TW201118377A - Method of evaluating and predicting dementia and biological index thereof - Google Patents

Abstract

A biological index of evaluating and predicting dementia and the method thereof are disclosed. The method includes steps of: (1) providing a sample under test separated from the body; (2) in vitro inspecting at least a target gene of gene transcripts of the sample under test to see the presentation degree thereof, the target gene is selected from a group consisting of GUCY1B3, ABCB1, ABCC4, ABCC6, NUDT5, GAMT, HSD3B7, BMPR2 (serine/ threonine kinase), SMAD family member 2 (SMAD2), GP1BA, LAMB2 message RNA of adhesion protein of human β 2, and TNXB; and (3) comparing the differences between the presentation degree of the target gene of the sample under test and the presentation degree of the same gene of a normal control group to determine whether or not the person under test has dementia or the risk of the person under test suffering from dementia.

Description

201118377 VI. Description of the invention: [Technical field to which the invention belongs] This is a biological indicator for the evaluation and prediction of dementia and its detection method, in particular, the target of white blood cells in the peripheral blood of the subject The degree of gene expression is used as a biological indicator for assessing and predicting dementia. [Prior Art] Alzheimer's disease (Alzheimer, disease, AD) is the most common cause of dementia in the elderly, accounting for 65-7〇% of all dementia patients [Blenn〇w乩, 2006], so Alzheimer's disease has been regarded as one of the major public health problems [Ferri, 2005]. According to studies in nerve cells and extra-neurai tissue, Alzheimer's disease has significant and specific pathological changes in the brain: amyloid deposition and neurofibrillary tangles. Therefore, the possible changes in peripheral blood cells such as lymphocytes have been proposed as a model for studying the biochemical and molecular biology of central nervous system degeneration, and as a study in different neuropsychiatric Possible patterns of systemic disorganization in neuropsychiatric disorders [Dolman, 1984]. According to a previous report by Κέΐιηέη et al., 'the lymphocyte cells of dementia patients were compared with lymphocytes of healthy people in 3200 gene sequences on a piece of cDNA microarray, of which 20 The gene expression of the sequences is different, and the expression of some genes related to the immune response is also significantly reduced [Κάΐιηάιι et al·, 2005]. In addition, the human mammalian gene database from the National Institute of Aging (NIA) was used (Mammalian 201118377)

Gene Collection, MGC) cDNA microarray was used to study the blood mononuclear cells of the patients with Alzheimer's disease, 〇〇dm〇n〇nuciear ceus, bmq and cytoskeleton maintenance compared with the normal elderly control group. (cytoskeletal maintenance), cyiiuk trafficking, cell stress response (ceiiuiarstress reSp〇nse), redox balance, transcription, and DNA repair-related gene performance were significantly reduced [Maeseta L2〇〇7]. Because cognitive and functional deterioration is a clinically important feature of Alzheimer's disease, a gradual decline in the expression of this gene may serve as an important marker for determining the severity of neurodegeneration in Alzheimer's disease. People with mild cognitive impairment (MCI) are at high risk for dementia [Morris et al., 2006]. In general, the proportion of elderly people with Alzheimer's disease is Γ2%, and the proportion of people with mild dysfunction to Alzheimer's disease is as high as 1〇~15% per year. [Petersen et Al., 2006]. Therefore, it is important that the m-group recognizes a group of patients with mild dysfunction who will progress to a high-risk group of Alzheimer's disease. Some scholars have begun to investigate the knowledge or biological indicators of these mildly dysfunctional people. The decline in memory is mild dysfunction and the main clinical symptoms of early Alzheimer's disease. Because Temporal Lobe is important for memory integration, pathological changes in temporal lobe structure are the earliest and most important in the pathological study of Alzheimer's disease [Hyman et al', 1984; Van Hoesenet al ., 1990]. Publicly published data on the clinical and radiological features of mild dysfunction, Da Xixi's research on Caucasians; and the Asian genus of mild dysfunctions are rarely studied [Maruyama et al., 2004] Meguro et al., 2004; Wang et al., 2002 and Yamaguchi et al., 2002]. In the study of mainland China, Wang et al. examined the neuropsychological manifestations of people with mild knowledge of 201118377 [Wang et al., 2GG2], and observed the coding and recall of episodic memory (episodicmemory) in these patients. The ability has been compromised. In addition, the inventor of the case (4) showed that the poor performance of the performance and the more severe hippocampal atrophy in the brain magnetic resonance imaging can be used in the next Pingxian rib, the higher the risk will be mild The deterioration of intellectual dysfunction has become a cause of death in patients with Alzheimer's disease. Individuals with the Ap〇E4 allele (age) have a smaller hippocampus in their brains. However, ApoE4 does not predict whether a correct patient will progress from mild dysfunction to Alzheimer's disease. The current medical treatment for Alzheimer's disease is based on the physiological changes of the disease. However, in the past, the study of abnormal gene expression in the central nervous system of Alzheimer's disease patients is mainly due to (4) the domain of the dead silk _ read, and the degradation of RNA and protein repair may occur in the brain tissue after the patient dies. (10) Changes, while changes in materials may cause misjudgment of histopathological changes. It can be seen that the above-mentioned methods for assessing neurological retreats are still missing, and they are not the designers of goodness, but need to be improved. As more and more evidence has emerged in recent years, some key functions in the patient's cells have been abnormal, including the scaled egg repeller idp__p_in, App). (anti〇xidant defenses) , (%(^#^m(acute Phase _, post-translational modification, etc., and these changes may be misinterpreted by analysis of the age of the Alzheimer's disease in the analysis of the tissue [Colcmghi et 纪. , 2〇〇4; Schipper over &, 2〇〇〇— Yu Pass, 2003] 〇5 201118377 The inventor of this case, in view of the shortcomings derived from the study of post-mortem brain tissue, is to improve and innovate After long-term painstaking research, I finally succeeded in researching and developing this method to evaluate and predict dementia with peripheral blood biological indicators. [Invention] The purpose of the present invention (4) is in the miscellaneous supply - cake-like dementia Finding, using the subject's target gene expression in the blood-white domain of the subject as the assessment and prediction of dementia's recording and subtraction' mf estimates and the occurrence of dementia, and can be used to assess the loss The course and development of the mental illness Reference for treatment or diagnosis. In order to achieve the above-mentioned purpose, the inventor of the present invention collected whole blood of patients with mild-domain disorders and Alzheimer's disease, and recruited nucleotide microarrays (〇lig〇nude〇 The tide microairay) examined the transcriptome of the blood samples of these patients, ie, all mRNAs, and compared these data with the normal control group. The inventor of this case is based on patients with Alzheimer's disease. Understanding of inflammatory response and immunomodulatory effects, in addition to cytokines, there are other immune factors involved in the development of this neurodegenerative disease; in order to confirm this speculation, the inventors confirmed the ex vivo by quantitative pCR The degree of expression of each gene in the sample; the results showed that the blood levels of the normal control group, mild dysfunction, and Alzheimer's disease in the two groups of subjects, the degree of expression of some genes related to the immune pathway in white blood cells did Statistically significant differences; the inventors of the present invention used the degree of expression of these genes as a biological indicator for assessing and predicting dementia, and provided the following assessments and predictions. Method of dementia. The methods and biological indicators for assessing and predicting dementia provided in this case are to test the degree of target gene expression in the tested cells of 201118377 as an indicator for evaluating and predicting dementia. The method comprises the following steps: Step 1 provides an isolated test subject; Step 2 detects the degree of expression of at least one target gene in the gene transcript of the test subject, wherein the target gene is selected from guanylate cyclization Enzyme j (Gaunylate cyclase 1, GUCY1B3), multidmg resistance P-glycoprotein 1, ABCB1; ATP-binding cassette transporter subfamily B, member 1), ATP-binding cassette transporter ABCC4 (subfamily) c, member • 4), ATP-binding cassette transporter ABCC6 (subfamily C, member 6), nucleoside diacid-linker X-type motif 5 [Nudix (nucleoside diphosphate linked moiety X)-type motif 5, NUDT5] , guanidinoacetate N-methyltransferase (GAMT), transbasic-d _ 5 _

Hydroxy-delta-5-steroid dehydrogenase (3 beta- and steroid delta-isomerase 7, HSD3B7) ' skeletal morphogenetic protein receptor BM BMPR2 Bone morphogenetic protein receptor (type II (serine/threonine kinase)], SMAD family member 2 (SMAD2), glycoprotein lb (platelet) a polypeptide (Glycoprotein ft (platelet), alpha polypeptide, GP IB A), human β2 adhesion protein LAMB2 message RNA (H. sapiens LAMB2 mRNA for beta2 laminin), and at least one of the group consisting of tenoscin XB (TNXB) And step 3 compares the degree of expression of the target gene in the test subject with the degree of expression of the same gene in the normal control group, and determines whether the subject has the risk of dementia or suffering from dementia. The test subject in the method includes, but is not limited to, peripheral blood leukocytes or other ex vivo samples suitable for use in the present invention; the method for detecting the degree of expression of the target gene includes quantitative polymerase chain reaction (Q PCR) or 疋The reverse transcription polymerase chain reaction (4) was performed by e reverse transcripti〇n PCR 'QRT-PCR) ' or microarray ^j (micr〇array) analysis. Applicants, probes, and the like used in the foregoing detection methods may, in addition to the disclosure of the present invention, modify, adjust, or design the applicable nucleotide sequence, or detection method (eg, fluorescent type), as needed. Eli detection. Wherein the target gene ornithine cyclase j (GUCY1B3) has a nucleotide sequence as set forth in SEQ ID NO: 39, and the target gene Ατρ binding cassette transporter abcc4 has SEQ ID No: 41 The nucleotide sequence of the target gene Ατρ binding cassette transporter ABCC6 has a nucleotide sequence as shown by SEq ID N〇: 42, and the target gene thioglycolate N-methyltransferase (GAMT) has SEQ IDN〇 The nucleotide sequence shown in Figure 44, wherein the target gene has a nucleoside as shown in SEQ ID No: 45 via a base-d-5-steroid dehydrogenase 3?- and a steroid d-isomerase 7 (HSD3B7) Acid sequence, target gene bone morphogenetic protein cleavage type BMPR2 (serine/threonine kinase) has a nucleotide sequence as shown in SEQ ID No: 46, and the target gene SMAD family member 2 (SMAD family member) 2, SMAD2) having the nucleotide sequence shown in SEQ ID No: 47, the target gene glycoprotein lb (platelet) a polypeptide (GP1BA) having the nucleotide sequence as shown in SEQ ID No: 48 The gene tendin protein XB (TNXB) has a nucleotide sequence as shown in SEQ ID No: 50 and the expression of these genes The degree increases with the onset of dementia. The target gene multidrug resistance P-glycoprotein 1 (ABCB1) has a nuclear bud sequence as set forth in SEQ ID No: 201118377. The target gene nuclear bud diphosphate linkage X-type subject 5 (NUDT5) A LAMB2 message RNA sequence having a nucleotide sequence as shown by SEq ID N〇: 43 and a target human human amylin has a nucleotide sequence as shown by SEq N〇: 49, and the degree of expression of these genes is The occurrence of dementia is reduced. ". The degree of gene expression, or related to it, means that the protein contains the gene expression of the gene. The term "dementia" includes, but is not limited to, neurodegenerative diseases such as mild dysfunction and Alzheimer's disease. • The case is further improved by providing a kit for assessing and predicting dementia, which includes a level of genetic performance of at least 5 months of estimation and prediction of dementia as a biological indicator. The present invention is exemplified by the following examples, but the present invention is not limited by the following examples. The materials used in the present invention are commercially available materials which are readily available, unless otherwise specified. [Embodiment] Evaluation of Neuropsychological The overall knowledge function is performed by the Mini_Mentai State Examination (MMSE). [F〇isteinetal., 1983]; The sub- δ 己 力 力 performance was evaluated by the Verbal Naming Test (9 items, 4 series, whether the test was remembered after 15 minutes). The scores of all remembered items (the sum of the scores of all the items in the six queues) and the delayed memory recalls are used as indicators of vocabulary memory. The performance of the visual space ability is evaluated using the score of the replicated graphic in the modified Rey-Ostemeth c〇mpiex figUre test (CFT). The visual singularity of the test uses the immediate memory score and the delayed 15-minute memory score for the 7-point analysis. All participants also accepted the verbal fluency test and the Boston naming test (B〇st〇n). Naming Test), Path Test A & B (Trail Making Test A & B) evaluation. The severity of dementia was analyzed by the ciinicai dementia rating (CDR) [Hughes et al., 1982] and the Chinese version of the geriatric depression scale short form (GDS-S) [Burke et al., 1991] 15 questions to assess depression symptoms. 1. Diagnostic criteria of the subject The clinical diagnosis is performed by a neurologist. All Alzheimer's patients are in compliance with the National Association of Neurological and Communication Abnormalities and Stroke and Alzheimer's Disease and Related Abnormal Associations (Nati〇nal

The Institute of Neurological and Communicative Disorders and Stroke and the Alzheimer disease and Related Disorders Association, NINCDS-ADRDA) is highly likely to be the standard for Alzheimer's disease [McKhann et al" 1984]; to distinguish between mild dysfunction and early The difference in Alzheimer's disease is that only patients with Alzheimer's disease with a Clinical Dementia Rating Scale (CDR) score of 1 can participate in the trial. All patients with mild dysfunction are eligible for petersen's forgetfulness. Barrier criteria [Petersen et al" 1999]: (1) Subjective memory compiaint, preferably confirmed by others; (2) objective memory impairment [selective reminder memory test port light) or

The Rey-Osterrieth Complex Graphic Test (CFT) has a delayed memory (fi*ee delayed recalls) score less than a normal person score greater than one standard deviation (3) normal cognitive function; (4 normal activities are normal; and (5) memory or Knowledge degradation does not meet the criteria for dementia. In addition, the clinical dementia (CDR) score for each mildly informed 201118377 - (4) patient is 0.5. ^ For ',,,,,,,,,,,,,,,,,,,,, Some participants in the control group had ^Heshen, h recorded (such as · headache, spinal nerves or spinal nerve abnormalities); some participants were rounded or (4) spouses suffering from Haimo face; all the participants in the Qiqi test were connected The same standard is used to exclude those with knowledge impairment, and no one has significant information-related memory impairment. All participants excluded evidence of other neurological, psychiatric, or systemic states that may cause cognitive impairment (eg··· Stroke, alcohol addiction, severe depression; participants with brain structural abnormalities (eg, tumors, strokes) or significant ventricle (4) white matter changes (p__ penventncular whke face Han Shenqin s) are also excluded. · Measured traits All subjects were assessed by Simple Intelligence Assessment (MMSE); all Alzheimer's patients were considered as sporadic, late-onset Alzheimer's disease (symptoms occurred in more than 6 domains, no The history of dementia silk family 'and no other gods return to God's abnormality.) The simple intelligent assessment of 28 Alzheimer's disease _SE) scores an average of 1SJ soil 3.9 points [4 of which are in the transcriptional change map _Scriptionalpr〇flling) The average is 19±33]. As a job, the simple intelligent assessment of patients with mild dysfunction is _SE) score average (the average of +5 for transcriptional change maps is 26±3·2) and 18 normal controls (4 of them) The mean of the transcriptional change maps were 28 ± 0'8) with an average of 25.9 ± 2.8 and 28.1 ± 1.5, respectively. One-way ANOVA was used to analyze the statistical differences of each ethnic group (IV).〇1). All the subjects were signed to participate in the trial by a consent form approved by the Board of Directors of the Human Body Medical Research Institute of 2011. 3· ▲•Liquid sampling and RNA extraction The peripheral blood (10 mL) was collected from a vacuum blood collection tube (10 mL K3EDTA, Becton Dickinson, USA) containing EDTA and treated within 4 hours of blood collection; on average, 8 ~ 12 mL of blood is processed at room temperature; in short, mixed with blood for 15 minutes with low-permeability buffer (EL buffer, QIAGEN), and rapidly oscillated to lyse red blood cells; then, at 4 ° C 'The mixture was centrifuged at 4 〇〇 xg for 15 minutes, and the white blood cells formed a mass after centrifugation' and the supernatant was completely removed. Total RNA was extracted with Trizol reagent (Invitrogen) and the extraction method was based on the manufacturer's instructions. Briefly, white blood cell pellets were solubilized with mL Trizol reagent' and then 〇. 2 mL of chloroform was added, followed by centrifugation, and the aqueous phase was mixed with isopropyl alcohol to precipitate total RNA. The RNA was pelleted by centrifugation and the pellet was washed with 75% ethanol and centrifuged again. The RNA pellet was air-dried and lysed with RNase-free water. The quality of the RNA was quantified by analysis with an RNA 6000 Pico LabChip kit and an Agilent 2100 bio-analyzer (Agilent Technologies Inc. USA). The extracted total RNA was used for microarray analysis and reverse transcription quantitative polymerase chain reaction. 4. Gene expression change map (eXpressi〇n profi丨ing) and data collection The Trizol-purified RNA line was quantified using a NanoDrop spectrometer (Rockland, Dallas, USA); the RNA integrity was achieved using the BioAnalyzer 2100 (Agilent, Palo Alto 'California, USA) confirmed. High-quality rna 201118377 (RIN>7) was reverse transcribed into complementary DNA (cDNA) by double reverse transcriptase PCr technology, and biotin-labeled complementary rna was generated by in vitro (!·η Wiro) transcription ( crnA), this biotinylated cRNA was then used for heterophilic reactions with HQ-U133 Plus2 GeneChips® wafers (Affymetrix, Inc., California, USA). The wafer contains 47,000 transcripts and variants containing 38,500 well-characterized human genes. The hybridized target cRNA was stained with streptavidin phycoerythrin (streptavidin_phyC〇erythrin) and the microarray was inhibited by a GeneArray® scanner (Affymetrix, Inc., California, USA) at an excitation wavelength of 488 nm. The gene expression analysis software (GeneSpring® GX software, version 7.3, Agilent Technologies, USA) was used to analyze differential gene expression profiles. 5. Data Analysis and Statistics Analysis of microarray data using GeneChip® operating software (GeneChip8 〇perating Software GC〇s, Affymetrix, California, USA) and GeneSpring(g) GX software (Affymetrix, Inc., California, USA); GeneChip8 operating software Statistical analysis can be performed on the degree of quantitative signal expression from Affymetrix microarrays, mainly to compare the changes in gene expression among single subjects in each group; GeneSpring® GX software is used to compare it to ethnic groups. A map of changes in gene expression differences. Measured by microarray scanning

Robust #t^^J^^(R〇bust Multi-Array Average, RMA) mathematical method calculates the average number of all gene probes on each microarray, Prizarry et al" 2003], and performs background corrections and The data obtained from the microarray gene wafer is de-stated. Then the 'scaiing fact〇r' is used to make the average intensity of all the probes on the microarray 13 201118377 5GQ. In order to enter the normal data of the county Turn all the signal values into the number of rides (base 2, log2), and the logarithm is less than (10), set to 〇〇1; each measurement data is made with the 5th percentile of all the measured data in the sample. To distinguish 'and each gene is distinguished by the median of the gene measurement data in all samples; and use Benjamini and H〇chberg false discovery rate multiple gene correction

Hochberg false discovery rate multiple gene correction). 6. Quantitative Reverse Transcription Polymerase Chain Reaction (Quantitatjve RTpCR) Validation Using Instant Thermal Cycler GeneAmp® PCR System 9700 (Applied

Bl〇SyStemS, Inc., USA, performed a quantitative reverse transcription polymerase chain reaction (QRT-PCR). The RNA purified by Trizol was reverse transcribed into cDNA by following the cDNA Archive Kit (Applied Biosystems, USA) and following the manufacturer's instructions. Briefly, the volume of 2 pg RNA was adjusted to 1 以 with nuclease-free pure water (nuclease-free 0), and 2 μι 10 times reverse transcription buffer (i〇x rt buffer), 〇·8 pL 25x dNTP, 2 pL 10 fold random primer (lOxrandomprimer), 1 pL MultiScribeTM reverse transcriptase (5 〇υ/μί), and 4·2 μί nuclease-free pure water. Amplification of the target (ampiificai; i〇n) in a 96-well plate using an instant thermal cycler GeneAmp8pCR System 9700 at 25 ° C for 10 minutes, 37 ° C for 120 minutes, and 85 ° C for 5 seconds. The temperature is kept low at 4 °C. Quantitative reverse transcription polymerase chain reaction (QRT-PCR) was performed to select genes with different fold changes and statistically significant differences in up-and down-regulation genes. The forward and reverse primers of QRT_PCR were designed using Primer Express 3.0 software, and the primer sequences are shown in Table 1. Actin and glyceraldehydes phosphate 14 201118377 dehydrogenase (GAPDH) were used as control genes. Table 1 Introduction sequence used in QRT-PCR reaction

Gene Name Primer Sequence SEQ ID No: Length of amplified product (bp) ABCB1-F (5'-GCTCATCGTTTGTCTACAGTTCGT-3,) 1 65 ABCB1-R (5'-ACAATGACTCCATCATCGAAACC-3') 2 ABCC4-F (5'-TGTGCTnTTAAGGCTTCACTCAAT -3') 3 79 ABCC4-R (5-.TTGTCCTTCGTATAGCAAGTTTTTTG-3') 4 -- - ABCC6-F (5'-AAAGTACACACAGCATGGCAGTTC-3r) 5 64 ABCC6-R (5'-GCTCCCGGCTAGACCCTTAA-3') 6 Actin- F (5'-GGCACCCAGCACAATGAAGATC-3') 7 182 Actin-R (5'-AAGTCATAGTCCGCCTAGAAGCAT-3') 8 BMP2-F (5,-AAAGGGCATCCTCTCCACAA-3') 9 62 BMP2-R (5'-AGGCGTTTCCGCTGTTTG-3' 10 BMPR2-F (5,-GTGTGAAAACTCCCTATTCTCTTAAGC-3,) 11 69 BMPR2-R (5'-CCAGCGATTCAGTGGAGATGA-3,) 12 CXCL5-F (5'-AATCTGCAAGTGTTCGCCATAG-3') 13 60 CXCL5-R (5'- CAGGGAGGCTACCACTTCCA-3,) 14 ——— GAMT-F (5,-GCCATCGCAGCGTCAAA-3,) 15 65 GAMT-R (5'-TCATTGCACTCGATGATCCAA-3.) 16 GAPDH-F (5'-TTGACGGTGCCATGGAATTT-3·) 17 88 GAPDH-R (5'-GCCATCAATGACCCCTTCATT-3') 18 --- GP1BA-F (5'-TCCCTGGCAACCCTGATG-3,) 19 68 GP1BA-R (5'-TTGGTGAGCTCGCACCTATCT-3') 20 ____ GUCY 1B3-F (5.-GCGTGTCCTGGGCTCTAATG-3·) 21 64 GUCY1B3-R (5,-MGGTGGTCGTGCAGAGCAT-3·) 22 ...---- HSD3B7-F (5,-CAGTCCCAAGACCATCCATGA-3,) 23 64 HSD3B7- R (5,-CACACAAGCCTCGATCACGTT-3,) 24 LAMB2-F (5.-GGAGGGAGATCCGAGAGAAGTAC-3,) 25 80 LAMB2-R (5'-GGCGTGTCCGTAGCAGAAG-3') 26 NUDT5-F (5,-CGCAGGGCAAGGCAGATA-3,) 27 56 NUDT5-R (5.-GATGCTCACCGAGCCACTGT-3.) 28 PDE1C-F P'-GACCATTGCCCAGTGTTTCC-S.) 29 64 PDE1C-R (5,-CATTCTCAGGCTAGCCCTCAA-3,) 30 _____1 SARDH-F (5.-TGGGAAAGTGTGCTGTGATGTT -3·) 31 62 SARDH-R (5,-GTTTGCCGAGGGCTTCCT-3,) 32 SMAD2-F (5'-GCCCCGTCCTAGTCATCmTTAT-3·) 33 74 SMAD2-R (5,-CATTTATTGATGCCTGTTGTTTGC-3,) 34 __ SMURF1-F (5,-GATAATGAAGATGCGACCGAAAG-3,) 35 67 SMURF1-R (5,-ACCTTCTTCCCCACGGAATT-3,) 36 ______ 15 201118377 Gene Name Primer Sequence SEQ ID No: Amplification Product Length (bp) TNXB-F (5'-TTCCTGACGGTCCCACACA -3·) 37 60 TNXB-R (5'-GCAGCACGGCGAATCC-3,) 38 cDNA samples used in microarray and validation tests were first SYBR Green I PCR protocol (Applied Biosystems Division, USA) QRT-PCR was performed and l〇 ng cDNA was taken from each sample for analysis. The normalization of the threshold cycle (Ct) in the reaction is carried out by a passive reference dye (passjve reference dye). The change in gene expression fold in Alzheimer's disease patients is determined by the Ct comparative method, which is the actin and glycerol phosphate dehydrogenase (GApDH) in the normal control group. The average Ct value was subtracted from the average Ct obtained from the mean Ct of patients with mild dysfunction and Alzheimer's disease as a baseline. This normalization takes into account the calculation of relative changes in gene expression; this method normalizes the threshold cycle number (Ct) and corrects it to the control sample, and then uses the result as an index with a base of 2 to determine gene expression. The change in multiples. Differences in the degree of transcriptional response between Alzheimer's disease patients and controls were examined using the Mann-Whitney U-test method; comparisons of multiple cohorts were also evaluated by the Kruskal-Wallis test method. The correlation between the performance of the target gene and the score of the dementia was tested by a non-parametric Spearman’s rank test. Results L subject data and neuropsychological assessment results The detailed background data of the subjects are shown in Table 2. Except that the subjects with the number LE100 are normal young individuals, the average age of the group is 70±13 years old; As expected, the average score of the simple intelligence assessment (MMSE) of the normal control group (N) was 28 ± 7.7, while the mean score of MMSE of mild dysfunction 201118377 was 2 ό soil 3; the mean score of mmse of patients with Alzheimer's disease It is 19 士3. The mean MMSE score was significantly lower in the Alzheimer's group than in the mild dysfunction group (/>=0.0104), and was also significantly lower than the normal control group (p=〇〇〇14). There was no significant difference in the amount of white blood cells from the normal control group, the mild dysfunction group, and the abbreviated group, and there was no significant difference in the quality of each group (Kru caught all the coffee, P= 〇.63). In addition, the M'li of the 18S and 28S _ body _ Α ▼ ▼ 填 填 填 填 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并 并

The quality of this RNA was not considered (results not shown).

Table 2 detailed background information and simple intelligence of the subject

Diagnosis result 3 Simple intelligent assessment score (MMSE: / normal LE039 normal LE045 normal LE078 normal LE100 mild dysfunction LE038 mild dysfunction LE046 mild dysfunction LE049 mild dysfunction LE085 mild dysfunction LE106 Alzheimer's disease LE086 Alzheimer's disease LE102 Alzheimer's disease LE110 Alzheimer's disease LE114 28 27 29 28 27 28 22 24 30 19 20 16 24 70.7 78.8 67.4 28.0 83.2 57.5 68.6 76.3 31.7 78.1 78.3 75.8 77.0 Men, women, men, women, men, women, women, men and women Male RNA integrity (μ§)ΰ 6.21 6.09 34.44 18.78 16.2 15.53 8.5 18.09 36.3 16.83 29.02 16.73 26.87 a According to the Simple Intelligence Assessment (MMSE), the test results b Simple Intelligence Assessment (MMSE) score is 0-30. 17 201118377 e Age (average, range) is (67.1, 28-83.2). dRNA integrity represents the quality of the total rnA of each sample.

The microarray test was designed to compare the genetic changes of white blood cells in the normal control group, the mild dysfunction group, and the Alzheimer's disease group. The inventors collected 13 samples and conducted 9 microarray experiments. The 9 HG-U133_Plus2 GeneChip microarrays (Affymetrix, California, USA) were equally divided into 3 groups, and each group was evenly distributed to 3 wafers (such as 3)). The three microarray experiments (normal control group, mild dysfunction group, Alzheimer's group) were performed with mixed RNA to reduce the difference between individuals; therefore, the GeneSpring® GX software was used to analyze the 9 The wafers were analyzed for analysis of variance (ANOVA) or fold change and combined with functional analysis in the path description. Table 3 Sample data used in microarray test group number of subjects Microarray wafer number Ν-1* LE039 LE045 6S133P020 Ν-2 LE078 6S133P021 Ν-3 LE100 6S133P042 LE038 MCI-1* LE046 6S133P029 LE049 MCI-2 LE085 6S133P022 MCI -3 LE106 6S133P030 AD-1* LE110 LE114 6S133P043 AD-2 LE086 6S133P023 AD-3 LE102 6S133P029 Original: Upper 4 — * — — L ______

The sample to be tested is RNA mixed with different subjects.

In the Caucasian population, 'mild dynamism is considered to be pre-Alzheimer's disease; however, in the Chinese community, only 15-2% of the mild knowledge of P premature disease The patient will become Alzheimer's disease, possibly with the Chinese ApAE Μ 等 因 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 18 According to the clinical observation of the inventor of the present invention, the analysis strategy can be divided into two directions; it is a gene having a significant difference in 'selection gene expression between the normal control group and the Alzheimer's disease group; and the other is to utilize the microarray The software was analyzed for the significant expression of the gene between the normal control group, the mild dysfunction group, and the Alzheimer's disease group. The gene expression profile of the normal control group and the Alzheimer's disease group was generated by Affymetrix's GeneChip® operating software (GC0S) for statistical comparison, and was generated by the Welch i-test. The rate of error correction by gene correction screened out 1,229 gene transcripts (corporate <〇〇5). Subsets are further analyzed for their functional annotations by route category. The results are shown in Table 4. Five functional categories were assembled by statistical differences, and 38 involved ATP-binding Cassette (ABQ Transporter), ascorbate and aldarate metabolism, and glycine (giycine). ), serine and threonine metabolism, transforming growth factor-β (TGF-β) signaling pathway, and extracellular matrix receptor interaction [extracellular matrix (ECM) )-receptor interaction] gene transcripts (34 unique genes) are considered to be putative features of these functions. As shown in Table 5, 'by comparing normal control group, mild dysfunction group, Alzheimer's disease group Three groups of gene expression were selected by analysis of variance (ANOVA) with false discovery rate correction (/><0.05) and 698 gene transcripts were selected, and 9 gene transcripts (8 unique genes) were selected. Functional categories: purine metabolism, ATP transporter, Alkaloid biosynthesis I, and ketone body synthesis/ Deterioration pathways). 201118377 [(InO.OVJf-1ΙΡΡΜ) guest 韹龙龙朱4rw-wr«/韬锑®suus3so xr]%^^-甽甽蘧杷荽澴¢0硇Ew^^¥®ri#? ?rGaN·^蚝寂扭-3Γ Inch<Intelligence ±1, 面面币饀 JIUBqso

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The primers for the characteristic genes were designed using Primer Express 3.0 software. The sequences of these primers are shown in Table 1. Further, cDNA dilution of the normal control group was used as a template' PCR and QRT-pcr were performed to confirm the specificity and efficacy of these primers (data not shown). • Three replicates for the QRT_PCR assay; combined with actin plus (iv) and glycerol acid dehydrogenase (GAPDH) as a reference for the normalization of target cDNAs. The result of QRT_pcR is the threshold value (10) and the inter-valued % difference (d-cT) comparing the target gene of each sample with the reference control group; such comparison makes the threshold cycle difference (d CT) can be explained The true amount of expression of the target gene in the cell. The inventor of the present invention converted the threshold cycle number (4)_CT) to the base β] nuclear degree and made the ship's calculations have a significant difference in performance. The results are shown in Figure 1. According to Mann-Whitney tZ-test, 13 of the 18 genes screened showed significant differences between the normal control group and Alzheimer's disease. Genes with increased gene expression as a result of Alzheimer's disease 疋·(1)Gaunylate cyclase 1, GUCY1B3, (3) ATP-binding•transporter ABCC4 (subfamily C' member 4 ), (4) ATP-binding cassette transporter ABCC6 (Asian C, member 6), (6) guanidin acetate N-methyltmnsfcrase (GAMT), (7) hydroxy-d - 5 _ steroid Dehydrogenase 3 and steroid d-isomerase 7 (Hydroxy-ddta-5-steroid dehydrogenase, 3 beta_ and st Γ <- 23 201118377 delta-isomerase 7, HSD3B7), (8) skeletal morphogenetic protein receptor Bone morphogenetic protein receptor (type II (serine/threonine kinase)], (9) SMAD family member 2 (SMAD2), (10) glycoprotein Lb (also known as Glycoprotein lb (platelet), alpha polypeptide, GP1 BA), 12) tenascin XB (Tenascin XB, TNXB). On the other hand, the genes whose gene expression decreases with the occurrence of Alzheimer's disease are: (2) multidrug resistance P-glycoprotein 1, ABCB1; ATP-binding cassette transporter subfamily b, member i), (5) Nucleic acid dinucleotide linkage X-type subject 5 [Nudix (nucleoside diphosphate linked moiety X)_type motif 5, NUDT5]' (11) human β2 adhesion protein LAMB2 message RNA ( H. sapiens LAMB2 mRNA for beta2 laminin) ° In addition, as shown in Table 6, the multidrug resistance P_glycoprotein i (ABCB1) was used as an example, using the Spearman's rank test statistical method in the normal control group and the mildly dysfunctional group. Between the two groups of Alzheimer's disease group, the multi-drug resistance p_glycoprotein J (ABCB1) was positively correlated with the simple intelligence assessment (MMSE) score ((4)-deletion, production α〇〇11), and Not related to the age between individuals. Therefore, when it is presumed that the amount of expression of the multidrug resistance protein 1 (ABCB1) is decreased in the test subject, the simple intelligence assessment (MMSE) score of the patient of the test subject is also lowered. trend. Therefore, it is possible to use the target of φ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ 24 201118377 Table 6ABCB1 performance and age, simple intelligence assessment (MMS between the MMS MMSE rs -0.0973 0.3858 p-value 0.2311 0.0011 N 60 60 where: rs represents the correlation coefficient; η represents the number of samples. Figure 2 shows that the microarray analysis results were confirmed by quantitative PCR (QRT-PCR) (Figure υ 'Multiple drug-resistant P-glycoprotein i (abcb) as an example, when the target gene is ABCB1 day-to-day, compared In the normal control group, the multidrug-resistant p-glycoprotein I (ABCB1) was detected in the subjects of the mild dysfunction group and the Alzheimer's disease group. The results of the microarray analysis were consistent. Therefore, from Table 6 and Figure 2, it can be seen that if the multi-drug resistance P-glycoprotein 1 (ABCB1) gene expression is low in the mild dysfunction group, the predictable and degraded patients The age of heterozygous age is deteriorated into Alzheimer's disease by mild knowledge. _ 试验 The results of the test in this example show that 'the degree of expression of white blood cells (10) in the tester is detected by PCR analysis of micro-aged mosquitoes. Can be used as an assessment and prediction limit for the disease and The method of risk, * the specific degree of gene expression can also be used as a biological indicator for evaluating and predicting dementia. The method for assessing and _ dementia provided by the present invention and its biological indicators, and other When the conventional techniques are compared with each other, the following advantages are obtained: The method for estimating and predicting dementia provided by the present invention and the biological indicator thereof are based on the degree of expression of a specific gene in the sample of the subject in vitro as an evaluation and prediction loss. The push of wisdom syndrome 25 201118377 ^ 'In contrast to the previous analysis of the remains of the patient after death, the α provided by the present invention, the isolated samples for living patients are analyzed, and appropriate treatment is performed as early as possible. DETAILED DESCRIPTION OF THE INVENTION The detailed description of the preferred embodiments of the present invention is not intended to limit the scope of the invention, and the equivalent implementation or modification of the present invention should be included. In the scope of the patent in this case. In summary, the case is not only innovative in terms of method, but also can improve the above-mentioned functions more than the conventional methods. It should be fully consistent with the success and progress. Set the patent requirements for inventions, and submit the application in accordance with the law. 'Please ask your office to approve the application for the invention patent, so as to invent the invention, and to feel the convenience. [Figure is a simple description] Figure 1 to verify the normal control group in the microarray analysis by 疋畺PCR Differences in gene expression among subjects in the Alzheimer's disease group. Financial error analysis (ANOVA) transferred 18 genes with different gene expression between the normal rhyme group and the Alzheimer's disease group, and then the root i/test knife The degree of qRT PCR analysis of these genes showed that there were 13 significant differences in the performance of the target genes between the normal dragons and the disease. The 13 target genes were different. (1) Avian acid cyclase 1 (Gaunyiate cyCiaSe 1, GUCY1B3), (2) multidrug resistance p_giyC〇protein 1, aBcbi; ATP-binding cassette transporter subfamily B, member UKWATP-binding cassette transporter ABCC4 (subfamily C 'Member 4', (4) ATP-binding cassette transporter ABCC6 (subfamily C, member 6), (5) nude〇side diph〇sphate linked moiety X) Type motif 5, NUDT5], (6) thioglycolic acid N-methyl Guanidinoacetate N-methyltransferase (GAMT), (7) trans-d-5-steroid detachment 26 201118377 Hydrogenase 3 β - and steroid d-isomerase 7 (Hydroxy-delta-5-steroid dehydrogenase, 3 beta- and Steroid delta-isomerase 7, HSD3B7), (8) Bone morphogenetic protein receptor (type U (serine/threonine kinase)] (9) SMAD Family member 2 (SMAD family member 2, SMAD2), (10) Glycoprotein lb (platelet), alpha polypeptide (GP1BA), (11) human β2 adhesion protein LAMB2

Message RNA (H. sapiens LAMB2 mRNA for beta2 laminin), (12) Myosin XB (TenascinXB, TNXB) 〇 Figure 2 was verified by QRT-PCR in normal control, mildly dysfunctional group and Alzheimer's disease group There was a difference in the amount of multidrug resistance P-glycoprotein 1 (ABCB1) between the three groups. According to the non-variable Mairn-Whimey tZ-Test statistical method, * indicates the gene of the normal control group and the Alzheimer's disease group. The performance was significantly different (0=0027), which indicated that there was a significant difference in the gene expression between the normal control group and the mild dysfunction group (IV) 〇29) 4 axis: degree of gene expression. [Explanation of main component symbols] None 27 201118377 Sequence Listing - <110>Wang Peining<12〇> - Method for evaluating and predicting dementia and its biological index <160> 50 <210> 1 <211&gt 24 <212> DNA <213>Artificial Sequence <220><223> Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Forward Primer for Detection of ABCB1 Gene Expression

<400> 1 gctcatcgtt tgtctacagt tcgt 24 <210> 2 <211> 23 <212> DNA <213> artificial sequence < 22 0 >< 22 3 > used to detect the quantitative expression of ABCB1 gene Reverse transcription polymerase chain reaction (QRT-PCIU reverse primer <400> 2 acaatgactc catcatcgaa acc

<211> 25 <212> DNA <213> artificial sequence < 22 0 >< 22 3 > quantitative reverse transcription polymerase chain reaction (QRT-PCR) forward primer for detecting ABCC4 gene expression <400> 3 tgtgcttttt aaggcttcac tcaat <210> 4 <211> 26 < 212 > DNA <213> Artificial sequence 201118377 <220><223> Quantitative reverse transcription for detecting ABCC4 gene expression Polymerase chain reaction (QRT-pcr) reverse primer <400> 4 ttgtccttcg tatagcaagt tttttg <210> 5 <211> 24 <212> DNA <213> artificial sequence <22 0>< 2 2 3>Quantitative reverse transcription polymerase chain reaction (qrt-pcr) forward primer for detecting abcc6 gene expression <400> 5 aaagtacaca cagcatggca gttc <210> 6 <211> 20 <212> DNA <213 〉Artificial sequence < 2 2 0 ><223> Quantitative reverse transcription polymerase chain reaction (QRT-PCR) reverse primer for detecting ABCC6 gene expression <400> 6 gctcccggct agacccttaa <210> 7 < 211 > 22 <212> DNA <213 > artificial sequence <22 0 ><22 3 &gt Quantitative Reverse Transcription Polymerase Chain Reaction (QRT_PCR) Forward Primer for Detection of Actin Gene Expression <400> 7 ggcacccagc acaatgaaga tc <210> 8 <211> 24 <212> DHA <213> 201118377 <22〇>< 22 3 > Quantitative reverse transcription polymerase chain reaction for detecting Act in gene expression (QRT-PCR> Reverse primer <400> 8 aagtcatagt ccgcctagaa gcat <210> 9 <; 211 > 20 <212> DNA <213>Artificial sequence <22 0 ><223> Quantitative reverse transcription polymerase chain reaction (QRT_PCR) forward primer for detecting BMP2 gene expression <400> 9

Aaagggcatc ctctccacaa <210 > 10 <211> 18 <212> DNA <213>Artificial sequence <220 >< 22 3>Quantitative reverse transcription polymerase chain reaction (QRT_PCR) for detecting BMP2 gene expression ) Reverse Primer <400> 10 aggcgtttcc gctgtttg

<211> 27 <212> DNA <2 13 > artificial sequence <220>< 223 > quantitative reverse transcription polymerase chain reaction for detecting BMPR2 gene expression <〇RT_PCR>Sub-lt;4〇〇> 11 tgtgaaaac tccctattct cttaagc <210> 12 <211> 22 <212> DNA <213> artificial sequence 201118377 < 22 0 ><223> for detecting BMPR2 gene expression Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Reverse Primer <400> 12 gccagcgatt cagtggagat ga <210> 13 <211>

<212> DNA < 213 > Artificial Sequence < 22 0〉 < 2 2 3 > Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Forward Primer for Detection of CXCL5 Gene Expression <400> 13 aatctgcaag tgttcgccat ag <210> 14 <211> 20 <212 > DNA <213> Artificial sequence < 2 2 0 < 22 3 > Quantitative reverse transcriptase polymerase for detecting CXCLS gene expression Chain reaction (QRT-PCR) reverse bow tweezers <400> 14 cagggaggct accacttcca <210> 15 <211> 17 <212> DNA <213> artificial sequence < 22 0 >< 2 2 3 > Quantitative Reverse Transcription Polymerase Chain Reaction (QRT.PCR) forward primer for detecting GAMT gene expression <400> 15 gccatcgcag cgtcaaa <210> 16 <211> 21 <212> DNA <213 Artificial sequence 201118377 <220> - < 22 3 > Quantitative reverse transcription polymerase chain reaction (QRT-PCR) reverse primer for detecting GAMT gene expression <400> 16 tcattgcact cgatgatcca a <210><211> 20 <212 > DNA <213> artificial sequence < 2 2 0 ><223> Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Forward Primer for Measuring GAPDH Gene Expression <400>

Ttgacggtgc catggaattt <210> 18 <211> 21 <212> DNA <213> artificial sequence <22〇><223> quantitative reverse transcription polymerase chain reaction (QRT) for detecting GAPDH gene expression - PCR) Reverse primer <400> 18 gccatcaatg accccttcat t <210> 19 <211> 18 < 212 > DNA <213>Artificial sequence <220>< 22 3 > Used to detect GP1BA Quantitative Reverse Transcription Polymerase Chain Reaction (ORT-PCR) Forward Primer for Gene Expression < 400 > 19 tccctggcaa ccctgatg <210> 20 <211> 21 <212> DNA <213>Artificial Sequence 201118377 < 220 >< 223> Quantitative reverse transcriptase polymerase chain reaction for detecting GP1BA gene expression (QRT-PCIO reverse primer <400> 20 ttggtgagct cgcacctatc t <210> 21 <211> 20 <212> DNA <213>Artificial sequence <220><223> Quantitative reverse transcription polymerase chain reaction (QRT-PCR) forward primer for detecting GUCYIB3 gene expression <400> 21 gcgtgtcctg ggctctaatg <210> 22 <;211> 20 <212> DNA <2 13 > Human Procedure Column < 2 2 0> <223> Quantitative reverse transcription polymerase chain reaction (QRT-PCR) reverse primer for detecting GUCY1B3 gene expression <400> 22 aaggtggtcg tgcagagcat <210> 23 <211><212> DNA < 213 > artificial sequence <220><223> quantitative reverse transcription polymerase chain reaction <QRT·PCR) forward primer for detecting HSD3B*7 gene expression <400> 23 cagtcccaag accatccatg a <210> 24 <211> 21 <212> DNA <213> artificial sequence 201118377 <220 > - <223 > quantitative reverse transcription polymerase chain linkage for detecting HSD3B7 gene expression Reaction IQRT-PCR) Reverse primer <400> 24 cacacaagcc tcgatcacgt t <210> 25 <211> 23 <212> DNA <213> Artificial sequence<2 20 ><223> Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Forward Primer for Detection of LAMB2 Gene Expression <400> 25

Ggagggagat ccgagagaag tac <210> 26 <211> 19 <212> DNA <2 13 > artificial sequence < 22 0 ><223><223>><223> (QRT-PCR) Reverse Primer <400> 26 ggcgtgtccg tagcagaag

<211> 18 <212> DNA <213>Artificial sequence <220>< 223 > Quantitative reverse transcription polymerase chain reaction (QRT-pCR) forward primer for detecting NUDT5 gene expression <400> 27 cgcagggcaa ggcagata <210> 28 <211> 20 <212> DNA <2 13 > artificial sequence 201118377 < 22 0 >< 223 > Quantitative reverse transcription polymerization for detecting NUDTS gene expression Enzyme chain reaction (QRT-PCR) reverse primer <400> 28 gatgctcacc gagccactgt <210> 29 <211> 20 <212> DNA <213> artificial sequence <220><223> Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Forward Primer for PDE1C Gene Expression <400> 29 gaccattgcc cagtgtttcc <210> 30 <211> 21 <212> DNA <213 > Artificial Sequence < 22 0 >< 2 2 3 > Quantitative Reverse Transcription Polymerase Chain Reaction (QRT-PCR) Reverse Primer for Detection of PDE1C Gene Expression <400> 30 cattctcagg c tag ccc tca a <210> 31 <;211> 22 <212> DNA <2 13 > Artificial Sequence < 220 >< 223 &g t; quantitative reverse transcription polymerase chain reaction (QRT-pCR) forward primer for detecting the expression of the SARDH gene <400> 31 tgggaaagtg tgctgtgatg tt <210> 32 <211> 18 <212> DNA <213 〉Artificial sequence 201118377 <220><223>Quantitative reverse transcriptase polymerase chain reaction (WRT_PCR) for detecting SARDH gene expression) Reverse primer <400> 32 gtttgccgag ggcttcct <210> 33 <211> 24 <;212>DNA<2 13 > artificial sequence < 2 2 0 ><223> Quantitative reverse transcription polymerase chain reaction (QRT_PCR) for detecting SMAD2 gene expression is positive W sub- <400>

Gccccgtcct agtcatcttt ttat <210> 34 <211> 24 < 212 > DNA <213>Artificial sequence <220 ><223> Quantitative reverse transcription polymerase chain reaction (QRT_PCR) for detecting SMAD2 gene expression ) Reverse Primer <400> 34 catttattga tgcctgttgt ttgc

<211> 23 <212> DNA <213>Artificial sequence <220>< 22 3>Quantitative reverse transcription polymerase chain reaction (QRT-PCR) forward primer for detecting SMURF1 gene expression <400> 35 gataatgaag atgcgaccga aag <210> 36 <211> 20 <212 > DNA <213> Artificial sequence 201118377 <220><223> Quantitative reverse transcription polymerase chain reaction for detecting SMURF1 gene expression (QRT, PCR > Reverse Primer <400> 36 accttcttcc ccacggaatt 20 <210> 37 <211> 19 <212> DNA <213>Artificial Sequence<220 >< 2 2 3> Quantitative reverse transcriptase polymerase chain reaction (〇RT_PCR) positive primer for detecting TNXB gene expression <400> 37 ttcctgacgg tcccacaca 19 <210> 38 <211> 16 <212> DNA <213 > artificial sequence < 22 0 >< 2 2 3>Quantitative reverse transcription polymerase chain reaction for detecting TNXB gene expression (QRT-pCR> reverse primer <400> 38 gcagcacggc gaatcc 16 <210> 39 <211> 544

< 212 > DNA < 213 > wise man (Hojtjo sapiens) <300 ><308> W93728 <309> 1995-12-07 <400> 39 tttttttttt cacgtgaaaa aaataattta t tacagac tc 111 tacaca t taa ca tggaa 60 catttataca tatatcgatg tgctgatatg aaatactaaa tttaaaggca aacattttta 120 cacaaaagta gttgcactct attttataaa gatagatatt aataagttat cagagacatt 180 taagagctag aggccaatta ttccaacagt aatgcattct a tg c tgaaag taaactaagt 240 tttctgaaca tgatgtcctg gatataatca cattcttcta age taaggaa agggagetea 300 10 201118377 tttctgggaa tacaaggcca agaagggctc taacagcagt atcccagcag tgtgtttcca 360 gatttattct tggatgtggt tggagcgccc aacatttagc ctgaactaat gtaacagctc 420 aatgtgaaac aatggcagct ttctggtaan agctgcctgt gggttaatga gatttaatac 480 agggggatac agttaccaat gatagnc111 taggaggaat tataattggc catatgattt 54 0 ggaa 544

<210> 40 <211> 4192 <212 > DNA <213> 智•慧人(Ko/no sapiens) <300 ><308> AF016535 <309> 1997-09-03

≪ 400 > 40 tccaagctca aagaagcaga ggccgctgtt cgtttccttt aggtctttcc actaaagtcg 60 gagtatcttc ttccaagatt tcacgtcttg gtggccgttc caaggagcgc gagg tcggga 120 tggatcttga aggggaccgc aatggaggag caaagaagaa gaactttttt aaactgaaca 180 ataaaagtga aaaagataag aaggaaaaga aaccaac tg t cagtgtattt tcaatgtttc 240 gctattcaaa ttggcttgac aagttgtata tggtggtggg aactttggct gcca tea tec 300 atggggctgg acttcctctc atgatgctgg tgtttggaga aatgacagat atctttgcaa 360 atgcaggaaa tttagaagat ctgatgtcaa acatcac taa tagaagtgat atcaatgata 420 cagggttctt catgaatctg gaggaagaca tgaccaggta tgcctattat tacag tggaa 480 ttggtgctgg ggtgctggtt gctgcttaca ttcaggtttc attttggtgc c tggeage tg 540 gaagacaaat acacaaaatt agaaaacag t tttttcatgc tataatgcga caggaga tag 600 gctggtttga tg tgcacga t gttggggagc t taacacccg acttacagat gatgtctcca 660 agattaatga aggaa t tgg t gacaaaattg gaatgttctt tcagtcaatg gcaacatttt 720 tcactgggtt ta tag tagga tttacacgtg gttggaagct aacccttgtg attttggcca 780 tcagtcctgt tcttggactg tcagctgctg tc tgggca aa gatactatct tcatttactg 840 ataaag ei act cttagcgtat gcaaaagctg gagcagtagc tgaagaggtc ttggcagcaa 900 ttagaactgt gattgcattt ggaggacaaa agaaagaac t tgaaagg tac aacaaaaatt 960 tagaagaagc taaaagaat ggga taaaga aagctattac agccaatatt tctataggtg 1020 ctgctttcct gctgatctat gcatcttatg ctctggcctt ctggtatggg accaccttgg 1080 tcc tc tcagg ggaa ta ttc t attggacaag tac tcac tg ta t 1111 c tg ta t taa t tgggg 1140 c 111 tag tg tggacaggca tctccaagca ttgaagcatt tgcaaatgca agaggageag 1200 cttatgaaat ct tcaagata attgataata agccaagtat tgacagc ta t tegaagag tg 1260 ggcacaaacc aga taa tat t aagggaaa 11 tggaa t tcag aaatgttcac ttcagttacc 1320 catctcgaaa agaagttaag atcttgaagg gtctgaacct gaagg tgcag t agtgggcaga 1380 cggtggccct ggttggaaac agtggctgtg ggaagagcac aacagtccag ctgatgcaga 1440 ggctctatga ccccacagag gggatggtca gtgttgatgg acaggatatt aggaccataa 1500 atgtaaggtt tctacgggaa atcattggtg tggtgagtca ggaacctgta ttgtttgcca 1560 ccacgatagc tgaaaacatt cgctatggcc gtgaaaatgt caeca tgga t gaga t tgaga 1620 11 201118377 aagctgtcaa ggaagccaat gcc ta tgact ttatca tgaa actgcctcat aaatttgaca 1680 ccctggttgg agagagaggg gcccag t tga gtggtgggca gaageagagg atcgccattg 1740 cacgtgccct ggttcgcaac cccaagatcc tcctgctgga tgaggccacg teageettgg 1800 acacagaaag cgaagcag tg gttcaggtgg ctctggataa ggccagaaaa ggtcggacca 1860 ccattgtgat agctcatcgt ttgtctacag ttcgtaatgc tgaegteate gctggtttcg 1920 atgatggagt cattgtggag aaaggaaatc atgatgaact ca tgaaagag aaaggcattt 1980 acttcaaact tgtcacaatg cagacagcag gaaatgaagt tgaattagaa aatgcagctg 2 04 0 atgaatccaa aagtgaaatt gatgccttgg aaatgtcttc aaatgattca agatccagtc 2100 taataagaaa aagatcaact eg taggagtg tccgtggatc acaagcccaa gacagaaagc 2160 ttagtaccaa agaggctctg ga tgaaagta tacctccagt ttccttttgg aggattatga 222 0 agctaaattt aac tgaa tgg cct ta 111tg ttgttggtgt attttgtgcc attataaatg 2280 gaggcc tgca accagcattt Gcaataatat tttcaaagat tataggggtt tttacaagaa 2340 ttgatgatcc tgaaacaaaa cgacagaata gtaac ttgtt ttcactattg tttctagccc 2400 ttggaattat ttctttta tt acatttttcc ttcagggttt cacatttggc aaage tggag 2460 agatcctcac caagcggc tc cgatacatgg ttttccgatc catgctcaga caggatgtga 2520 gttggtttga tgaccc taaa aacaccactg gageattgae taccaggc tc gccaatgatg 2580 ctgctcaagt taaaggggct ataggttcca ggcttgctgt aattacccag aatatagcaa 2640 atcttgggac aggaataatt a ta tee t tea tctatggttg gcaactaaca ctgttactct 27 0 0 tagcaattgt acccatcatt gcaatagcag gagttgttga aatgaaaatg ttgtctggac 2760 aagcac tgaa agataagaaa gaac tagaag gtgctgggaa gatcgctact gaagcaa tag 2820 aaaac ttccg aaccgttgtt tctttgactc aggageagaa gtttgaacat atgtatgctc 2880 agagtttgca ggtaccatac agaaactctt tgaggaaagc acacatcttt ggaattacat 2940 tttccttcac ccaggcaatg atgtattttt cctatgctgg atgtttccgg tttggagcct 3000 acttggtggc aca taaac tc atgagctttg aggatgttct gt tag ta 111 tcagctgttg 3060 tctttggtgc catggccgtg gggcaagtca gttcatttgc tcctgactat Gccaaagcca 3120 aaatatcagc agcccacatc atcatgatca ttgaaaaaac ccctttgatt gacagc taca 3180 gcacggaagg cctaatgccg aacacattgg aaggaaatgt •cacatttggt gaag t tg ta t 3240 tcaactatcc cacccgaccg gacatcccag tgcttcaggg actgagcctg gagg tgaaga 3300 agggccagac gctggctctg gtgggcagca gtggctgtgg gaagagcaca gtggtccagc 3360 tcctggagcg gttctacgac cccttggcag ggaaag tgc t gettgatgge aaagaaa taa 3420 agcgactgaa tgttcagtgg ctccgagcac acctgggcat cgtgtcccag gagcccatcc 3480 tgtttgactg cagcattgct gagaacattg cctatggaga caacagccgg gtggtgtcac 3540 aggaagagat tgtgagggca gcaaaggagg ccaacataca tgccttcatc gagteae tgc 3600 ctaataaata tagcac taaa g taggagaca aaggaac tea gctctctggt ggccagaaac 3660 aacgca t tgc catagctcgt gcccttgtta gacagcctca tattttgctt t tggatgaag 3720 ccacgtcagc tctggataca gaaagtgaaa aggttgtcca agaagccctg gacaaagcca 3780 gagaaggccg cacctgcatt gtgattgctc accgcctgtc caccatccag aa tgeagae t 3840 taatagtggt gtttcagaat ggeagagtea aggageatgg cacgcatcag cagctgctgg 3900 cacagaaagg catctatttt tcaatggtca gtgtccaggc tggaacaaag Cgccagtgaa 3960 ctctgactgt atgagatgtt aaatac tttt taatatttgt ttagatatga catttattca 4020 aagt taaaag caaacactta cagaa t ta Tg aagaggtatc tgtttaacat t tee tcag tc 4080 aagttcagag tcttcagaga ettegtaatt aaaggaacag agtgagagac atcatcaagt 4140 ggagagaaat catagtttaa actgcattat aaattttata acagaattaa ag 4192 12 201118377

<210> 41 <211> 2256 <212> DNA <213 > Homo sapiens <300>

≪ 308 > AY133679 < 309 > 2002 -12-31 < 400 > 41 gcgcccgccc cgcccggcaa gatgctgccc gtgtaccagg aggtgaagcc caacccgc tg 60 caggacgcga acctctgctc acgcgtgttc ttctgatgga gtctcgctgt gtcacccagg 120 ctggagtgca gtggcgcagt c tcagctcac tgcagcc tcc acctcctgtg ttcaagcagt 180 cctcctgcct cagccaccag actagcaggt ctcccccgcc tctttcttgg aaggacaett 240 gccattggat ttaggaccca cttggataat ccaggatgat gtcttcactc caacatcctc 300 agtttaattc ca tg tgcaaa tacccttttc ccaaataaca ttcaattctt taccaggaaa 360 ggtggctcaa tcccttgttt aaaattggcc ataaacggag attagaggaa gatgatatgt 420 attcagtgct gccagaagac cgctcacagc accttggaga ggagttgcaa gggttctggg 480 ataaagaagt 11 taagagc t gagaatgacg cacagaagcc ttctttaaca agagcaatca 540 taaagtggta c tggaaatc t tatttagttt tgggaatttt tacgttaatt gaggaaagtg 600 ccaaagtaat ccagcccata Tttttgggaa aaattattaa ttattttgaa aattatgatc 660 ccatggattc tgtggctttg aacacagcg t acgcc ta tgc cacggtgctg actttttgca 720 cgctcatttt ggctatactg catcacttat atttttatca cgttcagtgt gctgggatg a 780 ggttacgagt agcca tgtgc catatgattt atcggaaggc acttcgtctt agtaacatgg 840 ccatggggaa gacaaccaca ggccagatag tcaatctgct gtccaatgat gtgaacaagt 900 ttgatcaggt gacagtgttc ttacacttcc tgtgggcagg accactgcag gcgatcgcag 960 tgactgccct actctggatg gaga taggaa tatcgtgcct tgctgggatg gcagttctaa 1020 tcattctcct gcccttgcaa agctgttttg ggaagttgtt c tea tcac tg aggag taaaa 1080 ctgcaacttt cacggatgcc agga tcagga cca tgaa tga agttataact ggtataagga 1140 taa taaaaa t gtacgcctgg gaaaagtcat tttcaaatct egge tgaegg 1380 ttaccctctt cttcccctca gccattgaga gggtgtcaga ggcaatcgtc agcatccgaa 1440 gaa tccagac ctttttgcta cttgatgaga tatcacagcg caaccgtcag ctgccgtcag tattaccaat ttgagaaaga 1200 aggagatttc caagattctg agaagttcct gcc tcagggg gatgaatttg gcttcatttt 1260 tcagtgcaag caaaatcatc gtgtttgtga ccttcaccac ctacgtgctc ctcggcagtg 1320 tga tcacagc cagccgcgtg ttcgtggcag tgacgctgta tggggctgtg 1500 a tggtaaaaa ga tgg tgca t gtgcaggatt ttactgcttt t tggga taag gca tcagaga 1560 ccccaactct acaaggcctt tcctttactg tcaga cctgg cgaattgtta gctgtggtcg 1620 gccccgtggg agcagggaag tcatcactgt taagtgccgt gc tcggggaa ttggccccaa 1680 gtcacgggct ggtcagcgtg catggaagaa ttgcctatgt gtctcagcag ccctgggtgt 1740 tc tcgggaac tctgaggagt aatattttat t tgggaagaa a tacgaaaag gaacgatatg 1800 aaaaagtcat aaaggcttgt gc tctgaaaa aggatttaca gctgttggag gatggtgatc 1860 tgactgtgat aggagatcgg ggaaccacgc tgagtggagg gcagaaagca cgggtaaacc 1920 t tgcaagagc agtgtatcaa gatgc tgaca tc ta tc tcc t ggacgatcct ctcagtgcag 1980 13 201118377 tagatgcgga agttagcaga cacttgttcg aactgtgtat ttgtcaaatt ttgcatgaga 2040 agatcacaat tttagtgact catcagttgc agtacctcaa agctgcaagt cagattctga 2100 tattgaaaga tggtaaaatg gtgcagaagg ggacttacac tgagttccta aaatctggta 2160 tagattttgg ctccctttta aagaaggata atgaggaaag tgaacaacct ccagttccag 2220 gaactcccac actaaggaat cgtaccttct cagagt 2256

<210> 42 <211> 744 <212> DNA <213>智,慧人(Ho/no sapiens) < 3 0 0 ><308> AI074459 <309> 1998 -08-06 < 400 > 42 ttttttttta caggtaaaac gggaaatcga gcaagattgt tgtgtcaatg tcaacaccct 60 ggctgtgatg ctttatccta gagt tttgca agatgttacc attgggtgaa atggggtacc 120 aagtacacga atctctctat attattgttt ctttaactgc atgtgagtct gggattatcc 180 caaaataaaa gtgttaattt gtaaaaagta cacacagcat ggcagttccc agcc tcagag 240 attctgattt aagggtctag ccgggagcct gggcatgtgc cagagtacag cggggctgag agtcgc ttgaggcccc caggtgagtc 300 tgtt gacattggct gcaggg tgga cagggcgaga 360 tgggccc tgc ccgggcagac ctgtgtattg ctaggtcctt ccggctc tga tgctctgtga 420 taattggcca ctttctctgc cattttcctc ccagagagca aacacaggtc tagac tcaa t 480 atcgtgtgga gctatcgatg accacgggtc acttccatct ccagcactgc aggctgtgcg 540 ggctggtcca actggggtac ggttgagggt cctggctcag accaggcctg ac tcc tgggc 600 cag tc tg taa aacaggccct tctgggccag cagctgggcc gggc ttgcgc tc tc tgccac 660 ctgccccttg tccatgaccaGaacccgggc acagtccatc acggaacgca agcgg tgggc 720 aa tgagcagc acag tgcac t gtgc 744 <210> 43 <211> 565 <212> DNA <213> Homo sapiens < 300 >< 3 0 8 > AW237290 < 3 0 9 > 1999-12-13 < 4 0 0 > 43 ttttttaaca gtagacaata aacttttatt taagaaaact gattcagttg tgttggaaaa 60 aataaagaaa tctgatatta aacgttttct aagatcattt gtataggttc agtgtattca 120 14 201118377

taagaagtcc accctgagat gcctgtaaaa gtcaaatgta attacacttc aaactttaat 180 cctaaattat tgacagatag atagatagtg accaacttaa gggtaatcat atatgtgact 240 aacatttggg aggaaacagg aaaacag tgg tc tcaaaaca acaatatccc agtctccatt 300 tgaaagagca tagatcttgg taaatcattt tgaaaactat gtgctttatt tcccaaaaga 360 tcaaac t taa tttttaaaag acaccctttt cagaagtatg ggttttcatt gctgtttagt 420 ttcttaggtg tggtatgtct tacgtagctt atgcttacaa agccaaataa tcgcagggca 480 aggcagataa aggcaggcag gatagtgaca gtggctcggt Gagcatccgc caagggcctc 540 agaccactgt taccataaaa acatc 565 <210> 44 <211> 960 <212 > DNA < 2 1 3 > Homo sapiens < 3 00 >< 3 08 > NM_13 8 924 < 309 > 2009-05-10 < 400 > 44 ccccggccca caagcccctg cagggagcgg gcccgggcgg cgcgcgatcg aggtcgggtc 60 gccgtccagc c tgcagcatg agcgccccca gcgcgacccc ca tc t tegeg cccggcgaga 120 ac tgcagccc cgcgtggggg gcggcgcccg cggcctacga cgcagcggac acgcacctgc 180 gca tcc tggg caagccggtg at ggagcgct gggagacccc c ta ta tgcac gege tggccg 240 ccgccgcctc ctccaaaggg ggccgggtcc tggaggtggg ctttggcatg gcca tcgcag 300 cgtcaaaggt gcaggaggcg cccattgatg agcattggat catcgagtgc aa tgacggcg 360 tcttccagcg gc tccgggac tgggccccac ggcagacaca caagg tcatc ccc t tgaaag 420 gcctgtggga ggatgtggca cccaccctgc ctgacggtca ctttgatggg atcctgtacg 480 acacgtaccc ac tc tcggag gagacctggc acacacacca gttcaacttc atcaagaacc 540 acgcctttcg cc tgc tgaag ccggggggcg tcctcaccta ctgcaacctc acctcctggg 600 gggagc tgat gaagtccaag tac tcagaca tcacca teat gtttgaggtg cgcccacctg 660 aagttcccca tgggtctcca ggaagtgacc ttggatgggg gtgggaaggg gctgctggag 720 ccaccttgct acctggggag ggtcccttcc tgaccccctg ggtgggctgg actgtgctgg 780 ttcatttaga aa tcaaagtc ctttgcctgg cgcagtggct gccaggagca g tgge tcagg 840 tcta taatcc cagcac tgtg gaaggccgag Gtgggcagat tgettgagee taggagttca 900 agaccagcct gggctacaga gcaagacctc atctttccta aaaaaaaaaa atacaaaaaa 960 <210> 45 <211> 2189 <212> DNA <213> Wisdom (Homo sapiens) < <; 3 00 & gt 3 08 > BC004929 15 201118377 < 309 > 2006-07-15 < 400 > 45 gaaggacggt gtgcgggccg gccagccctg gacgaaagaa gagggcccct ccaggccagt 60 ctgggcaccc tgggatagcg gctgcagcca ggcatggccg actctgcaca ggcccagaag 120 ctggtgtacc tggtcacagg gggctgtggc ttcctgggag agcacgtggt gegaatgetg 180 c tgcagcggg agccccggct cggggagc tg cgggtctttg accaacacc t gggtccctgg 240 ctggaggagc tgaagacagg gcctgtgagg gtgac tgcca tccaggggga cgtgacccag 300 gcccatgagg tggcagcagc tgtggccgga gccca tgtgg tcatccacac ggctgggctg 360 gtagacgtgt ttggcagggc cagtcccaag accatccatg aggtcaacgt gcagggtacc 420 cggaacgtga tcgaggcttg tg tgcagacc ggaacacggt tcctggtcta caccagcagc 480 atggaagttg tggggcctaa caccaaaggt caccccttct acaggggcaa cgaagacacc 540 Ccatacgaag cagtgcacag gcacccctat ccttgcagca aggccctggc cgagtggctg 600 gtcctggagg ccaacgggag gaagg tccgt ggggggctgc ccctggtgac gtgtgccctt 660 cgtcccacgg gcatctacgg tgaaggccac cagatcatga gggacttcta ccgccagggc 720 c tgcgcc tgg gaggttggct ct tccgggcc a tc ccggcc t ctgtggagca tggccgggtc 780 tatgtgggca atgttgcc tg gatgcacgtg ctggcagccc gggagctgga gcagcgggca 840 accctgatgg gcggccagg t atacttctgc tacgatggat caccctacag gagetatgag 900 gatttcaaca tggagttcct gggcccctgc ggactgcggc tggtgggcgc ccgcccattg 960 ctgccctact ggctgctggt gttcctggct gccctcaatg ccctgctgca gtggctgctg 1020 cggccactgg tgctctacgc acccc tgctg aacccc taca cgctggccgt ggccaacacc 1080 acc ttcaccg tcagcaccga caaggc tcag cgccatttcg gc ta tgagcc cctgttctcg 1140 tgggaggata gccggacccg taccattctc tgggtacagg ccgc tacggg ttcagcccag 1200 tgacggtggg gc tgggg cct ggaggcccag atacagcaca tccacccagg tcccgagccc 1260 tcacaccc tg gacgggaagg gacagctgca ttccagagca ggaggcaggg ctctggggcc 1320 agaa tggc tg tccttgtcgt agagccctcc acattttctt tttctttttt gagacaggg t 1380 cttgctctgt cacccagac t ggagtgcagt ggtgtgatca tagctcactg caccc tcaac 1440 ctcctgggtt Caagcaa tcc tcctgcctca gcctcctgaa cage tgggac cacaggtgca 1500 cgccaccaca cctggctttt ttttgttgtt tttagagaca gggtctcact atattgetea 1560 ggc tgg tc 11 gaactcctgg gctcaagtga tcttcccacg tgggcctccc aaaacgctgg 1620 aac tacaagt gtgagccacc gcgcc tggcc caagccctcc acattttcaa tccaggagcc 1680 ttgagtctgt gttgtgtcct gacacctcca agttctaggg ccgtcaggac acgggaggg t 1740 ttggggacag agtgtccttc ctctgtcctc tcatcccagt cctgatggcc gcttggtgag 1800 tgtctggtgc cctggtggcc tgccccagct ctcttctggc tttctgagca ggaagegage 1860 agaggctcca caggcttacg ctgctctcct gacagccaca cgcgaccc tc ggtgcagagt 1920 gcagaggcgg ctctggttcc tccagccacc tcagtccctc tttgggaggt gatgttccca 1980 ttgtttttca aaggcc tcac cttcaactgt c tg tc t taga attcccctct ggagggctat 2 04 0 ggcc tccc ta tgctttcact tcccacctct c tacc taag t tee ttcccag cacatcgcca 2100 gccc tgggcc tggggatgtc c ccaa tgctg tacctggctg accccggatt aaaagcctca 2160 tccacgaccg aaaaaaaaaa aaaaaaaaa 2189 < 210 > 46 <211> 3167 16 201118377

<212> DNA <2 13 > Homo sapiens <300 >

≪ 308 > 020165 < 3 09 > 1995 - 03-10 < 400 > 46 ttttctttgc cctcctgatt cttggctggc ccagggatga cttcctcgct gcagcggccc 60 tggcgggtgc cctggctacc atggaccatc ctgctggtca gcactgcggc tgcttcgcag 120 aa tcaagaac ggctatgtgc gtttaaagat ccgtatcagc aagaccttgg gataggtgag 180 agtagaatct ctcatgaaaa tgggacaata ttatgetega aaggtagcac ctgctatggc 240 ctttgggaga aatcaaaagg ggacataaat cttgtaaaac aaggatgttg gtctcacatt 300 ggagatcccc aagagtgtca c ta tgaagaa tgtgtagtaa ctaccactcc tccctcaatt 360 cagaatggaa cataccgttt ctgctgttgt agcacagatt tatgtaatgt caactttact 420 gagaattttc cacctcctga cacaacacca ctcagtccac ctcattcatt taaccgagat 480 gagacaa taa teattgettt ggcatcagtc tctgtattag ctgttttgat agttgcctta 540 tgctttggat acagaatgtt gacaggagac eg taaacaag gtcttcacag tatgaacatg 600 atggaggcag cagcatccga accctctctt gatetaga Ta atctgaaact gttggagctg 660 attggccgag gtcgatatgg ageagtatat aaagge tec t tggatgagcg tccagttgct 720 gtaaaagtgt tttcctttgc aaaccgtcag aattttatca acgaaaagaa catttacaga 780 gtgcctttga tggaacatga caacattgcc egetttatag ttggagatga gagag tcac t 840 gcagatggac gcatggaata tttgcttgtg atggagtact atcccaatgg atctttatgc 900 aagtatttaa g tc tccacac aagtgactgg gtaagetett gccgtcttgc tcattctgtt 960 actagaggac tgge t tatc t tcacacagaa ttaccacgag gagatcatta taaacc tgea 1020 atttcccatc gagatttaaa cagcagaaat g tec tag tga aaaa tgatgg aacctgtgtt 1080 attagtgact ttggactgtc catgaggetg actggaaata gactggtgcg cccaggggag 1140 gaagataatg cagccataag cgaggttggc actatcagat atatggcacc agaagtgeta 1200 gaaggagc tg tgaaettgag ggactgtgaa tcagctttga aacaagtaga ca tg ta tgc t 1260 cttggactaa tctattggga gatatttatg agatgtacag acctcttccc aggggaatcc 1320 gtaccagagt accagatggc ttttcagaca gaggttggaa acca tcccac 111 tgagga t 1380 a tgcagg t tc tcgtgtctag ggaaaaacag agacccaagt tcccagaagc c Tggaaagaa 1440 aatagcctgg cagtgaggtc actcaaggag acaatcgaag actgttggga ccaggatgca 15 00 gaggc tcggc ttactgcaca gtgtgctgag gaaaggatgg ctgaacttat gatgatttgg 1560 gaaagaaaca aatctgtgag cccaacag tc aa tccaatgt ctactgctat gcagaatgaa 1620 cgcaacctgt cacataatag gcgtgtgcca aaaattggtc cttatccaga ttattcttcc 1680 tcc tea taca t tgaagac tc tatccatcat ac tgacagca tcgtgaagaa ta 11 tec tc t 1740 gageatteta tgtccagcac acc 11 tgac ta taggggaaa aaaaccgaaa t tcaa t taac 1800 tatgaaegae agcaagcaca agetegaate cccagccctg aaacaagtgt caccagcctc 1860 tccaccaaca caacaaccac aaacaccaca ggactcacgc caagtactgg catgactact 1920 atatetgaga tgccataccc aga tgaaaca aatctgcata ccacaaatgt tgcacagtca 1980 attgggccaa cccctgtctg cttacagctg acagaagaag acttggaaac caacaagcta 2040 gacccaaaag aagttgataa gaacctcaag gaaagctctg atgagaatet ca tggageae 2100 tctcttaaac agttcagtgg cccagaccca ctgagcagta etagttetag cttgctttac 2160 17 201118377 ccactcataa aacttgcagt agaagcaact ggacagcagg acttcacaca gactgcaaat 2220 ggccaagcat gtttgattcc tgatgttctg cctactcaga tctatcctct ccccaagcag 2 2 8 0 cagaaccttc ccaagagacc tactagtttg cctttgaaca ccaaaaattc aacaaaagag 2340 ccccggctaa aatttggcag caagcacaaa tcaaacttga aacaagtcga aactggagtt 240 0 gccaagatga atacaatcaa tgcagcagaa cctcatgtgg tgacagtcac catgaatggt 2460 gtggcaggta gaaaccacag tgttaactcc catgctgcca caacccaata tgccaatggg 2520 acagtactat ctggccaaac aaccaacata gtgacacata gggcccaaga aatgttgcag 2580 aatcagttta ttggtgagga cacccggctg aatattaatt ccagtcctga tgagcatgag 2640 cctttactga gacgagagca acaagctggc catgatgaag gtgttctgga tcgtcttgtg 2 7 0 0 gacaggaggg aacggccact agaaggtggc cgaactaatt ccaataacaa caacagcaat 2 7 6 0 ccatgttcag aacaagatgt tcttgcacag ggtgttccaa gcacagcagc agatcctggg 2820 ccatcaaagc ccagaagagc acagaggcct aattctctgg atctttcagc cacaaatgtc 2880 ctggatggca gcagtataca gataggtgag tcaacacaag atggcaaatc aggatcaggt 2940 gaaaagatca agaaacgtgt gaaaactccc tattctctta agcggtggcg cccctccacc 3000 tgggtcatct ccactgaatc gctggactgt gaagtcaaca ataatggcag taacagggca 3060 gttcattcca aatccagcac tgctgtttac cttgcagaag gaggcactgc tacaaccatg 3120 gtgtctaaag atataggaat gaactgtctg tgaaatgttt tcaagcc 3167

<210> 47 <211> 553 <212> DNA <213> wise man {/iomo sapiens) < 300 ><308> AI765747 <309> 1999-06-28 <400> tgggtattta atatctttta tttgatttca actagcaaga gactaagact gatgtttgtt 60 tgtttgtttt accttcaaag tcttagtctt aaattgacaa gggcacaaaa aaagtgaaat 120 taaaataatt ttaacaaaag ggaaagcatt tgacagtgtt taaaagacag acaaatcaag 180 caaaactcaa tgtggcttaa ggtaaaaaat acaactaggt attaaacaat ttgtaaaaac 240 ttacaaagaa aatttagctt tcaagaaaaa ttaggcagat ttccttctgc caaagtgaca 300 ggtcctgagg aaaaaaaaca caacctctac aagaccaagc acagctcaag ggtcagggcc 360 ccgtcctagt catcttttta ttattcaata tccatcacag tggctgggca aacaacaggc 420 atcaataaat Gtttactaac tgaatgtgta ttataaggta taatcggtat aactgatata 480 gctcaaatct aatcatgaaa gcttctttaa agtaggataa aaagagcaca atcagcggta 540 ttccagaagg tag 553

<210> 48 <211> 2463 <212> DNA 18 201118377 <213> wise man (Ηο/πο sapiens)

≪ 3 00 > < 308 > ΝΜ_000173 < 309 > 2009 - 06-28 < 400 > 48 gagagaagga eggagtegag tggcacccta gaagacgctc tgtgccttcg gaggtctttc 60 tgcctgcctg tcctcatgcc tctcctcctc ttgctgctcc tgc tgccaag ccccttacac 120 ccccacccca tctgtgaggt ctccaaagtg gccagccacc tagaagtgaa ctgtgacaag 180 aggaa tc tga cagcgctgcc tccagacc tg ccgaaagaca caaccatcct ccacctgagt 240 gagaacc tcc tgtacacctt ctccctggca accctgatgc cttacactcg cctcactcag 300 ctgaacctag ataggtgcga gctcaccaag ctccaggtcg atgggacgct gccagtgctg 360 gggaccctgg atctatccca caatcagctg caaagcc tgc ccttgctagg gcagacactg 420 cc tgc tc tea ccgtcctgga cgtctccttc aaccggc tga cctcgctgcc tcttggtgcc 480 ctgcgtggtc ttggcgaact ccaagagc tc tacctgaaag gcaatgagct gaagaccctg 540 Cccccagggc tec tgaegee cacacccaag c tggagaage tcagtctggc taacaacaac 600 ttgactgagc tccccgctgg gctcctgaat gggctggaga atctcgacac ccttctcctc 660 caagagaac t cgctgtatac aa taccaaag ggcttttttg ggtcccacct cctgcctttt 720 gcttttctcc acgggaaccc ctggt tatgc aactgtgaga tcctctattt teg tege tgg 780 c tgcaggaca a tgc tgaaaa tgtctacgta tggaagcaag gtgtggacgt caaggccatg Θ40 acctctaatg tggccagtgt gcagtgtgac aattcagaca agtttcccgt ctacaaatac 900 ccaggaaagg ggtgccccac ccttggtgat gaaggtgaca cagacctata tga t tac tac 960 c cagaag agg acac tgaggg cgataaggtg eg tgccacaa ggac tg tggt caagttcccc 1020 accaaagccc atacaacccc ctggggtcta ttctactcat ggtccactgc ttetetagae 1080 agccaaa tgc cctcctcctt gca tccaaca caagaatcca etaaggagea gaccacattc 1140 ccacctagat ggaccccaaa tttcacactt cacatggaat ccatcacatt ctccaaaact 1200 ccaaaatcca ctactgaacc aacc ccaagc ccgaccacct cagagcccgt cccggagccc 1260 gccccaaaca tgaccaccc t ggagcccact ccaagcccga ccaccccaga gcccacctca 1320 gagcccgccc ccagcccgac caccccggag cccacctcag agcccgcccc cagcccgacc 1380 accccggagc ccaccccaat cccgaccatc gccacaagcc cgaccatcct ggtgtctgcc 1440 Acacaccttaga tcactccaaa aagcacattt ttaactacca caaaacccg t atcactctta 1500 gaatccacca aaaaaaccat ccctgaactt gatcagccac caaagctccg tggggtgctc 1560 caa gggcatt tggagagctc cagaaatgac ccttttctcc accccgac 11 ttgctgcctc 1620 c tccccc tgg gettetatgt cttgggtctc t tc tgge tgc tctttgcctc tgtggtcctc 1680 atcctgctgc tgagctgggt tgggcatgtg aaaccacagg ccctggactc tggccaaggt 1740 gctgctctga ccacagccac acaaaccaca cacc tggage tgcagagggg acggcaagtg 1800 acagtgcccc gggcctggct gctcttcctt cgaggttcgc ttcccacttt ccgc tccagc 1860 ctcttcctgt gggtacggcc taatggccgt gtggggcctc tagtggcagg aaggaggccc 1920 tcagctctga gtcagggtcg tggtcaggac ctgctgagca cagtgagcat taggtactct 1980 ggccacagcc tctgagggtg ggaggtttgg ggaccttgag agaagagcct gtgggctctc 2040 ctattggaat ctagttgggg gttggagggg taaggaacac agggtgatag gggaggggtc 2100 ttagttcctt tttctgtatc agaagccctg tcttcacaac acaggcacac aatttcagtc 2160 ccagccaaag cagaaggggt aatgacatgg acttggcggg gggacaagac aaagctcccg 2220 19 201118377 atgctgcatg gggcgctgcc agatctcacg gtgaaccatt t tggcagaa t acagcatggt 2280 tcccacatgc atctatgcac agaagaaaat c Tggaaagtg atttatcagg a tg tgagcac 2 34 0 tcgttgtgtc tggatgttac aaatatgggt ggtttta Ttt tctttttccc tgtttagcat 2400 tttctagttt gga tccac ta t ta ttgtatatta tc tg ta taa t aaaaaataat tttagggttg 2460 2 4 63 <210> 49 <211> 5683 <212> DNA <213> wise man (Homo sapiens) < 300 > < 308 > X79683 < 309 > 2005- 04-18 < 400 > 49 ccgcccggtg ttgcgctcct tcccagaatc cgctccggcc tttccttcct gccgcgattc 60 ccaactttgc tcaaag tcgc cggactctaa gctgtcggag ggaccgctgg acagacctgg 120 gaactgacag agggcctgga gggaaatagg ccaaagaccc acaggatgga gctgacctca 180 accgaaagag ggaggggaca gcctctgccc tgggaac t tc gactgcccc t ac tgc taage 240 gtgctggctg ccacactggc acaggcccct gccccggatg tccctggctg ttccagggga 300 agctgctacc ccgccacggc cgacctgctg gtgggccgag ctgacagact gactgcctca 360 tccacttgtg gcctgaatgg ccgccagccc tac tgca teg tcagtcacct gcaggacgaa 420 aagaagtgct tcctttgtga c tcccggcgc cccttctctg ctagagacaa cccacacacc 480 catcgcatcc agaatgtagt caccagc ttt gcaccacagc ggcgggcagc ttggtggcag 540 tcacagaatg gtatccctgc ggtcaccatc cagctggacc tggagge tga gtttcatttc 600 acacacctca ttatgacctt caagaca ttt cgccctgctg ccatgctggt egaaege tea 660 gcagactttg gccgcacctg gcatgtgtac egatatttet cctatcactg tggggctgac 720 ttcccaggag tcccactagc acccccacgg cactgggatg atgtagtctg tgagtcccgc 780 tactcagaga ttgagccatc cactgaaggc gaggteatet atcgtgtgct ggaccctgcc 840 atccctatcc cagaccccta cagctcacgg attcagaacc tgttgaagat caccaaccta 900 cgggtgaacc tgactcgtct acacacgttg ggagacaacc tactcgaccc aeggagggag 960 a tccgagaga ag tac tac ta tgccctctat gagctggttg tacgtggcaa ctgcttctgc 1020 tacggacacg cctcagagtg tgcacccgcc ccaggggcac cagcccatgc tgagggcatg 1080 gtgcacggag ct tgca tc tg caaacacaac acacgtggcc tcaactgcga gcagtgtcag 1140 gat t tc ta tc g tgacc tgcc c tggcg tccg gctgaggacg gccatagtca tgcctgtagg 1200 aagtgtgatc ggcatgggca cacccacagc tgccac tteg acatggccgt atacctcgga 1260 tctggcaatg tgagtggagg tgtgtgtgat ggatgtcagc a taacacagc g tggcgccac 1320 tgtgagctct gtcggccctt cttctaccgt gacccaacca aggacctgcg ggatccggct 1380 gtgtgccgct cctgtgattg tgaccccatg ggttctcaag acggtggtcg ctgtgattcc 1440 catga tgacc ctgcactggg actggtctcc ggccag tgtc gctgcaaaga acacgtggtg 1500 ggcactcgct gccagcaatg ccgtgatggc ttctttgggc tcagcatcag tgacccgtct 1560 gggtgccggc gatgtcaatg taatgcacgg ggcacagtgc ctgggagcac tccttgtgac 1620 20 201118377

cccaacagtg ctgcctggcc gtgggtggtg cacatggttg cacc taa 111 gtgacccccg cagaccc tgg cgcttagagc gggcc tg tgc gggac tc tgc ggtatctcct ac tccc tac t gtgctagaga cgc taccaa t gcacccctcc aaccctcaag cctggagtgg acaggc tg tc accag tgggc cgtggccagt tgcaacaccc gaaaggtgca ccctgtccct gatgaatatt gaagcttgtg tgtgagtgca caa tgcc tgc ttccatggcc aatccgcagc ccatgcctcc c tcaccagtg acc tgcaacg tctgagtgcc gactctcgtg ccaggggtgt gcctgccatc gcccgtacac gcc t ttgaga ggtgcccgca cggcgtgaaa gtgcaagatg gcacttaatc ttcctgggtg cgtgccaata gatcctgtta ac tggggcc t ctttggatcc ggcgacgc tg gggaggctga gggaaactcc ag t tcctgg t cccaggtccc c tgcccacag aaccacatgc acaagctgca ctggacctgg tgtttagtgg gccatgagga tcatcagcct gttcactgag ttgggcgccg aagcctgcca aatgtctctg ggggattccc acacaggcgc ttgctggttt gtcctgaagg cccagcaga t cccc tgggca gtgggaaca t gctgtttaca aggc tgcccg agtgcccatc ccaatgtcca gccatggttg agttcacagg aagagc tcca gaatagatac ctggtgtgcg cctgccatgc agcgcctaga gcagcttctg acacctcagc ttggggaggc agaacttcaa tcacac tgcg cctatgacag cc tcagcc ct ctgcaaacgt gagcctcgac ccagtgtgat tgagcaggtg gaacacccga atcctggact ggcctctgtg tgagcaatgg cc tgtgtggg caggtac ttg tctgaagctg cc tgctcatt gggtgatgct gggtctggtg gtccaccctc ttctgagtgc ctgtgacacg gtgcagccca tcgaac tggt tagctgccgg ttgcctgggc ccacggggac ccc tgggagc tgtgtgccac gtttggggac tgacccaatg ccacacagag gcagagctgt tcc tgaccag ggccctagct ccagcc t tg t gcagtgccac ctggggagac acctcagtgt ctgtgaccag atgc ttcggg gcagcgggcg gcacatgcag cgcctccact cactgagcac tgccaacca t gcagctcgac catccggcat ggcagtacct c tag tgac tg c tgc tcggc t gagggcacag caacc tggc t gggcaggtgc ggctcaggct ccgaacgcga gcagagttgg catttggtgc atatttccta gtacggacag gactcgctgg gctgccctgg cccagcaaga atctacaatg aaccctcatg tgtgcccctg cgaggggcac gcctttgggc ccatgtgtct tgccgtgatc ccacggctgc caacggcact tgccgggcag ccatcaaggc gatcctgatg ggtccacact caccgctgca tgccactgtg gtagaccgct gcctgcctcc tgcctgtgcg cctgggttgc caccgcttca tgtgcccgtg gattgggacc Caggagttgc gagaagctgg gcacagcttg ctgactcagc gcactaagtg cagcatcttg gcccatagcc agccc tg tga gacgtgg atg gccgcccc tg gtcaatgcca acttccggcc tcga tgtgg t tcgtgcgact tggactatga aactgattgt ccagggatga atcctgtctg ggggaagtgc tgctgctgcc agcgccaggc cttctccctc gtgccctgcc gtggtcagtg gctactatgg tcagcagtct ttcgctgtga gcaatgggca tcacaggggg catatggggc ttgctacttc gctatacggg caggtggccg cctgtgaccc gtgcccactc catgcaacc t atccaagcag gtgcccccaa caagcccgga gctttggagg agtgccatgc caggtcactg gcttctcagg gagtggtgca aacagacggg gcattgtgca tggaggccac tcgaggcaga gtctggagcg acttgctcaa agtctgcaga gcaactcggc tgaccgc tgc tgactgcgac c tgccgccag cttcctggac ggagcgcctg acaggaaggt cc tgc tgc tg gcagcgtcca tcgca tccaa cc ttgagcct ccagcctgag ccgtgtcctg cacctttgaa tgaggcctgc atgtcagtgc cc tgtgcaag c 11 tggcccc ctgtgaaagg cgcctgccag tgcagatgag tgagcactgt gcagtgccgg ttgccaccag gc tgcgatgt gtgccaactg acaccccggg gaagcc tggc gctgggcaca tgggcagtgc cttctggaac agaaggcccc gcggacttgt ctgtgattgt cacgtgccgc aatctttcct ggac t tggca tgtgctgggt gggca Teg ta agaggagetg cctgacagat agataggett acattcaaac ggcagaacgt aagtgctcgg 1680 1740 1800 1860 1920 1980 204 0 2100 2160 2220 2280 2340 2400 2460 2S20 2580 2640 2700 27 60 2820 2880 2940 3000 3060 3120 3180 3240 3 300 3360 3420 3480 3540 3600 3660 3720 3780 3840 3900 3960 4020 4080 4140 4200 4260 21 201118377 catcggacag aggeae tga t ggatgctcag aaggaggac t tcaacagcaa acacatggcc 4320 aaccagcggg cacttggcaa gctctctgcc catacccaca ccctgagcct gacagacata 4380 aatgagctgg tgtgtggggc ccagggattg catcatgatc gtacaagccc ttgtgggggt 4440 gccggctgtc gagatgagga tgggcagccg cgctgtgggg gcctcagctg caatggggca 4500 gcggctacag cagacctagc actgggccgg gcccggcaca cacaggcaga getgeagegg 4560 gcactggcag aaggtggtag catcctcagc agagtggctg agactcgtcg gcaggcaagc 4620 gaggcacagc agcgggccca ggcagccctg gacaaggcta atgcttccag gggacaggtg 4680 gaacaggcca accaggaact tcaagaactt Atccagagtg tgaaggaett cctcaaccag 4740 gagggggctg atcctgatag ca ttgaaatg gtggccacac gggtgctaga gctctccatc 4800 ccagcttcag ctgagcagat ccagcacctg gegggegega ttgcagagcg agteeggage 4860 ctggcagatg tggatgegat cc tggcacgt actgtaggag atgtgcgtcg tgc cgagcag 4920 ctactgcagg atgeaeggeg ggcaaggage tgggctgagg atgagaaaca gaaggeagag 4980 acagtacagg cagcactgga ggaggcccag cgggcacagg gtattgccca gggtgccatc 5040 cggggggcag tggctgacac acgggacaca gagcagaccc tg taccagg t acaggagagg 5100 atggcaggtg cagagcgggc actgagctct gcaggtgaaa ggge teggea gttggatgct 5160 ctcctggagg ctctgaaatt gaaacgggca ggaaatagtc tggeagee tc tacagcagaa 5220 gaaacggcag gcagtgccca gggtcgtgcc caggaggetg ageagetget acgcggtcct 5280 ctgggtgatc agtaccagac ggtgaaggcc etagetgage gcaaggccca aggtgtgctg 5340 gctgcacagg caagggcaga acaactgccg gatgaggctc gggacctgtt gcaagccgct 5400 caggacaagc tgeagegget acaggaattg gaaggcacct a tgaggaaaa tgagcgggca 5460 ctggagagta aggcagccca gttggacggg ttggaggcca ggatgcgcag cgtgcttcaa 5520 gccatcaact tgcaggtgca gatctacaac acctgccagt gacccctgcc caaggcctac 5580 cccagttcct agcactgccc cacatgcatg tctgcctatg cactgaagag ctcttggccc 5 64 0 ggcagggccc ccaataaacc ag tg tgaacc cccaaaaaaa aaa 5 6 8 3

<210> 50 <211> 2656 <212> DNA <213 > Homo sapiens < 300 ><308> M25813 <309> 1993-04-27 <400> ggtctgtcag gcaggaaacg actgggcccc atctctgctg acagcaccac age tcccc tg 60 gagaaggage tacctcccca cctgggggaa ctgaccgtgg ctgaggagac ctccagctct 12 0 ctgcgcctgt cctggacggt agcccagggc ccctttgact ccttcgtggt ccagtacagg 180 gacacggacg ggcagcccag ggcagtgcct gtggccgcag accagcgcac agtcaccgta 240 gaggacctgg agcctggcaa gaaatacaag tttctgctct acgggctcct tgggggaaag 300 cgcctgggcc cggtctctgc cctgggaatg acagccccag aagaggacac accagcccca 360 gagttagccc cagaggcccc tgagcctcct gaagagcccc gcctaggagt Gctgaccgtg 420 accgacacaa ccccagactc catgcgcctc tcgtggagcg tggcccaggg cccctttgat 480 22 201118377

gttccagcgc cgcatggatg gacagacaga cttctggagg tggttttggg aacatctctg gagagttctg gctgggcaat ctaccacggc accgcagggg actccatgag ctaccacagc gtacaggaac tgccactacg ccaacctcaa cgggctctac tccttcgtgg gaccagagca tatggcctcc gctcctgctg gacgtgacca ttcctgctcc ctgcagcgcg tccgggactc atccagggaa agtgaaatca agcttcaaag ggccaggccc gtggtctcgg cctgacggtc tggaagcccc ccgcctctgc ctccacacca gccagcatca accccccgca ctcagcttcc tc tcaccagc tggggccaga cccttcccca accatcttcc gatgggggcg gac tgggagg gaggccctgc ggggacgagg taccgcctcc ggcagtgtct tcctaccgag gggagcacag gtgcccttca tgagctgctg egagagaaga a tc tcgcagc gaaac tgagg tccagtatga agatcctcat atgaagggaa gtcagacc tc ccagttcact gctttggggt agctgatggt tgtacagcct ccgcccgcac gggagacc tc tctcctacca ggacccagaa teegagge ccacacagtt cccagaatcc aggeggagae actacaccgc ccttcaccac ccgccctgct acacccctgg tee t tggee t gcctcctgcc gggac tgegg tcaacggcaa gctggctggt actatgccca acagcc tgac ctgtgttcgc acttggaggg tc tc tgcccg gggcctggtg tggaccatca tt Eggaaa tgaa cccacctctc gccagggtcc ac tg tg ttta ta cccg ggacacgaac ctcaggcctg gcgcctgggg agaggag tea gaggctcaac tccatcacca gccggggacg gacactgtat cctcagccca agccaaggtc gc tggeggae actccagggg tgaggagag t geg tgcac tg tgtggacacc cccaggcagc cacagtgcgt agggetagag cacttggact tggacagaac ctttgggtcc gcccgtgtcc ggaggagatg ccgcgagcgg acaggcagg t ccagtacgac tgatcgggac gggag tgage gctgagacca tcgcacccca ttcaccaccc caggggggag gggcagcccc gagcccagca cccc tctcag aggccccgcc tgggaggccc agcactctgg cggcactcgg ggge tgegag gttctggaga aactggatgc ggaggggagc ctgatcccag gagcctctca aacttgaccg tatgacgtcc gcggtggact ggcctgcggg gcccctcggg gagcccccag caggagatcc acctcctaca acctctttca cagaacggag cccctgaacg gactactcca tccttccacg cccaacagct tggtaccact agaaactttc gtatgactgc agccgctgga gggaggggtt aggeettget ccccctacag c tgagggcac tgtcccagct caccgggggc agccgcatcc ccgtgctccg gaccccacaa gcccccgtga ccccaccatc ctcagagtgt gcgctcgcta caggcttcct agggattege aggtcacagc accccctgca gccccaacct aettggagge tccggcccgc tgctcccagg atgeaegge t ccacggg tgg ccgg Tgee tc tgttttgcga tccgcgtgga ta gac tegge tgc tca tc tc ggaagggc tt gctccccagc egageae tga ggaagee t tc cgtacaggag eg tggaegge gttcctcctc cacagggctg gtctgtgact cttcgactcc gcgtccac tg ggacctgcgt ggccgacagc cctccaattc ccgggcggac gcaggtggat tgaggtgacc caccacggtt cgtgctgcac ccctggggcc tgaccttgtc cacttcccca caaggaagtg agge tacc tg agggatcaca ccaggccatg getgeggate caggaccagc catggagact cctgcgggct tgeggag tac ctgcgctgtc egagtteteg ggggggaggc ggggtcgccc Tctgccagcg caataaagga 540 600 660 720 780 840 900 960 1020 1080 1140 1200 1260 1320 1380 1440 1500 1560 1620 1680 1740 1800 1860 1920 1980 2 04 0 2100 2160 222 0 2 2 8 0 2 3 4 0 2400 24 60 2520 2 580 2 64 0 2 6 5 6 23

Claims (1)

201118377 VII. Scope of Application: 1. A method for assessing and predicting dementia, which is to test the degree of expression of a target gene in a test subject cell as an indicator for evaluating and predicting dementia. The method comprises the following steps: Step 1 provides an isolated test subject; Step 2: Ex vivo detection of the degree of expression of at least one target gene in the gene transcript of the test subject 'The target gene is selected from the group consisting of guanylate cyclase! (GUCY1B3), multi-drug resistant p-sugar Protein 1 (muitjdmg resjstanee P-glycoproteinl, ABCB1; ATP-binding cassette transporter subfamily B, member 1), ATP-binding cassette transporter ABCC4 (subfamily C, member 4), ATP-binding cassette transporter ABCC6 (subfamily C, Member 6), nucleoside diphosphate linked moiety X-type motif 5 [Nudix (nucleoside diphosphate linked moiety X)-type motif 5, NUDT5], guanidinoacetate N-methyltransferase (GAMT), Base-d-5-steroid dehydrogenase 3β- and steroid d-isomerase 7 (Hydroxy-delta-5-steroid Dehydrogenase, 3 beta- and steroid delta-isomerase 7, HSD3B7), bone morphogenetic protein receptor BMPR2 (Ber morphogenetic protein receptor, type II (serine/threonine kinase)] SMAD family member 2 (SMAD2), glycoprotein lb (platelet), alpha polypeptide (GP1BA), human β2 adhesion protein LAMB2 message RNA (H. sapiens LAMB2) mRNA for beta2 laminin), and at least one of the groups consisting of tenascin XB (TNXB); and step 3 comparing the degree of expression of the target gene in the test subject with the same base in the normal control group 28 201118377 The difference in performance is judged whether the subject has a risk of dementia or suffering from dementia. 2. The method of claim 1, wherein the subject to be tested is a peripheral blood leukocyte. 3. The method of claim 1, wherein the method for detecting the degree of expression of the target gene comprises a quantitative polymerase chain reaction (Q-PCR) or a quantitative reverse transcription polymerase chain reaction (quantitative) Reverse transcription PCR, QRT-PCR) ' or microarray analysis. 4. The method according to claim 1, wherein the target gene guanylate cyclase j (GUCY1B3) has a nucleotide sequence as shown in SEQ ID No: 39, and the target gene ATP-binding cassette transporter The protein ABCC4 line has the nucleotide sequence shown in SEQ ID No: 41, and the target gene ATP-binding cassette transporter ABCC6 line has a nucleoside 序列 sequence as shown by SEqIDN〇: 42 'target gene thioglycolic acid N-A The basal transferase (GAMT) has a nucleotide sequence as shown in SEQ ID No: 44, and the target gene is basally-d-5-steroid dehydrogenase 3 β- and steroid d-isomerase 7 (HSD3B7) Having the nucleotide sequence shown in SEQ ID NO: 45, the target gene bone morphogenetic protein receptor!! type BMpR2 (serine/threonine kinase) has a nucleus as shown in SEQroN〇:46 _ sequence, the target gene SMAD family member 2 (SMAAD2) has a nucleotide sequence as shown in SEQ, still N: 47, and the target gene glycoprotein lb (platelet) a polypeptide (GP1BA) has The nucleus-sequence shown in SEQ ID 48 48, the target gene 腱 腱 protein ΤΝχΒ (ΤΝχΒ) has the SEQn) N 〇: 5 〇 The nucleotide sequence is 'and the degree of expression of these genes increases with the onset of dementia. 5. The method according to claim 1, wherein the target gene multi-drug resistance p_glycoprotein r C'· 29 201118377 white 1 (ABCB1) has the nucleoside as shown in SEQ ID No: 40 Acid sequence, the target gene nucleoside diphosphate linkage X-type subject 5 (NUDT5) has a nucleotide sequence as shown in SEQ N: 43, and the target gene human β2 adhesion protein LAMB2 message RNA has The nucleotide sequence shown in SEQ ID No: 49, and the degree of expression of these genes decreases as the dementia occurs. 6. The method of claim 1, wherein the dementia comprises mild dysfunction, Alzheimer's disease. 7. The biological indicator for assessing and predicting dementia is based on the degree of expression of a target gene in the peripheral blood leukocytes of the subject in vitro, the target gene being selected from avian acid cyclization S# 1 (GUCY1B3), multidrug resistance p-audiosin 1 (abcbI), ATP-binding cassette transporter ABCC4, ATP-binding cassette transporter ABCC6, nucleoside diphosphate linkage χ_type theme 5 (NUDT5), thioglycolate- Methyltransferase (GAMT), trans-d-5-steroid dehydrogenase 3 β - and steroid d-isomerase 7 (HSD3B7), skeletal morphogenetic protein receptor Π BMPR2 (serine/threonine) Acid kinase), SMAD family member 2 (SMAD2), glycoprotein ib (platelet) a polypeptide (GP1BA), human β2 adhesion protein LAMB2 message RNA, and tendon protein xb (TNXB) At least one of them. 8. The biological indicator according to claim 7, wherein the target gene guanylate cyclase 1 (GUCY1B3) has a nucleotide sequence as shown in SEQ ID No: 39, and the target gene ατΡ binding cassette transporter The ABCC4 line has the nucleotide sequence shown in SEQ ID No: 41, and the target gene ATP-binding cassette transporter ABCC6 line has the nucleotide sequence shown in SEQ ID No: 42 and the target gene thioglycolic acid N-methyl The transferase (GAMT) has the nucleotide sequence 'target gene hydroxy_d_5_steroid dehydrogenase 3 β_ and steroid d-isomerase 7 (HSD3B7) as shown in SEQ ID 201118377 No: 44 having The nucleotide sequence shown in SEQ ID No: 45, the target gene bone morphogenetic protein receptor Π type BMPR2 (serine/threonine kinase) has the nucleotide sequence shown in SEQ ID No: 46, The target gene SMAD family member 2 (SMAD2) has a nucleotide sequence as shown in SEQ ID No: 47, and the target gene glycoprotein lb (platelet) a polypeptide (GP1BA) has SEQ ID NO: 48 The nucleotide sequence shown, the target gene tendon protein (ΤΝχΒ) has the same as SEQ The nucleotide sequence shown by ID No: 50, and the degree of expression of these genes increases with the occurrence of dementia. 9. The biological indicator according to claim 7, wherein the target gene multidrug resistance p_ vinegar protein 1 (ABCB1) has a nucleotide sequence as shown in SEQ ID No: 40, the target gene nucleoside The diphosphate linkage X-type subject 5 (NUDT5) S has a nucleotide sequence as shown by SEq ID N〇: 43 and the target gene human β2 adhesion protein LAMB2 message has the SEQ ID No: 49 Nucleotide sequences are shown, and the degree of expression of these genes decreases with the onset of dementia. 10. The biological indicator as described in claim 7 of the patent scope, wherein the dementia comprises mild dysfunction, Alzheimer's disease. 11. A kit for assessing and predicting dementia, comprising at least one biological indicator for assessing and predicting dementia as described in Section 7 of the patent application. 12. In the case of the invention described in claim 11, the dementia comprises mild dysfunction, Alzheimer's disease. 31