Biological impacts of "hot-spot" mutations of hepatitis B virus X proteins are genotype B and C differentiated

AIM: To investigate the biological impacts of 'hot-spot'mutations on genotype B and C HBV X proteins (HBx).METHODS: Five types of'hot-spot' mutations of genotype B or C HBV X genes, which sequentially lead to the amino acid substitutions of HBx as I127T, F132Y, K130M+V131I,I127T+K130M+V131I, or K130M+V131I+F132Y, respectively,were generated by means of site-directed mutagenesis.To evaluate the anti-proliferative effects, HBx or related mutants' expression vectors were transfected separately to the Chang cells by lipofectamine, and the cells were cultured in hygromycin selective medium for 14 d, drug-resistant colonies were fixed with cold methanol, stained with Giemsa dyes and scored (increase of the colonies indicated the reduction of the anti-proliferation activity,and vice versa). Different types of HBx expression vectors were co-transfected separately with the reporter plasmid pCMVβ to Chang cells, which were lysed 48 h post-transfection and the intra-cellular β-galactosidase activities were monitored (increase of the β-galactosidase activities indicated the reduction of the transactivation activity, and vice versa). All data obtained were calculated by paired-samples t-test.RESULTS: As compared to standard genotype B HBx,mutants of I127T and I127T+K130M+V131I showed higher transactivation and anti-proliferative activities, while the mutants of F132Y, K130M+V131I, and K130M+V131I+F132Y showed lower activities. As compared to standard genotype C HBx, I127T mutant showed higher transactivation activity, while the other four types of mutants showed no differences. With regard to anti-proliferative activity,compared to standard genotype C HBx, F132Y and K130M+V131I mutants showed lower activities, and K130M+V131I +F132Y mutant, on the other hand, showed higher activity,while the mutants of I127T and I127T+K130M+V131I showed no differences.CONCLUSION: 'Hot-spot' mutations affect the antiproliferation and transactivation activities of genotype B and/or C HBx, and the biological impacts of most 'hot-spot' mutations on HBx are genotype B and C differentiated.

High Eckardt score and previous treatment were associated with poor postperoral endoscopic myotomy pain control:A retrospective study

BACKGROUND Peroral endoscopic myotomy(POEM)is a safe and effective endoscopic treatment for achalasia.However,postoperative pain management for these patients is often neglected by anesthesiologists because of the short operative time,short hospital stay and the minimally invasive nature of the procedure.AIM To assess the pain and sleep quality of achalasia patients receiving the POEM procedure and investigate factors that affect postoperative pain.METHODS This observational study included patients with achalasia who underwent POEM at Zhongshan Hospital from December 2017 to March 2018.General anesthesia was performed with endotracheal intubation.The postoperative visual analog scale(VAS),postoperative sleep quality,basic patient information,and surgical parameters were collected.Depending on whether the 12-h post-POEM VAS score was less than 4,patients were divided into two groups,a well-controlled pain group and a poorly controlled pain group.Univariate,multivariate,and stepwise logistic regression analyses were used to investigate risk factors for poor pain control.A prediction model of post-POEM pain risk was established in the form of a nomogram.The calibration curve and receiver operating characteristic curve were used to evaluate the clinical usage of the prediction model.Repeated measures analysis of variance and simple effect analysis were used to verify whether differences in the VAS and sleep scores of the high-and low-risk groups,divided by the model from the raw data,were statistically significant.RESULTS A total of 45 eligible patients were included.Multivariate logistic regression and further stepwise logistic regression analysis found that the preoperative Eckardt score[odds ratio(OR):1.82,95% confidence interval(CI):1.17-2.84,P<0.001],previous treatment(OR:7.59,95%CI:1.12-51.23,P=0.037)and the distance between the end of the muscle incision and the cardia(OR:1.52,95%CI:0.79-293.93,P=0.072)were risk factors for post-POEM pain.Repeated measures analysis of variance demonstrated that VAS(P=0.0097)and sleep scores(P=0.043)were higher in the highrisk group,and the interactions between the two main effects were obvious(VAS score:P=0.019,sleep score:P=0.035).Further simple effect analysis found that VAS scores were higher in the high-risk group at 2 h,6 h and 12 h(P=0.005,P=0.019,P<0.001),and sleep scores were higher in the high-risk group at day 1(P=0.006).CONCLUSION Achalasia patients who underwent POEM experienced serious postoperative pain,which may affect sleep quality.A higher Eckardt score,previous treatment,and a longer distance between the muscle incision ending and the cardia were risk factors for poor post-POEM pain control.

Efficacy of tenofovir alafenamide in treatment of hepatitis B virus:A meta-analysis and non-inferiority evaluation

Objective:To evaluate the effect of tenofovir alafenamide versus tenofovir disoproxil fumarate on antiviral efficacy in patients with hepatitis B virus infection.Methods:Randomized controlled trials were searched on CNKI,Wanfang,VIP,China Biomedical Literature Database,PubMed,Cochrane Library,Embase,ClinicalKey,Chinese Clinical Trial Registry and ClinicalTrials.gov from the date of inception to April 2020.The literature was screened according to the inclusion and exclusion criteria,and the efficacy evaluation index of the included RCT was set as the success rate of reaching the endpoint of viral suppression and achieving normalized ALT values at 48 weeks of treatment.Intentionality analysis was adopted and the analysis results were taken as the final conclusion.RevMan 5.3 software was used for this Meta-analysis.Meanwhile,VassarStats was used to evaluate the non-inferiority of TAF and calculate the difference of virus inhibition efficiency rate and 95%confidence interval between experimental group and the control group of each RCT.Results:After the literature search,411 potential articles were found,5 studies were finally included according to the criteria,and 2,120 patients were included.Intentionality analysis showed that TAF regimen and TDF regimen had similar viral suppression success rates(RR=0.97,95%CI:0.94~1.01,P=0.19).The ALT normalization rate in the TAF treatment group was higher than that in the TDF treatment group,and the difference was statistically significant(RR=1.35,95%CI:1.20-1.53,P<0.00001).The non-inferiority margin was set at 10%,and it was found that three RCT studies in the international multi-center all showed that TAF was not inferior to TDF in controlling HBV viral load,while two RCT studies in China's Mainland failed to achieve non-inferiority after calculation.Conclusions:At 48 weeks of treatment,TAF was similar to TDF in controlling HBV viral load.However,the efficacy of TAF in controlling HBV viral load may vary among different populations,which requires further confirmation by more clinical trial evidence.Based on AASLD criteria,the ALT normalization rate of the TAF group was higher than that of the TDF group at 48 weeks of treatment,showing an obvious advantage.