Research advances in the clinical effect of tenofovir alafenamide in treatment of chronic hepatitis B / 临床肝胆病杂志

Tenofovir alafenamide (TAF) is a novel nucleoside reverse transcriptase inhibitor used in the treatment of human immunodeficiency virus (HIV) infection and chronic hepatitis B virus (HBV) infection. Compared with tenofovir disoproxil fumarate, TAF has better plasma stability and stronger liver-targeting ability and can significantly reduce the adverse events of renal injury and reduced bone mineral density. This article summarizes the research advances in the pharmacological characteristics, metabolic pathways, drug interactions, drug resistance, and renal safety of TAF and its role in patients with chronic HBV infection.

A phase II, single-arm, open-label, multicenter clinical study to evaluate the efficacy and safety of sofosbuvir combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection / 中华肝脏病杂志

Objective@#To evaluate the efficacy and safety of sofosbuvir (Nanjing Zhengda Tianqing Pharmaceutical Co., Ltd.) combined with ribavirin in patients with genotype 2 chronic hepatitis C virus infection.@*Methods@#Treatment-naïve or treatment experienced genotype 2 chronic hepatitis C patients from sixteen research centers of China were screened. All subjects received once-daily dose of sofosbuvir (400 mg) combined with ribavirin (body weight < 75 kg, 1 000 mg/day, 400 mg in the morning and 600 mg in the evening; body weight > 75 kg, 1 200 mg/d, 600 mg in the morning and 600 mg in the evening) for 12 weeks. Patients were followed-up for a period of 12 weeks after discontinuation of treatment. Continuous variables were expressed as mean ± standard deviation. The proportion of subjects with virologic response at different follow-up time points and 95% confidence intervals were estimated by maximum likelihood ratio and Clopper-Pearson interval.@*Results@#132 cases with genotype 2 chronic hepatitis C virus infection from sixteen research centers of China were included, 12 cases of whom were associated with cirrhosis, and the remaining 120 cases were not associated with cirrhosis. One hundred and thirty-one cases completed the study, and one patient lost to follow-up at week 4 after the end of treatment. The sustained virological response rate was 96.2% (95% confidence interval: 92.37% - 99.16%) after 12 weeks of drug withdrawal. Virological relapse occurred in four cases. Of the 132 subjects enrolled in the study, 119 (90.2%) reported 617 adverse events during treatment, of which 359 (76.5%) were TEAE related to sofosbuvir and/or ribavirin. There were nine TEAEs of grade 3 and above, and six cases (4.5%) of them had six severe adverse events. Only one serious adverse event was associated with sofosbuvir and ribavirin (unstable angina pectoris). There were no adverse events leading to drug discontinuation or death.@*Conclusion@#Sofosbuvir combined with ribavirin has a high SVR rate in the treatment of genotype 2 chronic hepatitis C virus infection, and most of the adverse events occurred were mild with acceptable safety profile.

Efficacy and safety of yimitasvir phospha combined with sofosbuvir in patients with chronic hepatitis C virus infection / 中华传染病杂志

Objective To assess the efficacy and safety of 100 mg or 200 mg yimitasvir phosphate combined with sofosbuvir in patients with non-cirrhotic chronic hepatitis C virus ( HCV) genotype 1 infection who were treatment-na?ve or had a virologic failure to prior interferon-based treatment.Methods A multicenter, randomized, open-label, phase 2 clinical trial was conducted.The patients were randomly assigned to yimitasvir phosphate 100 mg+sofosbuvir 400 mg group (Group 100 mg) and yimitasvir phosphate 200 mg+sofosbuvir 400 mg group ( Group 200 mg) in a 1∶1 ratio with the stratified factors of " treatment-naive" or"treatment-experienced" for 12 weeks and followed up for 24 weeks after the end of treatment.During the clinical trial, HCV RNA was tested in all patients.Resistance of virus in patients who didn′t achieved sustained virological response (SVR) was monitored.Safety and tolerability were assessed by monitoring adverse events , physical examination , laboratory examination, electrocardiogram, and vital signs during the study.The primary end point was SVR12 after the end of therapy.Descriptive statistics were used for categorical variables and eight descriptive statistics were used for continuous variables.Descriptive statistics were used and summarized according to HCV genotypes and treatment groups.Safety data were presented using descriptive statistics and summarized according to treatment groups.Results A total of 174 subjects were screened from July 31, 2017 to September 26, 2018.One hundred and twenty-nine patients were successfully enrolled and received treatment , and 127 completed the study.There were 64 patients and 65 patients assigned to Group 100 mg and Group 200 mg, respectively.Among the 129 patients who underwent randomization and were treated , 18.6% were treatment-experienced and: 100%were HCV genotype 1b infection.The total SVR rate was 98.4%(127／129), with 98.4%(63／64, 95%confidence interval [CI]: 91.60%-99.96%) in the Group 100 mg, and 98.50%(64／65, 95%CI: 91.72%-99.96%) in the Group 200 mg.There was no significant difference between the two groups (χ2 ＝0.000 2, P＝0.989 2).The SVR rates in treatment-naive group and treatment-experienced group were 98.10%(95%CI: 93.29%-99.77%) and 100.00%(24／24, 95%CI: 85.75%-100.00%), respectively.Virological failure during treatment ( including breakthrough , rebound and poor efficacy) and relapse after treatment did not occur during the trial.By Sanger sequencing , 11.6%(15／129) patients had baseline NS5A Y93H／Y or Y93H resistance-associated substitutions ( RAS), 1.6%( 2／129) patients had baseline NS5A L31M RAS.No mutation was observed in NS5B S282 at baseline.There was no S282 mutation in HCV NS5B.A total of 100 (77.5%) subjects had adverse events.No adverse events ≥Grade 3 or severe adverse events related to the study treatment.No patient prematurely discontinued study treatment owing to an adverse event.No life-threatening adverse event was reported.Conclusion Twelve weeks of yimitasvir phosphate 100 mg or 200 mg combined with sofosbuvir 400 mg daily is a highly effective and safe regimen for patients without cirrhosis with HCV genotype 1b infection who had not been treated previously or had a virologic failure to prior interferon-based treatment.

Evaluation of training course of hepatic disease in grass roots clinical hepatic physicians in some poverty-stricken counties in Shanxi and Shaanxi provinces / 中华流行病学杂志

Objective To understand the awareness of hepatic disease related knowledge among hepatic physicians in poverty-stricken counties in China,assess the effectiveness of training and provide a reference for the training in the future.Methods The training was conducted in 90 clinical hepatic physicians selected from county hospitals in poverty-stricken counties (or cities) in Shanxi and Shaanxi provinces.An examination was conducted before the training,immediately after the training and at 5th month after the training,respectively.One-way analysis of variance and x2 test were conducted to evaluate the score and the correct rate.Results The knowledge score was (42.96± 14.02) before the training,(62.86 ± 13.28) immediately after the training and (59.03 ± 17.92) at 5thmonth after the training,and the differences were significant.After the training,the awareness of all aspects of related knowledge was improved,the difference was significant compared to knowledge score before training,and at 5th month after the training,the difference was still significant.Conclusion After the training,the awareness of liver disease related knowledge of clinical hepatic physicians in poverty-stricken counties (cities) in Shanxi and Shaanxi provinces was improved,and the improvement could be maintained for nearly halfa year.

Efficacy and safety of ombitasvir/paritaprevir/ritonavir and dasabuvir combined with ribavirin in Asian adult patients with chronic HCV genotype 1b infection and compensated cirrhosis / 中华肝脏病杂志

Objective@#To evaluate the efficacy and safety of ombitasvir/paritaprevir/ritonavir (OBV/PTV/r) 25/150/100 mg once daily and dasabuvir (DSV) 250 mg twice daily combined with ribavirin in adult patients of Mainland China with chronic HCV genotype 1b infection and compensated cirrhosis. @*Methods@#An open-label, multicenter, phase 3 clinical trial study was conducted in mainland China, Taiwan, and South Korea. Adult patients with compensated cirrhosis (Metavir score =F4) who were newly diagnosed and treated for hepatitis C virus genotype 1b infection with ombitasvir/paritaprevir/ritonavir and dasabuvir combined with ribavirin for 12 weeks were included. Assessed SVR rate of patients obtained at 12 and 24 weeks after drug withdrawal. Efficacy and safety were evaluated in patients who received at least one time study drugs. @*Results@#A total of 63 patients from mainland China were enrolled, 62 of whom (98.4%) had a baseline Child-Pugh score of 5 points. The overall rate of SVR12 and SVR24 in patients was 100% (95% CI: 94.3% to 100.0%). Most of the adverse events that occurred were mild. The incidence of common (≥10%) adverse events and laboratory abnormalities included elevated total bilirubin (36.5%), weakness (19.0%), elevated unconjugated bilirubin (19.0%) and conjugated bilirubin (17.5%), and anemia (14.3%). Three cases (4.8%) of patients experienced Grade ≥ 3 adverse events that were considered by the investigators to be unrelated to the study drug. None patients had adverse events leading to premature drug withdrawal. @*Conclusion@#Mainland Chinese patients with chronic HCV genotype 1b infection and compensated cirrhosis who were treated with OBV/PTV/r plus DSV combined with RBV for 12 weeks achieved 100 % SVR at 12 and 24 weeks after drug withdrawal. Tolerability and safety were good, and majority of adverse events were mild.

Evaluation of training course of hepatic disease in grass roots clinical hepatic physicians in some poverty-stricken counties in Shanxi and Shaanxi provinces / 中华流行病学杂志

Objective To understand the awareness of hepatic disease related knowledge among hepatic physicians in poverty-stricken counties in China,assess the effectiveness of training and provide a reference for the training in the future.Methods The training was conducted in 90 clinical hepatic physicians selected from county hospitals in poverty-stricken counties (or cities) in Shanxi and Shaanxi provinces.An examination was conducted before the training,immediately after the training and at 5th month after the training,respectively.One-way analysis of variance and x2 test were conducted to evaluate the score and the correct rate.Results The knowledge score was (42.96± 14.02) before the training,(62.86 ± 13.28) immediately after the training and (59.03 ± 17.92) at 5thmonth after the training,and the differences were significant.After the training,the awareness of all aspects of related knowledge was improved,the difference was significant compared to knowledge score before training,and at 5th month after the training,the difference was still significant.Conclusion After the training,the awareness of liver disease related knowledge of clinical hepatic physicians in poverty-stricken counties (cities) in Shanxi and Shaanxi provinces was improved,and the improvement could be maintained for nearly halfa year.

Progress in treatment of hypersplenism in patients with post-hepatitis cirrhosis / 临床肝胆病杂志

Hypersplenism is a common complication of post-hepatitis cirrhosis, and may not need to be treated. The treatment of hypersplenism is not considered if it is not proven clinically beneficial to patients. However, there are still some patients with hypersplenism who should be treated for some reasons. This article reviews the latest progress in the therapies for hypersplenism, including splenectomy, splenic artery embolization, splenic radiofrequency ablation, liver transplantation, medication, and transjugular intrahepatic portosystemic shunt. It is demonstrated that all the above treatments have advantages and disadvantages. Minimally invasive methods, through currently preferred for hypersplenism, have not yet been clinically applied for a long period, and remain to be technically improved and supported by relevant evidence for standardization.

Retrospect and reflection: the past 30 years of research on hepatitis C prophylaxis and treatment / 临床肝胆病杂志

The research on prophylaxis and treatment of hepatitis C has been performed for more than 30 years, with outstanding achievements. The discovery and confirmation of hepatitis C virus (HCV) was a milestone for how humans discovered new life. Emphasis on HCV molecular biology, infection immunity, and pathogenesis is the basic rule for scientific research on infectious diseases. PEG-IFN combined with ribavirin as the standardized antiviral treatment has been a great success; this combination therapy achieves a sustained viral response more than 80% in Chinese people, which is a typical example for successful clinical application of cytokines. Direct-acting antiviral agent(DAA) or combined application has made it possible to cure all patients infected with hepatitis C, which is the most successful example for reference. Persistent viral infection and maintenance of immune homeostasis under certain conditions are the results of the interaction between the host and the virus, and the development of vaccines will be continued.

Research advances in cellular immunotherapy for primary hepatocellular carcinoma / 临床肝胆病杂志

The present therapy for primary hepatocellular carcinoma (HCC)consists of surgery as well as local radiotherapy and chemother-apy.However,the majority of patients are susceptible to recurrence after comprehensive treatment,and the overall treatment outcome is not ideal due to the lack of effective drugs and strategies.Increasing evidence has demonstrated that the immune system is closely related to the development,progression,metastasis,and recurrence of HCC.Thus,immune therapy,especially cellular immunotherapy,could regulate immune function and induce specific antitumor immunity to achieve the goal of controlling HCC and reducing its recurrence and metastasis, which has become an essential part in the comprehensive treatment of HCC.The findings in preclinical and clinical studies on cellular immu-notherapy for HCC data are reviewed,and the current problems are discussed.

Improving understanding of liver immune function and enhancing research on cell therapy for liver diseases / 临床肝胆病杂志

As an important immune organ,the liver has a unique immune microenvironment,and is one of the important places for innate immunity and adaptive immunity.The liver is closely connected with the immune system through the gut-liver axis and is also the target or-gan of immune injury.However,the defensive role of the liver for Hepadnaviridae remains unclear.Cell therapy for liver diseases includes input of liver cells or stem cells for functional replacement,therapy by immune cells for antiviral and antitumor purposes,and genetic therapy for hereditary hepatopathy with cells as the carrier.The features of mesenchymal stem cells,multi-directional differentiation potential and immunomodulatory property,become the hot spots of cell therapy for liver diseases,and efficient in vitro amplification of cells makes it possi-ble for the use of NK cells in the treatment of hepatocellular carcinoma.

Persistent hepatitis virus infection and immune homeostasis / 临床肝胆病杂志

Homeostasis between the host and viruses is naturally maintained.On the one hand,the immune system activates the immune re-sponse to kill or eliminate viruses;on the other hand,the immune system controls the immune response to maintain immune homeostasis. The cause of persistent infections with hepatitis viruses such as HBV and HCV is that viral molecules damage the immune system of the host and their variants escape immune clearance.Long-term coexistence of the host and viruses is the process involving various immune cells and molecules and is the result of homeostasis maintenance in antiviral immune response.The immune homeostasis maintained during persis-tent infections with hepatitis viruses is analyzed by the cellular and molecular mechanisms.

Practice of training for postgraduates in infectious diseases and its reflection / 中华医学教育探索杂志

Clinical medicine is a comprehensive discipline integrating natural science and hu-manities and social science. Lemology is closely related with basic medicine and medical microbiology and medical immunology are the basis of lemology. Therefore, in the process of cultivating postgradu-ates of lemology, we should not only should attach importance to the cultivation of basic medical knowl-edge and clinical professional quality, but also pay more attention to the development of the intelligence factors and non-intelligence factors. Meanwhile education on humanity, social sciences and relevant laws and regulations should be enhanced to cultivate doctors' professional quality. Reverse thinking and lateral thinking in the clinical diagnosis should be strengthened to achieve the training objectives of cultivating international medical talents.

Life quality analysis of hepatitis C patients with liver cirrhosis undergoing antiviral therapy following splenectomy / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the effect of antiviral therapy on the quality of life (QOL) of patients with chronic hepatitis C (CHC) and cirrhosis during the 5-year period following splenectomy to treat hypersplenism.</p><p><b>METHODS</b>Data of patients with CHC and cirrhosis who had undergone treatment for hypersplenism were retrospectively selected from the hospital database of medical records. The patients were first grouped according to the hypersplenism treatment: splenectomy (group A, 28 cases) and conservative/non-operative (group B, 30 cases). Sub-grouping was carried out according to the CHC treatment: interferon-alpha-2a and ribavirin (15 cases in the A1 group, and 19 cases in the B1 group) and non-antiviral (13 cases in the A2 group, and 11 cases in the B2 group). To determine the intergroup differences in QOL during the 5-year period following the hypersplenism treatment, the QOL was assessed by chronic liver disease questionnaire (CLDQ), listing of specific symptoms (SS), and the World Health Organization QOL scale (WHOQOL-BREF).</p><p><b>RESULTS</b>Between-group statistical comparison of the subjective feeling, physiological status, mental state, and social life relationship of the patients showed no significant differences among the patients who received splenectomy compared to those who received the conservative treatment. However, the QOL of splenectomy-treated patients who received non-antiviral CHC treatment was worse than that of the patients who were given conservative treatment for the hypersplenism and antiviral therapy for the CHC. The patients who received splenectomy and antiviral therapy had better QOL than the other patient group(3.69 +/- 0.75 vs 2.15 +/- 0.98, P = 0.0003).</p><p><b>CONCLUSION</b>Splenectomy followed by antiviral therapy may improve the QOL of patients with CHC-related cirrhosis and hypersplenism.</p>

Observation of chimeric hepatitis C virus In infected Huh7.5 cell through transmission electron microscopy / 中华微生物学和免疫学杂志

Objective To observe the morphological characteristics of HCV particles and intracel-lular ultrastructure changes in Huh7. 5 cells which was infected with chimeric HCV via transmission electron microscopy. Methods Plasmid J6/JFH encoding the full length HCV chimeric genome was transcribed to HCV RNA in vitro and the RNA was transfected into Huh7.5 cells by electroporation. Quantitative real-time PCR(qRT-PCR) was used to assay HCV copies of the supernatant of transfected cells. Indirect immunofluo-rescence was used to detect the expression of HCV proteins. The cell culture superoatant were used to infect narve Huh7.5 cells, transmission electron microscopy was used to observe morphological characteristics of vi-rus particles and intracellular ultrastructure changes in infected Huh7. 5 cells. Results qRT-PCR showed high level virus copies in supernatant of transfected cells collected in different times, indirect immuno-fluo-rescencc proved high expression of HCV NS5A proteins in the transfected cells. Large numbers of enveloped or unenveloped virus-like particles (VLPs) were observed in infected Huh7. 5 cells via transmission electronmicroscopy. We also found hyperplasia of some membrane-enclosed organelles in the cytoplasm. Several fea-tures characterizing flavivirus infected cells and a cytoplasmic inclusion of unknown origin were observed. Conclusion The chimeric HCV from in vitro cell culture system is proved to be intact virus particles which can efficiently infect Huh7.5 cells.

Toll-like receptor-4 siRNA protects mice from acute liver injury induced by lipopolysaccharide and D-galactosamine / 中华传染病杂志

Objective To observe the protective effects of Toll-like receptor(TLR)-4 siRNA against acute liver injury in mice induced by lipopolysaccharide(LPS)and D-galactosamine(D-GalN).Methods One hundred and fifty C57BL/6 male mice were divided into 5 groups: phosphate buffered solution(PBS)pretreatment group,negative control plasmid pretreatment group,TS4 pretreatment group,TS6 pretreatment group and TS7 pretreatment group.Acute liver injury was induced in mice by intraperitoneal coinjection of LPS(10 ng/g)and D-GalN(1 mg/g).In vivo delivery of siRNA was performed via the tail vein by hydrodynamic injections(50 μg siRNA dissolved in 1 mL PBS)24 h and 48 h before coinjection of LPS and D-GalN. Expression of TLR-4 in liver tissues was measured by immunohistochemistry.The changes of TLR-4,tumor necrosis factor(TNF)-α and macrophage nflammatory protein(MIP)-2 mRNA levels in liver tissues were determined by reverse transcriptasepolymerase chain reaction(RT-PCR)analysis.MIP-2 and TNF-α concentrations in the sera of mice were determined by enzyme-linked immunosorbent assay(ELISA). Levels of alanine transaminase (ALT) and aspartate transaminase(AST) in serum were measured by standard autoanalyzer techniques. Liver pathological changes were observed by haematoxylin-eosin staining, while cell apoptosis levels in liver were determined by terminal deoxynucleotidyl-mediated-dUTP nick end labeling (TUNEL)assay. The difference of survival rates in 5 groups was analyzed by Fisher's exact probability test.ResultsPretreatment with TLR-4 siRNA down-regulated the TLR-4 mRNA and protein expressions,and significantly decreased the mortality and liver injury caused by coinjection of LPS and D-GalN in C57BL/6 mice.TLR-4 siRNA significantly down-regulated the TNF-α and MIP-2 mRNA expression and cytokine levels as determined by RT-PCR and ELISA,respectively. TLR-4 siRNA abrogated hepatocyte necrosis and inflammatory infiltration and also remarkably reduced serum concentrations of transaminases. The percentage of TUNEL-positive hepatocytes was significantly reduced in TLR-4 siRNA pretreatment group(TS4 pretreatment group: 0.065±0.015 vs PBS pretreatment group; 0.346±0.062,P＜0.05).ConclusionIt suggest that inhibition of TLR-4 expression by TLR-4 siRNA may provide potential application value for preventing liver injury.

The expression of protein fused HCV envelope protein E2 with His tag and its implication / 解放军医学杂志

Objective To construct the eukaryotic expression vector coding HCV gene E2 fused with His-Tag, and to express fused protein in CHO cells for investigating the function of HCV envelope protein E2. Methods The gene encoding HCV envelope protein E2 was amplified from pBRTM/HCV1-3011, a plasmid containing the cDNA of HCV's ORF, by polymerase chain reaction (PCR) method and cloned into the vector pET28(a) containing His-Tag to obtain the fused HCV envelope protein E2 gene fused with His-Tag. The fused gene was cloned into pcDNA3.1 to construct the recombinant plasmid pcDNA3.1-His-E2, which will express the E2 protein, fused with His tag. This recombinant plasmid was transfected into CHO cells by Lipofactamine 2000 reagent. The fused protein was identified by indirect immunofluorescence (IIF) and Western-blot (WB) methods. Result The positive results were obtained when the fused protein of HCV E2 with His-Tag were identified by IIF and WB methods. Conclusion The eukaryotic expression vector pcDNA3.1-His-E2 was constructed successfully and the fused proteins were expressed in cells.