Objective 　Determination of Wilson disease gene mRNA expression in human fibroblast cell strain (Me32aT22/2L) by reverse transcription-polymerase chain reaction (RT-PCR). Methods　Using lipofection reagent, the plasmid vector carrying the Wilson disease gene (pRc/CMV-WD) was transferred into Me32aT22/2L cultured in serum free complement medium. RT-PCR was used to determine WD mRNA expression in Me32aT22/2L. Results　 Wilson disease gene expression was detected in Me32aT22/2L, while no specific signals were detected in untransfected fibroblast. Conclusions　It demonstrated that Me32aT22/2L strain could express the Wilson disease gene, suggesting that Wilson disease gene transfer might develop a new approach to study Wilson disease.

Objective:To investigate the early warning effect of two polymorphisms of AKT1 gene (rs1130214, rs2494744) on the risk of atherosclerotic cerebral infarction in Hainan province.Methods:In this study, totally 243 patients with atherosclerotic cerebral infarction who were hospitalized of Hainan Province from January 2019 to October 2020 were selected as the cerebral infarction group, including 148 Han nationality people and 95 Li nationality people. And 272 healthy people who received physical examination in the same hospital during the same period were selected as the control group, including 197 Han nationality people and 75 Li nationality people.All participants signed informed consent. The peripheral anticoagulant DNA was collected, and the genomic DNA was extracted and amplified by PCR.The genotypes of rs1130214 and rs2494744 were analyzed by mass spectrometry, and the distribution frequency of genotypes in cerebral infarction group and control group was analyzed by Chi-square test with SPSS 25.0 software.Results:Regardless of nationality, there was no significant difference in the distribution frequency of rs1130214 and rs2494744 of AKT1 gene between cerebral infarction group and control group (both P>0.05). The frequencies of AA, AG and GG genotypes of rs2494744 locus were 44.59%, 51.36% and 4.05% in the cerebral infarction group of Han nationality, while they were 47.21%, 42.13% and 10.66% in the control group of Han nationality, with significant difference between the two groups(χ 2=6.396, P<0.05). The independent effects of the three genotypes were analyzed by regression analysis. The results showed that GG genotype might be a resistance factor of cerebral infarction in Han population ( P=0.024, OR=0.354, 95% CI: 0.139-0.901). The frequency of AA, AG and GG genotypes of rs2494744 was 58 (61.05%), 25 (26.32%), 12 (12.63%) in the control group of Li nationality, and 28 (37.33%), 39 (52.00%), 8 (10.67%) in the control group of Li nationality. The results showed that the distribution of AA was significantly higher than that of the control group ( P<0.05, OR= 2.631, 95% CI=1.410-4.09), while AG was on the contrary ( P<0.05, OR=0.330, 95% CI=0.173-0.627). Conclusion:AA genotype of rs2494744 in AKT1 gene polymorphism is a risk factor for cerebral infarction in Li nationality group, which has potential early warning value for cerebral infarction in Hainan Li nationality group, while AG has protective effect on cerebrovascular health in Hainan Li nationality group.

Objective　To investigate the pathogenesis of Wilson's disease(WD). Methods　 Hepatocytes were isolated from WD patients' bioptic hepatic samples and cultured in vitro; WD proteins, the gene putative encoded products, were detected by SDS-PAGE in conjunction with Western blotting in liver samples of three patients and two controls. Their genomic DNAs were analyzed by means of direct DNA sequencing of WD gene (ATP7B) on exon 8. Results　 The WD proteins lanes from two WD patients were found to be much weaker than that from the control, from which one WD patient was proven as heterozygote of 778 position CGG→CTG(Arg778Leu) and 770 position CTC→CTG change of ATP7B. Conclusion　 WD is highly heterogeneous in clinical manifestations and inheritance pattern. Abnormally expressed putative WD proteins in WD patients might be the results of ATP7B mutations, and the study of ATP7B products would help to probe into the pathogenesis of WD.

Objective:To investigate the association between rs2494739 polymorphism of the AKT1 gene and cerebral infarction in the Han and Li ethnic groups in Hainan.Methods:A total of 143 cases of cerebral infarction and 197 healthy controls from the Han ethnic group, and 93 cases of cerebral infarction and 75 healthy controls from the Li ethnic group were enrolled.Genomic DNA was collected from peripheral blood.Rs2494739 polymorphisms were detected with the Snapshot technology, and SPSS 25.0 was used for statistical analysis.Results:The CC, CT and TT genotypes of rs2494739 were found in both the Han and Li ethnic groups, and their frequencies of distribution were 19.6%, 51.7% and 28.7% in the Han cases and 20.3%, 51.8% and 27.9% in the Han controls, whereas the frequencies were 19.3%, 45.2% and 35.5% in the Li cases and 33.3%, 42.7% and 24.0% in the Li controls, conforming to the Hardy-Weinberg equilibrium( P>0.05). The detection rate of genotype CC in the Li healthy controls was significantly higher than that in the Han controls( P<0.05), while no statistical difference was found between the Han and Li cases( P>0.05). The codominance hereditary model showed rs2494739 polymorphisms were not associated with cerebral infarction in the Han and Li ethnic groups( P=0.981 and 0.081, respectively)in Hainan.Further analysis revealed that the distribution frequency of genotype CC in the Li control group was significantly higher than that in the Li patient group( P=0.039, OR=0.48, 95% CI: 0.237~0.970). The distribution frequency of the T allele in the cases was significantly higher than that in the controls( P=0.020, OR=1.67, 95% CI: 1.082-2.576). Conclusions:Healthy Han and healthy Li ethnic groups in Hainan show differences in polymorphisms of rs2494739, which are not associated with the occurrence and development of cerebral infarction in the Hainan Han population.The CC genotype is a protective factor for the Li ethnicity from cerebral infarction and the T allele may be a susceptibility factor for cerebral infarction for the Li ethnicity in Hainan.