V intermittent explosive disorder
Total points
		The branch (3 * 2.5) that diagnosis needs	????7
RDS level: 7-9 moderate 10-12 severe
		VI divides sample/avoidance personality obstacle

Total points (4 that each part is the highest)
		The branch (8 * 2.5) that diagnosis needs	????20
RDS level: (division sample+avoidance) 20-26 moderate 27-32 severe
		The VII substance use disorders
Substance depilatory
		Total points (the highest 3)
The branch (3 * 2.5) that diagnosis needs	????7
		RDS level: 7-9 moderate 10-12 severe
Mark the type that relies on material: ethanol G, with amphetamine correlative G, carbohydrate G, crack/cocaine G, heroin G, Fructus Cannabis G, nicotine G
		Substance abuse
Total points (the highest in 4)
		The branch (1 * 3) that diagnosis needs	????3
RDS level: 3 moderates, 4 severes
		Mark the type of the material of abuse: ethanol G, with amphetamine correlative G, carbohydrate G crack/cocaine G, heroin G, Fructus Cannabis G, nicotine G
The VIII pathological gambling
		Total points (the highest 5)
The branch (5 * 2.5) that diagnosis needs	????12
		RDS level: 12-16 moderate 17-20 severe
The IX posttraumatic stress disorder
		Total points (the 1st of first, 3 of second portions, the highest in 2 of the third parts)
Branch (1 * 3)+(3 * 2.5)+(2 * 2.5) that diagnosis needs	????13
		RDS level: 13-18 moderate 19-24 severe

R.D.S. RDS test and appraisal scale (adult with) John G.Cull, Ph.D. with Kenneth Blum, Ph.D.
		Name: date of birth on date: address: sex:
Marital status: _ _ married _ _, divorce _ _, live alone as a widow _ _, separation _ _, how long pointed out:
		Education (marking the most well educated): senior middle school's diploma _ _, some institutes _ _, commerce or technical school _ _, university's educational background _ _, graduate degree _ _,
Please point out to draw a circle " 1 "=not on all numerals that are described in expression intensity that meet you or you kinsfolk or be not suitable for, " 2 "=slight intensity, " 3 "=moderate strength, " 4 "=serious, and " 5 "=extremely strong
		Be suitable for your behavior now or	Be suitable for your mother's behavior now or
Drink or indulge in excessive drinking 12345	Drink or indulge in excessive drinking 12345
		Autism 12345	Autism 12345
Carbohydrate gluttony 12345	Carbohydrate gluttony 12345
		Controlled have an attacking crime behavior 12345	Controlled have an attacking crime behavior 12345

Long-term lower back portion pain 12345	Long-term lower back portion pain 12345
		Use or abuse cocaine 12345	Use or abuse cocaine 12345
Because of the attacking crime behavior is determined a crime 12345	Because of the attacking crime behavior is determined a crime 12345
		Domestic violence agitator 12345	Domestic violence agitator 12345
Domestic violence victim 12345	Domestic violence victim 12345
		Many moving 12345	Many moving 12345
Spirit is subjected to cruel 12345	Spirit is subjected to cruel 12345
		Use or abuse nicotine 12345	Use or abuse nicotine 12345
Pathological gambling 12345	Pathological gambling 12345
		Pathologic violence 12345	Pathologic violence 12345
Health is subjected to cruel 12345	Health is subjected to cruel 12345
		Stress 12345 after the wound	Stress 12345 after the wound
Premenstrual syndrome 12345	Premenstrual syndrome 12345
		Sexual activity too much 12345	Sexual activity too much 12345
Property indiscriminate 12345	Property indiscriminate 12345
		Tourette obstacle 12345	Tourette obstacle 12345

Be suitable for or be applicable to your father's behavior	Be suitable for or be applicable to compatriot's behavior
		Drink or indulge in excessive drinking 12345	Drink or indulge in excessive drinking 12345
Autism 12345	Autism 12345
		Carbohydrate gluttony 12345	Carbohydrate gluttony 12345
It is controlled for aggressive criminal behavior 12345 is arranged	It is controlled for aggressive criminal behavior 12345 is arranged
		Chronic lower back portion pain 12345	Chronic lower back portion pain 12345
Use or abuse cocaine 12345	Use or abuse cocaine 12345
		Because of the attacking crime behavior is determined a crime 12345	Because of the attacking crime behavior is determined a crime 12345
Domestic violence agitator 12345	Domestic violence agitator 12345
		Domestic violence victim 12345	Domestic violence victim 12345
Many moving 12345	Many moving 12345
		Spirit is subjected to cruel 12345	Spirit is subjected to cruel 12345
Use or abuse nicotine 12345	Use or abuse nicotine 12345
		Pathological gambling 12345	Pathological gambling 12345
Pathologic violence 12345	Pathologic violence 12345
		Health is subjected to cruel 12345	Health is subjected to cruel 12345

Stress 12345 after the wound	Stress 12345 after the wound
						Sexual activity too much 12345	Sexual activity too much 12345
Property is subjected to cruel 12345	Property is subjected to cruel 12345
						Property sadist 12345	Property sadist 12345
Tourette obstacle 12345	Tourette obstacle 12345
						I thirst for a kind of material or activity or behavior
I change my emotion with things such as food, wine or loosen
						For regulating pressure and problem, I pretend to have no problem
I abuse coffee, aspirin, medicine and wait and attempt to make me to handle thing better
						I have sensible attitude, and I complain and criticism (something)
Emotion	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						The skill of here director's thing
The ability that I break away from from pressure and trouble
						I control the hope or the demand of other people or situation
My self awareness be (do not supervised and defend under the ability of doing things)
						The ability that my calmness acts with composure

The ability that I act with composure under the pressure and the state of emergency
		I under the pressure or the state of emergency to other people patient ability
I am to the noise on every side and the toleration of troubled waters
		I am to the toleration of glistening light and troubled waters on every side
My emotion is felt bad number of times
		The ability that I concentrate one's energy in each sharp environment
The ability that I fall asleep
		The ability that I all sleep whole evening
I sleep well and heavy ability
		I dream the number of times of glad dream
I remember the ability that I have a dream
		Clear-headed and energetic number of times when I awaken
My daylong energy level
		My property energy level
My libido
		The level that I am uncontrollably angry
The level that my calmness is loosened

My impulsion
						The number of times of my headache
The number of times that feels like jelly in my myalgia, pain, misery, joint
						The general background level of my pain
The stability of my appetite (appetite of every day is approximately identical)
						The number of times of my nerve stomachache
My the tendency that accidents happened easily
						I feel uncomfortable amount
Spirit	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						The sensation of my inanition
I lose the sensation of meaning of life
						I suspect and to control oneself and the sensation of the meaning of the thing that I am done
I feel that I resemble a martyr (perceptual image victim) number of times
						I find that I control oneself and wish or seek a number of times that the solution (method) of " charming " is arranged
I have a few " stiff " and merciless number of times to other people

I lose the sensation of direction
						I prove the demand that I control oneself
The attitude of I cynical (bored, pessimistic, suspection)
						The attitude of I cold and detached (indifferent, shortage is concerned about)
Memory	Far below on average	Be lower than average	On average	Be higher than average	Far above on average
						My impermanent memory
I recall rapidly, and (remembering the ability on a word, name, incident, date etc.) is
						The ability that I concentrate one's energy and learn
I remember that I see or the ability of the thing heard
						The ability that I learn like a cork
I am to the interest of reading
						I am to the school curricula of studying me or the interest of my work

Embodiment 24

Trinucleotide (GGC) the repetition polymorphism of androgen receptor gene (ARO) and twitch-

ADHD, conduct disorder and oppositional defiant disorder is related in the obscene words syndrome

Many behaviors and cognitive disorder comprise ADHD, CD, ODD, antisocial personality disorder, reading difficulty, other cooperation obstacle, autism etc., and be more common in the male, is women three to five times (DSM-IV, 1994).Though be commonly referred to be because hormone and environmental factors may also comprise inherited genetic factors.If certain gene is an X-linkage, the function of this special gene just is very easy to understand.Like this, the percentage rate of the variation that relevant allelic frequency of dependence and gene cause, and suppose a dominant hereditary pattern of recessive inheritance, the chain gene of such X may account for excessive ratio in the male.Also playing an important role in the replying of testosterone as fruit gene, it is highly suitable in the ratio of male/female and works so.The gene that androgen receptor gene (AR) comes to this.It is positioned the Xq 11-12 (Migeon etc. 1981, and Brown etc. 1989) of X chromosome.

Lack in the normal allele of AR CAG and GGC repetitive sequence, with androgen receptor level and related to increasing of the sensitivity of testosterone, the inventor supposes that it may be related with one or several disruptive behavior disorder of child, especially conduct disorder and oppositional defiant disorder; These diseases of possible explanation dominant reason in the male also, in order to detect this hypothesis, the GGC trinucleotide that the inventor has detected in 267 Tourette syndrome patients and 59 normal persons (326 people) first exon of AR gene totally repeats polymorphism (Sleddens etc., 1992,1993).Because dopamine is normal relevant with aggression with 5-hydroxy tryptamine, the inventor is to influence and the dopamine D of AR gene pairs conduct disorder and ODD ₂The influence of acceptor gene (DRD2) and serotonin transporter gene (HTT) compares.This research be whether related with conduct disorder (CD), oppositional defiant disorder (ODD) or attention deficit hyperactivity disorder (ADHD) in order to determine that CAG and GGC in the human androgen receptor gene (AR) repeat the short allele of polymorphism.

Method is determined allelic frequency in 326 experimenters.This 326 philtrum, the people who suffers from the Tourette syndrome is 267, and 59 people of normal control, 237 people are the male, and 89 people are the women, are non-Hispanic white people.Detected the dependency of the short allelic heterozygote of male or short allelic homozygote of women and quantity behavior variable simultaneously with multivariate analysis of variance.The percentage rate of the difference that causes by the AR gene with linear regression analysis decision.And together by dopamine D ₂Acceptor gene (DRD ₂) and serotonin transporter gene (HTT) cause that the difference percentage rate of (the two causes separately and jointly) compares.

The experimenter is made up of 59 normal controls and 267 Tourette syndrome patients.Relating to the details of patient and matched group and the evaluation of behavior provides in the text.The inventor has detected following quantitative trait in those researchs: Inattention, impulsion property, many moving, CD, ODD, learning disorder (LD) and one learns in grade school show (GSAP).Follow in the research present research and one, the inventor has increased a quantity twitch scoring.

The GGC of AR gene repeats polymorphism and increases with described polymerase chain reaction technologies in 1992,1993 such as Sleddens.It is modal allele that Irvine equals nineteen ninety-five discovery 16 repetitions (0.57), is that 17 repetitions (0.32) and 15 repeat (0.08) subsequently.Only other allele is 10 repetitions.For study between AR allele and the individual quantity behavior scoring may be related, the AR gene is given a mark as follows: to the male:＞16 repeat=0, 〉=16 repeats=1; To the women,＞16/＞16 repeat=0; Heterozygote=0; 〉=16/ 〉=16 repeat=1.

In conduct disorder, dopamine D ₂Acceptor gene (DRD ₂) Taq A polymorphism (Grandy etc., 1989) be used to effect with this locus with the AR genetic comparison.Based on positive heterotic evidence (Comings and MacMurray, 1998) in this polymorphism, its scoring is as follows, and 11,22=0,12=1.

In the serotonin transporter gene promoter region, utilized insertion/deletion polymorphism (.1995 such as Hells, 1996).The scoring of HTT gene is as follows, SS=0, SL, LL=1 (Kauck etc., 1997; Lesch etc., 1996).

In patient TS and normal control, the possible difference of AR allele or allele class frequency is analyzed with X 2 test.Proofread and correct for fear of Bonferroni, check eight behavior scorings simultaneously with the multivariate analysis of variance method.In order to check the additive effect of three genes, generated the AR+DRD2+HTT scoring, herein the scoring of each individual gene of each individuality is added up, like this its scope for from 0 (any related gene type that does not have these three genes) to 3 (the related gene types of all three genes).With linear regression analysis AR, DRD ₂(independent or associating) accounts for the percentage rate of CD, ODD and the variation of ADHD score with the HTT gene.In order to check these three genes that the hypothesis of accumulative action is arranged, with the linear equation analytical control mean of each score, comprise that amounts to an ADHD score.

The result has detected the allelic frequency of AR.In 59 normal controls, it is as follows that difference repeats allelic frequency: 10-0.01,12-0.09,15-0.03,16-0.47,17-0.36,18-0.01,19-0.01,20-0.01.Compare the frequency of normal control group and the frequency of TS group, comprised chi-square value=19.69 of all allelic matched groups and TS group, d.f.=11, p=0.0731.In the potential related research that repeats allele length and phenotype effect, the inventor finds that the most thrifty method is that allele is divided into short a group and one long group, and makes two groups identical in size as much as possible.According to the method, the inventor is divided into AR allele 〉=16 repeating groups and＞16 repeating groups, and in patient TS, 〉=16 same matched groups of allelic frequency (0.66) (0.60) are compared does not have significant trend, x2=1.06, d.f.=1, p=30.Thereby, o'clock there is not difference in α=0.05 when matched group and TS group relatively the time.

The result of multivariate analysis of variance according to the sequence arrangement that significance successively decreases, lists in Table I.The maximum p=0.005 of the significance of difference of CD score, the significance of the score of ODD takes second place (p=0.022), and many moving scores also have significance (p=0.032).The significance of difference of twitch, GSAP and LD score minimum (p＞0.6).In order to check this preferential relevant whether existing only among the male, the inventor has compared male (n=237) and women's (n=89) result, because sample size is too little, has lost the ability of check, and the two difference is not remarkable.Yet when pressing the ordering of p value, the difference of CD all is the most significant in two kinds of sexes.

Because the variation of AR gene and sexual behaviour have potential dependency, the inventor has checked the possible dependency of AR gene with sexual behaviour score (Comings, 1994) with post hoc analytic process in the people more than 14 years old, finds not have significance.Also to have measured short allele in the women be recessive or show one's color with post hoc analytic process.Since the average of all scores of heterozygote similar in appearance to or less than＞16/＞16 homozygotes, show 〉=16 to repeat allelic effect be recessive in the women, to have only 〉=16/ 〉=16 homozygote could the demonstration effect.

Regression analysis the results are shown in table 74.To the scoring of CD, the AR gene accounts for 2.4% (p=0.005) of difference, DRD ₂Gene accounts for 1.3% (p=0.005), and HTT accounts for 0.5%.AR gene and DRD ₂Gene is united 3.3% (p=0.0009) that accounts for difference, accounts for 3.5% (p=0.0007) when adding HTT.The AR gene accounts for 1.6% (p=0.022) in the difference of ODD score.Herein, the AR gene has more comparability with the DRD2 gene of explaining 1.4% variance and 0.7% HTT.All three genes account for 3.2% different (p=0.001) of ODD score variance, and the AR gene accounts for 1.1% (p=0.053) of ADHD score variance.All three genes account for 2.7% (p=0.0027) of variance.

Whether with the add up linear result of AR+DRD2+HTT score of linear equation analytic process check, measuring score increases along with the increase of related gene type number.In α=0.05 o'clock, the score of Inattention, impulsion, many moving, ADHD, ODD and CD all has significant statistical significance.After Bonferroni proofreaied and correct, many moving, impulsions, ADHD, ODD and CD score still had significant statistical significance.

Through multivariate analysis of variance check, the allelic existence of short GGC of AR gene and CD (p=0.005), ODD (p=0.022) and move between the symptom of (p=0.023) more significant dependency is arranged.In masculinity and femininity, the dependency of CD is all maximum.The AR gene accounts for 2.4% of CD score difference.AR, DRD2 and HTT gene join together to account for 3.5% (p=0.0007) of conduct disorder score difference, account for 3.2% (p=0.001) of ODD, account for 2.7% (p=0.0027) of ADHD.

Table 73

The multivariate analysis of variance of behavior scoring

(n＝326)

A. 8 all behavior scores

Scoring F value p

CD????????????????????????8.03??????????????????????.005

How moving 5.20 0.23 impulsion property, 3.18 .075 Inattentions, 1.76 .181 twitch 0.22 .634 to ODD 5.32 .022

GSAP??????????????????????0.22??????????????????????.643

LD????????????????????????0.95??????????????????????.923

Add up to (Wilkes) 1.48 .171

B. has only male (m=237)

Scoring F value p

CD????????????????????????2.57??????????????????????.110

Many moving 1.50 .221

Twitch 1.18 .277

ODD???????????????????????1.13??????????????????????.288

GSAP??????????????????????0.38??????????????????????.534

Learn 0.34 .559 impulsion property, 0.15 .695 Inattention, 0.11 .736

Add up to (Wilkes) 1.14 .334C. to have only women (n=89)

Scoring F value p

CD????????????????????????1.63??????????????????????.205

ODD???????????????????0.61??????????????????.433

Inattention 0.54 .463

Impulsion property 0.48 .489

GSAP??????????????????0.41??????????????????.523

Many moving 0.15 .703

Twitch 0.13 .712

Learn 0.03 .853

Amount to (wilkes) 0.38 .929

Table 74

To CD, the linear regression analysis of ODD and ADHD scoring

(n=326) scoring CD AR .155 .024 2.83 .005

DRD2?????????????.113??????.013??????2.05??????.041

HTT??????????????.073??????.005??????1.34??????.183

AR+DRD2??????????.182??????.033??????3.35??????.0009

AR+HTT???????????.158??????.025??????2.89??????.0041

AR+DRD2+HTT??????.187??????.035??????3.43??????.0007ODD?????AR???????????????.127??????.016??????2.31??????.022

DRD2?????????????.119??????.014??????2.17??????.031

HTT??????????????.086??????.007??????1.57??????.118

AR+DRD2??????????.167??????.028??????3.06??????.0024

AR+HTT???????????.146??????.021??????2.66??????.0081

AR+DRD2+HTT .181 .032 3.32 .0010 scoring ADHD AR .106 .011 1.93 .053

DRD2???????????.091????.008????1.64????.101

HTT????????????.109????.012????1.97????.049

AR+DRD2????????.134????.018????2.44????.015

AR+HTT?????????.146????.021????2.66????.0081

AR+DRD2+HTT????.165????.027????3.02????.0027

Embodiment 25

Chromium picolinate replenishes the influence to organization: one is to contrast at random with the placebo

Double blind experiment

Preface: the inventor also attempts to answer the problem of SOME METHODS.At first; can additional CrP influence the absorption of heat and increase the consumption of heat by increasing metabolism or daily routines level by influencing appetite? second; if inventor's control is also got rid of the heat absorption of experimental group and matched group and/or the difference of energy expenditure; can also obtain same result? the 3rd; detect method (as the double energy X-ray absorption measuring method) that health forms when testing with other that more accurately and still less depends on experimenter performance than test under water; can these results repeat? the 4th; although three all basic test all similar that research group is formed about human body; but seen in other research to the high relatively rate that withdraws from (Anderson; 1995) (29.7%) is offset the result by the selectivity loss? if with reducing the rate that withdraws from someway or if with the statistical analysis of " deliberately processing ", these same results can also occur?

In order to answer these problems, the inventor has taked some measures to control the difference that body movement and heat are taken in, and uses the composition that the DEXA method is measured health, uses a kind of method and reaches and be close to perfect compliance and finally finish test.

Material and method have been studied 130 patients altogether in this research, have 122 (93.8%) to finish all detections, and wherein the male is 17, and the women is 105, and the mean age is 42.3 years old.The patient is recruited from the health of the San Antonio of Texas and Xiu Sidun and sport club by fitness specialist and sales force in the research, these coaches and sales force offer the clubber with the information of relevant research, they or take part in person or introduce its kith and kin participation.Under most situation, in the carrying out of research, employ the fitness specialist and supervise the experimenter, follow the tracks of in the material amount that their level of activation of record and heat are taken in weekly to guarantee them, and finishing final test.Before filling in letter of consent, all experimenters all are required to seek advice from their private doctor.

A large amount of studies show that DEXA can accurately measure in the meat and spoil in fat and content (Evans, 1989 of lean meat; Evans and Meyer, 1992; McCarty, 1993), have dependency highly with the actual skeleton weight of neutron activity analysis method mensuration and the total content of whole body calcium, measure whole body skeleton content of mineral substances with the neutronic analyse method, its error rate is less than 1% (Felig, 1975).Show also that simultaneously it is a kind of accurate method (.1993 such as Page that the health of assessing fat and non-obese patient is formed this; Eckel etc., 1992).Original research about DEXA method accuracy is in the news in nineteen ninety (Mooney and Cromwell, 1993) and is studies confirm that (Page etc., 1993 by subsequently three; Hasten etc., 1994; Evans, 1989), though show DEXA and weigh under water, dependency (Page etc., 1992) that heavy hydrogen dilution and the total potassium of whole body have height, the mistake in DEXA detects is less than half of the mistake of using whole body moistures or detecting under water.The coefficient of variation (CV%) to the DEXA of LunarCorporation has following report: the quality=500g ± 2.5g of fat; FFM=600 ± 1.3g; Total tissue quality=400g ± 0.6g.Except being used to estimate the variation of clinical disease, the reliability of DEXA makes it can monitor short-term limit food and/or motion to influence (Page etc., 1993 local and the whole body composition; Eckel, 1992).In one piece of nearest summary about DEXA, the author concludes that DEXA the most accurately analyzes one of instrument that health forms (Lindemann etc., 1993) so far.

The three-compartment model that DEXA provides health to form: fat, lean tissue quality and skeleton inanimate matter content.Method is the consistent light beam that obtains a branch of stable, dual energy with the constant voltage energy of 78kVp and K-edge filter (cerium), and its effective energy reaches 40keV to 70keV.This Shu Guang carries out a series of transversal scanning from top to bottom, is spaced apart 1 centimetre; The area of scanning approximately is 200 centimetres of 60 cm x.120 pixel elements, the about 5 * 10min of each pixel are collected in each transverse section.Whole body was measured and was finished in 10-20 minute when scanning speed was 16 cels, needed 20 minutes when scanning speed is 8 cels.The scope of R value (ratio of mental retardation decay and high energy decay in soft tissue) is from 1.2 to 1.4 (Lindemann etc., 1993).

Except the variation of relatively body weight, body fat percentage rate, fat mass and FFM, the inventor has also used the health composition of describing in the research the inventor before and has improved index (BCI) (Glinsmann and Mertz, 1966).The basis of BCI is following hypothesis, and promptly the acquisition of losing with FFM of body fat is the positive treatment result, and the acquisition and the losing of FFM of fat are negative therapeutic outcomes.Thereby the acquisition of the minimizing of fat mass and/or FFM when scoring for just, the acquisition of fat and/or FFM lose for negative, the BCI value is the comprehensive net result of these scores.BCI measures than the advantage of body weight as variation and has detected during participating in the supervision exercise event in the research that health forms and be proved in the patient accepts or do not accept the study medication of behavior revision program simultaneously.Compare between two groups, the hypothesis variance is identical in placebo group, carries out two tail t checks, makes each group balance on heat is taken in and consumed with covariance analysis in experimental group.

Withdraw from a kind of method of rate as minimizing, the patient is required 100 dollars check is provided, if the patient can not finish final DEXA test and the research application form at end eventually, check can not be returned, and this requirement is attached in the same expectation of signature.It is to finish test that the patient is apprised of the anti-also unique conditional of their check, and no matter they are many good or how bad doing according to research approach, as long as report strictly according to the facts.After finishing initial DEXA test, the patient will obtain the report of a test result and by random number, scope is from 1 to 130, and each numbering also may be the capsule of placebo corresponding to a same 400mcg chromium picolinate that may be equipped with of profile.Which patient's correspondence the investigative technique personnel of investigator, dispense medicament and patient all do not know is that placebo or which patient number are activated medicines.An independently local pharmacists also distributes to the experimenter with medicine bottle at random as the Trustee who studies, and these medicine bottles are used " X " or " Y " labelling in advance, correspond respectively to activated medicine and placebo.Finish and all material collected and after calculating finished, the Trustee took apart by what manufacturer provided and shows which is the envelope of active drug and notifies senior investigator (GRK) in research.All information is all analyzed by the computational resource system at University of Texas health science center under the supervision of second author (KB).During the summary of test, the person that finishes the final test will obtain the check of test result and mortgage and be required to answer actually taking capsular amount every day, with the amount of the used medicine of cross check.Subsequent analysis to these data shows the CrP average out to 357mcg that the experimenter takes every day.

Offer service manual of patient, wherein summarized general procedure, the nutritional information and the calculating of common food and the daily record of writing down daily heat balance that the assessment heat is taken in.In order to monitor and the difference of energy expenditure when adjusting body movement, when walking, all patients all wear exhausted pedometer (Evans and Press, 1989 in the former research; Kitchalong etc., 1993), these pedometers can be reflected in the step number of their walking in every day, and perhaps being reflected in can't be with the value such as step number in the activity of these utensils.The total step number of patient's record their walking every day in the same log that the heat that is used for writing down them is taken in, the net change that is used for regulating patient's body fat subsequently, i.e. 1 pound of the every variation of body fat+or-3500 cards.

The statistical table that 122 patients' finishing research baseline is described is provided in the table 75 as a result.Do not finish researcher and finish parameter that any health of comparison shows that of researcher forms there are no significant difference.

Table 75

122 patient's random screenings enter accept placebo or every days 400 MCG

The chromium picolinate group, the comparison Y of these patients' baseline demographic statistics

Body-mass index

Age (y) ^*Body weight (kg) ^*The % body fat ^*(kg/m ²) ^*Activated (n=62) 41.1+-10.5 85.5+23.0 42.4%+8.3% 30.2+7.1 placebo (n=60) 43.5=-7.6 79.9=-20.4 41.8%=-6.7% 28.4+5.4p level (two-tailed test)) p=0.24 p=0.16 p=0.65 p=0.13 ^*Mean value SD

Table 76 provides a comparison in the variation of test period generation in 90 days.

Table 77

At 90-days test periods, accept health between the participant of chromium picolinate of placebo or 400 mcg

The comparison of composition parameter mean change.All data are all controlled the difference that heat is taken in and consumed

Body weight is fat-free

(kg) ^*% body fat fat mass ^*Quality ^*BCI ^*Activated-7.8+-9.7-6.3%=8.5%-7.7+-9.5-0.1+-2.2+7.6+-4.5 (N=62) placebo-1.9+-4.0-1.2%+-5.7%-3.4+-6.8-0.3+-2.0+3.1+-7.6 (n=60) P level ^*P=＜.001 p=＜.001 p=.004 p=.568 p=.004

^*Average+-SD

^*Two tail student t-checks

The baseline data of discussion list 76 shows any health composition parameter of two groups, and difference is not remarkable statistically, illustrates that random packet is successful, provides two identical groupings of patient's number.The data of table 77 shows that the heat of proofreading and correct two groups is taken in and energy expenditure make two groups quite after, taking CrP can the appreciable impact body weight, the percentage rate and the BCI of body fat.Also merit attention is that the variation of medication group is consistent with the previous result of study of inventor, comprises the remarkable minimizing (p=0.02) of body fat even without correction heat absorption and energy expenditure.

It should be noted that the main improvement that health is formed in this research is the minimizing of body fat.The DEXA detection method is that a few health that directly detects the body fat tissue is formed one of detection technique, hydraulic pressure detects and the fat of the detection method assess patient health that other many healths are formed all depends on a hypothesis, and the fatty percentage rate of promptly supposing their the body density reflection result who finds in the research of some corpse that coexists is identical.And, hydraulic pressure detect can not actual measurement be used for calculating body density body volume-it infers it from the body weight that obtains in water and outside the water.Even thereby the weight of the body fat tissue that detects with hydrostatic(al) process also obtains by two kinds of different suppositions, rather than from the measurement of body fat tissue, obtain.Certainly, being detected to obtain assessing by hydraulic pressure and continue the influence of expiration ability when also being subjected to the experimenter under water, is the consistent influence that yet will be subjected to lung volume difference even exhale.The DEXA detection method has solved these difficulties, because this detection only need be when health be scanned on open monitor station static 15 to 20 minutes.The inventor thinks that the changeableness of control detection technology is indispensable when attempting to detect the effect of the material that causes the very little variation of health composition.

Require the patient to provide the deposit of according to condition returning that experimenter's number of finishing final test has been produced very big influence, no longer need to set up statistical contrast, as " deliberately handling ".Comment after the research discloses the experimenter and thinks that the requirement of paying deposit (not implementing) is rational.In 130 experimenters that participate in this research, have only 8 people not finish final test.Wherein a people is conceived and be required to withdraw from experiment; Three people move from this locality; One people is sick at the later stage test period; Three people's reasons are unclear.Thereby, concerning all intention and purpose, do not have under study for action can migration result withdraw from.Though inventor's material is not conclusive, requiring deposit it seems is worth further research.

Because the deposit that requires the patient to hand over is the application form when finishing research and research and finish based on the experimenter fully, and do not consider that the patient is that much degree ground is done according to former scheme, the equal number of the patient that do not take medicine in placebo group and active drug group can not influence between the balance group.The patient who does not take medicine in the placebo group detects the result does not have influence, because do not contain activated component in the placebo.Yet the patient who does not take medicine in the active drug group will reduce the former effect that should play of medicine.In fact, the patient who disobeys fully the active drug group of experimental program is actually placebo patients.Thereby what lack that compliance will be very natural reduces difference between two groups, so will emphasize to obtain the actual accurate material of taking dose of experimenter.Registration and personnel supervise to the inventor weekly provides more fully data and has reduced owing to lacking the skew to the result that compliance causes.

These data of conclusion show, when health form index (BCI) be used as result standard represent in the non-fat tissue have a net increase of value added add body fat lose value the time, take chromium picolinate and can significantly improve the health composition.

Embodiment 26

Polygenic inheritance and little/moonlet

Little/moonlet polymorphism moonlet in the psychogeneties is defined as the repetitive sequence (Wright, 1994) that length reaches 65 base pairs.Little satellite is made up of shorter repetitive sequence, and its length is that 2-5 bp does not wait.In the present embodiment, the inventor is referred to as the two with this term of little/moonlet unless stated otherwise.

Because the altofrequency of little in whole genome/moonlet repetitive sequence, inventor and other people utilize these polymorphisms in the association study of being everlasting.The initial hypothesis of inventor is the polymorphism as neutral and reticent single base pair, and microsatellite allele should be linkage disequilibrium with other " key " sudden change that influences gene function.Yet through after the years of researches, the inventor begins to suspect that microsatellite itself may be exactly " key " sudden change.Though inventor's research does not comprise the extremely long multiple polygenic inheritance of triplet, but provided such notion really in these researchs, promptly repeat allelic length variations,, can pass through the machine-processed widely function that influences gene even within " normally " scope.Two aspects relevant with the polymorphism of little/moonlet are their mutation rate and size.

The mutation rate of little/moonlet is than the mutation rate of non repetitive sequence higher (Jeffreys etc., 1987).The exchange that lacks the flank labelling shows that new sudden change may be owing to duplicate slip or complex conversion sample incident (Wolff etc., 1989).High mutation rate makes these polymorphisms diminish to the value of association study, because number has been sneaked into too many Confounding Factor in the linkage disequilibrium relation after generation, and this linkage disequilibrium relation requires to keep in the homophase two kinds of different polymorphisms in many generations.Yet if little/moonlet sudden change itself is " key " allele, in fact they may play bigger effect in association study, although mutation rate is higher.

If linkage disequilibrium is involved in little/moonlet allele and concrete phenotype is interrelated, the new sudden change of satellite sequence takes place so each time, should there be specific probability to occur on the identical chromosome with the allele of " key ".Yet, on average, in having these " key " allelic linkage disequilibriums, should not have the advantage tendency of long mutation allele to short mutation allele.Yet if the size of repetitive sequence works in the adjusting of gene (seeing below), allele of different sizes in each group so, rather than concrete allele or allelic sequence just should have certain tendentiousness with the dependency of quantitative trait.If there is the main peak of a reiterated genes frequency, so long with short allele all may be relevant with phenotypic effect.

The probability that exists length and sequence all to be related to, and different length little/moonlet allele may be influential to the function of gene.If different little/moonlet allele is shown gene transcription efficient or translation are worked, this hypothesis may be more meaningful.In fact many such examples are arranged.

The various aspects of the little/moonlet hypothesis of the general hypothesis polygenic inheritance of polygenic inheritance are as follows: many little/moonlet works to associated expression of gene; Same or the more little/moonlet of a lot of genes is associated; As a result, a lot of genes occur with the function polymorphic allele variant of certain limit.Those with several little/the relevant gene of moonlet polymorphism (each has a plurality of allele), will present the especially function haplotype of big figure.Though most gene and gene activity average level ± 10-15% interrelates, its scope can be up to ± 40%, and if relate to a plurality of little/their effect of moonlet polymorphism can adding up property.

In addition, the important function variant is general in the general population, and its prevalence rate scope is 1% to 100%.If have allelic two minutes forms of distribution of forming by high function allele (＞average 10%) and low-function allele (＜average 10%) fully, just 100% prevalence rate may occur.These functional varieties only play slight (0.5-8% of variation) to phenotype, seldom cause disease, that is, they are not autosomal dominance or recessive inheritance's disease of typical " gene-a disease " usually.Because most little/the moonlet polymorphism is in exon, the sudden change in the multigenic disease often is outside exon and outside the transcription sequence of being everlasting.When given disease relates to gene more than six, the Log chain research of marking will lose effectiveness (Risch and Merkangas, 1996; Weeks and Lathrop, 1995).Chain research is with the following basis that is assumed to be, i.e. given allelic an appearance and certain disease association connection, a given allelic disappearance is to be associated with the disappearance of certain disease.On the contrary, disease is related with the threshold value of several heterogeneic allele numbers in polygenic inheritance.So, the many members in family may carry a mutant gene, but not ill, because lack the threshold number of other necessary mutant gene.Thereby, many members of one family may carry a specific genes and also do not have ill, other ill members may not carry this sudden change allele (Comings, 1996f, l, m).

High function and low-function allele all can influence phenotype.For example, in psychiatric genetrics, the expression increase of a given acceptor gene causes receptor hypersensitivity or acceptor gene to be expressed descending the receptor hyposensitivity that causes all relevant with the variation of phenotype.As an individual inheritance when influencing the threshold number of low or high functional gene of identical phenotype, a kind of multigenic disease just can take place.Like this, if in a given quantitative variation, there are 20 kinds of different polygenes to work, anyly have more than 10 or 10 these people low or high functional variety and obvious phenotypes will occur and change.The value of threshold number will change according to allelic degree low or high functionality, and the order of severity that phenotype changes depends on the scope that polygenic number exceeds crucial threshold number.

Because little/moonlet is very general, so the probability that multigenic disease takes place is also quite high.Thereby multigenic disease is more more common than single-gene disorder.In the general population, a lot of polygenic frequency ranges are to surpassing 50% from 25%, the chance of heredity threshold number is than high many of the chance of single-gene disorder, to a given little/moonlet, allele may play positive effect or negative effect to phenotype, and this depends on the genetic background of other gene and the characteristic of phenotype.Related between the paranoid that real example is caused by (1994) observed cocaines such as Gelernter and 9 allele of dopamine receptor transporter gene (DAT1).On the contrary, Cook etc. (1995) is related with attention deficit hyperactivity disorder with 10 allele that (1996j) such as Comings observes the DAT1 gene.Someone may conclude from these dispersive reports that the observed result of or other must be incorrect.Yet, have the two all correct rational probability here, one given little/the not iso-allele of moonlet may with different phenotypic correlations.The rational method of research multigenic disease is to select a candidate gene and detect the influence of nearest little/moonlet polymorphism allele to the correlated measure characteristic.

The characteristic of the polygenic inheritance that proposes above also has a lot of other implications.If many little/the moonlet polymorphism has and changes the gene transcription nearest with it or the probability of translation rate, if a lot of genes same at least little/moonlet is relevant, the polygenic inheritance of the phenotype that so a lot of genes or most gene might be to a certain extent controlled them is worked.

In many researchs about special gene latent effect in psychosis and other disease, all the sudden change with research candidate gene exon begins.When negative findings appearred in this research, this gene usually was considered to the not effect of detected disease.If the great majority that relate in complicated polygenic character sudden change is in non-exon, such conclusion will be invalid.Present hypothesis is not got rid of the effect that gene function is only played the exons mutation of very little effect, although and just emphasize to exist variant important on the function of that gene, these researchs still negative findings may occur.

If each candidate gene only plays 0.5-8% to the difference of a given quantitative trait, according to the phenotype and the size of research, the possibility of result is slight, the boundary line or negative significance.Yet the accumulative action that detects two or more candidate genes may show a more significant effect.

Certain special allelic frequency that many association studies only detect the patient simply compares with matched group.Yet, when diagnosis is when being the basis with the complicated symptom of a cover, the SEVERAL QUANTITATIVE CHARACTERS significant correlation that specific allele may relate to this diagnosis, but be not diagnosis itself.For example, if schizophrenia is a kind of multigenic disease, the allele of specific candidate gene may show and negative symptoms (as abepithymia) significant correlation, this can occur in some case but be not all can occur in all cases, but not relevant with the positive symptom (as hallucination or illusion) or schizoid diagnosis itself.Limited analysis is carried out in simple two branch=property diagnosis, rather than detected secondary symptom quantitative trait, may miss the gene that some play an important role to multigenic disease.When single allelic frequency did not have difference in normal control and patient, hybrid vigor also may make a gene pairs phenotype play an important role (Comings and MacMurray, 1977).

If a kind of hypothesis has direct inspiration to be worth, it is just very useful.When each allele in a large amount of or even the allele genotypic more big figure of possibility is detected, the effectiveness of research may be badly damaged and makes it useless, this be in association study, use highly polymorphic little/shortcoming of moonlet polymorphism.Yet, if the size of repetitive sequence is key variables, even if the most complicated polymorphism also can be reduced three genotype, short allelic homozygote, long allelic homozygote and heterozygote.Do the time spent at some genes of research (OB, MAOA, MAOB, CNR1, GABRA3, GABRB3, DBH, FRAXA and NOS1) in the behavior character, the inventor has set up this valuable method.

Though many potential significant genes are cloned and are checked order, concerning major part, in genome database, all do not report its polymorphism, make association study impossible.Based on present hypothesis, be known as infructescence, a kind ofly identify that the method for useful little/moonlet is the big genomic clone that obtains carrying gene of interest from the commercial channel.Screening may provide the repetitive sequence of more information for association study in these clones.

Present model shows in the research of polygenic inheritance, and the repetitive sequence that forms Z-DNA is proved to be most worthy.This shows with Z-hunt-II program (Schroth etc., 1992) examination known array, can provide about the localized important clue of the most useful polymorphic regions.In order to verify this point, the inventor utilizes this program to detect the sequence (Hall etc., 1994) of NOS1.With to NOS1 knock out mice (Nelson etc., 1995) research of aggressive behavior is the basis, the inventor has detected people's various actions character with the repetition polymorphism, the Sequence Identification of inventor by actual detected NOS1 gene repetition polymorphism (Hall etc., 1994).Owing to observed some associations, the inventor wants to know: whether the polymorphism that the inventor utilizes can be identified by the Z-hunt-II program; In this gene, whether also have other zones to contain useful polymorphism.These two predictions all are correct.NOS1 (CA) based on different length _nThe hypothesis that allele may work in the function of NOS1 gene, the inventor is divided into two groups with allele, comprises the shortest by 50% allelic (＜199bp) group and the longest by 50% (〉=199bp) group.Initial research prompting and some dependencys of behavior phenotype.Subsequently the inventor with Z-hunt-II algorithm (Wang etc., 1997) examination the Z-DNA district of the NOS1 gene order announced, the result shows that the polymorphism that the inventor uses is one of three high Z-DNA content districts.The new polymorphic regions that second higher zone of Z score do not found before being.

In fact these three examples have illustrated the prediction of this model is how to cause accelerating to identify the gene that relates to multigenic disease.This model with combine application with regard to the material of the utilizable Human Genome Project soon, by improving the ability of identifying near the little/moonlet the known candidate gene, the knowledge of the inventor being understood multigenic disease has very big effect.

Ammoxidation single enzyme A gene (MAOA) and VNTR polymorphism allele are divided into four groups, and the many quantitative variations relevant with characteristic symptom have significant association.Shown appraisal result to the mania shape of the proband of 351 coprolalia syndrome and their relatives and normal control.Carrying, the longest allelic experimenter's score significantly improves (Gade etc., 1997).(b) in substance abuse person's group polymorphism identical with The control group.Carry significant association (Gade etc., 1997) is also arranged between the longest allele and the drug dependence score.(c) quantity score and the allelic dependency of ob gene microsatellite polymorphism based on the anxiety of SCL-90 test be divided into＜experimenter that 208bp allele (left side) isozygotys and 〉=208bp (right side) allele isozygotys or the individuality of heterozygosis.(d) event related potential (ERP) P300 ripple (with microvoltometer) little with CNR1 Fructus Cannabis ester acceptor gene/moonlet polymorphism allelic related being divided into 〉=5 repeat experimenter that allele (right side) isozygoty and＜5bp (left side) repeats that allele isozygotys or the individuality (Johnson etc., 1997) of heterozygosis.(e) Brown AD in adults D scoring and GABAA, the allelic association of B3 (GABRB3) acceptor gene microsatellite polymorphism is divided into 〉=experimenter that 185bp allele (left side) isozygotys and＜185bp allele (right side) isozygotys or the experimenter of heterozygosis.(f) in the adult at random of assessment IQ, related (Comings etc., 1997a, b or the c) between the normal allele of behavior IQ and FRAXA locus.

Embodiment 27

Three adrenergic genes (ADRA2A, ADRA2C, DHB) are to following and not following

Study can not ADHD patient's adding up property effect

The inventor has detected three adrenergic genes, epinephrine α 2A receptor (ADRA2A) gene, epinephrine (ADRA2C) receptor α 2C gene and dopamine gene (DBH) with follow and the dependency of the ADHD that not concomitant learning can not (LD) or its adding up property effect.

Two types ADHD and inventor are specified may to be summarized in table 78 by candidate gene.

Table 79

Follow and do not follow the ADHD that cognition can not (CD)

The secondary neurotransmitter NA of cerebral nerve nuclear lid district's elementary neurotransmitter dopamine norepinephrine of veutro locus coeruleus that does not follow the ADHD of CD to follow the ADHD cognitive disorder of CD not have to exist the normally low brain region prefrontal lobe prefrontal lobe/temporal lobe that relates to of language IQ to relate to, GABA, five hydroxytryptamine and other dopamine, GABA;

Five hydroxytryptamine and other functional analysis information and start acknowledge adapt to and handle new stimulate the extensive selectivity of attention deficit type carry out dysfunction exist or not usually candidate gene DRD2, DRD4, DRD5, ADRA1A-ID, ADRA2A, ADRA2C,

DAT1、DBH??????????DBH、NT、PNMT

Halperin etc. (1997) report, the same young criminal's who is caught because of violence or great crime of the dependency between blood plasma MHPG and the language IQ (but not being behavior IQ) lower discovery closely similar (Hirschi and Hindelang, 1977 of language IQ; Miller, 1988; Shulman, 1951; Moffitt and Silva, 1988).Negative correlation explanation between blood plasma MHPG level and the low language IQ may relate to the NA gene in antisocial behavior.

Because the epinephrine alpha-2 receptor is the site of action of clonidine, be rich in forehead and parietal attention center, and in the NA of synapse conversion, play regulating action, whether thereby the inventor attempts to confirm whether the difference of epinephrine α 2A (ADRA2A) gene or epinephrine α 2C (ADRA2C) gene or dopamine gene genotype is relevant with ADHD itself, and preferentially relevant with ADHD+ cognitive disorder hypotype.The dinucleotide that the inventor has used MspI polymorphism (Lario etc., 1997) in the ADRD2A gene promoter region, ARDA2C gene repeat polymorphism (Riess etc., 1992) and DBH gene the TaqIB1/12 polymorphism (d ' Amato etc., 1989; Wu etc., 1997).

Method determines single base pair polymorphism among ADRA2A and the DBH and the dinucleotide among the ADRA2C to repeat the genotypic frequency of polymorphism in 325 experimenters, and wherein 267 people are the coprolalia syndrome patient, and 58 people are normal control.ADHD and LD behavior variable are estimated by the mode of father and mother or application form of oneself filling in and directly meet.In order to assess and may the getting in touch of antisocial behavior, also estimated the symptom of conduct disorder and oppositional defiant disorder.

Seminar is made up of 325 incoherent experimenters.Wherein 267 people meet DSM-III-R and DSM-IV coprolalia syndrome diagnostic criteria and all directly meet by D.E.C..58 remaining people are contrast.Everyone is non-Hispanic white people.TS patient is from wishing coprolalia syndrome clinic, medical centre city (City of Hope Medical Center).The inventor before had been divided into TS patient following several groups, slight (one-level, chronic twitch, very light need not the treatment), moderate (secondary needs treatment) and severe (three grades, some aspect during they are lived have seriously influence very much) (Comings, 1990; Comings and Comings1987b; Comings and Comings, 1984).In TS patient, 25% is three grades, and 12% is one-level, and remaining 71% is secondary.TS is similar obstacle with ADHD, counts to the patient who goes to a doctor the clinic mostly and suffers from ADHD (Comings, 1990 simultaneously; Coming and Comings, 1987b; Comings and Comings, 1984).Contrast, do not follow the TS patient of ADHD and follow the TS patient's of ADHD existence, make this group research be particularly suitable for detecting the dependency of heterogeneic allele and ADHD (as the continuous trait variable).TS patient and detailed description (Comings etc., 1996j are all arranged to impinging upon other place; Comings, 1994a; Comings 1994b; Comings, 1995a; Comings, 1995b).TS patient's mean age is 18.0 years old (S.D.13.2).But great majority are bigger child and teenager, and 29% is 21 and more than 21 years old.The mean age of matched group is 46.3 years old (S.D.15.38).

Each of matched group and TS group all needs to fill in an application form, comprising all DSM-III, DSM-III-R and the DSM-IV diagnostic criteria to ADHD.Bigger experimenter is required they hour marquis's situation is answered.ADHD scoring be based among the DSM-IV (1994) to the diagnostic criteria of ADHD.Problem is to ask child and juvenile years whether have these symptoms, be not have or rare (scoring=0), there be (scoring=1) once in a while, be still there (scoring=2) always? all these three answers are used to distinguish fully the patient of three orders of severity.The ADHD scoring is the summation of Inattention among the DSM-IV, impulsion and hyperkinetic syndrome shape.

If child less than 14 years old, is filled out a questionnaire by father and mother, the experimenter more than 14 years old or 14 years old is filled in by me or fills under father and mother's help.See together that with experimenter and family members thereof application form is to guarantee its correctness in person.

In order to estimate the existence of cooperation problem on obstacle, were the experimenter is required to answer following three problems: 1) you once assigned to the special class of education difficulty (EH), learning difficulty (LH) or learning disorder (LD)? were 2) you once assigned to intelligence class? did were 3) you once apprised of you and have learning disorder? each problem must be divided into: no, got 0 fen; Be to get 1 fen.The score addition is formed the score of LD.Any one of participating in the above-mentioned special class in California all needs the comprehensive assessment of one or more educational psychologists, promptly estimates its companion of this student's ratio and falls behind more than 2 years or 2 years.

In order to estimate school grade in primary school, the experimenter be required to answer " your a) mathematics, b from one grade to six grades) read, c) the writing achievement be on the whole average, average following, or on average more than? " average above getting 0 fen, average getting 1 fen, average following getting 2 fens obtains the GSAP score with each section's score addition.

0,1 and 2 classifications of similar conduct disorder and oppositional defiant disorder DSM-IV diagnostic criteria have been used.

If how moving the experimenter meet DSM-IV to attention deficit/obstacle associative form, Inattention advantage type or the how diagnostic criteria of moving advantage type, then be considered to serious ADHD problem.If the experimenter was assigned to above-mentioned special education class or once was apprised of and has learning disorder, then be considered to serious learning disorder.The experimenter who meets ADHD and learning disorder diagnostic criteria is divided into the A+LD+ group; The experimenter who does not meet ADHD and meet the diagnostic criteria of learning disorder is divided into the A-LD+ group; The experimenter who meets ADHD and do not meet the diagnostic criteria of learning disorder is divided into the A+LD-group; The two all incongruent experimenter is divided into the A-LD-group.Similar being grouped in the GSAP score used, and score 0-4 person is GS-, and must be divided into 5-6 person is GS+.

Application form (Comings, 1990) also provides the method for a highly structural, and this method can provide the quantitative trait relevant with ADHD: the secondary score of ADHD scale, problems such as study, one learns in grade school achievement, conduct disorder and opposition behavior.Use the situation of same structure chemical industry tool by more this instrument and the person of seeing and treating patients, confirmed (Gadow and Sprafkin, 1994 by other people as accuracy, practicality and the sensitivity of this application form on symptom appraisal procedure basis; Grayson and Carlson, 1991).The inventor has consulted each experimenter and has got application form, shows that they can reflect accurately that those are by the resulting information of seeing and treating patients in person.

ADRA2A utilizes single base pair MspI polymorphism of promoter region to determine genotypic (.1997 such as Lario).Utilized their PCR ^TMCondition and primer.ADRA2C utilizes dinucleotide to repeat polymorphism and PCR ^TMProgram (Riess etc., 1992).With the fluorescently-labeled primer of 0.1uM, by polymerase chain reaction (PCR ^TM) the amplification target DNA.PCR ^TMProduct is with the dilution of 100ul deionized water, gets in the mixture of the smart dyestuff of 75ul Methanamide+9.5ul ROX titer+9.5ul blue paste that 0.5ul joins 2.5ul.92 ℃ of 2 seconds of degeneration, sample is added Applied Biosystem 373 dna sequencing instrument, and ((ABI) Forster City among 6%PAGE CA), and ran glue 4 hours under the condition of 1200 volts and constant 30W for AppliedBiosystem, Inc..Gel is handled and is analyzed with internal standard (ROX 500).According to color and clip size, the peak by Genotyper (1.1 editions) (AppliedBiosystem, Inc.) identification. produce main allele at 183bp and 185bp,, also have several allele very in a small amount in addition than the 181bp allele of small frequency.These are divided into three genotype :≤183/≤183bp=1, heterozygote=2 ,≤183/≤183bp=3.

Determine DBH gene genotype (d ' Amato etc., 1989) with the TaqIB1/B2 polymorphism.But this originally need carry out the southern blotting technique analysis with the DBH clone of labelling, and the inventor has taked with PCR ^TMDetection (Wu etc., 1997) for the basis.

The gene type of dopamine gene (DRD2 and DAT1) formerly is described (Comings etc., 1996), and the inventor determines DRD5 gene genotype (Sherrington etc., 1994) with microsatellite polymorphism and technology thereof.

Carried out three types statistical analysis.At first, in order to estimate the genotypic effect of ADRA2A and ADRA2C, (each gene must be divided into 11=1, and 12=2 22=3) has carried out variance analysis to the ADHD score.In case the genotype method of dividision into groups is chosen, whether each gene is designated as existence that two fens property variable shows the related gene type.The A2A+A2C score is used to estimate the adding up property effect of two genes.Here, ADRA2A must be divided into 0 and ADRDA2C must be divided into 0 experimenter and be designated as 0 fen.ADRA2A must be divided into 1 and ADRA2C must be divided into 1 experimenter and be designated as 2 fens, the 1 fen person that must be divided into of any one gene is designated as 1 fen.The score of NA gene is to be got by the single score addition of all three genes.

Proofread and correct for fear of Bonferroni, the score of six behaviors is checked with the multivariate analysis of variance method as a group.Each gene reaches each gene separately and joins together to carry out this inspection.In order to assess each gene separately and join together to account for the percentage rate of score difference, carried out the linear regression analysis between individual gene score, A2A+A2C score and NA gene score and the behavior score.Based on the effect of previous each NA gene the hypothesis of adding up property effect is arranged, in order to assess the size of this effect, the result of the adding up property effect of ADRA2A and ADRA2C or ADRA2A, ADRA2C and DBH has been carried out linear equation analysis (multinomial subcommand=1).To being to have the mean of significant difference to carry out post hoc at 0.5 o'clock to analyze at α.All statistical analysiss all with the SPSS statistical package handle (SPSS, Inc, Chicago, IL).

The result has carried out variance analysis in order to check the effect of ADRA2A genotype to the ADHD score.The average of carrying 11 genotypic experimenters is 17.4, and (n=180, SD=11.1), 12 genotype are 19.05, and (n=125, SD=10.52), 20 genotype are 22.05, and (n=20, SD=11.67), F is than=2.05, p=0.13.In multivariate analysis of variance, the score of ADRA2A gene is as follows: 11=1,12=2,22=3; In regression analysis, it must be divided into: 11,12=0,22=1.

Six scores have been carried out multivariate analysis of variance (table 80A).ADRA2A gene and LD, GSAP, impulsion and total MANOVA have significance related.ADHD, LD, GSAP, CD and ODD score have been carried out linear regression analysis (table 3).It has significance related with LD and GSAP score, if proofread and correct (α is 0.05/5 or 0.01) through Bonferroni, the LD score still has significance.

Table 80

The multivariate analysis of variance of ADRA2A, ADRA2C and DBH gene

(N＝325)

Scoring F-Duo how moving 2.82 .061 GSAP, 1.91 .149 LD, 1.78 .169 conducts, 1.44 .239 ODD, 1.64 .194 summations (Wilkes), 1.09 .360C.DBH gene Inattentions, 2.02 .134 impulsion property, 1.28 .278 1.12 .325 GSAP, 2.48 .086 LD, 1.40 .506 conducts, 1.40 .247 ODD, 1.71 .182 of moving of 1.23 .295 GSAP, 3.42 .034 LD, 3.80 .023 conducts, 0.27 .763 ODD, 1.21 .299 summations, 1.81 .034B.ADRA2C gene Inattentions, 5.18 .006 impulsion property, 4.11 .017 than pA.ADRA2A gene Inattention 1.45 .236 impulsion property 3.67 .027 more

Scores F-ratio p sum total (Wilkes) 0.53 .915D.ADRA24+ADRA2C gene Inattention 5.20 .006 impulsion property 5.51 .004 move 3.73 .025 GSAP, 5.26 .006 LD, 5.40 .005 conducts, 0.35 .709 ODD, 3.06 .050 summations (Wilkes), 1.62 .070E.ADRA2A+ADRA2C+DBH gene Inattentions, 5.25 .002 impulsion property, 4.21 .006 more, and how moving 3.76 .011 GSAP, 5.01 .002 LD, 3.62 .013 conducts, 0.85 .465 ODD, 3.38 .018 summations (wilkes), 1.49 .070F.ADRA2A+ADRA2C+DBH gene+DRD2+DAT1+DRD5 Inattention 4.81＜.001 impulsion property 5.31＜.001 moves 4.44＜.010 GSAP, 4.41＜.001 LD, 2.91 .009 conduct 2.80 .011 ODD, 4.73＜.001 summations (Wilkes), 1.58 .01 more

Table 81

ADRA2A, the linear regression analysis of ADRA2C and DHB gene

(N＝325)

R r ²T pADHD ADRA2A .086 .007 1.57 .117 ADRA2C .161 .026 2.94 .0035 DBH .107 .011 1.94 .054 A2A+A2 .181 .033 3.32 .0010 All3NA .200 .040 3.67 .0003 All3DA .157 .025 2.85 .0047 NA+DA .247 .061 4.56＜.0001LD ADRA2A .152 .023 2.78 .0056 ADRA2C .077 .006 1.40 .162 DBH .057 .003 1.04 .291 A2A+A2C .136 .019 2.49 .013 All3NA .136 .018 2.48 .0135 All3DA .028 .001 0.51 .61 NA+DA .111 .012 1.97 .049GSAP ADRA2A .128 .016 2.32 .020 ADRA2C .102 .010 1.86 .065 DBH .122 .015 2.22 .027 A2A+A2C .148 .022 2.70 .007 All3NA .185 .034 3.40 .0008 All3DA .097 .009 1.73 .085

r??????????r ²????????T???????????p

NA+DA gene .203 .042 3.66 .0003

The conduct score

ADRA2A gene .0008 .0000 0.01 .989

ADRA2C gene .045 .002 0.83 .409

DBH gene .088 .008 1.59 .113

A2A+A2C?????????.041???????.002???????0.74???????.462

All3NA gene .085 .007 1.54 .125

All3DA gene .071 .005 1.25 .211

NA+DA gene .109 .012 1.92 .054

The ODD score

ADRA2A gene .085 .007 1.55 .122

ADRA2C gene .093 .009 1.70 .091

DBH gene .100 .010 1.81 .070

A2A+A2C?????????.121???????.015???????2.20???????.028

All3NA gene .151 .023 2.77 .006

All3DA gene .162 .026 2.89 .004

NA+DA gene .221 .049 4.00 .0001

In order to detect of the effect of ADRA2C genotype, carried out variance analysis to the ADHD score.The average of carrying 11 genotypic experimenters is 18.9, and (n=122, SD=10.41), 12 genotype are 15.90, and (n=112, SD=10.99), 22 genotype are 20.54, and (n=91, SD=11.1), F is than=4.90, p=0.0080.The mean that detects 12 heterozygotes by post hoc Tukey significantly is lower than 11 and 22 homozygotes, illustrates to have negativity hybrid vigor (Commings and MacMurray, 1998).Thereby in linear regression analysis, the score of ADRA2C gene is as follows: 12=0,11 or 22=1.

The multivariate analysis of variance (table 80-B) that the relation of ADRA2C gene score and six behavior scores has been carried out.ADRA2C and Inattention and impulsion property have significant dependency.In linear regression analysis (table 81), with ADHD significant dependency is arranged, be 0.01 o'clock at α, still have significance.

The inventor had before studied dopamine-B '-hydroxylase gene in TS-ADHD patient (Comings etc. 1996j), and find the allelic existence of TaqB1 relevant with ADHD (d ' Amato etc., 1989).Thereby the inventor is defined as DBH gene score: 22=0,11 or 12=1.By multivariate analysis of variance (table 80-C), find that the neither one score has significance.In linear regression analysis, it is significantly relevant that DBH gene and GSAP score are, and α did not then have significance at 0.01 o'clock.

In order to detect the adding up property effect of two epinephrine alpha-2 receptor genes, the score addition of two individual genes is obtained the A2A+A2C score.In 325 experimenters, must be divided into 0 106 people (32.6%) are arranged, must be divided into 1 205 people (63.1%) are arranged, must be divided into 2 14 people (4.3%) are arranged.The results are shown in of multivariate analysis of variance to A2A+A2C score and six behavior scores shown 80-D.The result has significance to Inattention, impulsion property, many moving, GSAP, LD and ODD score.At α is 0.01 o'clock, and ADHD and GSAP score still have significance.Linear equation to other behavior score except the CD score the analysis showed that the score that carries the experimenter of 0,1 or 2 mutant gene increases gradually.ADHD, Inattention, impulsion, how score moving and GSAP all has significance, p＜0.01.

For checking possible the adding up property effect of three epinephrine genes, the score addition of ADRA2A, ADRA2C and DBH gene is obtained NA gene score.In 325 experimenters, must be divided into 0 36 people (11.1%) are arranged, must be divided into 1 132 people (40.6%) are arranged, must be divided into 2 145 people (44.6%) are arranged, must be divided into 3 12 people (3.7%) are arranged.The score of NA gene and the results are shown in of multivariate analysis of variance of six behavior scores are shown 80-E.The score of Inattention, impulsion property, many moving, GSAP, LD and ODD all has significance.In linear regression analysis (table 81), ADHD, LD, GSAP and ODD score all have significance, are 0.01 o'clock at α, and ADHD, GSAP and ODD still have significance.

The linear equation of the secondary score of ADHD, LD, GSAP and ODD score the analysis showed that the score that carries the patient of 0,1,2 or 3 mutant gene increases gradually.The score of ADHD, many moving, Inattention, GSAP and ODD has significance, p＜0.01.

The inventor has carried out X 2 test with the linear equation analysis to the relation between NA gene score and ADHD-LD-, ADHD+LD-, four groups of ADHD-LD+, ADHD+LD+, the results are shown in table 82.The result shows that patient's frequency of carrying all three NA genetic mutations increases gradually in above-mentioned four groups: be 0.6% in the ADHD-LD-group, be 5.4% in the ADHD+LD-group, be 7.7% in the ADHD-LD+ group, be 11.4% in the ADHD+LD+ group.In above-mentioned four groups, the frequency of not carrying the patient of NA mutant gene reduces gradually, is respectively 14.4%, 8.9%, 7.7%, 2.9%.When complete 4 * 4 forms are carried out linear trend X 2 test (Cohran, 1954), p=0.0005.If A-LD+ and A+LD+ are merged Pearson chi-square value=17.6, d.f.=6, p=0.007, linear trend chi-square value=12.9, d.f.=1, p=0.0003.

Table 82

The scoring of NA gene is to ADHD-LD-, ADHD+LD-, ADHD-LD+, ADHD+LD+ group

The X 2 test analysis

[case load (%)]

NA score A-LD+ A+LD-A-LD+ A+LD-T

0?????????24(14.5)??????10(8.9)?????1(7.7)?????1(2.9)??????36(11.1)

1?????????73(44.2)??????41(36.6)????4(30.8)????13(37.1)????131(40.3)

2?????????67(40.6)??????55(49.1)????7(53.8)????17(48.6)????146(44.9)

3?????????1(0.6)????????6(5.4)??????1(7.7)?????4(11.4)?????12(3.7)

T?????????165(51.1)?????112(34.3)???13(4.0)????35(10.7)????325(100.0)

Pearson card side 18.3, df=9, p=0.03

Linear trend card side 12.0, df=1, p=0.0005

Because the NA gene is bigger with the dependency of LD score with the dependency ratio of GSAP score, the inventor has also detected the intersection form of GSAP score and NA gene score.The result shows that the frequency range that the NA gene must be divided into 3 minutes patient is: it is 1.6% that GSAP must be divided into 0 to 2 patient, and it is 0.8% that GSAP must be divided into 3 to 4 patient, and it is 11.8% that GSAP must be divided into 5 to 6 patient.In the gamut of the two score, the linear trend X 2 test has significance, p=0.004.Thereby, not relying on the existence of ADHD or do not exist, those 2 or 3 sub-average children of purpose achievement of section most possibly carry the NA gene of variation.

In order to detect the effect whether ADHD exists, the experimenter is divided into four groups again, and here GS+ is that GSAP must be divided into 5 to 6, and GS-is that must to be divided into 0 to 4, four group be respectively A-GS-, A+GS-, A-GS+ and A+GS+ to GSAP.The table 83 that the results are shown in the X 2 test of NA score.In these four groups, the frequency that the NA gene must be divided into 3 experimenter increases gradually, is respectively 0.6%, 2.4%, 4.8%, 12.5%.The linear trend X 2 test of 4 * 4 forms has significance, p=0.0003.

Table 83

The scoring of NA gene is to the X 2 test analysis of A-GS-, A+GS-, A-GS+ and A+GS+ group

[case load (%)]

NA score A-GS+ A+GS-A-GS+ A+GS-T

0?????????23(14.7)?????7(8.4)??????2(9.5)?????4(6.3)??????36(11.1)

1?????????71(45.2)?????31(37.3)????6(28.6)????23(35.9)????131(40.3)

2?????????62(39.5)?????43(51.8)????12(57.1)???29(45.3)????146(44.9)

3?????????1(0.6)???????2(2.4)??????1(4.8)?????8(12.5)?????12(3.7)

T?????????157(48.3)????83(25.5)????21(6.5)????64(19.7)????325(100.0)

Pearson card side 25.89, df=9, p=0.002

Linear trend card side 12.97, df=1, p=0.0003.

The inventor has detected mathematics, reading and the writing that is lower than average achievement respectively, and NA gene score and three section's purpose scores have been carried out X 2 test, and three section's purpose score standards are: average or average above=0, average following=1.In the mathematics subject, PearsonX ²=9.17, p=0.027, linear X ²=4.18, p=0.04.In reading subject, PearsonX ²=15.14, p=0.0017, linear X ²=13.14, p=0.0003.In the writing subject, Pearson X ²=13.18, p=0.004, linear X ²=10.54, p=0.001.Thereby, though in these three subjects, be lower than the score of average achievement and NA gene significant dependency is arranged, the strongest to the effect of reading, writing is taken second place, the mathematics minimum.

In order to compare, the inventor has also detected the effect of dopamine gene score to each subject achievement.In reading subject, Pearson X ²=12.08, p=0.007, linear X ²=8.24, p=0.004.In the mathematics subject, Pearson X ²=2.90, p=0.41, linear X ²=2.21, p=0.137.In the writing subject, Pearson X ²=11.11, p=0.011, linear X ²=0.02.Thereby the dopamine gene also works in reading achievement, but role is less in reading and mathematics.

In order to detect the importance of the relative dopamine of NA gene (DA) gene, the inventor will comprise the score (table 80-F) of DA in multivariate analysis of variance.To all six genes, Inattention, impulsion property, many moving, GSAP, LD, conduct and ODD score all have significance.In to the linear regression analysis of all six genes (table 81), ADHD, LD, GSAP and ODD score have significant dependency.As (Comings etc., 1996) that before shown, the dopamine gene plays an important role in ADHD.This is confirmed by three genes in this application.They account for 2.5% of ADHD score difference, and when with the addition of NA gene, six all genes account for 6.1% (p＜0.0001) of difference.In contrast, AD gene role in LD and GSAP is very little.Yet, must assign to from GSAP, the percentage rate of difference is 3.4% 4.2% when being increased to the combined effect of DA gene when three NA genes work alone.Though DA gene role in the behavior score is very little, plays an important role in the ODD score, account for 2.6% (p=0.004) of difference, all six genes account for 4.9% (p=0.0001) of ODD score difference.

The inventor has detected the frequency of three DA genes of four ADHD:LD groups, and all three genes are in 10% ADHD-LD-group, in the 19.8%ADHD+LD-group, exist in the 15.48%ADHD-LD+ group and in the 17.1%ADHD+LD+ group.Thereby opposite with the NA gene, the phenomenon that does not increase gradually between four groups occurs.In complete 4 * 4 forms, Pearson chi-square value=10.48, d.f.=9, p=0.31, linear chi-square value=0.90, d.f.=1, p=0.34.

Multivariate analysis of variance has shown between the quantity score of Inattention, impulsion property, many moving, learning disorder, one learns in grade school achievement and opposition disobedience behavior and detected three genes significant dependency.The dependency maximum be the additive effect (p=0.0003) of three genes.The number of variation NA gene significantly increases in the following patient's grouping successively: no ADHD (A-) or do not have learning disorder (LD-), A+LD-, A-LD+, A+LD+ (p=0.0005), but in the dopamine gene, there is no comparable effect.

Two ADHD hypotypes-follow are discussed or are not followed cognitive defect, relate to the zones of different of brain, different neurotransmitter and different several groups of genes (table 80)-discriminating may sizable value be arranged to ADHD child's diagnosis and treatment.Yet, must remember be ADHD be the polygenes disease (Comings, 1996a), from parents heredity a plurality of mutant genes (Comings, 1996b).In most children that these two types of compositions are arranged and adult, exist sizable overlapping.

Yet as what show in table 82 and table 83, the NA metabolic deficiency is compared the ADHD child that may include cognitive defect more with the ADHD child of NA gene with no cognitive defect that make a variation.

Adding up property of observation score also has such advantage, if gene is inoperative in given experimenter or one group of experimenter, but another gene with identity function works, and this may comprise the effect of the two.This is proved in this research well.For example, when the phenotype effect of three NA genes during by single independent detection, the significance level of multivariate analysis of variance is lower (table 80-A, table 80-B, table 80-C), and with five scores of linear regression analysis check the time, correlation coefficient can not support Bonferroni to proofread and correct.Yet when with ADR2A, ADR2C gene or ADR2A, ADR2CD, the addition of DBH gene, its result is just more reliable.When doing the time spent, also be like this with the progression of linear equation analyzing and testing.With the ADHD score as an example, single-factor regressioning analysis shows, the independent effect of each gene only accounts for 0.7% to 2.6% of quantity score difference, and when ADR2A and the addition of ADR2C gene, account for 3.3% (p=0.001) of difference, when with three NA gene additions, then account for 4.0% (p=0.0003) of difference.Three dopamine genes account for 2.5% of ADHD score difference, when with all six gene additions, then account for 6.1% (p＜0.0001) of difference.This explanation NA and AD gene work adding up property effect in ADHD.

Twin study shows that the additive inheritance effect accounts for 70% or more (Sherman etc., the 1997a of ADHD; Sherman etc., 1997b; Gillis etc., 1992).Three NA genes of this explanation account for 6% of ADHD genetic constitution greatly, and NA adds the DA gene and accounts for 10% greatly.Though this seems a bit little, when increasing gene is added up, percentage ratio will continue to raise (Comings, etc., 1998).Only account for 6.1% of ADHD score difference although it should be noted that these six genes, the correlation coefficient based on 0.25 relies on other variable, and the predictability of these six gene pairs ADHD reaches 25%.

The adding up property effect of NA gene pairs GSAP is also given deep impression, and the percentage rate that individual gene accounts for difference is 1.0% to 1.6%, and the adding up property effect of three genes accounts for 3.4%.

The result of table 83 shows, in A-LD-, A+LD-, A-LD+ and A+LD+ group, patient's frequency of carrying 2 or the NA gene that more makes a variation increases gradually.Yet another kind of hypothesis thinks, ADHD+LD or ADHD+ other to send out the obstacle group altogether only be a group of the representative genetic load that has more ADHD gene.Whether the inventor has same adding up property effect by detecting three different dopaminergic genes in three groups, can check this hypothesis.In the experimenter, the inventor observes in six different dopamine genes (DRD1, DRD2, DRD3, DRD4, DRD5 and DAT), DRD2, DAT and DRD5 have very significantly adding up property effect to the ADHD score, they account for 0.5% to 1.0% of difference as the time spent separately, and account for 2.5% of difference when adding together.Thereby the inventor has worked out a dopamine gene scoring index, and scope also is from 0 to 3.Patient's the frequency range of carrying all three mutant genes is from 10% of A-LD-group, 19.8% of A+LD-group, 17.1% of 15.4% to A+LD+ group of A-LD+ group.Like this, opposite with this hypothesis, the A+LD+ group compares with the A+LD-group, does not have to increase the genetic load to three dopamine genes.In addition, concerning the score scope of whole dopamine gene, there are no significant for the result of Pearson and linear trend X 2 test.This linear X 2 test p=0.0005 with the NA score is obviously different.These results further support Halerin-Pliszka hypothesis b, and promptly the NA gene is easier in the ADHD+LD individuality is related to, and no matter has or not in the ADHD individuality of cognitive disorder, to relating to of dopamine gene be equality.

Another feature is in the differentiation by the number decision of variation NA gene, and the inquiry individual is the same good in the achievement of mathematics, reading and the writing of the school effect that they cross special LD study class on whether with inquiry simply.When independent detection, to reading the effect maximum of achievement, to the effect of writing achievement much at one, to the effect minimum of mathematics achievement.These presentation of results are in primary school, and the child who suffers from ADHD that reading, writing and mathematics achievement are lower than average achievement most possibly carries the NA gene of variation.This also shows need do secular double blind experiment, and the checking clonidine is to the therapeutic efficiency of these child's problems concerning study.In the clinical experiment that schedules to last some months that one of inventor (D.E.C) carries out, the bad subcutaneous independent application clonidine of child of school grade can improve school grade significantly.

The dependency of low language IQ and teen-age aggressivity antisocial behavior is repeated (Hirschi and Hindelang, 1977 in many researchs; Miller, 1988; Shulman, 1951; Moffitt and Silva, 1988; Moffitt, 1990).After having controlled factors such as socioeconomics, race, school grade and motivation, still there are (Moffitt, 1993) in the dependency of low language IQ and juvenile delinquency.The research of Moffitt and Silva (Moffitt and Silva 1988) has shown that also the dependency of low IQ and juvenile delinquency is not an illusion that is easier to the low wit crime caught by the police, because the undiscovered juvenile delinquent of confirming by interview also has lower IQ.The artificial dependency of finding with conduct disorder and oppositional defiant disorder of invention has detected the NA gene.Though any one of three NA genes all do not have tangible dependency with conduct disorder, slight dependency is arranged with oppositional defiant disorder.

The probability of studying ADHD with the patient who suffers from TS promptly has advantage that shortcoming is arranged again.The advantage of a practicality is that the inventor has collected 1500 parts of DNA samples in TS patient, they have all finished an application form of highly structural widely, these application forms are to be the basis with " diagnosis see and treat patients program " (Robins etc., 1981) and DSM-IIIR standard.This makes the inventor can select the serious relatively object of study of symptom, and they more likely have higher genetic load than those symptoms the lighter.Than being applied in the genetics research of extreme phenotype by extensive review (Risch and Zhang, 1996; Plomin etc., 1994).Since clinically, great majority (but being not whole) TS patient also suffers from ADHD, and this provides one group of patient who suffers from and do not suffer from ADHD.This is particularly useful to the method that the inventor detects complete ADHD quantitative trait scope because it with only study ADHD patient compare provide the scope of marking more widely (Comings etc., 1996j).TS patient's number also is useful, because the disease of suffering from altogether in proband and their relatives such as the frequency of ADHD, conduct disorder, oppositional defiant disorder and learning disorder etc. are high (Knell and Comings, 1993; Comings and Comings, 1987; Comings, 1995a, Comings 1995b).Biederman and colleague's report ADHD proband and their relatives have common occurring degree (Biederman etc., 1990a, the b of same high level; Biederman etc. 1993).Although using TS patient's a potential shortcoming and being has sizable overlappingly in clinical and genetic aspect, have the ADHD patient of twitch and compare with the ADHD patient who does not have twitch, may carry different slightly mutant gene groups.Thereby it is important repeating these discoveries in the patient who does not have ADHD.The further restriction of this research is the application form that depends on father and mother and oneself report, and it is optimal that application WISC-R, WRAT-R and other directly detect.But based on the experience that all patients that are studied that see and treat patients in person draw, these application forms have reflected patient's behavior separately exactly.In addition because the inventor never observes the example that the child who does not in fact have significant problem concerning study once was assigned to EH, LD, LD or special education class, the inventor believe by this way the assessment to the LD score showed to cognition can not reliable test.Ironically the assessment whether achievement of two or more subjects (mathematics, reading, writing) of a child is lower than average achievement with whether went up LD class and compared, the difference of a better NA gene score is provided.This show actual school grade can provide cognition can not reliable assessment, and illustrate that many study have the child of obstacle never to be placed into special class.

Because 22 homozygotes of ADRA2A gene seldom, and patient's number of carrying two epinephrine α 2 genes or three NA genes simultaneously also seldom, and the someone may think that present result gets from the occasionality that the homozygotic high score of these minorities exists.In order to check this point, the inventor has also carried out following scoring: 11=0 with the ADRA2A gene, and 12 or 22=1.When two α 2 genes being united with this methods of marking, multivariate analysis of variance still has significance to Inattention (p=0.010), impulsion property (p＜0.001) and move (p=0.021) more, when detecting with linear regression analysis, this 3.1% (p=0.004) that accounts for difference that unite.When the ADRA2A gene of scoring by this way when being added to ADRA2C and DBH gene, then have 66 patients the NA gene must be divided into 3.Multivariate analysis of variance has significance to Inattention (p=0.001), impulsion property (p=0.001) and many moving (p=0.004) scores, and in the linear regression analysis, the NA score accounts for 4.1% (p=0.0002) of difference.The present discovery of these presentation of results is not because the accidental of minority ADRD2A 22 patients' high score occurs.

Inventor's supposition is increased in the several other NA gene that is not related in this research, and this result is further confirmed.These genes comprise norepinephrine transporter (NT), epinephrine α 1 receptor (ADRA1A is to ADRA1D) and PNMT gene, and all these all work in the NA of synapse horizontal adjustment.

Embodiment 28

Neuronic nicotine-like acetylcholinergic receptor α 4 (CHRNA4) gene and attention deficit are many

Move the related of obstacle and tourette's syndrome

The preface nicotine is a kind of addictive drug, and its stimulates dopamine recompense path, improves attention, awakening, learning and memory, in the treatment of tourette's syndrome and ADHD by subcutaneous application successfully.The neuron form of the most outstanding nicotine-like acetylcholinergic receptor is α 4 β 2.The inventor proposes hypothesis and thinks that neuronal nicotinic sample α 4 receptor CHRNA4 genes may be relevant with ADHD or tourette's syndrome.

Neuronal nicotinic sample receptor is made up of α (α 2-α 9) and β subunit (β 2-β 4), and they arrange (Unwin, 1993) around a centre gangway.Alpha subunit has a pair of cysteine, and this muscularity α 1 subunit to nicotine-like acetylcholinergic receptor is similar, and the β subunit does not have this to cysteine.Main neuronal nicotinic sample receptor subtype is formed (Whiting etc., 1991 by α 4 and β 2; Schoepfer etc. 1988).Because α 4 and β 2 are subunit (Wada etc., 1989 of wide expression in brain; Whiting and Lindstrom, 1988; Whiting etc., 1991), the inventor attempts to detect the dependency between the allele of the allele of CHRNA4 gene and CHRNB2 gene in tourette's syndrome and ADHD.Because have only the CHRNA4 gene that available suitable polymorphism (Weiland and Steinlein, 1996) is arranged, so the gene that the inventor detects is CHRNA4.

Method is in 282 uncorrelated non-Hispanic white people tourette's syndrome patients and 63 contrasts, and the inventor has detected the allelic dependency of chromosome set VNTR polymorphism of CHRNA4 gene.Seminar is made up of 345 uncorrelated patients, and wherein 282 people meet the DSM-IV diagnostic criteria and see and treat patients in person by D.E.C., and 63 remaining people are normal control.TS patient is from the tourette's syndrome clinic of wishing the medical centre city.These people and narration in embodiment 27 be used for that to study the noradrenaline plain gene be from same group the patient of the effect of ADHD and learning disorder.It is the same that behavior scoring and ADHD and the differentiation whether learning disorder exists coexist and narrate in that piece article.ADHD scoring is that the standard with ADHD among the DSM-IV (diagnosis, 1994) be basic.Problem is to ask child and juvenile years whether have these symptoms, and being does not have or rare (scoring=0), has (scoring=1) once in a while, be still there (scoring=2) always? all these three answers are used to distinguish fully three orders of severity.The ADHD scoring is the summation of Inattention among the DSM-IV, impulsion property and hyperkinetic syndrome shape.

The polymorphism (Welland and Steinlein, 1996) of first intron of CHRNA4 gene that application is described by Welland and Steinlein, and use the PCR of their report ^TMPrimer and reaction condition.In 88 incoherent white researchs, define 14 allele, length range is 196 to 264bp (seeing Table 84).

Use the polymerase chain reaction (PCR) amplification target DNA, the amount of every kind of used fluorescent dye primer is 0.2uM.PCT ^TMProduct is with 10 times of deionized water dilutions, gets in the mixed liquor that the 0.5ul diluent is added to 2.5ul, and the latter is set to by the smart dye ligand of 75ul Methanamide+9.5ulROX titer+9.5ul blue paste.92 ℃ of degeneration 2 minutes, sample is added to Applied Biosystems 373DNA sequence instrument, and ((ABI) Foster City on 6%PAGE CA), ran glue 5 hours under 1200 volts and stable 30W condition for Applied Biosystems, Inc..Gel is analyzed with internal standard (ROX 500).According to segmental color and size, and peak gene type instrument identification (Applied Biosystems, Inc.).

The inventor has identified the genotype of 345 individualities in this research as a result, has also identified a hundreds of individuality in other research.Except Welland and Steinlein report (Welland and Steinlein, 1996), the inventor has also found many other allele in all research, totally 21.17 are observed in the experimenter of this research.Because Welland and Steinlein be (Welland and Steinlein, 1996) utilized silver to dye, and begun to count allele, up to the bottom from the top of glue, so the allelic bp number of their low numbering is the highest, and the maximum allele of numbering is minimum.Because ABI sequence instrument is to begin the scanning area band when their arrive the bottom of glue, so in inventor's program, low numbering allele is represented the allele of minimum bp number and the highest numbering allele represent the allele of maximum.Though the maiden work of inventor much make great efforts to attempt to use with Welland and Steinlein (Welland and Steinlein, 1996) Yi Yang numbering, but because the result of order-checking shows that allelic bp number varies in size (especially endways), this is very difficult.Thereby the inventor has used the ABI numbering system, and in order to compare, the inventor lists in table 84 with two kinds of nomenclatures.

Table 85 has been listed four kinds of PCR ^TMThe sequencing result of product.This has confirmed the complexity of this VNTR polymorphism.

In main allele, 9,12,14 and 16 allelic frequencies are higher in TS patient, and 13,15 and 18 allelic frequencies are higher in matched group.Analyzing 17 allelic results with X 2 test is: X ²=33.65, d.f.=16 p=0.0061.When analysis is defined to frequency in matched group or the TS group is 0.05 or during bigger patient, the result is: X ²=71.6, d.f.=6, p＜0.0000001.Gene frequency difference maximum be #9 allele.

Easy for what keep analyzing, for the effect of amount of inspection variable and homozygote and heterozygote, in order to produce the regression analysis variable, the standards of grading of CNRNA4 gene are as follows: x/x genotype (non--9/ non--9)=0,9/x=2,9/9=3.

Beyond the one group of behavior variable that in embodiment 28 noradrenaline plain gene (Comings etc., 1998) is detected, the inventor has detected total twitch score (Comings etc., 1996).Detect the dependency of CNRNA4 gene score and these variablees simultaneously with the multivariate analysis of variance method.The result shows that the dependency of study performance (GSAP), learning disorder (LD), oppositional defiant disorder (ODD) and many moving scores of CNRNA4 gene and primary school is significant.

In order to estimate the influence of different genotype, the variance test of total ADHD score and the polynary variance test (in p＜0.1) (table 87) of each significance of difference of detection have been carried out to these scores.Presentation of results all is the highest to each score of 9/9 homozygote.

Because also have the another one problem is whether CNRNA4 gene and smoking have some dependencys, the inventor has detected two variablees in the patient more than 17 years old: " inhaling cigarette " and " bag every days 7 ", this does not have significance, 9/9 homozygote patient smoking capacity every day is 0.5 bag, and other genotype is 0.10 to 0.20 bag.9 9/9 homozygotic patients are here only arranged, and bigger sample number may obtain significant result.

When analyzing the concerning of four groups of CNRNA4 genotype and ADHD-LD-, ADHD+LD-, ADHD-LD+ and ADHD+LD+ with X 2 test, the frequency of 9/9 genotype in above-mentioned four groups increases, and is followed successively by 5.6%, 6.0%, 7.1% and 13.5%.During the Pearson X 2 test, total dependency is inapparent (p=0.385), is critical relevant (p=0.051) during linear X 2 test.Thereby not as detected norepinephrine (Comings etc., 1998), the evidence that the CNRNA4 gene works in the ADHD-LD type relevant with brain calvarium leaf seldom.

The genotype variable to the result of the single-factor regressioning analysis of total ADHD score is: r=0.123, r ²=0.015, T=2.30, p=0.022.In TS patient, 9,12,14 and 16 allelic frequencies increase, and 9 allele increase maximum; 13,15 and 18 allelic frequencies increase in matched group, and 13 gene frequencies increase maximum.When with X 2 test all allele being compared, they have significant difference, p=0.002.When comparison frequency only during, significant difference is arranged, X between each group greater than 0.05 allele ²=71.6, d.f.=6, p＜0.0000001.Checked eight relevant quantity scores of ADHD, twitch, conduct and study with based on the relation between the 9 allelic genotype groupings (9/9,9/x, x/x) with the multivariate analysis of variance method, disobeyed and how in the moving score significant dependency is arranged in one learns in grade school performance, learning disorder, opposition.Variance test shows that 9/9 homozygotic score is the highest in all scores.The linear regression analysis of genotype score shows that the CNRNA4 gene accounts for 1.5% (p=0.022) of ADHD score difference.

These presentation of results of conclusion CHRNA4 locus be with the dangerous relevant several genes of ADHD and TS in one.

The PCR that in this research, uses ^TMThe sequence analysis of amplified fragments shows that this is a very complicated polymorphism, relates to the difference of size and the difference of sequence.Repeating polymorphism itself may work in the adjusting of the gene relevant with them.This is the tendentiousness that forms Z-DNA based on these repetitive sequences.The length of repetitive sequence and the variation of sequence are all worked in the decision of Z-DNA total amount, and Z-DNA participates in the adjusting (Comings, 1997) of gene by several approach.Show on size and sequence in the complicated polymorphism of this detection significant variation has all taken place.Therefore it may be suitable for association study uniquely.

For fear of the reduction to the independent evaluation capacity of many other scores, the inventor is defined in the same behavior score that detects with their analysis and adds a total twitch score in embodiment 27.Twitch because nicotine has been used to treatment, increase again whether studied this gene relevant with twitch itself.Multivariate analysis of variance is used for estimating the score as a group, avoids Bonferroni to proofread and correct.This shows that four have significant dependency (table 86) in eight behavior scores.When each variable being carried out the research of post hoc variance, 9/9 homozygotic score is the highest.The presentation of results CHNRA4 that merges is one of genetic risk factor gene that works in ADHD and ST.Its percentage rate of accounting for ADHD score difference of regression analysis presentation of results may reach 1.5%.

Though the dependency of the demonstration of CHRNA4 gene and learning disorder and one learns in grade school performance than with the dependency of ADHD time grade of score more greatly, but not as the noradrenaline plain gene of detection, having or do not having ADHD and having or do not have in four groups of learning disorder, the sign that 9/9 genotypic frequency does not increase gradually.

Though the inventor does not also know to be with the increase of CHRNA4 gene expression or to reduce relevantly that in the grouping of the genotype of this narration this research support nicotine is to the clinical effectiveness of ADHD and TS symptom.In other crowd, the difference between contrast and TS or ADHD also might relate to other allele beyond 9 allele.When a plurality of pattern detection, the most repeatably the possibility of result is to organize the comparison of common allele frequency among the patient with what X 2 test was analyzed at matched group and TS group or ADHD.

Table 84 Comings and Wu with Applied Biosystems, Inc DNA sequence instrument be the basis:

Allele is with Welland and the allelic comparison of Steinlein

(Weiland and Steinlein, 1996)

Comings and Wu Welland and the Steinlein allele frequency allele frequency that is harmonious that is harmonious

14??????????????196??????.04

13??????????????198??????.011????????????????203??????.0082?????????207?????????????.000???????12??????208??????????????.013????????????????215??????.0244????????????????217??????.0635?????????221?????????????.000???????11??????208??????????????.016?????????223?????????????.008???????10??????220??????????????.047?????????225?????????????.238???????9???????222??????????????.018????????????????227??????.0249?????????231?????????????.015???????8???????228??????????????.0610???????????????233??????.00811???????????????235??????.03212???????????????237??????.06313????????239?????????????.302???????7???????236??????????????.4514????????241?????????????.02415????????243?????????????.238???????6???????240??????????????.1916????????245?????????????.008???????5???????242??????????????.0117????????253?????????????.000???????4???????248??????????????.0118????????255?????????????.063???????3???????250??????????????.1119????????257?????????????.024???????2???????252??????????????.0320????????259?????????????.000

1????????????????264?????.0121????????272?????????????.008

Multiple 4 PCR of table 85CHRNA4 VNTR ^TMSequence allele ABI sequence nucleotide sequence # size 9 231.6 231 CCGGGCGTGCGC (TG) of product ₈(CG) ₀CCGGGCGTGCGC (TG) ₈CCGGGCGTGCGC (TG) ₁₁13 239.5 239 CCGGGCGTGCGC (TG) ₈(CG) ₂CCGGGCGTGCGC (TG) ₈CCGGGCGTGCGC (TG) ₁₃15 243.8 243 CCGGGCGTGCGC (TG) ₈(CG) ₂CCGGGCGTGCGC (TG) ₁₀CCGGGCGTGCGC (TG) ₁₃18 254.9 254 CCGGGCGTGCGC (TG) ₉(CG) ₁CCGGGCGTGCGC (TG) ₁₀CCGGGCGTGCGC (GTG) ₁CCGGGCGTGCGC (T (

Table 86

The multivariate analysis of variance of 8 behavior scores

CHRNA4?9/9＝3，9/x＝2，x/x＝1

F-is Duoed 3.10 .046 Inattentions, 2.92 .055 than pGSAP 3.60 .028LD 3.60 .028ODD 3.46 .033 and is twitched 1.63 .198 impulsion property, 0.92 .401CD, 0.55 .573 summation (Wilks), 1.22 .246

Table 87

Variance analysis to the individual score of ADHD factor

CHRNA4 (n=345) scoring genotype N mean S.D. F p F ₁p ₁ADHD x/x 204 17.28 11.12

13/x????113????18.94????11.05

9/9 23 22.61 ^*9.09 2.83 .060,5.27 .022 move x/x 209 5.75 4.43 more

9/x?????113????6.18?????4.18

9/9 23 8.08 ^*3.86 3.11 .045,4.85 .028 Inattention x/x 209 8.72 5.53

9/x?????112????9.73?????5.55

9/9?????23?????11.08????4.88?????2.69????.069????5.34????.021ODD?????????x/x?????209????3.45?????3.21

9/x?????113????3.83?????3.19

9/9?????23?????5.26 ^*???2.87?????3.46????.032????5.60????.0185LD??????????x/x?????209????0.61?????0.93

9/x 113 0.84 1.03 scoring genotype N mean S.D. F p F ₁p ₁

9/9??????23????1.04????1.10????3.61????.028???7.21???.0076GSAP???????????x/x??????209???2.78????1.99

9/x??????113???3.35 ^*??1.90

He once smoked x/x 91 1.39 0.49 (age＞17 year old) 9/x 49 1.31 0.46 9/9 23 3.40 1.92 3.61 .028,6.49 .011 base

9/9 6 1.66 0.52 1.66 .192,0.000 .995 bag/sky x/x 91 0.12 0.44

9/x??????49????0.10????0.37

9/9??????6?????0.50????0.84???2.24?????.110???1.02???.314

^*Tukey check has marked difference with x/x in α=0.05 o'clock

F ₁The analysis of=linear equation,

P ₁The p that linear equation is analyzed

Embodiment 29

MAOA gene VNTR allele length and TOURETTE that X is chain

The dependency of the phenotypic effect of syndrome and drug dependence

Preface. ammoxidation single enzyme (MAO) level unusual relevant with many psychiatric disorders.Do the inventor has detected the polymorphism of the chain MOA gene VNTR of X to check two kinds of hypothesis: the variation of (1) MAOA gene work in the part behavior disorder relevant with Tourette syndrome or drug dependence? (2) if like this, between allele length and phenotypic effect, is there dependency? the inventor has detected two independently groups: 375 patients TS, relatives and contrasts, and 280 substance abuse persons and contrast.Allele is divided into ever-increasing four groups of length.Have significant dependency between the behavior phenotype of MAOA gene and two groups, and the longest allele is relevant with the strongest phenotypic effect in two groups.The strongest effect is the diagnosis (p=0.00003) of drug dependence.Allelic these groups of VNTR and the Fnu4H1 polymorphism significant linkage disequilibrium relevant that showed in the past with MAO-A activity.The probability that the different length allele of the multiple polymorphism of these results and short weight itself may work in Gene regulation is corresponding to.

Because age (Devor etc., 1994), ethanol, antidepressants, medicine, sex, experimental technique, diet and other variable are to thrombocytozyme level (Fowler etc., 1982) potential effect, the application of MAO gene genetic polymorphism may provide than enzyme level and more many repeatably result.The clone of MAOA and MAO-B gene makes with the evaluation of order-checking and relevant polymorphism can carry out this genetics research now.

For the hereditary variation of clear and definite whether MAOA gene relevant with TA or ADHD, technology (the Comings etc. that the inventor utilizes DRD2 among the TS, D β H and DAT1 gene to report, 1996a), detected the MAOA VNTR polymorphism (Hinds etc., 1992) in a series of contrasts, TS proband and their relatives.Since contrast with respect to the simple comparison of the different gene frequencies of TS proband may omit the MAO gene only with the relevant probability of some special behaviors that in all examples, does not occur, the inventor has checked these genes may act in 27 different behavior variablees.

The inventor may the hypothesis relevant with phenotypic effect produce interest to the equipotential mrna length.Its theoretical basis is that most of simple repeating sequences has caused the formation of Z-DNA and its quantity to depend on multiple length (Schroth etc., 1992).The Z-DNA structure has been opened the dna double spiral and has been exposed single base, makes its can interact with nucleoprotein (Rich etc., 1984).Owing to these and other reason, Z-DNA relevant (Comings, 1996 a with Gene regulation; 1996b).If little or microsatellite polymorphism works in the gene function variation that relates to polygenic inheritance really, they can cause individual gene rather than polygenes obstacle.

The chain gene of X-has formed the carrier of a uniqueness to check this hypothesis and to study meticulous effect, this is because at least in the male, each allele occurs with hemizygosity, so just eliminated the complicated factor of heterozygote, and the quantity of heterozygote is a lot of when the allele of a plurality of different sizes occurs.In order to check the repeat length may the hypothesis relevant with phenotypic effect, the inventor be divided into ever-increasing four groups of length with VNTR allele.This allow the inventor can be clear and definite whether shorter or longer allele is preferential relevant with bigger phenotypic effect.

Method. the experimenter comprises 57 contrasts, 229 TS proband, and proband's major part is subjected to have a strong impact on (Comings etc., 1990) with a plurality of behavior associated disorders, and 90 ill and not ill TS proband relatives.All derive from West Europe more than experimenter's right and wrong Spain white man and 90%.The contrast of TS group contains TS proband's foster parents and stepparent, comes from the other than psychotic obstacle experimenter who wishes other clinics, city and wishes occupation of medical center, city and the non-professional employee of hospital.TS experimenter and contrast all have detailed description (Comings etc., 1995b elsewhere; Comings etc., 1996a; 1997b).

Each TS contrast and TS proband or relatives are required to fill out an application form based on diagnosis meet program (Robins etc., 1981) or DSM-III-R (mental disorder diagnostic and statistical manual, 1987) standard.This provides the structural description about a large amount of psychotic symptoms.These symptoms are divided into 27 different behaviors, comprise ADHD, substance abuse, emotion, anxiety, school's performance, stutter, twitch and other.The problem that is used for these behavior scorings is described (Comings, 1995a elsewhere in detail; 1994a; 1994b; 1995b; Comings etc., 1996a; Rubins etc., 1981; Comings, 1995c).Two kinds of behavior scorings are used to assess ADHD.At first, why be referred to as ADHD, be based on half of 22 ADHD variable series occurring DSM-III and DSM-III-R standard at least.The second, ADHD-R is based on the DSM-III-R diagnostic criteria.Three of usefulness QTV were not Inattention, impulsion property and moved obstacle more in the past.There is the accumulation of these three inferior scorings can obtain the ADHD scoring.QTV abbreviation comprise the CD of conduct disorder, the ODD of oppositional defiant disorder (Comings etc., 1995a) and MDE (Comings, 1995c) symptom of major depression outbreak.

The theoretical basis that detects the behavior of falling ill altogether is former observation, special genes may with the special behavior of morbidity altogether that occurs among the TS than diagnosis itself have stronger relevant (Comings etc., 1996a).This application form does not also mean that the highly structural method that provides DSM-III-R or DSM-IV to diagnose and provide generation behavior different aspect QTV.The advantage of continuous trait is can provide than the bigger seriousness grade classification of two fork diagnosis.Accuracy, practicality and the sensitivity of the symptom appraisal procedure of application form approach is by other people (Gadow and Sprafkin, 1994; Grayson and Carlson, 1991) use the situation of same structure chemical industry tool to be illustrated by relatively using the such instrument and the person of seeing and treating patients.The inventor consults explanation they has accurately reflected the information that obtains by individual's talk for hundreds of experimenter's application forms.

Second group of experimenter contained 120 non-Spain white man male, they have come from the California since in October, 1994, Loma Linda, Jerry L Pettis VeterransAdministration Hospital inpatient addiction therapy unit (ATU), the person that newly is not admitted to hospital that all agree to enter ATU has participated in the research about drug dependence/dependence inherited genetic factors that National Institute of Drug Abuse is sponsored.

All ATU experimenters are by Michigan alcoholism level evaluation (Davis etc., 1987) (24 oneselfs of revision carry out application form, comprise drug dependence (MAST-R)), diagnosis meet program (DSM-III-R the version) (Robins etc. that carry out of clinicist, 1981) come diagnostic substances to rely on the appearance of obstacle, assess a series of ethanol and drug use variable with the 5th edition (ASI) (Hodgins and Guebaly, 1992) of addiction severity index that the clinicist carries out.

The inventor utilizes the statutory status part of medicine/ethanol use and ASI.This aspect has comprised with the lower part:

A) the concrete material of Shi Yonging. in order to assess the use of concrete material, asked about using (year number) all one's life of drunken, heroin, other Opium/analgesic, barbiturate, other analgesic/hypnotic/tranquilizer, cocaine, amphetamine, Fructus Cannabis, hallucinogenic agent and inhalant.

B) approach of Shi Yonging. for each above-mentioned medicine, in due course, the experimenter may I ask about using the approach problem.Selection has mouth, nose, smoking and IV injection.This continuous variable of #IV drug use is calculated by different IV injectable drugs is counted addition.Variable IV drug use is two distortion amounts, the 0th, there is not drug use, and≤1 is to use one or more medicines IV.

C) problem. have you had ethanol DT how many times? overdose? is there there the ethanol problem that how many days run in the past 30 days? the medicine problem?

D) Hua Fei money. how much have you spent on ethanol in the past 30 days? on medicine?

E) seriousness. based on the level evaluation of interviewer, (there is no need treatment) to 9 (under life-threatening situation, needing the treatment intervention) from 0 to the treatment desirability.Alcohol abuse? drug dependence?

F) statutory status. also asked different laws aspect about medicine and alcohol abuse.Be there in life at you that how many times is controlled drives when intoxicated? does non-hibernating eggs at you have how many times because drug issue arrested and charge? is there the such charge of how many times to cause determining a crime?

G) generalized scoring. can be when answering from 0 when any number change, they are 0 is that other any several then scorings are ' 1 ' by scoring for ' 0 '.Use relevant problem to count total ethanol scoring with ethanol, the problem relevant with drug use counts total medicine scoring.

The contrast of substance abuse group and patient's TS contrast are irrelevant.They comprise two parts.First is 45 older male, from the non-Spain student of the state university in California of San Bernardino (30.1 years old mean age).Those have being got rid of of significant substance abuse problem on the basis of MAST-R check.Second portion comprises gemellary father in the Minnesota geminus family study project.Because these people come from a state clearly, merely based on having 11 or 17 years old geminus, they than the student delegate of university more at random part in all social economies and the education colony.Though all scorings to impinging upon substance abuse variable aspect are negative, owing to the result who gemellary contrast is carried out the substance abuse assessment is invalid, so part may be male.Yet, be main cross-over experiment at random because this is agriculture state, inventor's imagination, false-negative quantity is very little in this group.

Select the theoretical basis of MAOA gene VNTR polymorphism as follows.The series connection of selecting a weak point repeats polymorphism and is used for this hypothesis of special detection: multiple length may be relevant with phenotypic effect.Select the chain gene of X to be owing at least when the research male, avoided explaining the complexity of heterozygote.Selecting the MAO gene is that X is chain because of it.Selecting the MAOA gene is because reported that two different repetition polymorphisms are relevant with it.The inventor select VNTR polymorphism (Hinds etc., 1992) because it allelic length (40+bp) go up than (CA) n repeat (16bp) wide ranges many.

This complicated polymorphism contains directly and repeats the adjacent little satellite of GT of new 23-bp VNTR motif with incomplete two times, and wherein dinucleotide repeats all different with the multiple number of VNTR in the allele.The VNTR polymorphism appears on the 2.9-Kb SalI-EcoRI fragment of the phage 6.12 that contains MAOA gene first exon (Hinds etc., 1992).From whole blood, extract DNA by standard method.Target DNA passes through PCR ^TMAmplification (Mullis etc., 1986).For labelling PCR ^TMProduct, labelling fluorescence HEX or FAM Amidite (applying biological system, Foster City, CA, every primer 0.1 μ M USA) are used for reaction (table 88).The PCR of 10 times of dilutions of two microlitres ^TMProduct adds ROX 500 standards (applying biological system) of 2.5 μ l deionized formamides and 0.5 μ l and 92 ℃ of degeneration 2 minutes, goes up sample then to the polyacrylamide gel of applying biological system 373 sequenators 6%.Gel electrophoresis 5 hours under 1100V and constant 30W.Utilize inner ROX500 standard that gel is carried out laser scanning and analysis.The peak is discerned based on the color fragment size that is determined by base pair length by gene type instrument (1.1 version) (applying biological system).The full detail of each sample prints from the file of every clotting glue, and the data of compilation are analyzed.

Table 88

MAOA VNTR polymorphism: by the different more different row of allele grouping of variance analysis

Be scoring.(mean and standard deviation) (n=287) behavior passes through the allele F of bp size packets than P F ₂

＜320???????320-333????????334??????????≥335

(n=82) (n=43) (n=110) (n=52) manic 1.43* 1.8 1.36* 1.9 1.59* 2.27 2.59 2.8 3.59 0.014 6.39 ^aOCD 2.18 2.8 2.18 2.4 2.80 2.8 3.31 3.2 2.16 0.093 5.89 ^aProperty 0.62* 1.1 0.47* 0.9 0.66* 1.2 1.25 1.6 3.91 0.009 5.82

^aSleep 0.36 0.7 0.38 0.8 0.49 0.9 0.76 1.1 2.31 0.075 5.56 ^aPrimary school 2.60 1.7 2.90 2.0 2.98 2.1 3.46 2.1 1.89 0.131 5.14 ^aGambling 0.13* 0.9 0.27 0.8 0.22 0.8 0.61 1.6 2.49 0.060 4.82 ^aStutter 0.13 0.3 0.11 0.3 0.25 0.4 0.23 0.4 2.33 0.084 4.41 ^aStudy 0.52* 0.9 0.65 1.0 0.54* 0.9 1.00 1.1 3.32 0.020 4.37 ^aInattention 6.69 5.2 7.37 4.9 7.67 4.7 8.48 5.3 1.42 0.235 4.15ADHD 19.43 14.9 20.95 13.5 21.18 13.9 24.80 15.3 1.53 0.206 3.74 ^aADDR 4.63 4.9 4.84 4.6 5.18 4.7 6.42 5.2 1.57 0.196 3.74 ^aTable 88 (continuing) behavior is passed through the allele F of bp size packets than P F ₂

＜320????????320-333????????334?????????≥335

(n=82) (n=43) (n=110) (n=52) gets excited 5.74 4.9 6.30 4.8 6.49 4.7 7.46 5.0 1.35 0.257 3.68 ^a 0.58 1.3 1.00 2.2 1.32 2.8 1.09 2.4 1.61 0.186 3.23MDE 3.00 2.8 3.29 3.1 3.45 3.1 3.92 3.1 1.01 0.385 2.90CD 2.78 2.4 2.90 2.1 2.97 2.2 3.54 2.6 1.15 0.328 2.54 6.91 5.6 7.27 5.2 7.00 5.4 8.86 5.9 1.59 0.190 2.29 2.00 2.7 2.15 3.0 2.45 2.7 2.65 3.3 0.71 0.548 2.10 1.31 2.3 0.68* 1.3 1.34 2.2 1.84 2.3 2.26 0.081 1.92 0.21 0.4 0.18 0.3 0.28 0.4 0.29 0.4 0.85 0.463 1.60 2.13 3.0 1.58 2.4 2.17 3.0 2.90 3.7 1.16 0.325 1.34 0.36 1.2 0.45 1.6 0.40 1.3 0.75 2.0 0.81 0.489 1.28 1.86 1.9 1.36 1.8 1.78 2.1 2.34 2.5 1.75 0.156 1.270DD 3.07 3.3 3.02 2.7 3.16 3.1 3.73 3.4 0.57 0.633 1.02 2.81 3.7 3.04 3.4 2.84 3.5 3.57 4.1 0.55 0.642 0.73 0.51 2.3 0.81 2.9 0.31 1.8 1.11 3.2 1.41 0.241 0.45 2.91 2.0 3.09 2.1 3.18 2.2 2.98 2.2 0.27 0.845 0.20 0.07 0.3 0.11 0.3 0.05 0.2 0.77 0.3 0.53 0.662 0.10^aSignificance＜0.05, F ₂The F ratio of=linear equation.* significance be lower than＞335, pass through Tukey in α=0.05 level.

In order to check the MAOA allele length may the hypothesis relevant with phenotypic effect, allele be divided into four groups (seeing the result).1 to 4 labelling is arranged, the shortest in the longest formation MAOA genotype variable.Only in the TS group, use the women.Only those that isozygoty in given allele grouping just are included in the analysis.

The inventor has utilized two principle to divide into groups: enough groups a) must be arranged to detect a series of length; B) for the maximized with statistics, the quantity of experimenter in every group must be similar.Like this, if gene frequency has only single peak, just should be divided into the shortest by 1/3, medium by 1/3, the longest by 1/3.Yet the allelic size distribution of MAOA VNTR is two peaks.For having only the male, small peak contains 32% allele, and size is long from 299 to 314bp.Big peak contains 68% allele, size from 323 to 338bp.Because allelic main body is in this peak, the inventor with it as existing single peak to separate (promptly short, medium, long allele grouping).One 334bp group is contained at the center at this peak, and it comprises 31% the allele that can not divide into groups again.The application of these regulations caused four group＜320bp, 320-333bp, 334bp, 〉=335bp contains 32,23,31,14% allele respectively.

It is impossible utilizing the grouping of (1992) descriptions such as Hinds, because three gene frequency is very low in their five groups.In fact, the inventor does not have the experimenter in this three accessory group.

The Fnu4H1 polymorphism is to the check of this polymorphism method based on Hotamisligil and Breakefield (1994).The inventor is their '-' called after inventor's ' 1 ' allele, their '+' called after inventor's ' 2 ' allele.In their research ,+allele has higher MAOA activity.

For Tourette syndrome group, with the relative size of each QTV in four not homoallelic groupings of variance analysis check.Remarkable progressivity with average between four groups of linear equation analytical control increases.Use SPSS (SPSS, Inc, Chicago, IL, USA) statistical package.For the linear equation analysis, the subcommand multinomial is set at 1.Whether these QTV when all variablees are checked simultaneously are significant with the multivariate analysis of variance check.With the multiple linear regression analysis as the check when all variablees are checked simultaneously second whether remarkable method of QTV.The MAOA genotype is set at non-independent variable and 27 QTV progressively enter as independent variable.

Chi-square analysis explanation has the highest meansigma methods to main QTV for the longest allelic group.The potential progressivity of 〉=335bp allele class frequency descends and compares between four groups that symptom progressivity at TS reduces: have the TS proband of ADHD, no ADHD the TS proband, the relatives of TS are arranged and do not have the relatives of TS.

For the substance abuse group, multivariate analysis of variance whether be used to check between the variable ethanol of four allelic groupings of MAOA and two simplifications and medicine scoring, exist significant relevant.Average with four allele grouping ethanol of variance analysis check and medicine scoring.

The potential progressivity of frequency between three groups with linear X 2 test 〉=335bp allele group increases: the substance abuse person's (ATU has) who contrasts, do not have substance abuse person's (ATU does not have) of behavior and behavior is arranged.ATU does not have group and is included to get rid of because the probability of the frequency increase of this equipotential genome that the common morbidity of different behaviors causes in substance abuse person.In order to help to get rid of it, the genomic frequency of equipotential will be at least exceeds 20% than no behavior in the substance abuse person that behavior is arranged.Because these allelic frequencies of hypothesis progressively increase, therefore used linear chi-square statistics in these three groups.

In order to detect the maximum variation ratio of the medicine correlated variables that the MAOA gene causes, be that 1 carry＜allelic experimenter of 335bp and scoring are that 2 carry 〉=allelic experimenter of 335bp has carried out regression analysis in scoring.This analysis is carried out for the drug dependence variable, because this is and the maximally related chi-square variable of MAOA gene (the contrast scoring is 1, and it is 2 that ATU does not have scoring, and it is 3 that ATU has scoring).

The allelic distribution of .MAOA VNTR polymorphism as a result (allele sum=768).Because this is complicated VNTR, allele can not become the dividing mode of base pair even number or odd number.It is such that result and gene type instrument program produce.Between 316bp and 323bp, do not have allele, so just produced two clear and definite important group＜320bp and＞320bp.Yet, may the hypothesis relevant for the check table effect with size, allelic bigger grouping 323-339bp is divided into three subgroups, comprises the main peak of allele 320-333bp, the 334bp shorter than main peak and the allele 〉=335bp longer than main peak.In TS group 219 male and 156 women 375 experimenters are altogether arranged.88 is heterozygote among the women.After these have been got rid of, be left 287 experimenters in the research, wherein 36 is contrast.In last group, the male compares four allele groupings with the women frequency distribution does not have significant difference.

Each The results of analysis of variance with respect to four allele groupings of TS group QTV sees Table 88.Conventional The results of analysis of variance is below F ratio and P value.The F ratio that linear equation is analyzed is at F ₂Below the hurdle, subscript ^aExpression significance＜0.05.QTV passes through F ₂The F ratio magnitude on hurdle falls progressively and arranges.In≤0.05 horizontal detection, mean significantly is lower than 〉=those allele groupings of 335bp group mark with an asterisk by Tukey check.Except stutter, shopping and terrified (minimum F ratio is arranged), 24 remaining QTV those carry 〉=335 allelic experimenters in mean be the highest.

The multivariate analysis of variance result of all 27 QTV is significant for property (P=0.012), problem concerning study (P=0.023), gambling (P=0.025) and manic (P=0.025).

When all 27 QTV simultaneously when progressively multiple regression analysis detects, variable primary school problem (P=0.012) and gamble (P=0.038) are significant.Based on r ²Value, MAOA gene have only caused 3.9% of these QTV variations.

Utilize chi-square analysis, carry 〉=335bp allele experimenter's ratio exists significant progressivity to reduce, from the TS proband (24% of ADHD is arranged, n=129), to the TS proband of no ADHD (20.0%, n=50), arrive TS relatives (12.5%, n=16) to non-TS relatives (5.6%, n=56) (P=0.003).

Contrast and ATU experimenter have been compared in the substance abuse group.Unite contrast for 160, the distribution of four allele groupings is as follows:＜320 34.4%, 320-333 38.1%, 334-33521.3%, 〉=335 6.3%.For 120 ATU experimenters, frequency is as follows:＜320 39.2%, 320-333 18.3%, 334 20.8%, 〉=335 21.7%.There is significant difference, x ²=22.17, P=0.00006.The frequency of 〉=335bp group is similarly at two matched groups, 8.9% and geminus father and mother's of SanBernardino group 5.2% (x ²=0.774, P=0.38).

The multivariate analysis of variance explanation of ethanol and medicine scoring, though all with MAOA gene VNTR allele significant correlation, medicine scoring (P=0.001) is than ethanol scoring (P=0.012) more remarkable (table 89).The result of associating multivariate analysis of variance also is significant (P=0.007).257 n is than the sum of+120 ATU of 160 contrasts or 280 little, because have only 97 experimenters to finish ASI.By contrast, all finished the DIS that is used for ethanol and/or drug dependence DSM diagnosis validation for all 120.

Table 89 substance abuse group ethanol and medicine scoring are to the multivariate analysis of variance of MAO allele grouping

(n=257 has only the male) variable F ratio P ethanol 3.72 0.012 medicines, 5.85 0.001 sums (Wilks) 2.99 0.007 of marking of marking

Represent the variance analysis of two groups of scorings of each allele grouping mean, see Table 90.For TS group, the highest mean appears at 〉=the allelic grouping of 335bp in.For medicine scoring, by the Tukey check, other three allele groups significantly than 〉=335bp allele group is low.

Table 90

The ethanol of substance abuse group and medicine scoring are to variance analysis allele n mean standard deviation F ratio P grouping ethanol scoring＜320 94 2.27 3.1320-333 81 1.49 of MAOA allele grouping ^a3.0334 56 1.94 2.7 〉=335 26 3.73 3.52 3.72 0.012 medicines scoring＜320 94 3.59 ^a5.2320-333 81 1.94 ^a3.8334 56 3.34 ^a4.9 〉=335 26 6.42 6.2 5.85 0.0007

^aSignificantly be lower than in α=0.05≤mean of 335 allele grouping by Tukey check.

Whether preferential relevant in order to detect the MAO gene with the particular type of substance abuse, utilize chi-square analysis to detect 14 variablees relevant with the material type of using.The experimenter's (ATU does not have) who contrasts relative no behavior ATU has the frequency of experimenter's (ATU has) of behavior ATU 〉=335bp allele grouping to see Table 91 relatively.Owing to checked material to use 14 types of variable, have only those P values be lower than 0.0036 (0.05/14) be considered to significant by the Bonferroni rectification.Have only being listed on the table of those P value＜0.01.Having only alcohol dependence is exception.This has illustrated and the comparing of drug dependence or medicine and alcohol dependence, the alcohol dependence experimenter 〉=only there is a spot of increase in the 335bp gene frequency.By contrast, have only the variable of drug dependence to obtain peak (x ²=17.4, P=0.00003).

Table 91 contrast with respect to the ATU experimenter who does not have behavior with respect to the ATU experimenter that behavior is arranged carry 〉=

The linear chi-square analysis behavior contrast ATU of 335bp allele experimenter quantity does not have ATU card side P

N % n % n % n % has only drug dependence 160 6.3 58 15.5 62 27.4 17.4 0.0000

3IV drug use 160 6.3 56 14.3 27 29.6 13.5 0.0002

2ODed 160 6.3 71 12.7 25 28.0 11.00 0.0009 barbitals use 160 6.3 61 13.1 32 25.0 11.56 0.0011DUIs, 160 6.3 34 8.8 62 21.0 9.92 0.0016 amphetamines to use 160 6.3 19 5.3 77 19.5 9.37 0.0022OSH to use " 160 6.3 67 14.9 26 23.1 8.96 0.0027 cocaines use 160 6.3 25 12.0 67 19.4 8.86 0.003 hemps to use 160 6.3 14 7.1 82 18.3 8.35 0.004 heroin to use 160 6.3 68 14.7 28 21.4 8.05 0.004 opium to use 160 6.3 58 15.5 38 18.4 6.88 0.009 to only have alcohol dependence 160 6.3 98 24.5 22 9.1 non-linear

^aOther opiates of OSH=(non-heroin or methadone), tranquilizer and sleeping pill.

Allele grouping (＜335 pairs 〉=335) has obtained following result: r=0.25, r with respect to the regression analysis of drug dependence diagnosis ²=0.0625, T=4.305, and P=0.0001.

In order to detect the allelic potential linkage disequilibrium of VNTR and Fnu4H1, the inventor has also carried out VNTR polymorphism gene type to 273 male of gene type.It is because clearer than women's result that the inventor is defined as the male with analysis.There is the nonrandom relevant (x of highly significant in two polymorphisms ²=132.91, P＜0.000001).＜320 VNTR allele grouping is relevant with uncommon Fnu4H1 2 allele, and remaining three VNTR grouping is relevant with Fnu4H11 allele.

Based on the comparison of data, can predict, if VNTR allele be divided into＜320bp and＞320bp, just can obtain the result similar, Fnu4H1 2 ≈＜320 and Fnu4H1 1 ≈＞320 to the Fnu4H1 polymorphism.The TS group has 71 experimenters that two kinds of polymorphisms have been carried out gene type.Because the manic the most significant result (table 88) that obtained, so this variable is used for comparison.Carrying Fnu4H1 1 allelic 53 experimenters, to get mean be 2.01 (standard deviations 2.17), and 2 allele are 1.55 (standard deviations).The VNTR similar data is＞1.94 (standard deviations 2.12) of 320bp and＜1.75 (standard deviations 1.98) of 320bp.(16 〉=335 experimenter's mean is 2.43 (standard deviations 2.42)).Because quantity is few relatively, the neither one group is significant.Because it is active relevant that the research (1994) of Hotamisligil and Breakefield shows Fnu4H1 allele (inventor's I allele) and low MAOA, the allelic grouping of inventor imagination 〉=335VNTR is active relevant with minimum MAOA.

Because the Tourette syndrome is highly heritable and usually impulsion, attack, emotion, the property with a large amount of is excessively relevant with other behaviors, it is to be fit to very much this research.Present result shows that the MAOA gene is one that plays in a small amount of effect gene in the etiology of many TS corelation behaviours.

Though multivariate analysis of variance has illustrated MAOA allele and ethanol and medicine all significant correlations of marking, there is the morbidity property altogether in a large number of these two kinds of substance abuse forms.Shown in table 91, when drug dependence and alcohol dependence detect respectively, relevant stronger with the correlation ratio of alcohol dependence and drug dependence.

Though the outstanding of male may be partly owing to hormone and environmental factors, the chain gene of X also may be a factor.For TS group, the r that utilizes regression coefficient to determine ²Illustrate that for different QTV, the MAOA gene causes 2.5% or QTV still less variation at most, shows that the chain MAOA gene of X can not cause the outstanding of TS among the male, ADHD and associated disorders.By contrast, 〉=335bp allele lacks or occurs the r of relative drug dependence diagnosis ²Illustrate that the variation up to 6.2% may be because the MAOA gene.This may play moderate in the male of drug dependence is outstanding.

The inventor begins to guess that the different length allele of little satellite and moonlet polymorphism may work in the Gene regulation relevant with them.Though the dependency of the long moonlet allele and the special QTV of Tourette syndrome group is not strong, shown in table 89, there is significant concordance in the trend between all QTV.Because it is relevant that this may be an accidental random, whether the inventor attempts to prove conclusively them can repeat these results in complete independent moral experimenter and matched group.The allele that this group (substance abuse group) and MAOA VNTR are long especially 〉=335bp allele, presents stronger dependency than observed in the TS group.This pattern of two groups is very similar, and 〉=335bp allele has the highest scoring, and minimum allele (＜320) has higher scoring, scoring in the middle of 334-335bp allele has.

For whether see clearly 〉=335bp allele may be active relevant with higher or lower MAO-A, the inventor has also carried out Fnu4H1 polymorphism gene type to 273 male subject of inventor.With the linkage disequilibrium of VNTR allele grouping be very significant (p＜0.000001).Uncommon Fnu4H1 2 allele and＜320 VNTR grouping is relevant, and more common I allele and 320-333,334 and＞335 VNTR organize relevant.Owing to there is the people to illustrate that a series of behavior disorders and low MAO-A are active relevant, and because the maximum phenotypic effect that the inventor observes the VNTR polymorphism and 〉=335bp organize relevantly, this shows that this group is also relevant with minimum MAO-A activity.These presentation of results, when carrying Fnu4H1 1 allelic experimenter when placing subgroup according to the VNTR polymorphism, be carry 〉=the allelic experimenter of 335bp promotes Fnu4H1 result.Though the last demonstration of these suggestions needs VNTR allele to serum among the experimenter or the active research of fibroblast MAO-A, these discoveries are consistent with following probability: the reason that Fnu4H1 polymorphism and MAO-A activity difference are relevant is: 1 allele and 〉=335VNTR allele linkage imbalance,＜320 allele (active relevant) with high MAO-A relatively 〉=the big difference of the repetition number of 335bp allele (infer with minimum MAO-A is active and be correlated with) works in the adjusting of MAO-A gene.

Consistent with the probability that this dependency that repeats the allele size and moonlet itself may work in the adjusting of MAO gene.Yet, be clear that very this can not prove this hypothesis, because still may take place with the linkage disequilibrium in unidentified another site.Need prove this hypothesis of MAOA gene to the possible Study of Interaction of expression vector and longer allele and transcription factor.

The presentation of results of TS group, a series of behavior of MAOA gene pairs have the effect of low relatively amount.Though four variablees of multivariate analysis of variance explanation are significant, two of multivariate regression analysis explanations are significant, 12 in 27 of the linear equation analysis explanations is significant, but has the people can propose that neither one is significant at 0.05/27 or 0.0018 place when complete Bonferroni rectification is used for The results of analysis of variance.Yet this is main points exactly, although promptly in the lot of documents MAO relevant with different behaviors, when in special gene pleiomorphism horizontal detection, the MAOA gene seems only many behavior variablees to be played a small amount of effect.Though strong many of effect in drug dependence, even the variation ratio that causes of MAOA allele remains a spot of like this.Repeated experiments is an importance of association study, and these results appear in two diverse subject group.These discoveries are consistent with this notion: many genes participate in different behaviors in the polygenic inheritance, and each gene only plays a part less; Also consistent with this hypothesis: moonlet polymorphism itself may work among the important function equipotential polymorphic variation to polygenic inheritance providing very much.

Embodiment 30

Predictive scheme about aminoacid treatment and premenstrual dysphoric disorder (PMDD)

Premenstrual dysphoric disorder (PMDD) is a kind of moon premenstrual dysphoric disorder that takes place in most of menstrual cycle circulation.It is included in DSM-IV " non-special depressive disorder " category in.Yet many factors (biology and Cognitive Study, therapeutic response) make PMDD distinguish (Yonkers, 1977) mutually with all the other dysthymic disorders.

Although there is the indication of symptom performance luteal phase, the etiology of this obstacle is not established.May be relevant or irrelevant about the theory that hormone and vitamin defective are relevant with PMS with PMDD.Yet, progesterone, estrogen, prostaglandin, insulin, vitamin B ₆, or thyroxin (Severino, and Moline, 1989; Rickels etc., 1990; Bancroft and Rennie, 1993) absolute and relative defective does not all have to establish in PMS or patient's PMDD group.Similarly, functional hormone test is as result unusual (Casper etc., 1989 in patient PMDD to the thyroid releasing hormone and the glucose tolerance test of thyrotropin reaction; Girdler etc., 1995; Haskett etc., 1984; Roy-Byrne etc., 1987).

For symptom before the menstruation is to give up this hypothesis that causes by the endogenous opiate thing, and several research groups have been estimated the level that the β of symptom women and contrast endorphins is arranged.One comprise retrospective report have menstruation before in symptom women's the research, Giannini and partners thereof find, have the decline (Giannini etc., 1984) of β endorphins level in the luteal phase of menstrual cycle; Yet, in this research, do not have matched group.But, four other discovering compared with the control, have β endorphins level luteal phase lower (Tulenheimo etc., 1987 of patients with symptom; Facchinetti etc., 1987; Chuong etc., 1985; With Giannini etc., 1990).Discover that in folliculus phase and luteal phase low-level (Tulenheimo etc., 1987) are all arranged for above-mentioned one, in another research, Symptomatic women β of ovum cycle endorphins level low (Chuong etc., 1994).Have only two to comprise that expection determined the crowd of symptom, this strict standard of determining to meet or not meet the PMDD diagnosis in the research of mentioning before it should be noted that.The size of the difference between patient population, small sample or both may be the bases that causes nearest research to draw different conclusions, and they can not find the difference (Bloch etc., 1996) in two stages of menstrual cycle patient PMDD and contrast in any one.Yet in this research, two groups premenstrua β endorphins level has all descended.β endorphins level is also found in primate in pylic variation in the menstrual cycle, though in ovum cycle differentiation the most remarkable (Wehrenberg etc., 1982).

The fluctuation of the β endorphins that descends rapidly in menstrual cycle can increase the Adrenergic activity of PMDD women and can explain the result of adrenoreceptor in conjunction with research.The symptom women was in the bonded increase of the imidazoline receptor luteal phase of menstrual cycle before Halbreich and partner thereof (Halbreich etc., 1993) found menstruation.According to Grunhaus and partner (1990) thereof summary, the bonded change of adrenoreceptor is also relevant with MDD and panic disorder, though the direction that changes (increasing and the reduction affinity) depends on the aglucon of hematoblastic preparation and use in analysis.

In addition, β endorphins and estrogenic level have shown and have changed.Between puerperal and premenstrua, both levels change rapidly and remarkable (Halbreich and Endicott, 1981), and other people illustrate that narcotic antagonists has reduced the PMS symptom.

Halbreich and partner thereof find level women of anxiety symptom before menstruation of serum r aminobutyric acid (GABA) descend luteal phase (Halbreich etc., 1996).Also in patient MDD, found serum GABA low-level (Petty etc., 1992), though how relevant unknown with above-mentioned discovery it is.

The etiologic etiological theory of PMS almost all concentrates on the secretion of estrogen, progesterone or prolactin antagonist.By contrast, Labrum (1983) proposes first that at the mid-80 the symptom that betides PMS has common etiology basis, comprises 5-hydroxyl color ammonia, GABA and the unusual fluctuations of the brain level of the endocrine process that connects each other.The estrogen feedback may be a factor of undue fluctuation, especially 5-hydroxyl color ammonia.

About 5-hydroxyl color ammonia, many diverse ways have been used to assess this system of PMS+PMDD women, comprise the absorption and the neuroendocrine challenge that detect 5-hydroxyl color ammonia, platelet 5-HT in the whole blood.Based on primate and other evidence: low 5-hydroxyl color ammonia is relevant with sleep, appetite and irritable variation, the 5-hydroxyl color ammonia that Rankin (1992) has studied in the whole blood of anxiety women before the serious menstruation is found, compare with asymptomatic contrast, have symptom women's 5-hydroxyl color ammonia level lower.Part Study person (Ashby etc., 1988; Taylor etc., 1984; Ashby etc., 1990) and be not all research group (Mahngren etc., 1987; Rojansky etc., 1991) find that compared with the control, PMS or PMDD women's platelet luteal phase have descended to the absorption of 5-HT.Compared with the control, the imipramine binding site that clearly is evaluated as PMDD women luteal phase morning (Rojansky etc., 1991) and menstrual cycle two stages (Steege etc., 1992) all reduced.In one research of back, only reached significance,statistical in the folliculus phase.

PMDD women is used tryptophan produced growth hormone and the hydrocortisone reaction (Bancroft etc., 1991) that weakens, the character difference between patient PMDD and the contrast has been described in two stages of menstrual cycle.Yet in two same groups, the reaction of the prolactin antagonist of tryptophan has only weakened (Bancroft etc., 1991) in the menstruation last stage of menstrual cycle relatively.On the other hand, when the incomplete analeptic buspirone of 5-HT1A is used for folliculus phase patient PMDD and normal healthy controls, produced the prolactin antagonist reaction (Yotham, 1993) that weakens.About Fenfluramine being weakened the data of prolactin antagonist reaction, find the reaction (FitzGerald etc. that weaken with one group of PMDD experimenter who clearly identifies compared with the control, 1996) and other one group find that as broad as long (Bancroft and Cook, 1995) mix mutually.At last, compare with asymptomatic women, the precursor tryptophan that exhausts 5-hydroxyl color ammonia obviously more likely causes the preceding symptom (Menkas etc., 1994) of menstruation of patient's PMDD luteal phase and folliculus phase.

Alleviation and the antidepressant of some psychoactive drug for PMDD assessed in many nearest researchs, comprises clonipramine (Sunblad etc., 1993), fluoxetine (Stone etc., 1991; Wood etc., 1992; Steiner etc., 1995; Brandenberg etc.; Pearlstein and Stone, 1994), amfebutamone (Pearlatein etc., 1995), paroxetine (Eriksson etc., 1995; Yonkers etc., 1996a), maprotiline (Eriksson etc., 1995), Sertraline (Yonkers etc., 1996b), nefazodone (Girdler etc., 1995) and Fenfluramine (Brzezinski etc., 1990).

Inventor expection, to single or even two discrete neurotransmitteies single medicine of having only limited effect for overcome women's premenstrua, during and to change " recompense " system exception state that causes by hormone after the menstruation be not enough.In addition, though above-mentioned biological evidences can not belong to any one neurobiology system clearly, adrenoreceptor is unusual relevant with the PMDD expression in conjunction with the variation prompting neurobiology of, GABA level and 5-HT system.Also in single-phase MDD, found the variation of these signs.The positive Study on Effect of PMDD is not reported about enkephalinase inhibitor.In fact, the use of narcotic antagonists such as Trexan (Dupont Delaware) has shown and has reduced rather than strengthened PMDD, with the inventor do in the present invention opposite.

The nearest discovery (deLourdes-Figuerola etc., 1997) of this and Figuerola and co actor thereof has shown that menstruation migraine group is in the increase of the 22nd day blood plasma ME and the decline and blood plasma ME, the NE increase during pain of plasma norepinephrine (NE) level.The author infers, changes betiding on blood plasma ME and the sympathetic nerve adrenal gland function, not only during pain also in middle luteal phase.

The inventor proposes, and the application of aminoacid treatment will be to suffering PMDD misery person useful.Utilize the work that the special compositions that this obstacle is advised is finished in many dipsopathy people in the table 11 significantly to reduce typical PMS symptom, comprise irritability, anxiety, mammary gland pain, headache and depression.The instantiation that utilizes is being listed in the PMX of table 11 ^TMDescribe in detail in the prescription.

The double blinding placebo-controlled study will be at Dallas, implements among the PMDD outpatient of Texas clinic.To study 100 patients altogether.The inventor will propose the PMDD yardstick and offer 100 all participants.This yardstick will be prior to accepting any therapy (placebo or PMX ^TM) mark.For this research, at least three cycles are the bottom lines that enter this research.Patient must be the women of child-bearing age and not have pregnancy.With the comparison that obtains between two groups, statistical analysis is undertaken by healthy the learning under the guidance that the center calculation resource ties up to Robert Wood of Texas university.Only just can study via the ill PMDD candidate of DSM-IV criterion evaluation.After three months, each patient will utilize the MAA technology that proposes among the present invention to carry out gene type.Therefore will assess 29 genes at least.

The relevant allelic carrier of expection DRD2 A1 allele and other RDS disclosed herein will be to PMX ^TMSound response is arranged, under the placebo situation, will feel bad especially.Further contemplate that individual's gene type will provide extra information to prediction targeted therapy result.

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(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:13:ATTAATCCTC TGGGTATTGT 20 (2) SEQ ID NO:14: (i) sequence signature:

(A) length: 23 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:14:TAGTCTTATC CCTCTTTTTC TTA 23 (2) SEQ ID NO:15: (i) sequence signature:

(A) length: 24 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:15:CTCTTACAAT AGAAGAAACC ATTT 24 (2) SEQ ID NO:16: (i) sequence signature:

(A) length: 20 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:16:TCTCCTCTCT TTCCCTTCCC 20 (2) SEQ ID NO:17: (i) sequence signature:

(A) length: 19 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:17:GCATGGCTGG AAGAACTCC 19 (2) SEQ ID NO:18: (i) sequence signature:

(A) length: 22 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:18:TCTTCCAGGC CTCTGGTCAT AT 22 (2) SEQ ID NO:19: (i) sequence signature:

(A) length: 20 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:19:CACCCATTTA CAAGTTTAGC 20 (2) SEQ ID NO:20: (i) sequence signature:

(A) length: 22 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:20:CACTGATTAC TAATTCAGGA TC 22 (2) SEQ ID NO:21: (i) sequence signature:

(A) length: 21 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:21:AGTGGGCAGG GCGGGGCAGG T 21 (2) SEQ ID NO:22: (i) sequence signature:

(A) length: 23 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:22:CGCTGCCTCC CTTCCACCTG TTG 23 (2) SEQ ID NO:23: (i) sequence signature:

(A) length: 25 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:23:AGGTGGCACG TCGCGCCAAG GTGCA 25 (2) SEQ ID NO:24: (i) sequence signature:

(A) length: 25 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:24:TCTGCGGTGG AGTCTGGGGT CGGAG 25 (2) SEQ ID NO:25: (i) sequence signature:

(A) length: 23 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:25:GACACTTCTG GAATTAGTGG AGG 23 (2) SEQ ID NO:26: (i) sequence signature:

(A) length: 21 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linearity

(xi) sequence description: the information of SEQ ID NO:26:GAAGTTAAAT CCATGTGGCT C 21 (2) SEQ ID NO:27: (i) sequence signature:

(A) length: 101 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:27:TCATTAATCC TCTGGGTATT GTAAATGTGG ATTTAGGTTA ATATATTATATATAATGCCA AATAATGGCA TAGATAAGGA ATAGGGAGAA AAAGGGAATT A 101 (2) SEQ ID NO:28: (i) sequence signature:

(A) length: 25 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:28:TAGTCTTATA TCCCTCTTTT TCTTA 25 (2) SEQ ID NO:29: (i) sequence signature:

(A) length: 20 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:29:GCTGCTTCTG TTAACCCTGC 20 (2) SEQ ID NO:30: (i) sequence signature:

(A) length: 21 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:30:TACATCTCCG TGTGATGTTC C 21 (2) SEQ ID NO:31: (i) sequence signature:

(A) length: 27 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:31:CTCTTGTTCC TGTTGCTTTC AATACAC 27 (2) SEQ ID NO:32: (i) sequence signature:

(A) length: 25 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:32:CACTGTGCTA GTTTAGATTC AGCTC 25 (2) SEQ ID NO:33: (i) sequence signature:

(A) length: 18 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: the information of SEQ ID NO:33:CATCTCCTGG GACGTAGC 18 (2) SEQ ID NO:34: (i) sequence signature:

(A) length: 20 base pairs

(B) type: nucleic acid

(C) chain: strand

(D) topology: linear (xi) sequence description: SEQ ID NO:34:AGAGAAGGGA AGGAGGGAAG 20

Claims (94)

1. compositions that is used in experimenter treatment RDS behavior, its basic comprising is as follows:

A) the destructive material of enzymolysis of at least a inhibition neuropeptide acyl opium of inhibition opium destruction amount, said material is selected from: aminoacid, peptide and analog thereof or derivant;

B) at least a precursors of neurotransmitters of the synthetic promotion amount of neurotransmitter, it is selected from: dopamine precursor L-tyrosine, L-phenylalanine and L-DOPA, 5-hydroxy tryptamine precursor L-tryptophan and 5-hydroxyryptophan, and gamma aminobutyric acid (GABA) precursor L-glutaminate, L-glutamic acid and L-glutamate, Glu; And

C) chromium picolinate or the nicotinic acid chromium of tryptophan concentration enhancing amount,

The amount of above-mentioned material or precursors of neurotransmitters and chromium compound is effectively to reduce experimenter's RDS behavior.

2. compositions that is used to prevent or treat unnecessary weight increase, its basic comprising is as follows:

A) the destructive material of enzymolysis of at least a inhibition neuropeptide acyl opium of inhibition opium destruction amount, said material is selected from: aminoacid, peptide and analog thereof or derivant;

B) at least a precursors of neurotransmitters of the synthetic promotion amount of neurotransmitter, it is selected from: dopamine precursor L-tyrosine, L-phenylalanine and L-DOPA, 5-hydroxy tryptamine precursor L-tryptophan and 5-hydroxyryptophan, and gamma aminobutyric acid (GABA) precursor L-glutaminate, L-glutamic acid and L-glutamate, Glu; And

C) chromium picolinate or the nicotinic acid chromium of tryptophan concentration enhancing amount,

The amount of above-mentioned material or precursors of neurotransmitters and chromium compound is the unnecessary body weight that can effectively prevent or reduce the experimenter.

3. compositions that is used to prevent or treat attention deficit hyperactivity disorder, its basic comprising is as follows:

A) the destructive material of enzymolysis of at least a inhibition neuropeptide acyl opium of inhibition opium destruction amount, said material is selected from: aminoacid, peptide and analog thereof or derivant;

B) at least a precursors of neurotransmitters of the synthetic promotion amount of neurotransmitter, it is selected from: dopamine precursor L-tyrosine, L-phenylalanine and L-DOPA, 5-hydroxy tryptamine precursor L-tryptophan and 5-hydroxyryptophan, and gamma aminobutyric acid (GABA) precursor L-glutaminate, L-glutamic acid and L-glutamate, Glu;

C) chromium picolinate or the nicotinic acid chromium of tryptophan concentration enhancing amount; And

D) the synthetic material that promotes of at least a neurotransmitter of the synthetic promotion amount of neurotransmitter, it is selected from Rhodila or huperzine,

Select the synthetic material that promotes of above-mentioned material, precursors of neurotransmitters, inorganic compound and neurotransmitter, make said composition effectively reduce attention deficit hyperactivity disorder, improve attention processing or memory.

4. method for the treatment of experimenter's RDS behavior, described behavior is selected from substantially: stress syndrome after SUD, obesity, smoking, Tourette syndrome, ADHD, division sample/elusive behavior, the wound, PMS or Nicotiana tabacum L. use, this method comprises and gives the experimenter following compositions that it comprises:

A) the destructive material of enzymolysis of at least a inhibition neuropeptide acyl opium of inhibition opium destruction amount, said material is selected from: aminoacid, peptide and analog thereof or derivant;

B) at least a precursors of neurotransmitters of the synthetic promotion amount of neurotransmitter, it is selected from: dopamine precursor L-tyrosine, L-phenylalanine and L-DOPA, 5-hydroxy tryptamine precursor L-tryptophan and 5-hydroxyryptophan, and gamma aminobutyric acid (GABA) precursor L-glutaminate, L-glutamic acid and L-glutamate, Glu; And

C) chromium picolinate or the nicotinic acid chromium of tryptophan concentration enhancing amount.

5. the method for claim 4, wherein said administration is to contain 32 to 10 in the diet consumption of every day, 000mg DL-phenylalanine, 5 to 5,000mg L-tryptophan, 3 to 30,000mg L-glutaminate, described compositions also further contain 1 to 300mg 2-methyl-3-hydroxy-4-formyl-5-hydroxymethylpyridine .-5 '-phosphoric acid.

6. the method for claim 4, wherein said RDS behavior are fat.

7. the method for claim 6, wherein said administration is to contain 460mg DL-phenylalanine, 25mg L-tryptophan, 25mg L-glutaminate in the diet consumption of every day, this mixture also further contains 5mg 2-methyl-3-hydroxy-4-formyl-5-hydroxymethylpyridine .-5 '-phosphoric acid.

8. the method for claim 6, wherein the experimenter has the family history that chemicals relies on, and said family history prompting possibility of success improves.

9. the method for claim 6, wherein said administration suppresses the gluttony behavior.

10. the method for claim 6, wherein said administration suppresses addiction.

11. the method for claim 6 wherein exists at least a following allele to point out the probability of successfully reaction to improve in the experimenter: D ₂TaqI A1, B1, C1 or exon ^6-7Haplotype, HTR2A-C allele isozygotys, and the OB-1875 dinucleotide repeats polymorphism＜208bp allele homozygosity, human No. 2 chromosome microsatellite polymorphisms, APO-D-TaqI2.2 or 2.7bp, or OB gene D7S1875 allele.

12. the method for claim 6, wherein said administration comprises the nicotinic acid chromium of taking effective agent, and exists the probability of DRD2 A1 allele prompting reaction to improve among the experimenter.

13. the method for claim 11, wherein said administration comprises the chromium picolinate of taking effective dose, and exists the probability of DRD2 A2 allele prompting reaction to improve among the experimenter.

14. the method for claim 4, wherein said RDS behavior is the Nicotiana tabacum L. use.

15. the method for claim 14, wherein in the experimenter, exist at least a following allele to point out the probability of successfully reaction to improve: D1 (Dde A1 homozygosity), D2 (TaqI A1), D4 (VNTR2), D5 (dinucleotide 13 allele, scope is at 135-159bp), DAT1VNTR (10/10), D β H (TaqI B1 allele).

16. the method for claim 4, wherein said RDS behavior further comprises: autism, Tourette syndrome or ADHD, and wherein said experimenter is contained at least a following allele: D (Dde A homozygosity), D (Taq), D (VNTR), D (dinucleotide allele scope-bp), DAT VNTR (/), D β H (Taq B allele), MAOA (X), at least a allelic existence points out the probability of successfully reaction to improve.

17. the method for claim 16, wherein said administration further comprises Rhodila or the huperzine of taking effective dose.

18. the method for claim 4, wherein said RDS behavior is a pathological gambling, and exists at least a following allele to point out the probability of successfully reaction to improve in the experimenter: D (Dde A homozygosity), D (Taq A, B, C).

19. the method for claim 4, wherein said RDS behavior further comprises pathologic violence, division sample/avoidances disease (SAB), attack, gets angry, hostility or posttraumatic stress disorder, wherein in the experimenter, exist at least a following allele to point out the probability of successfully reaction to improve: D (Taq A, B, C, exon), DAT (VNTR/), mNOSIa-is for≤allelic the homozygosity of BP.

20. the method for claim 4, wherein said RDS behavior is PMS, wherein exists at least a following allele to point out the probability of successfully reaction to improve: DAT1 VNTR (10/10), D ₂TaqI A1, B1, C1, exon ^6-7Haplotype, or come from the allele of DRD1, DRD2, DRD4, HTT, HTRIA, TDO2, D β H, MAO, COMT, GABRAB, GABRB3, PENk, ADRA2A or ADRA2C gene.

21. the method for claim 4, wherein said RDS behavior further comprises the substance abuse obstacle.

22. the method for claim 21, wherein said RDS behavior is a substance use disorders.

23. a definite experimenter, comprises at least a allele that detects in the group down to the method for the heredodiathesis of at least a RDS behavior: DRD1, DRD2, DRD3, DRD4, DRD5, DAT1, HTT, HTRIA, TDO2, D β H, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, HTRID β, HTR2A, HTR2C, gamma interferon, CD8A, or PSl gene.

24. the method for claim 23, wherein said RDS behavior is selected from: relate to manic, OCD, property, sleep, primary school's behavior, gambling, study, Inattention, ADHD, ADDR, impulsion, MED,, CD, many moving, terrified, division sample behaviors, the extensive obstacle that uses of anxiety, somatization, medicine, intravenous drug, reading, ODD, twitch, ethanol or Nicotiana tabacum L..

25. the method for claim 24, the VNTR polymorphism that wherein said allele is the MAOA gene.

26. the method for claim 23, wherein said RDS behavior are the division samples or avoid disease.

27. the method for claim 26, wherein said allele is selected from: DRD ₂Gene A ₁Allele, DAT ₁Gene VNTR10/10 allele, D β H gene B ₁Allele.

28. the method for claim 23, wherein said RDS behavior is drug use.

29. the method for claim 29, wherein said allele are in the CNR1 gene (AAT) _nThree repetitive sequence numbers increase.

30. the method for claim 23, wherein said RDS behavior is selected from: obesity, and anxiety, depression, psychosis, hostility, paranoid idea forces, the symptom summation, general SI is pursued novelty, whole summation, neuroticism and careful.

31. the method for claim 30, wherein said allele is selected from: D7S1873, D7S1875, D7S514 or D7S680 dinucleotide repetitive sequence number increase in the OB gene.

32. the method for claim 31, the number of wherein said D7S1875 dinucleotide repetitive sequence in two copies of CNR1 gene is all greater than 225bp length.

33. the method for claim 32, wherein said allele are the D of DRD2 gene ₂A1 allele.

34. the method for claim 30, wherein said detection are to detect the D of DRD2 gene ₂A1 allele and an allele that chooses from following group, this group comprises: D7S1873, D7S1875, D7S514 or D7S680 dinucleotide repetitive sequence number increase in the OB gene.

35. the method for claim 34 wherein saidly determines to be used for obesity.

36. the method for claim 23, wherein said RDS behavior is selected from: twitch-obscene words syndrome, manic symptoms, opposition disobedience, property, ADHD-R, division sample, ADHD, twitch, major depression, conduct, stutter, force, somatization, alcohol abuse, study and sleeping problems.

37. the method for claim 36, wherein said allele are the D of DRD2 gene ₂A1 allele.

38. the method for claim 23, wherein said RDS behavior is selected from: twitch-obscene words syndrome, ADHD, smoking, study, primary school's behavior, ADHD-R, opposition disobedience, twitch, manic, ethanol, reading, drug dependence, sleep, stutter, force, somatization and major depression.

39. the method for claim 38, wherein said allele are the Taq A1 allele of D β H gene.

40. the method for claim 36, wherein said RDS behavior is twitch-obscene words syndromes, wherein saidly determines to count increase and carry out by detecting in the D β H gene Taq B1 and Taq A1 allele.

41. the method for claim 23, wherein said RDS behavior is selected from: twitch-obscene words syndrome, autism, somatization, ethanol, ADHD-R, major depression, fear, force, generalized anxiety disorder, manic, opposition disobedience, property, reading and ADHD.

42. the method for claim 41, wherein said twitch-obscene words syndrome are to be undertaken by at least one the 10 allele number increase that detects the DAT1 gene.

43. the method for claim 23, wherein said RDS behavior is selected from: ADHD, stutter, ADHD-R, opposition disobedience, twitch, conduct, force, manic, ethanol, generalized anxiety disorder, fear, division sample, sleep, property, medicine and major depression.

44. the method for claim 43, wherein said RDS behavior is undertaken by detecting at least a allelic number increase, and these allele are selected from: 10 allele of DAT1 gene, the Taq A1 allele of D β H gene, the D2A1 allele of DRD2 gene.

45. the method for claim 23, wherein said RDS behavior is selected from: ethanol, smoking, force feed, twitch, gambling, medicine, reading, shopping, opposition are disobeyed, major depression outbreak, division sample, ADHD, conduct disorder, obsession and manic.

46. the method for claim 45 is wherein saidly determined to be undertaken by the allelic homozygosity of DdeI that detects the DRDI gene.

47. the method for claim 23, wherein said RDS behavior is selected from: opposition disobedience, conduct disorder, feed, smoking, gambling, ADHD, obsession, manic and ethanol.

48. the method for claim 47 is comprising the TaqI A1 and the TaqIA2 allele that detect the DRD2 gene.

49. the method for claim 23, wherein said RDS behavior is selected from: twitch-obscene words syndrome, smoking and gambling.

50. the method for claim 49 is wherein saidly determined to be undertaken by 11 or 22 genotype that detect the DRD1 gene.

51. the method for claim 50, the wherein said Dde1 allele of determining the DRD1 gene by detecting two copies of each genome carries out.

52. the method for claim 23, wherein said RDS behavior is selected from: opposition disobedience, conduct disorder, impulsion feed, smoking, gambling, ADHD, manic, stutter, obsession and the behavior of division sample.

53. the method for claim 52, comprise said determine to increase by the 11 genotypic numbers that detect the DRD1 gene carry out.

54. the method for claim 23, wherein saidly determine to be undertaken, and said RDS behavior is selected from by detecting DRD2 A1 allele: gambling, smoking, force feed, disobedience, major depression outbreak, ADHD, conduct disorder, division sample, obsession, manic and ethanol oppose.

55. the method for claim 54, the wherein said Dde1 allele of determining the DRD1 gene by detecting two copies of each genome carries out.

56. the method for claim 23, wherein said RDS behavior is selected from: ethanol, smoking, force feed, twitch, gambling, medicine, reading, shopping, gambling and primary school's problem.

57. the method for claim 56 is wherein saidly determined to be undertaken by 11 or 22 genotype that detect the DRD1 gene.

58. the method for claim 23, wherein said RDS behavior is twitch-obscene words syndromes.

59. the method for claim 58 wherein saidly determines it is that intron 6G → A polymorphism by detecting tryptophan 2,3 dioxygenase genes is carried out.

60. the method for claim 23, wherein said RDS behavior is selected from: ADHD, alcohol dependence, drug dependence, pathological gambling.

61. the method for claim 60 wherein saidly determines it is that intron 6G → T polymorphism by detecting tryptophan 2,3 dioxygenase genes is carried out.

62. the method for claim 23, wherein said RDS behavior is selected from: ADHD, alcohol dependence, drug dependence, pathological gambling and depression.

63. the method for claim 62 wherein saidly determines it is that intron 6DGGE polymorphism by detecting tryptophan 2,3 dioxygenase genes is carried out.

64. the method for claim 23, wherein said RDS behavior is drug use.

65. the method for claim 64 is wherein saidly determined the low number base pair allele by detecting ADRA2C dinucleotide polymorphism (≤181bp) polymorphism is carried out.

66. being ethanol, the method for claim 23, wherein said RDS behavior use.

67. the method for claim 66 is wherein saidly determined the high number base pair allele by detecting ADRA2C dinucleotide polymorphism (〉=183bp) polymorphism is carried out.

68. the method for claim 23, wherein said RDS behavior is selected from: ethanol and Nicotiana tabacum L. are used.

69. the method for claim 68 is wherein saidly determined to be undertaken by two homology allele that detect senilism element-1 (PS1) polymorphism.

70. the method for claim 68 is wherein saidly determined to be undertaken by two the homology allele greater than 80bp that detect PENK gene C A dinucleotide polymorphic repetitive sequence.

71. the method for claim 23, wherein said RDS behavior is selected from: CD, ODD or hyperkinetic syndrome.

72. the method for claim 71 is wherein saidly determined to be undertaken by the allelic existence of short GGC that detects the AR gene.

73. the method for claim 23 wherein saidly determines it is the quality of determining the type B behavior among alcoholic, cocainomaniac or the RDS proband, is included in that detection is relevant with said quality among the above-mentioned patient the allelic existence of DRD2 carries out.

74. the method for the heredodiathesis of a definite polygenic character, comprise at least a allele that detects with following group gene-correlation, this group comprises: DRD1, DRD2, DRD5, DAT1, HTT, HTRIA, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, DRD3, DRD4, HTR1D β, HTR2A, HTR2C, gamma interferon, CD8A or PS1 gene.

75. the method for claim 74, wherein said heredodiathesis is the heredodiathesis to ADHD, and said determine to comprise detect at least a allele relevant with following gene, these genes are: DRD1, DRD2, DRD5, DAT1, HTT, HTRIA, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABKB3, CNR1, CNRA4, NMDAR1, PENK, AR or CRF gene.

76. the method for claim 74, wherein said heredodiathesis is the heredodiathesis that ADHD is lacked susceptibility, and said determine to comprise detect at least a allele relevant with following gene, these genes are: DRD3, DRD4, NTR1D β, HTR2A, HTR2C, gamma interferon, CD8A, PS1 gene.

77. the method for claim 74, wherein said heredodiathesis is the heredodiathesis to ODD, and said determine to comprise detect at least a allele relevant with following gene, these genes are: DRD1, DRD2, DRD3, DAT1, HTT, HTR1A, HTR2A, HTR2C, DBH, ADRA2A, ADRA2C, MAOA, GABRA3, GABRB3, CNR1, CHRNA4, NMDAR1, PENK, AR or CD8A gene.

78. the method for claim 74, wherein said heredodiathesis is the heredodiathesis to twitching, and said determine to comprise detect at least a allele relevant with following gene, these genes are: DRD1, DRD5, HTR1A, HTR1D β, NTR2C, TDO2, DBH, ADR2C, COMT, GABRA3, CNR1 or CHRNA4 gene.

79. the method for claim 74, wherein said heredodiathesis is the heredodiathesis to LD, and said determine to comprise detect at least a allele relevant with following gene, these genes are: DRD1, HTR2C, TDO2, DBH, ADR2A, ADR2C, MAOA, CNR1 or CNRA4 gene.

80. the method for claim 74, wherein said heredodiathesis are the heredodiathesiies that the LDL level is raise, and said determine to comprise detect at least a allele relevant with following gene, these genes are: HTT, OXYR, DRD2 or PS1 gene.

81. the method for claim 74, wherein said heredodiathesis is the heredodiathesis that cholesterol levels is raise, and said determine to comprise detect at least a allele relevant with following gene, these genes are: HTT, OXYR, DRD2 or PS1 gene.

82. the method for claim 74, wherein said heredodiathesis is to the life-prolonging heredodiathesis, and said determine to comprise detect at least a allele relevant with following gene, these genes are: PS1, OXYR or APOE gene.

83. one kind develops the method that the diagnostic polygenes detects, comprises the following steps:

A). identify the character of being studied,

B). create measuring of a kind of seriousness that is used to measure the character of studying,

C). select at least a may be to the contributive candidate gene of said character,

D). identify the related polymorphism of at least a and said candidate gene,

E). with the allele pattern of described polymorphism with said measure related,

F). the dependency of the association of said allele pattern and described candidate gene and described character compared and

G). the allele pattern that those and said characteristics have positive association is added to come in, and forms the diagnostic polygenes and detects.

84. the method for claim 83, wherein candidate gene comprises: DRD1, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABRB3, CNR1, CNRA4, NMDAR1, PENK, AR, CRF, DRD3, DRD4, HTR1D β, HTR2A, HTR2C, gamma interferon, CD8A or PS1 gene.

85. the method for claim 83, wherein polygenic character comprises: ADHD, ADHD lack, and ODD, CD, LD, twitch, drug dependence/dependence, smoking, osteoarthritis, cholesterol levels raise, the LDL level raises or longevity.

86. the method for claim 85, wherein the described polygenes to described ADHD detects at least a allele that comprises that detection is relevant with following gene, and these genes have: DRD1, DRD2, DRD5, DAT1, HTT, HTR1A, TDO2, DBH, ADRA2A, ADRA2C, NET, MAOA, COMT, GABRA3, GABRB3, CNR1, CNRA4, NMDAR1, PENK, AR or CRF gene.

87. the method for claim 85, wherein the described polygenes that described ADHD is lacked detects at least a allele that comprises that detection is relevant with following gene, and these genes have: DRD3, DRD4, HTR1D β, HTR2A, HTR2C, gamma interferon, CD8A or PS1 gene.

88. the method for claim 85, wherein said polygenes to said ODD is identified at least a allele that comprises that detection is relevant with following gene, and these genes have: DRD1, DRD2, DRD3, DAT1, HTT, HTR1A, HTR2A, HTR2C, DBH, ADRA2A, ADRA2C, MAOA, GABRA3, GABRB3, CNR1, CHRNA4, NMDAR1, PENK, AR or CD8A gene.

89. the method for claim 85, wherein the polygenes to described twitch detects at least a allele that comprises that detection is relevant with following gene, and these genes have: DRD1, DRD5, HTR1A, HTR1D β, HTR2C, TDO2, DBH, ADR2C, COMT, GABRA3, CNR1 or CHRNA4 gene.

90. the method for claim 85, wherein the polygenes to described LD detects at least a allele that comprises that detection is relevant with following gene, and these genes have: DRD1, HTR2C, TDO2, DBH, ADR2A, ADR2C.MAOA, CNR1 or CNRA4 gene

91. the method for claim 85 wherein detects at least a allele that comprises that detection is relevant with following gene to the polygenes that described LDL level raises, these genes have: HTT, OXYR, DRD2 or PS1 gene.

92. the method for claim 85 wherein detects at least a allele that comprises that detection is relevant with following gene to described life-prolonging polygenes, these genes have: PS1, OXYR or APOE gene.

93. the method for claim 85, wherein the polygenes to described osteoarthritis detects at least a allele that comprises that further detection is relevant with following gene, and these genes have: COL2A1, COL2A1, COL2A1, COL9A1, COL9A1, AGC1, IGF1, IGF1, IGF1R, IGF1R, IGF2, IGF2R, TGFB1, TGFB2, IL1A, IL1B, IL1R1, IL1RN, MMP9, TIMP1 or vitamin D3 gene.

94. a compositions that is used in experimenter's treatment RDS behavior, its basic comprising is as follows:

A) suppress at least a destructive material of enzymolysis that can suppress neuropeptide acyl opium of opium destruction amount, said material be selected from aminoacid, peptide with and analog or derivant;

B) a kind of enkephalinase h substance, it discharges neuronic enkephalin or endorphins, and said material is selected from polypeptide or aminoacid; With

C) chemical compound of the effect of at least a retardance opium on δ, μ, κ, σ or epsilon receptor of opium antagonism amount, said material is selected from narcotic antagonist such as nalopotone or IC1154129,

The amount of above-mentioned material, precursors of neurotransmitters, chromium compound and opiate antagonist is effectively to reduce experimenter's RDS behavior.

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