CN101726580A - Use of endogenous formaldehyde in preparation of diagnostic reagent for treating Alzheimer's disease - Google Patents
Use of endogenous formaldehyde in preparation of diagnostic reagent for treating Alzheimer's disease Download PDFInfo
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Abstract
The invention relates to the use of endogenous formaldehyde in the preparation of a diagnostic reagent for treating Alzheimer's disease. In the method, the endogenous formaldehyde is used as a biomarker of Alzheimer's disease; and by detecting the formaldehyde concentration of a sample of the urine of the person receiving the test and comparing the formaldehyde concentration of the sample of the urine of the person receiving the test with the formaldehyde concentration of the urine of a healthy person receiving the test, whether a person receiving the test has Alzheimer's disease can be determined and the state of the Alzheimer's disease in the person receiving the test can be initially judged.
Description
Technical field
The present invention relates to the new purposes of formaldehyde in the diagnosticum pharmaceutical field, in particular to the new purposes of endogenous formaldehyde in the diagnosticum of preparation senile dementia, wherein with the biomarker of endogenous formaldehyde as senile dementia.
Background technology
Senile dementia (Alzheimer ' s disease, be called for short AD) be a kind of nerve degenerative diseases (Francis et al., 1999, Varadarajan et al., 2000), along with the continuous prolongation of human longevity, the senile dementia incidence of disease is also rising year by year, 65 years old and more than 85 years old the incidence of disease be respectively about 5% and 8% (executing Anguo etc., 2000).Senile dementia is the fourth-largest cause of disease that causes the elderly's death after tumour, heart disease and cerebrovascular disease, thereby causes the extensive concern of Chinese scholars.WHO has been decided to be senile dementia one of twentieth century five big emphasis diseases (Wei Hongmei, 2008), because of AD pathogenesis complexity, characterizes with other illnesss and easily obscures, and clinical being difficult to made a definite diagnosis.Present fixed AD type has: 1. vascular dementia: i.e. multi-infarct dementia is the dementia due to the brain tissue blood supply insufficiency due to the various reasons, and modal is cerebral infarction and cerebral arteriovenous malformation; 2. Alzheimer: mainly cause by the atrophy of brain cortex, cause neurocyte reduce in a large number (reaching more than 30%), the ventricles of the brain enlarge, so that the sex change disintegration of nerve net.The pathological change of Alzheimer has two distinguishing features: senile plaque expelling, neurofibrillary tangles sex change, neuron loss etc.General senile dementia behavioural characteristic shows as the change of memory, calculating, thinking, language, directive force and the disturbance of emotion and personality, and go down (Ni Qiwei, continental rise ancestor, 2007) of social activities ability occur.The rising of the senile dementia incidence of disease has had a strong impact on the elderly's quality of life and has become a serious social concern.
Studies show that in a large number, do not find a kind of practicable clinically, easy to operate and diagnostic method that diagnosis rate is high up to now as yet, the diagnostic method of senile dementia still rests on the AD stage that just is diagnosed as at present when serious sign obviously appears in the patient, it is reported, nonetheless, the diagnosis rate of this moment by afterwards biopsy or postmortem on neither 100%, still have the basic and AD of 20% left and right sides case have nothing to do (McKhann et al., 1984).Therefore when being diagnosed out AD clinically, missed best occasion for the treatment in fact already.In a single day the AD disease develops into the order of severity and just is difficult to retrieve, but if make a definite diagnosis the development that still can control the state of an illness with relative medicine and treatment means in early days, prolong patient's life-span, raising patient life quality.Because the factor that is associated with the AD disease is intricate, still do not find feasible clinically associated characteristic biomolecule at present, although report is arranged in the recent period, scientist is through making great efforts to find to exist 15 kinds of (Simonsen et al. of non-existent specific proteins in control group in patient's cerebrospinal fluid, 2008), but if be used as the clinical detection means with this, no matter in the heart of on the complicated operation degree or can the patient accept, probably all be difficult to promote, therefore find the method for the AD disease of just clarifying a diagnosis in early days just to become important and meaningful.
Although, and had breakthrough progress in some aspects about the existing a large amount of reports of the research work of senile dementia clinical specificity label.For example: wear by waist and to get cerebrospinal fluid, utilize the SELDI-TOF-MS technology, measure the variation of wherein a lot of albumen relevant, find out the characteristic biomarker, can distinguish AD and normal healthy controls (Simonsen et al., 2008) with AD.By the elisa technology, detect A β in patient's cerebrospinal fluid in addition
1-42,, Ptau and tau/A β
1-42, content, can find clocklike to change (Femke H.Bouwmana, 2008).Regrettably, the method for clinical diagnosis senile dementia is not resolved from practical application at present, and concentration of formaldehyde has the research work of specific biological labeled molecule also not appear in the newspapers at home and abroad to find clinically in patient's AD body fluid by detecting.
Summary of the invention
The present invention relates to the new purposes of formaldehyde in the diagnosticum of preparation senile dementia, wherein with the biomarker of endogenous formaldehyde as senile dementia.Endogenous formaldehyde can be used for determining as the biomarker of senile dementia whether the experimenter suffers from the danger of senile dementia or development senile dementia, can also be used to estimating the drug effect of the medicine for the treatment of senile dementia.
In the present invention, we carry out MMSE (Folstein et al. by the senile dementia patient to 81 routine contrast crowds (non-dementia patients) and 141 routine different dementia degree, 1975) test (general in the world simple and easy spirit intelligence is checked scale), be divided into normally according to the score value difference, slight dull-witted, moderate dementia and severe dementia.The urine of collecting the tester is simultaneously carried out fluorescence-HPLC and is analyzed (Luo etal., 2001).Interpretation of result is by experiment found the endogenous formaldehyde concentration in AD patient's body fluid and compares to present certain rules.This discovery is expected to the reference index of endogenous formaldehyde as clinical generaI investigation.
In research work of the present invention, the inventor finds that by analysis AD patient urinates concentration of formaldehyde and generally is higher than contrast (contrast: 0.082mM, n=38, AD:0.303 ± 0.032mM, n=141, report healthy human urine concentration of formaldehyde is that 0.08mM is similar on contrast and the document), and also present the trend that increases progressively and (see Fig. 3 a) along with dementia degree increases the weight of to urinate concentration of formaldehyde.
The present invention can be used for preparing the diagnosticum of senile dementia with the biomarker of endogenous formaldehyde as senile dementia.Embodiment of the present invention are described the relation between endogenous formaldehyde and the senile dementia in detail, and it is used for preparing the purposes of the diagnosticum of senile dementia.
In a preferred embodiment of the invention, by detecting the concentration of formaldehyde in experimenter's urine, concentration of formaldehyde in itself and the health volunteer's urine is compared, judge whether described experimenter suffers from senile dementia, and tentatively judge the degree of described experimenter's senile dementia.
In another preferred embodiment of the present invention, change by the concentration of formaldehyde of vertical monitoring experimenter in life process different time stage urine, judge that described experimenter develops the danger of senile dementia.
In another preferred embodiment of the present invention, by the monitoring experimenter before the medicine of taking the treatment senile dementia and afterwards the concentration of formaldehyde in the urine in different time stage change, estimate the drug effect of described medicine at senile dementia.
In application mentioned above, the urine concentration of formaldehyde can be measured by high performance liquid chromatogram fluorescence chromatogram (HPLC-fluorescence), but it should be appreciated by those skilled in the art that detection method is not limited to this, can also utilize any other suitable method well known in the art to detect.
Compared with prior art, the invention has the advantages that:
1. provide formaldehyde to be used to prepare the new purposes of the diagnosticum of senile dementia.Endogenous formaldehyde is as the biomarker of senile dementia, can be used for not only judging whether the experimenter suffers from senile dementia, and can tentatively judge the degree of described experimenter's senile dementia, thereby in time adopting suitable treatment that clinical foundation is provided;
2. endogenous formaldehyde is as the biomarker of senile dementia, can also be by vertically monitoring the variation of concentration of formaldehyde in the different time stage urine of described experimenter in life process, can predict in time that described experimenter develops the danger of senile dementia, in time take medical science prevention and intervention, in order to avoid affect the best opportunity of treatment adversely;
3. endogenous formaldehyde can also be used to estimating the drug effect of the medicine for the treatment of senile dementia as the biomarker of senile dementia, and evaluation method is simply quick, and cost is low, and is economical and practical;
4. detection method of the present invention is simple and practical, and the experimenter is not caused physical impairment, and therefore, this method has extraordinary practicality.
Description of drawings
From the detailed description below in conjunction with accompanying drawing, above-mentioned feature and advantage of the present invention will be more obvious, wherein:
Fig. 1 shows the typical curve of formaldehyde (FA) concentration of fluorescence high-performance liquid chromatogram determination, and equation of linear regression between concentration of formaldehyde and the chromatographic peak area and linearly dependent coefficient R
2Y-axis: chromatographic peak area; X-axis: concentration of formaldehyde.Equation of linear regression: Y=15357X+364.34; R
2=0.9997.
Fig. 2 shows the fluorescence high-efficient liquid phase chromatogram of 5 kinds of different representative samples.2a. it is blank: water; 2b. contrast: standard formalin (1ppm); 2c. the urine sample of slight dementia; 2d. the urine sample of moderate dementia disease; 2e. the urine sample of severe dementia disease.The retention time of formaldehyde is about 8 minutes.
Fig. 3 a shows the concentration determination result of urine formaldehyde in 4 kinds of different crowds, and concentration and patient's dementia degree of dementia patients urine formaldehyde are proportionate as seen from the figure, and with contrast significant difference are arranged, and used data all adopt mean value ± standard error among the figure.Described 4 groups of different crowds are that 1 group of normal healthy controls experimenter (does not suffer from any disease, n=38) and the AD patient of 3 groups of 3 kinds of degree (slight (n=50), moderate (n=39), severe (n=52)).3b. the concentration determination result of urine formaldehyde comprises vascular dementia in the different dementia classification, the Alzheimer dementia, Combination dull-witted and other, as seen from the figure, several dementia patients are urinated formaldehyde all significant difference compared with the control.3c. show AD patient's (suffering from hypertension or diabetes simultaneously) and corresponding non-AD patient (do not suffer from the health volunteer of any disease and suffer from the non-AD patient of hypertension or diabetes respectively) urine concentration of formaldehyde comparison, AD patient's urine concentration of formaldehyde and non-AD patient have utmost point significant difference as can be seen; 3d. the urine concentration of formaldehyde of suffering from non-AD patient's the urine concentration of formaldehyde of hypertension or diabetes and healthy people (not suffering from any disease) relatively.
Embodiment
Come further to illustrate the present invention by the following examples.But should be appreciated that described embodiment is illustrational purpose, and be not intended to limit the scope of the invention and spirit.
1. materials and methods
1.1 material and instrument
Ampicillin, and formaldehyde (37%, W/V) (top grade is pure, available from sigma company), trichloroacetic acid, sodium chloride (analyze pure, available from ancient cooking vessel state biotech firm), acetonitrile, ether (liquid chromatography is pure, available from Tedia company), water is the Milli-Q ultrapure water.Urine is picked up from Shantou, Guangdong medical college, Beijing Shunyi County resident community, the old experimenter in geriatric hospital and the apartment for elderly people.(HP1100, agilent), nitrogen dries up instrument (Shanghai is chemical industry company limited beyond one's expectations) to fluorescence-high performance liquid chromatograph.
1.2 the mensuration of simple intelligent state of mind scale (MMSE)
Randomly drawing 112 routine healthy people in community makes investigation, healthy people's investigation is under the physician guidance of Zhong Guan-cun hospital Geriatrics Dept. and psychiatry department, in resident community (community is without any the living environment different with other resident communities) door-to-door survey sampling at random, investigation sampling compliant scientific methods, the true representative and ubiquity of sample.The clinical investigation number is no less than 30 people, has statistical significance.Removed minority no statistical significance and crowd with illness except that hypertension and diabetes own at last, remaining 81 people do the contrast of dull-witted investigation.The investigator of dementia patients is in hospital or the outpatient at apartment for elderly people and geriatric hospital, the investigation of carrying out again after the medical practitioner diagnosis is made a definite diagnosis.
Dementia patients is totally 141 people, and at regular hospital (geriatric hospital, Beijing) and several homes for the aged sampling survey, this process is to finish under the guidance that the disease of old people house-owner appoints fully, according to scientific and normal method sampling, the representative and ubiquity of sample.
1.3 the collection of urine and processing
The tester prohibited against alcoholic drinks and high fat diet the same day, and stay urine second day early morning on an empty stomach, was temporary in-70 ℃, and was to be measured.
1.4 the derivative reaction of sample
Standard curve making: the formaldehyde water is made into the mother liquor of 1mM, and the water serial dilution becomes the standard 0,0.01,0.02,0.05,0.1,0.2,0.3 of eight variable concentrations, and 0.6mM uses water as blank.Variable concentrations formalin 0.8ml is added in 8 2ml that contain 200 μ l healthy human urine liquid glass tube with cover (needs that add urine and be deduction background for the back working sample time), add 0.1ml ampicillin (aqueous solution of 2.5mg/ml) and 0.25ml trichloroacetic acid (20% aqueous solution) more respectively, fully vibration is 30 seconds, places 90 ℃ of water-baths immediately 1 hour.Then, reduce to room temperature, and solution in the pipe is transferred in the 10ml glass centrifuge tube, just now 2ml pipe is washed respectively twice and merged in this 10ml pipe with the 1ml ether, and in every pipe, add small amount of sodium chloride (about 0.5g), fierce concussion 1 minute, centrifugal 5 minutes of 1000g, upper organic phase is transferred in the glass tube of 5ml, used 2ml extracted with diethyl ether water twice again, merge with organic phase.Dry up instrument at 40 ℃ with nitrogen and dry up organic phase,, shook 30 seconds, filter in the special-purpose sample bottle of liquid chromatography, seal to be measured with the organic filter of 0.45 μ m with the dissolving of 800 μ l acetonitrile-aqueous solutions (volume ratio: 1: 1).
Analyte derivativeization: the same operation of analyte derivativeization, prepare as follows: reagent blank: 1ml water; Urine blank: 800 μ l water+200 μ l urines; Urine+standard: 300 μ l water+500 μ l formaldehyde standard (1ppm)+200 μ l urines; Sample to be tested: 800 μ l water+200 μ l urine to be measured.Processing procedure is used the dissolving of 500 μ l acetonitrile-aqueous solutions (volume ratio: 1: 1) at last with above-mentioned.Reagent blank, the urine blank, urine+standard, sample to be tested, all do at every turn two parallel, get average again and be used for calculating.
Calculate: the concentration of formaldehyde of sample (ppm of unit)=(peak area of each sample deducts the peak area of reagent blank) ÷ (peak area of urine+standard deducts the peak area of urine blank) * 0.5 (normal concentration) * 5 (200 μ l urine).Be converted into mM at last.
1.5 the condition of high performance liquid chromatogram fluorescence chromatogram
Fluorescence-HPLC post: HP Zorbax StableBondSB-C18,5m, 250mm * 4.6mm (Germany), 40 ℃ of column temperatures, sample size 10 μ l, moving phase: acetonitrile: water (volume ratio: 25: 75), flow velocity: 1ml/min, fluorescence detector.
2. result and discussion
2.1 the typical curve of liquid chromatogram measuring formaldehyde (FA)
See that from Fig. 1 concentration of formaldehyde and chromatographic peak area have good linear relationship, regression equation has shown that linearly dependent coefficient is 0.9997.Therefore can measure the concentration of trace formaldehyde in blood or the urine fully with the method.
2.2 high-pressure liquid phase look fluorogram, clinical investigation and urine formaldehyde determination result
As can be seen from Figure 2, to go out the retention time at peak be quite stable (being about 8 minutes) to formaldehyde in the chromatogram, and it all is feasible that the method is used for qualitative and quantitative.The recovery of standard items (＞95%), in a few days deviation and poor in the daytime (＜5%) all reach requirement.
The result of Fig. 2 shows that the concentration of endogenous formaldehyde increases the weight of along with dementia degree and increases progressively, and the expression endogenous formaldehyde can be as the biomarker of diagnosis and judgement senile dementia degree.
2.3 different senile dementia patient clinical investigation situations and urine concentration of formaldehyde measurement result
Table 1 has been summed up the investigation of clinical epidemiology result of 141 patients of senile dementia in various degree of picked at random and the measurement result of their urine concentration of formaldehyde.
Table 2 has been summed up contrast crowd (comprising 81 individualities) investigation and their urine concentration of formaldehyde measurement result.Described contrast crowd comprises the healthy individual of not suffering from any disease and suffers from the individuality of one or more diseases except that AD, described disease except that AD comprises, but be not limited to hypertension, hyperglycaemia, diabetes, apoplexy, heart disease, thrombus, cerebral thrombus, hemiplegia and decrease of memory etc.
The different senile dementia patient of table 1. clinical investigation situation complete list and urine concentration of formaldehyde measurement result
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??1 | The man | ??82 | Vascular dementia, cerebral hemorrhage | ??0 | ??0.033333 |
| ??2 | The man | ??76 | Senile slow-witted, cerebral infarction, Parkinson, hypertension | ??0 | ??0.066667 |
| ??3 | The man | ??80 | Vascular dementia, cerebral infarction, hypertension | ??0 | ??0.1 |
| ??4 | The man | ??74 | Senile dementia, type ii diabetes | ??0 | ??0.133333 |
| ??5 | The woman | ??78 | Parkinson's, hypoxic-ischemic restoring consciouness disease | ??0 | ??0.166667 |
| ??6 | The man | ??83 | The Combination dementia, multiple brain more | ??0 | ??0.2 |
| ??7 | The woman | ??80 | Cerebral infarction, high fat of blood | ??20 | ??0.233333 |
| ??8 | The woman | ??77 | Senile dementia, cerebral infarction | ??0 | ??0.266667 |
| ??9 | The man | ??74 | The Combination dementia, Parkinson's | ??6 | ??0.3 |
| ??10 | The woman | ??76 | Senile dementia, cerebral infarction | ??0 | ??0.333333 |
| ??11 | The woman | ??83 | Senile dementia, high fat of blood | ??0 | ??0.366667 |
| ??12 | The man | ??78 | The Combination dementia, cerebral infarction, high fat of blood | ??20 | ??0.4 |
| ??13 | The woman | ??79 | High fat of blood, hypertension, gout | ??0 | ??0.433333 |
| ??14 | The woman | ??81 | The Combination dementia, cerebral infarction | ??0 | ??0.466667 |
| ??15 | The woman | ??64 | The Combination dementia, cerebral infarction, cerebral hemorrhage | ??0 | ??0.5 |
| ??16 | The man | ??79 | Senile dementia | ??0 | ??0.533333 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??17 | The man | ??71 | The Combination dementia, cerebral infarction | ??0 | ??0.6 |
| ??18 | The man | ??58 | Senile dementia, cerebral infarction, hypertension | ??17 | ??0.666667 |
| ??19 | The man | ??78 | Vascular dementia, cerebral infarction, high fat of blood | ??0 | ??0.7 |
| ??20 | The man | ??85 | Senile dementia, cerebral infarction, hypertension | ??0 | ??0.733333 |
| ??21 | The man | ??71 | Encephalatrophy | ??0 | ??0.766667 |
| ??22 | The woman | ??73 | Senile dementia, diabetes | ??0 | ??0.8 |
| ??23 | The woman | ??79 | Senile dementia, type ii diabetes, high fat of blood | ??17 | ??0.833333 |
| ??24 | The man | ??70 | Senile dementia, cerebral infarction | ??0 | ??0.866667 |
| ??25 | The man | ??74 | The Combination dementia, cerebral infarction, hypertension | ??0 | ??0.9 |
| ??26 | The man | ??78 | Senile dementia | ??14 | ??0.933333 |
| ??27 | The man | ??86 | Senile dementia, hypertension, high fat of blood | ??0 | ??0.966667 |
| ??28 | The woman | ??82 | The veteran form dementia is read again malnutritive | ??14 | ??1 |
| ??29 | The woman | ??78 | Senile dementia, cerebral infarction, hypertension | ??20 | ??1.033333 |
| ??30 | The woman | ??69 | The E Nieye dementia, cerebral infarction | ??0 | ??1.066667 |
| ??31 | The woman | ??76 | Vascular dementia, cerebral infarction, diabetes | ??20 | ??1.133333 |
| ??32 | The man | ??70 | The Combination dementia, cerebral hemorrhage sequelae | ??18 | ??1.166667 |
| ??33 | The man | ??79 | Encephalatrophy, senile dementia | ??9 | ??1.233333 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??34 | The woman | ??60 | Senile dementia, cerebral infarction, Meniere's disease | ??0 | ??1.333333 |
| ??35 | The man | ??64 | Encephalatrophy, senile dementia | ??0 | ??1.366667 |
| ??36 | The man | ??64 | Senile dementia, cerebral infarction | ??0 | ??1.4 |
| ??37 | The woman | ??82 | Vascular dementia, cerebral infarction | ??14 | ??1.433333 |
| ??38 | The man | ??88 | Vascular dementia, cerebral infarction, encephalatrophy | ??0 | ??1.533333 |
| ??39 | The woman | ??86 | Cerebral infarction, depression | ??12 | ??1.6 |
| ??40 | The woman | ??70 | Encephalatrophy, cerebral infarction, senile dementia | ??0 | ??1.666667 |
| ??41 | The man | ??81 | Vascular dementia, cerebral infarction | ??0 | ??1.7 |
| ??42 | The man | ??65 | Hypertension | ??6 | ??3.3 |
| ??43 | The man | ??80 | Cerebral infarction, vascular dementia | ??10 | ??3.4 |
| ??44 | The woman | ??76 | Do not have | ??13 | ??3.433333 |
| ??45 | The woman | ??74 | Dull-witted | ??8 | ??3.466667 |
| ??46 | The woman | ??77 | Multiple cerebral infarction, encephalatrophy | ??7 | ??3.566667 |
| ??47 | The woman | ??72 | Cerebral infarction | ??13 | ??3.633333 |
| ??48 | The man | ??71 | Encephalatrophy | ??0 | ??0.766667 |
| ??49 | The man | ??71 | Senile dementia, cerebral infarction | ??0 | ??1.466667 |
| ??50 | The man | ??57 | Hypertension, cerebral apoplexy, diabetes | ??13 | ??1.9 |
| ??51 | The woman | ??75 | Do not have | ??22 | ??1.933333 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??52 | The woman | ??90 | Do not have | ??12 | ??1.966667 |
| ??53 | The woman | ??82 | Diabetes, coronary heart disease | ??20 | ??2 |
| ??54 | The woman | ??84 | Cerebral infarction | ??6 | ??2.033333 |
| ??55 | The woman | ??73 | Cerebral thrombus, heart stalk, hypertension | ??4 | ??2.066667 |
| ??56 | The man | ??67 | Cerebral infarction | ??18 | ??2.1 |
| ??57 | The woman | ??76 | Do not have | ??4 | ??2.133333 |
| ??58 | The man | ??71 | Encephalatrophy | ??15 | ??2.166667 |
| ??59 | The man | ??54 | Congenital cerebral agenesis | ??6 | ??2.2 |
| ??60 | The woman | ??73 | Deaf | ??0 | ??2.233333 |
| ??61 | The man | ??71 | Diabetes | ??19 | ??2.266667 |
| ??62 | The man | ??85 | Do not have | ??0 | ??2.3 |
| ??63 | The woman | ??76 | Hypertension, cerebral blood supply insufficiency, cerebral infarction | ??13 | ??2.333333 |
| ??64 | The man | ??88 | Cerebral hemorrhage sequelae | ??6 | ??2.366667 |
| ??65 | The woman | ??81 | Hypertension, coronary heart disease, cerebral apoplexy | ??13 | ??2.4 |
| ??66 | The woman | ??59 | Depressed | ??11 | ??2.433333 |
| ??67 | The man | ??94 | Senile dementia | ??10 | ??2.466667 |
| ??68 | The woman | ??71 | Cerebral ischemia | ??14 | ??2.5 |
| ??69 | The woman | ??38 | Moronism, intelligence is slow | ??8 | ??2.533333 |
| ??70 | The woman | ??88 | Do not have | ??0 | ??2.566667 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??71 | The woman | ??91 | Do not have | ??6 | ??2.6 |
| ??72 | The man | ??91 | Do not have | ??15 | ??2.633333 |
| ??73 | The man | ??54 | Coronary heart disease, cerebral apoplexy | ??20 | ??2.666667 |
| ??74 | The man | ??61 | Cerebral apoplexy | ??25 | ??2.7 |
| ??75 | The woman | ??85 | Do not have | ??14 | ??2.766667 |
| ??76 | The woman | ??106 | Hypertension, coronary heart disease | ??11 | ??2.8 |
| ??77 | The woman | ??68 | Do not have | ??2 | ??2.833333 |
| ??78 | The woman | ??55 | Hydrocephalus | ??11 | ??2.866667 |
| ??79 | The man | ??69 | Coronary heart disease | ??9 | ??2.9 |
| ??80 | The woman | ??91 | Do not have | ??6 | ??2.933333 |
| ??81 | The woman | ??80 | Hypertension, encephalatrophy | ??4 | ??2.966667 |
| ??82 | The woman | ??88 | Do not have | ??13 | ??3 |
| ??83 | The woman | ??66 | Hypertension, cerebral apoplexy, diabetes | ??13 | ??3.033333 |
| ??84 | The man | ??68 | Cerebral apoplexy | ??14 | ??3.066667 |
| ??85 | The woman | ??80 | The blind person | ??15 | ??3.1 |
| ??86 | The man | ??95 | Do not have | ??0 | ??3.133333 |
| ??87 | The man | ??82 | Do not have | ??12 | ??3.166667 |
| ??88 | The man | ??75 | Hypertension | ??24 | ??3.2 |
| ??89 | The man | ??82 | Do not have | ??23 | ??3.233333 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??90 | The woman | ??81 | Do not have | ??0 | ??3.266667 |
| ??91 | The man | ??65 | Hypertension | ??6 | ??3.3 |
| ??92 | The woman | ??83 | Cerebral infarction, Combination dementia, hypertension | ??20 | ??3.333333 |
| ??93 | The woman | ??79 | Senile dementia, type ii diabetes, high fat of blood | ??17 | ??0.833333 |
| ??94 | The man | ??78 | Hypertension, cerebral infarction | ??16 | ??2.6 |
| ??95 | The man | ??58 | Hypertension, cerebral apoplexy | ??22 | ??1.933333 |
| ??96 | The woman | ??76 | High fat of blood, cerebral infarction | ??19 | ??2.533333 |
| ??97 | The man | ??80 | Cerebral hemorrhage | ??20 | ??2.666667 |
| ??98 | The man | ??89 | Cataract | ??23 | ??2.966667 |
| ??99 | The man | ??55 | Hypertension, cerebral hemorrhage | ??19 | ??1.833333 |
| ??100 | The woman | ??64 | Hypertension, coronary heart disease | ??19 | ??2.133333 |
| ??101 | The man | ??83 | Coronary heart disease | ??26 | ??2.766667 |
| ??102 | The woman | ??84 | Hypertension | ??20 | ??2.8 |
| ??103 | The woman | ??80 | Hypertension, cerebral infarction | ??20 | ??2.666667 |
| ??104 | The man | ??55 | Hypertension, cerebral apoplexy, diabetes | ??25 | ??1.833333 |
| ??105 | The woman | ??77 | Hypertension 40 years | ??23 | ??2.566667 |
| ??106 | The man | ??55 | Hypertension, cerebral apoplexy, diabetes | ??25 | ??1.833333 |
| ??107 | The man | ??63 | There is not record | ??23 | ??2.1 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??108 | The woman | ??78 | Hypertension, coronary heart disease | ??21 | ??2.6 |
| ??109 | The man | ??73 | Hypertension, coronary heart disease | ??20 | ??2.433333 |
| ??110 | The man | ??71 | Hypertension 10 years | ??25 | ??2.366667 |
| ??111 | The man | ??72 | Hypertension, cerebral thrombus | ??23 | ??2.4 |
| ??112 | The woman | ??62 | Polio | ??25 | ??2.066667 |
| ??113 | The man | ??86 | Coronary heart disease, diabetes | ??26 | ??2.866667 |
| ??114 | The man | ??74 | Hypertension | ??19 | ??2.466667 |
| ??115 | The woman | ??55 | Hypertension, cerebral hemorrhage | ??26 | ??1.833333 |
| ??116 | The man | ??88 | Hypertension | ??22 | ??2.933333 |
| ??117 | The man | ??66 | Hypertension, cerebral hemorrhage | ??20 | ??2.2 |
| ??118 | The woman | ??77 | Parkinson | ??21 | ??2.566667 |
| ??119 | The man | ??60 | Cerebral apoplexy | ??25 | ??2 |
| ??120 | The woman | ??44 | Phrenoblabia | ??29 | ??1.466667 |
| ??121 | The man | ??59 | Hypertension | ??19 | ??1.966667 |
| ??122 | The woman | ??49 | Polio | ??25 | ??1.633333 |
| ??123 | The woman | ??55 | Cerebral hemorrhage | ??18 | ??1.833333 |
| ??124 | The man | ??86 | Hypertension | ??21 | ??2.866667 |
| ??125 | The woman | ??44 | Do not have | ??23 | ??1.466667 |
| Numbering | Sex | Age | Medical history | ??MMSE | Concentration of formaldehyde in the urine (mM) |
| ??126 | The man | ??86 | Not quite clear | ??23 | ??2.866667 |
| ??127 | The man | ??66 | Polio | ??24 | ??2.2 |
| ??128 | The man | ??72 | Hypertension, cerebral apoplexy | ??23 | ??2.4 |
| ??129 | The woman | ??82 | Pneumonia | ??21 | ??2.733333 |
| ??130 | The man | ??56 | Hypertension, cerebral apoplexy, coronary heart disease | ??26 | ??1.866667 |
| ??131 | The woman | ??66 | Hypertension | ??22 | ??2.2 |
| ??132 | The man | ??74 | Schizophrenia 30 years | ??23 | ??2.466667 |
| ??133 | The man | ??54 | Cerebral apoplexy, coronary heart disease | ??19 | ??1.8 |
| ??134 | The man | ??53 | Cerebral apoplexy | ??22 | ??1.766667 |
| ??135 | The man | ??72 | Cervical spondylopathy | ??25 | ??2.4 |
| ??136 | The man | ??61 | Diabetes | ??21 | ??2.033333 |
| ??137 | The man | ??64 | Hypertension | ??18 | ??2.133333 |
| ??138 | The woman | ??83 | Not quite clear | ??22 | ??2.766667 |
| ??139 | The woman | ??82 | Diabetes, coronary heart disease | ??20 | ??2.733333 |
| ??140 | The man | ??53 | Hypertension | ??23 | ??1.766667 |
| ??141 | The man | ??75 | Hypertension, hypertrophy of the prostate 14 years | ??24 | ??2.5 |
Table 2 contrast crowd's * investigation complete list and urine concentration of formaldehyde measurement result
| Numbering | Sex | Age | Medical history | ??MMSE | Urine concentration of formaldehyde (mM) |
| ??1 | The woman | ??69 | Do not have | ??27 | ??0.064596 |
| ??2 | The woman | ??63 | Do not have | ??28 | ??0.076415 |
| ??3 | The man | ??68 | Do not have | ??27 | ??0.198749 |
| ??4 | The man | ??66 | Hypertension | ??26 | ??0.02448 |
| ??5 | The woman | ??60 | High blood pressure | ??27 | ??0.090435 |
| ??6 | The man | ??73 | Do not have | ??23 | ??0.119551 |
| ??7 | The woman | ??63 | Do not have | ??27 | ??0.30344 |
| ??8 | The man | ??60 | Diabetes | ??26 | ??0.17951 |
| ??9 | The man | ??64 | Do not have | ??26 | ??0.024954 |
| ??10 | The man | ??71 | Hypertension | ??25 | ??0.140127 |
| ??11 | The woman | ??70 | Do not have | ??27 | ??0.062137 |
| ??12 | The woman | ??65 | Do not have | ??28 | ??0.08776 |
| ??13 | The man | ??71 | Do not have | ??28 | ??0.15915 |
| ??14 | The woman | ??65 | Do not have | ??27 | ??0.019131 |
| ??15 | The woman | ??64 | Hypertension | ??29 | ??0.083576 |
| ??16 | The woman | ??69 | Hypertension | ??28 | ??-0.00524 |
| ??17 | The woman | ??60 | High blood pressure | ??30 | ??0.10165 |
| ??18 | The woman | ??71 | Hyperglycaemia, hypertension | ??24 | ??0.006966 |
| Numbering | Sex | Age | Medical history | ??MMSE | Urine concentration of formaldehyde (mM) |
| ??19 | The man | ??70 | Do not have | ??28 | ??0.697876 |
| ??20 | The man | ??73 | Hypertension | ??22 | ??-0.01016 |
| ??21 | The woman | ??62 | Cerebral blood supply insufficiency | ??28 | ??0.018441 |
| ??22 | The woman | ??65 | Hypertension, diabetes | ??27 | ??0.019433 |
| ??23 | The woman | ??70 | Diabetes | ??27 | ??0.048118 |
| ??24 | The man | ??67 | Do not have | ??28 | ??0.050879 |
| ??25 | The woman | ??75 | Do not have | ??28 | ??0.131371 |
| ??26 | The man | ??72 | Do not have | ??27 | ??0.263626 |
| ??27 | The woman | ??67 | Hypertension, minor stroke | ??27 | ??0.044538 |
| ??28 | The woman | ??66 | Heart disease | ??21 | ??0.068306 |
| ??29 | The woman | ??73 | Diabetes | ??25 | ??0.003904 |
| ??30 | The woman | ??61 | Do not have | ??28 | ??0.177397 |
| ??31 | The man | ??71 | Hypertension | ??27 | ??0.035048 |
| ??32 | The man | ??73 | Do not have | ??28 | ??0.28308 |
| ??33 | The woman | ??72 | Diabetes, hypertension | ??14 | ??0.197628 |
| ??34 | The woman | ??75 | Do not have | ??28 | ??0.043891 |
| ??35 | The man | ??63 | Hypertension | ??13 | ??0.1868 |
| ??36 | The woman | ??62 | Do not have | ??27 | ??0.08487 |
| Numbering | Sex | Age | Medical history | ??MMSE | Urine concentration of formaldehyde (mM) |
| ??37 | The woman | ??62 | Hypertension | ??28 | ??0.02944 |
| ??38 | The man | ??68 | High blood pressure, hemiplegia | ??15 | ??0.082239 |
| ??39 | The man | ??66 | Do not have | ??30 | ??0.032584 |
| ??40 | The woman | ??66 | Do not have | ??30 | ??0.166597 |
| ??41 | The woman | ??69 | Do not have | ??30 | ??0.119397 |
| ??42 | The woman | ??78 | Decrease of memory | ??24 | ??0.073779 |
| ??43 | The man | ??66 | Do not have | ??27 | ??0.004422 |
| ??44 | The woman | ??60 | Diabetes | ??27 | ??-0.01052 |
| ??45 | The man | ??64 | Diabetes, hypertension | ??25 | ??0.139413 |
| ??46 | The man | ??85 | Do not have | ??27 | ??0.037704 |
| ??47 | The woman | ??56 | Do not have | ??25 | ??0.06973 |
| ??48 | The woman | ??65 | Do not have | ??27 | ??0.371875 |
| ??49 | The man | ??69 | Hypertension | ??30 | ??0.130429 |
| ??50 | The woman | ??77 | Do not have | ??27 | ??0.16292 |
| ??51 | The man | ??66 | Hypertension | ??30 | ??0.109761 |
| ??52 | The woman | ??84 | Do not have | ??30 | ??0.086813 |
| ??53 | The woman | ??66 | Do not have | ??27 | ??0.148815 |
| ??54 | The woman | ??60 | Do not have | ??28 | ??0.088349 |
| Numbering | Sex | Age | Medical history | ??MMSE | Urine concentration of formaldehyde (mM) |
| ??55 | The woman | ??71 | Diabetes | ??24 | ??0.053112 |
| ??56 | The woman | ??65 | Hypertension | ??29 | ??0.050226 |
| ??57 | The man | ??65 | Do not have | ??28 | ??0.127543 |
| ??58 | The woman | ??64 | Do not have | ??21 | ??0.045571 |
| ??59 | The woman | ??66 | Hypertension | ??29 | ??0.03267 |
| ??60 | The woman | ??72 | Do not have | ??26 | ??2.45E-05 |
| ??61 | The woman | ??70 | Do not have | ??23 | ??0.159827 |
| ??62 | The woman | ??72 | Diabetes, hypertension | ??25 | ??0.076524 |
| ??63 | The woman | ??69 | Hypertension | ??28 | ??0.062918 |
| ??64 | The woman | ??68 | Hypertension | ??28 | ??0.048871 |
| ??65 | The woman | ??65 | High blood pressure | ??29 | ??0.012114 |
| ??66 | The woman | ??72 | Hypertension | ??29 | ??-0.0152 |
| ??67 | The man | ??68 | Hypertension | ??27 | ??0.201772 |
| ??68 | The man | ??64 | Diabetes, three height | ??29 | ??0.016714 |
| ??69 | The woman | ??70 | Hypertension, diabetes, thrombus | ??30 | ??-0.03928 |
| ??70 | The woman | ??66 | Do not have | ??28 | ??-0.01691 |
| ??71 | The woman | ??69 | Do not have | ??26 | ??0.196094 |
| ??72 | The woman | ??72 | Hypertension | ??23 | ??0.092773 |
| Numbering | Sex | Age | Medical history | ??MMSE | Urine concentration of formaldehyde (mM) |
| ??73 | The woman | ??63 | Hypertension | ??23 | ??0.337445 |
| ??74 | The man | ??66 | Do not have | ??30 | ??0.105597 |
| ??75 | The woman | ??62 | Do not have | ??30 | ??0.048773 |
| ??76 | The man | ??66 | Diabetes, cerebral thrombus | ??27 | ??0.236375 |
| ??77 | The woman | ??56 | Hypertension, hyperglycaemia | ??27 | ??0.093214 |
| ??78 | The woman | ??80 | Do not have | ??27 | ??0.12214 |
| ??79 | The man | ??58 | Hypertension | ??23 | ??-0.0128 |
| ??80 | The woman | ??68 | Diabetes | ??28 | ??0.088662 |
| ??81 | The woman | ??68 | Diabetes, hypertension | ??21 | ??0.054646 |
* contrast the crowd: comprise the health population of not suffering from any disease and suffer from the non-AD patient colony of one or more diseases except that AD.
2.4 discuss
The inventor's result of study shows, endogenous formaldehyde concentration and healthy people have significantly differently in patient's AD urine, and so to clinical diagnosis, this is the reference index that Practical significance and application prospect are arranged very much beyond doubt.In the inventor's research, senile dementia pathology section shows that pathology is that mat-ups and senile plaque expelling etc. appear in some albumen in brain tissue, the experiment in vitro result has proved that formaldehyde can cause that external a kind of Protein tau sex change that exists in the nervous system forms the gathering of Protein tau and the entanglement of formation spiral fiber in the brain tissue slice among the similar senile dementia patient, so, the inventor infers that courageously the concentration of endogenous formaldehyde may exceed the normal person in the dementia patients nervous system, and has performance in the metabolic process afterwards.Result of study of the present invention has confirmed this supposition, and the inventor has finished the present invention on this basis.Unusual about concentration of formaldehyde in the senile dementia patient urine, the inventor infers that this may be because formaldehyde synzyme and digestive enzyme unbalance causing in senile dementia patient body.
The AD disease is divided into a lot of types by pathogenic factor: vascular dementia, simple senile dementia, dull-witted and other (Wei Hongmei, 2008) (the seeing Fig. 3 b) of Combination all has extremely significant and significant difference compared with the control by the urine levels of formaldehyde of seeing various types of AD patients among Fig. 3.In order to get rid of the interference of other diseases, to also there be the non-AD patient of corresponding disease relatively to find (seeing Fig. 3 c) with the while with the AD patient of hypertension or diabetes, AD patient's urine levels of formaldehyde relatively still has significant difference with the non-AD patient who suffers from these two kinds of corresponding diseases respectively, and from Fig. 3 d, find out, the non-AD patient who suffers from hypertension or diabetes urinates concentration of formaldehyde and compares not significant difference with normal healthy controls, increasing at least with common hypertension and the diabetes of the elderly of explanation urine concentration of formaldehyde do not have positive correlation thus, shows that promptly the rising of urinating concentration of formaldehyde will be specific label in AD patient's urine.This is for providing the simple and easy to do reference index of the patient not being had any injury on clinical early diagnosis senile dementia, and for providing an evaluation method on the dull-witted curative effect of medication of research treatment, this result of study will be a brand-new discovery in this research field.
Should be appreciated that, although with reference to its exemplary embodiment, the present invention is shown particularly and describe, but will be understood by those skilled in the art that, can carry out the variation of various forms and details therein, and not deviate from by the defined the spirit and scope of the present invention of accompanying Claim.
List of references:
1Femke?H.Bouwmana(CSF?biomarker?levels?in?early?and?late?onsetAlzheimer’sdisease.Neurobiology?ofAging?7.2008).
2Folstein?MF，Folstein?SE，McHugh?PR(″Mini-mental?state″.A?practicalmethod?for?grading?the?cognitive?state?of?patients?for?the?clinician.JPsychiatr?Res?12：189-198.1975).
3Francis?PT，Palmer?AM，Snape?M，Wilcock?GK(The?cholinergichypothesis?of?Alzheimer′s?disease：a?review?of?progress.Journal?ofNeurology，Neurosurgery&Psychiatry?66：137-147.1999).
4Luo?W，Li?H，Zhang?Y，Ang?CYW(Determination?of?formaldehyde?inblood?plasma?by?high-performance?liquid?chromatography?withfluorescence?detection.Journal?of?Chromatography?B：BiomedicalSciences?and?Applications?753：253-257.2001).
5McKhann?G，Drachman?D，Folstein?M，Katzman?R，Price?D，Stadlan?EM(Clinical?diagnosis?of?Alzheimer′s?disease：report?of?theNINCDS-ADRDA?work?group.Neurology?34：939-944.1984).
6Simonsen?AH，McGuire?J，Podust?VN，Davies?H，Minthon?L，Skoog?I，Andreasen?N，Wallin?A，Waldemar?G，Blennow?K(Identification?of?anovel?panel?of?cerebrospinal?fluid?biomarkers?for?Alzheimer′s?disease.Neurobiol?Aging?29：961-968.2008).
7 execute the Anguo expense autumn gorgeous (the senile dementia Progress in Medication. pharmacy progress 24:338-341.2000).
8 Ni Qi are bright, the continental rise ancestor (senile dementia (alzheimer disease) and drug therapy thereof. China health care 15:59-60.2007).
9 Wei Hong plums (the pathogenetic research of senile dementia. practical preventive medicine 15:621-623.2008).
Claims (4)
1. the application of formaldehyde in the diagnosticum of preparation senile dementia, wherein with the biomarker of endogenous formaldehyde as senile dementia, by detecting the concentration of formaldehyde in experimenter's urine sample, concentration of formaldehyde in itself and the health volunteer's urine is compared, determine whether described experimenter suffers from senile dementia, and tentatively judge the degree of described experimenter's senile dementia.
2. according to the application of claim 1, wherein use the concentration of formaldehyde in the described experimenter's urine sample of high performance liquid chromatogram fluorescence chromatogram (HPLC-fluorescence) detection.
3. according to the application of claim 1 or 2, it is also by the vertical urine concentration of formaldehyde in monitoring described experimenter different time stage in life process, is used for judging that described experimenter develops the danger of senile dementia.
4. according to the application of claim 1 or 2, it also by monitoring described experimenter different time stage urine concentration of formaldehyde before taking the medicine that suppresses senile dementia and afterwards, is used for estimating the drug effect of described medicine.
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| CN115436494A (en) * | 2021-06-03 | 2022-12-06 | 中国科学院生物物理研究所 | Application of ratio of formaldehyde content to ribose content in urine in early screening or diagnosis of cognitive dysfunction |
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| CN115436494A (en) * | 2021-06-03 | 2022-12-06 | 中国科学院生物物理研究所 | Application of ratio of formaldehyde content to ribose content in urine in early screening or diagnosis of cognitive dysfunction |
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