Research Of Genetic Susceptibility To Primary Biliary Cholangitis And Anti-mitochondrial Antibody Type M2 Screening Results In Yangzhou

Objective:1.To explore the correlation between single nucleotide polymorphism genotyping and biochemical indexes in primary biliary cholangitis.2.To evaluate the value of anti-mitochondrial antibody type M2 for early screening of PBC,we investigate the positive rate of AMA-M2 in healthy people and analysis of test results of positive subjects.Methods:1.we finally select 25 SNPs combined with the results of genome-wide association study of PBC patients and the single nucleotide polymorphism which associated with the pathogenesis of PBC in Eastern and Western populations in the literature.To explore the correlation between different genotypes and serum biochemical indexes including ALT,AST,ALP,GGT,TBIL,DBIL,ALB and TP,we analyzed the polymorphism of related loci in 53 patients with PBC and 59 healthy controls using time-of-flight mass spectrometry.2.A triple hybrid clone was designed,which coexpresses the three immunodominant lipoyl domains of PDC-E2,OGDC-E2 and BCOADC-E2 from human sources.We established a specific immunological diagnostic method for PBC in our laboratory.Since November 1,2017 to August 31,2019,we have collected 11 024 cases of physical examination in Subei People's Hospital.The general data,clinical manifestations and serum biochemical indexes of AMA-M2 positive subjects were collected and statistically analyzed.Results:1.We analyzed the gene polymorphism in 53 Patients with PBC and 59 healthy controls,found that there were significant differences in genotype distribution and allele frequencies of STAT4 rs10168266 between PBC group and control group(P=0.008,0.017).Significant differences was revealed in genotype distribution and allele frequencies of IL21 rs17005934 between the two groups(P=0.018,0.004).There were significant differences in genotype distribution and allele frequencies of IL21 rs925550 between the two groups(P=0.004,P=0.001).There were significant differences in genotype distribution and allele frequencies of TNFSF15 rs4979462 between the two groups(P=0.001,0.000).There were significant differences in genotype distribution and allele frequencies of TNFSF8 rs2295800 between the two groups(P=0.027,0.011).There was no statistical difference in genotype distribution of IKZF3 rs9635726 between the two groups(P=0.179),but there was significant difference in allele frequency between the two groups(P=0.036).2.Using the ANOVA method,we analyzed the relationship between the SNP and clinical indicators of PBC.We found that the mean of ALT exist significant differences in IL21 rs17005934 and rs925550 loci different genotypes(P=0.039,0.045)and the mean of DBIL exist significant differences in STAT4 rs10168266 loci different genotypes.3.83 of 25713 healthy physical examination people were detected AMA-M2 antibody,and the positive rate were 0.334%.The positive rate of the females was higher than males(P<0.05).The average age of positive subjects was 53.12±17.10 years.The positive rate of healthy physical examination people was 0.297%?0.190%?0.316%?0.639% and 0.334% in <30,30?45,46?60 and <60 years groups,respectively.We found with the increase of age,the proportion of AMA-M2 positive rate tended to increase.The group who older than 60 had the highest positive rate,and the difference was statistically significant(P <0.05).At same time,we found that 3 of 86 AMA-M2 positive subjects had increased ALP and GGT,but none had clinical manifestations related to PBC.Conclusion:1.STAT4 rs10168266,IL21 rs17005934 and rs925550,TNFSF15 rs4979462,TNFSF8 rs2295800 and IKZF3 rs9635726 polymorphism were significantly associated with PBC,which may related to the pathogenesis of PBC.2.STAT4 rs10168266 allele T,IL21 rs17005934 allele C,IL21 rs925550 allele A,TNFSF15 rs4979462 allele T,TNFSF8 rs2295800 allele C and IKZF3 rs9635726 allele C may increase the risk of PBC.3.IL21 rs17005934 and rs925550 polymorphisms are associated with the level of ALT in PBC patients.STAT4 rs10168266 polymorphism is associated with the level of DBIL in PBC patients.4.The positive rate of AMA-M2 in Yangzhou is 0.334%,females(0.421%)is higher than that of males(0.258%).And we found the positive rate of AMA-M2 tend to increase with age.5.Anti-M2 mitochondrial antibody screening found that 3 healthy physical examination patients were diagnosed with PBC.We provided clinical ideas for the prevention and treatment of early PBC patients.