The Association Of Tyrosine Hydroxylase Gene Polymorphology With Psychosis Also In Medical Jurisprudence And Human Genetics Studies

PrefaceSince we have finished the human gene project(HGP),we have known about all the gene order.We are studying the functional gene,as well as finding the differences among the specific gene order.Single nucleotide polymorphisms(SNPs) as the most comon differences play an important part in the research of disease,medicine,and evolution.Tyrosine hydroxylase(TH) as an important rate limiting enzyme in the human body.It had a gene polymorphism caused by the differences among mRNA coding. More attention has been paid to the TH polymorphism for forensic research in these years.It has been proved that the mental disorders had some heritage by many research. And the heritage had some connections with the TH polymorphism.Some other researchers thought that the TH polymorphism was different among the different people,so there may be an important signification in telling different people.It may be useful to individual identification and paternity test.For the limiting of the technology, we have no systemic research results.With the development of technology,TH polymorphism research had become more and more popular.ObjectiveThe aim of this study is to evaluate the relationship between the TH gene and mental disorder by comparing the two TH gene SNP points(extron 3 and intron 9) of patients with mental disorder and those of human without mental disorder.And the significance of TH gene polymorphology in human genetic and in forensic medicine is also discussed. Materials and Methods1,specimen1.1 Normal contract group:127 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.1.2 173 tibetans' blood from Nepal offered by JAPAN university.1.3 130 normal japaneses' blood offered by JAPAN university.1.4 133 cases with chizophrenia or Schizophrenia,and 7 Schizophrenia family history offered by Kaiyuan madhouse.1.5 73 cases with depression offered by china medical university.2,methods2.1 Template DNA is made by hydroxybenzene/chloroform method from venous anti -thrombin DNA.2.2 Use PCR to amplify the target DNA fragment:Inducer sequence of G334A:F:5'-CAGCGCGGCTCCAGCCCGT-3',R:5'-AGT -CCCGGGCGGACAGCAGGC-3';Inducer sequence of C5162G:F:5'-CAGTGCA -CCCAGTATATCCG-3',R:5'-CCGTCGCACCCCCCACCCTC-3'。2.3 DNA sequencing:we use BigDye Terminator v.3.1 Cycle Sequencing Kit and ABI 3130 automatic sequencing machine.primer are TH-exon10-L: 5'-CAGCATGGG- CACGTGCCCCA-3',TH-exon9-U: 5'-CGCGTGGCCCCTTGCAGAGC-3'.2.4 Statistic analysis:Record all individual classification result;calculate each allele and gene type frequency;perform Hardy-Weinberg balance test..Using spss software.Results1,Naming of gene mutation of human TH.We have make the order of TH gene,which named G334A(Thr314Met) and rs12419447(C/G) or C5162G.2,G334A and C5162G distribution data.We found CC,CT,and TT in all three groups.Most common allele frequency is C=Japanese＞north chinese＞tibetan in nepal(0.269＞0.214＞0.205);T=tibetan in nepal＞north chinese＞Japanese(0.795＞0.786＞0.731).In C5162G there are CC and CG,but no GG,which proves that the GG is very rare.Most common allele frequency is C=Japanese＞north chinese＞tibetan in nepal(0.977＞0.961＞0.922);G=tibetan in nepal＞north chinese＞Japanese(0.078＞0.039＞0.023).3,G334A and C5162G signifitance in forensic medicine.Probability of discrimination power(DP):Japanese＞north chinese＞tibetan in nepal(0.5593＞0.5012＞0.4916);Excluding probability of paternity(EPP):Japanese＞north chinese＞tibetan in nepal(0.1580＞0.1399＞0.1364);Heterozygosity,H:north chinese＞Japanese＞tibetan in nepal(0.3629＞0.3538＞0.3509).In C5162G,DP:tibetan in nepal＞north chinese＞Japanese(0.2633＞0.1455＞0.0878);EPP:tibetan in nepal＞north chinese＞Japanese(0.0667＞0.0361＞0.0220);H:tibetan in nepal＞north chinese＞Japanese(0.1561＞0.0787＞0.0462).4,G334A distribution In north china:C=0.133,T=0.867 in Schizoph- renia,and C=0.116,T=0.884 in depression.5,C5162G distribution:C=0.962,G=0.038 in Schizophrenia,and C=0.959,G=0.041 in depression.6,SNPs of TH in the Schizophrenia family.For G334A spot,6 filial generations and 5 parents are all T/T gene type,and in the other two cases,father is T/T,mother is C/T,both filial generations are C/T;for C5162G spot,7 filial generations and 6 parents are all C/C gene type,and in the other case,father is C/C,mother is C/G,filial generation is C/G.7,The relationship between familial disease and the SNPs of TH.Any of the fathers is the families had no Schizophrenia history,and all the mothers had Schizophrenia history.In G334A and C5162G spots of the TH only one is T/T-C/G,the other 6 cases are T/T-C/C.The five mothers with Schizophrenia is T/T-C/C,and the other two cases is C/T-C/C and C/T-C/G.Conclusion1,Depending on the data from the internet,we discript the gene order of TH of human being,and a name method.2,First report G334A and C5162G.Describe the gene type distribution in north chinese,Japanese,and tibetan in nepal.3,Depending on the TaqMan real time PCR technology,we determinate G334A and C5162G gene distribution in Schizophrenia and depression,which will be very useful to genetic research.4,Compare G334A and C5162G in the mental disease.We find that prohand usually is the same gene type with the parents with the mental disease.5,Compare G334A and C5162G between normal people and the patients with mental disease.Finding that in G334A there are significant difference between normal people and patients with mental disease,and there are no significant difference in C5162G.