The Association Of DA2,DA3,DA4 Receptor Gene Polymorphology With Schizophrenia Also In Medical Jurisprudence And Human Genetics Studies

IntroductionSchizophrenia is a familiar psychotic disease with unclear etiology and is the most representative and the most common syndrome in psychotic disease. There is 80% -90% Schizophrenia patient in psychotic hospital and among cases with psychotic jurisprudence identification it occupied the first position. The etiology of this disease is not very clear, and has some relation with inheritance. Environmental factor may induce Schizophrenia. Research about central nervous system biochemical metabolism suggests that in the brain of Schizophrenia patient Dopamine is hyper - functioned at neural synapse. The relationship between Schizophrenia and dopamine receptor has been always paid attention to. It is generally accepted that dopamine hyper - function may lead to Schizophrenia. Sololoff,Van et al divided dopamine receptor into five kinds including DRD1, DRD2,DRD3,DRD4 and DRD5. Present studies mainly focus on DRD2, DRD3and DRD4.ObjectivePresent studies in always limited in single gene group comparison with Schizophrenia regardless of comparison among gene group. First we separately use Dopamine D2,D3 and D4 receptor gene group to analyze a same sample, then gather the pertinence data of each gene group poly - morphology with Schizophrenia , finally we adopt statistic software to analyze them in order to find three facilitated gene group type. Besides, we also acquire the application data of Dopamine D2,D3 and D4 receptor gene group poly -morphology in medical jurisprudence and human genetics.Method1. Process of object1.1 Normal contract group: 105 cases all come from blood donors in Shenyang blood center with no Schizophrenia or Schizophrenia family history or suicide disease. They are all Han nationality, 53 male, 52 female, and aging from 21 -58, 25y for average.1.2 Patient group: 101 cases with identified Schizophrenia. The diagnosis of Schizophrenia must according to both the fourth edition of American mental disturbance diagnosis and statistic manual ( DSM - Ⅳ) and the third edition of Chinese psychotic disease classification and diagnosis standard ( CCMD - 3). There are 51 male, 50 female in this group. They are aging from 22—55, 26y for average, and are Han nationality. These cases all come from Shenyang psychotic medical center. The age of the two groups are analyzed by t - test, and there is no significant difference between them (P <0.05).2. Experimental method2. 1 Template DNA is made by hydroxybenzene/ chloroform method from venous anti -thrombin DNA.2. 2 Use PCR to amplify the target DNA fragment, and use restriction endo - nuclease to act at the SNP site of Dopamine D2 and D3. DRD2 production fragment is 304bp, inducer sequence F 5'ACT GGC GAG CAG ACG GTG AGG ACC -3; R 5'- AAG TCT ACT CAC CTC CAG GTA -3' Then use the method of polypropylene amine gelatin electrophoresis for separating combined with Argentine - stained revealing method.2. 3 Adopt VNTR technique to process Dopamine D4, amplified inducer sequence is F 5'- GCG ACT ACG TGG TCT ACT CG -3', R 5'- AGG ACC CTC ATG GCC TTG -31 After amplification of PCR, use the method of polypropylene amine gelatin electrophoresis for separating combined with Argentine -stained revealing method.2.4 Statistic analysis: Record all individual classification result; calculate each allele and gene type frequency; perform Hardy - Weinberg balance test.Data processing is using statistic software SPSS10.0 to make factorial analysis. Allele frequency is analyzed by direct calculation. P value < 0. 05 represents significant difference. Calculate Power of Discrimination ( DP) according to Fisher method; calculate Power of Exclusion ( PE). Allele frequency is analyzed by direct calculation.Result1. Association of Dopamine D2 gene poly - morphology with Schizophrenia, Analyze DRD2 gene poly - morphology of both patient group and contract group; there is no significant difference in comparing gene type between two groups (X2= 1.949, P =0. 377) ; there is also no significant difference in comparing allele gene between two groups (X2 = 1. 67, P = 0. 34 ).2. Result analysis of Dopamine D3 receptor gene amplification, Analyze DRD3 gene poly - morphology of both patient group and contract group; there is significant difference in comparing gene type between two groups (X2 = 15.96, P <0.05) ; there is significant difference in the distribution of Ser/Ser and Gly/ Gly between two groups ( P = 0. 040, OR = 5. 8) , and suggests that sex is a jumble factor.3. Result analysis of Dopamine D3 receptor gene amplification, Analyze DRD4 gene poly - morphology of both patient group and contract group; there is an absent association between Dopamine D4 receptor gene VNTR poly - morphology and Schizophrenia (P >0.05).4. Dopamine D2N Dopamine D3 and Dopamine D4 receptor combined gene type Ins/Ins + Ser/Ser + 4/4 N Ins/Del + Ser/Gly compound has strong relationship with schizophrenia, Ins/Ins + Ser/Gly ^ Ins/Ins + Gly /Gly +4/4 that suggest the three gene group have no susceptibility.5. This study acquires medical jurisprudence application data of Dopamine DA2^DA3^DA4 receptor gene type^gene frequency^ Power of Discrimination and Power of Exclusion in Chinese Han nationality people.6. This study utilizes the acquired data of Dopamine DA2>DA3NDA4 receptor gene genetic poly - morphology to compare with Negro >> white people a-broad, and makes into genetic data. Thus acquire human genetics data which can compare Chinese Han nationality with other nationality.Conclusion1. This study finds that there is higher Power of Discrimination and Power of Exclusion of Dopamine D3 receptor gene frequency distribution in North China' Han nationality. Dopamine D3 receptor gene has no linkage relation with genetic marker applied in the traditional medical jurisprudence, instead it is an independent genetic system, and should belong to the valuable genetic marker in medical jurisprudence, which is especially important for future study of medical jurisprudence DNA chip.2. This study verifies by experimental data that single gene group Dopamine D2 receptor has no obvious association with schizophrenia; single gene group Dopamine D3 receptor has obvious association with schizophrenia; single gene group Dopamine D4 receptor has no obvious association with schizophrenia.3. It is the first time inland that experiment is performed to the same object for analyzing Dopamine D2 N D3 and D4 receptor gene poly — morphology which avoid error caused by experimental object and different layer. Through experimental data study acquire that Dopamine D2N Dopamine D3 and Dopamine D4 receptor combined gene type Ins/Ins + Ser/Ser + 4/4 N Ins/Del + Ser/Gly compound has strong relationship with schizophrenia, Ins/Ins + Ser/Gly ^ Ins/Ins + Gly /Gly +4/4 that suggest the three gene group have no susceptibility.4. Through experimental data analysis acquire the human genetic data for comparison between Chinese Han nationality and other nationality in Dopamine D2^D3 and D4 receptor gene group.Innovative spot in this study1. By data analysis of Dopamine D2^D3 and D4 receptor gene poly - morphology , it is reported the first time inland that Ser9Fly site has better frequency distribution in Chinese Han nationality. Dopamine D3 receptor gene has no link-age relation with genetic marker applied in the traditional medical jurisprudence, instead it is an independent genetic system, and should belong to the valuable genetic marker in medical jurisprudence, which is especially important for future study of medical jurisprudence DNA chip.2. It is the first time inland that combined gene group including Dopamine D2N Dopamine D3 and Dopamine D4 receptor is brought out to screen a same sample at same time, and acquire a now concept of schizophrenia facilitated gene group type. At present, no one use this method to study schizophrenia and avoid the result of incompatible conclusion from front to back which is easily induced by single gene group study.3. It is the first time to perform comparison of Dopamine D2ND3 and D4 receptor genetic data between Chinese Han nationality and other nationality, and acquire human genetics data.