The Correlation Research Between WD Carriers’ Genotyping And Laboratory Index

Background:Hepatolenticular Degeneration(HLD),also known as Wilson’s disease(WD),is an autosomal recessive disease because of ATP7 B gene mutation, and it’s clinical features is caused by abnormal deposition of copper in liver, brain and even the kidneys, heart, bones and other multi-system damage. WD genes with high heredity specificity, has a large range of mutation sites. While among them are mainly few hot spots mutations, and the compound heterozygous mutations take a high percentage. The high frequency gene mutation sites is different in different countries and regions. A research shows: WD carriers has not yet found the presence of neurological symptoms,but there is some copper metabolism, such as increased urinary copper after giving decopper drugs and cerebrospinal fluid is slightly higher than normal copper and so on.It may be due to the presence of ATP7 B heterozygous mutation of WD carriers,resulting in ceruloplasmin dysfunction, and causing mild copper accumulation in vivo.It also mean WD carriers have subclinical copper retention, but have not yet reached the level of clinical symptoms. At present, many scholars conducting WD genotype---phenotype studies while the relationship between WD carriers genotype and clinical laboratory parameters have no relevant research and reporte. Now it needs further study.Objective:By detecte the 21 exon of ATP7 B gene in 271 cases of WD carriers(WD patients diagnosed as parents) and 240 cases of WD patients(family personnel of WD carriers) and 132 cases of control group. And then testing liver function, copper biochemical and liver fibrosis, analyzing the law of WD carriers and gene mutation of ATP7 B and its correlation with clinical and biochemical indexes. Discussion the meaning of differential diagnosis and pathogenesis by checking the gene mutations of ATP7 B in WD carriers and patients. Further study of patients with genotype carriers and gene frequency and correlation with clinical phenotype.Methods:Selecting patients’ father or(and)mother as the experimental group who was treated in our hospital from 2013 to 2015. WD patients and ordinary healthy people in the control group. Specimens from peripheral blood samples, and recording name,gender, age, duration of disease, the first symptom as clinical datas. And detecting the liver function,copper biochemistry,liver fibrosis in each group,collecting and extracting the gene specimens.Then use the DNA sequencing methods to analyze the gene mutations of ATP7 B in three group and statistics gene mutation. Using statistical software analyze the correlation between hypermutation hotspots with clinical phenotype of age, sex and biochemical indicators.Results:1.23 cases were detected as compound heterozygous and homozygous mutation from the 271 cases in carreers group. In part of the rest 248 cases, 68 cases detected no pathogenic mutations. Heterozygous mutations detected in a total of 178 cases of 22kinds(which four cases detected equivocal point mutation).Chromosome mutation frequency was 35.89%. Hypermutation exon are the 8th, the 13 th and the 11 th.Hypermutation site is 2333G>T, 2975C>T and 2621C>T mutation, the mutation frequencies were 16.73%, 7.46%, 2.62%.2.36 types disease-causing mutations were detected from the 240 cases in WD group.10 cases detected equivocal mutation points, 12 cases haven’t detected pathogenic mutations, only one type of heterozygous mutation was detected from 90 cases.The main hypermutation outside exon was the 8th, the 13 th, the 11 th and the 16 th.3.4 cases were detected heterozygous mutations in 132 cases of healthy control group.4.There were no significant differences in gender in WD carriers between with and without mutations sites; and between detected two mutations sites and not; and between detected 2975C>T mutation sites and not; and between detected 2333G>T mutation sites and not; and btween detected 2975C>T mutation and detected 2333G>T mutation sites.5.The liver function, copper and biochemical and liver fibrosis indexes were no significant differences between cases who with 2333G> T mutation, C2975 T mutation and no mutations in WD carriers group.6.There were no significant differences in cases between who without mutation sites, with one mutation site and with more than one mutation sites in WD carriers group.7.WD group,carriers group and control group have significantly difference in TBA.WD group compared with carriers group have no significantly difference in TBIL and DBIL.The control group compared with WD group and carriers group have significantly difference in TBIL,DBIL and ALB.Carriers group compared with control group have no significantly difference in ALT,AST and TP.The WD group compared with carriers group and control group have significantly difference in ALT,AST and TP.8.The WD group and carriers group and control group have significantly difference in Cu and CP.The WD group compared with control group and carriers group have significantly difference in Sco.9.The WD group and carriers group and control group have significantly difference in LN and IV/C.The WD group compared with carriers group have no significantly difference in HA.The control group compared with WD group and carriers group have significantly difference in HA.Conclusion: The major mutation hot spots are located on the 8th, the 13 th, the 11 th exon outside in WD group and carriers group. Liver function, copper biochemistry and liver fibrosis indexes have significantly difference in three groups. Biochemical indicators can be considered as a auxiliary standards in diagnoses and differential diagnosis of WD carriers. The clinical phenotype of gene hypermutation has no significant difference.