ABSTRACT
ObjectiveThe genotypes and drug-resistance mutation of hepatitis B virus (HBV) are closely related to the progression of chronic hepatitis B (CHB) and effects of medication. This study was to identify the genotypes and drug-resistance mutation genes of HBV in West Guangxi and provide some laboratory evidence for anti-HBV treatment.MethodsWe collected serum samples from 869 CHB patients with HBV DNA >1.0×103 IU/mL, all from West Guangxi, between January 2016 and March 2018. We detected the genotypes and drug-resistance mutation genes in the patients by PCR and reverse dot blot (PCR-RDB).ResultsA total of 7 genotypes were identified in the patients, including genotypes B in 304 cases (34.98%), C in 268 (30.84%), D in 147 (16.92%), B+C in 28 (3.22%), B+D in 22 (2.53%), C+D in 1 (0.12%) and B+C+D in 1 (0.12%). Drug-resistance mutation was observed in 63 cases (7.25%), including 31 cases with genotype C (49.21%) and 19 with genotype B (30.16%). Fourteen gene mutation patterns were found in all, including rt204I, rt181V, rt236T, rt180M+rt204V, and rt180M+rt204V+rt204I, among which, rt180M+rt204V exhibited the highest mutation frequency (17.46%). There was a statistically significant difference between the mutation rates of genotypes B and C (19% vs 31%, P < 0.05).ConclusionCHB patients in West Guangxi have a wide variety of genotypes of HBV, mainly including genotypes B and C, with a high rate and complicated patterns of drug-resistance mutation, which has provided some reference for anti-HBV treatment in West Guangxi.
ABSTRACT
Objective There is still a lack of effective treatment for hepatitis B. The article aimed to investigate the inhibito-ry and antiviral effects of hepatitis B virus(HBV)S gene expression by anti-gene locked nucleic acid(LNA)in vivo in transgenic mice. Methods The HBV transgenic mice were divided into 5 groups by random number,6 in each group. They were blank group, irrelevant sequence group,lamivudine group,antisense locked nucleic acid group,and anti-gene locked nucleic acid group respective-ly. The lamivudine group was treated by oral gastric lavage,with 2mg/kg dose 2 times per day for continuous 7d,and the rest groups were injected with 500 mL by tail vein injection at 1,3,5 d after ad-ministration. HBV DNA was tested by real-time quantitative polymer-ase chain reaction(qPCR);HBsAg was tested by ELISA;mRNA of HBV S gene was detected by reverse transcription PCR(RT-PCR);the positive rate of HBsAg in liver cells was detected by immunohisto-chemistry. Results After the treatment of anti-gene locked nucleic acid on 3rd,5th,7thday,the inhibition rate of HBV DNA were 37.18%,50.27%,61.46%,and HBsAg were 30.17%,44%,57.76%. The inhibitory effect on 7thday was more obvious than those of the blank group,the unrelated sequence group,the lamivudine group and the antisense lock nucleic acid group,and the difference was statistically significant(P<0.05).The expression of mRNA S gene HBV(0.33)and liver cells HBsAg positive rate(31%)was signif-icantly reduced compared with control group(P<0.05).No abnormal change was found in the function of liver and kidney tests and tis-sue HE staining. Conclusion Anti-gene lock nucleic acid targeting to S gene has strong inhibitory effects on HBV replication and expression in transgenic mice,which provides theoretical and experimental knowledge for HBV gene therapy.