Knockout of C6orf120 in Rats Alleviates Concanavalin A-induced Autoimmune Hepatitis by Regulating Macrophage Polarization

Objective The effect of the functionally unknown gene C6orf120 on autoimmune hepatitis was investigated on C6orf120 knockout rats(C6orf120^(-/-))and THP-1 cells.Method Six–eight-week-old C6orf120^(-/-)and wild-type(WT)SD rats were injected with Con A(16 mg/kg),and euthanized after 24 h.The sera,livers,and spleens were collected.THP-1 cells and the recombinant protein(rC6ORF120)were used to explore the mechanism in vitro.The frequency of M1 and M2 macrophages was analyzed using flow cytometry.Western blotting and PCR were used to detect macrophage polarization-associated factors.Results C6orf120 knockout attenuated Con A-induced autoimmune hepatitis.Flow cytometry indicated that the proportion of CD68^(+)CD86^(+)M1 macrophages from the liver and spleen in the C6orf120^(-/-)rats decreased.C6orf120 knockout induced downregulation of CD86 protein and the mRNA levels of related inflammatory factors TNF-α,IL-1β,and IL-6 in the liver.C6orf120 knockout did not affect the polarization of THP-1 cells.However,rC6ORF120 promoted the THP-1 cells toward CD68^(+)CD80^(+)M1 macrophages and inhibited the CD68^(+)CD206^(+)M2 phenotype.Conclusion C6orf120 knockout alleviates Con A-induced autoimmune hepatitis by inhibiting macrophage polarization toward M1 macrophages and reducing the expression of related inflammatory factors in C6orf120^(-/-)rats.

Relationship between Phenotypic Changes of Dendritic Cell Subsets and the Onset of Plateau Phase during Intermittent Interferon Therapy in Patients with CHB

Objective This study aimed to evaluate whether the onset of the plateau phase of slow hepatitis B surface antigen decline in patients with chronic hepatitis B treated with intermittent interferon therapy is related to the frequency of dendritic cell subsets and expression of the costimulatory molecules CD40,CD80,CD83,and CD86.Method This was a cross-sectional study in which patients were divided into a natural history group(namely NH group),a long-term oral nucleoside analogs treatment group(namely NA group),and a plateau-arriving group(namely P group).The percentage of plasmacytoid dendritic cell and myeloid dendritic cell subsets in peripheral blood lymphocytes and monocytes and the mean fluorescence intensity of their surface costimulatory molecules were detected using a flow cytometer.Results In total,143 patients were enrolled(NH group,n=49;NA group,n=47;P group,n=47).The results demonstrated that CD141/CD1c double negative myeloid dendritic cell(DNmDC)/lymphocytes and monocytes(%)in P group(0.041[0.024,0.069])was significantly lower than that in NH group(0.270[0.135,0.407])and NA group(0.273[0.150,0.443]),and CD86 mean fluorescence intensity of DNmDCs in P group(1832.0[1484.0,2793.0])was significantly lower than that in NH group(4316.0[2958.0,5169.0])and NA group(3299.0[2534.0,4371.0]),Adjusted P all<0.001.Conclusion Reduced DNmDCs and impaired maturation may be associated with the onset of the plateau phase during intermittent interferon therapy in patients with chronic hepatitis B.

Association of Cytokines with Clinical Indicators in Patients with Drug-Induced Liver Injury

Objective To explore characteristics of clinical parameters and cytokines in patients with drug-induced liver injury(DILI)caused by different drugs and their correlation with clinical indicators.Method The study was conducted on patients who were up to Review of Uncertainties in Confidence Assessment for Medical Tests(RUCAM)scoring criteria and clinically diagnosed with DILI.Based on Chinese herbal medicine,cardiovascular drugs,non-steroidal anti-inflammatory drugs(NSAIDs),antiinfective drugs,and other drugs,patients were divided into five groups.Cytokines were measured by Luminex technology.Baseline characteristics of clinical biochemical indicators and cytokines in DILI patients and their correlation were analyzed.Results 73 patients were enrolled.Age among five groups was statistically different(P=0.032).Alanine aminotransferase(ALT)(P=0.033)and aspartate aminotransferase(AST)(P=0.007)in NSAIDs group were higher than those in chinese herbal medicine group.Interleukin-6(IL-6)and tumor necrosis factor alpha(TNF-α)in patients with Chinese herbal medicine(IL-6:P<0.001;TNF-α:P<0.001)and cardiovascular medicine(IL-6:P=0.020;TNF-α:P=0.001)were lower than those in NSAIDs group.There was a positive correlation between ALT(r=0.697,P=0.025),AST(r=0.721,P=0.019),and IL-6 in NSAIDs group.Conclusion Older age may be more prone to DILI.Patients with NSAIDs have more severe liver damage in early stages of DILI,TNF-αand IL-6 may partake the inflammatory process of DILI.

Inhibition of hepatitis B virus via selective apoptosis modulation by Chinese patent medicine Liuweiwuling Tablet

BACKGROUND Liuweiwuling Tablet(LWWL)is a Chinese patent medicine approved for the treatment of chronic inflammation caused by hepatitis B virus(HBV)infection.Previous studies have indicated an anti-HBV effect of LWWL,specifically in terms of antigen inhibition,but the underlying mechanism remains unclear.AIM To investigate the potential mechanism of action of LWWL against HBV.METHODS In vitro experiments utilized three HBV-replicating and three non-HBV-replicating cell lines.The in vivo experiment involved a hydrodynamic injectionmediated mouse model with HBV replication.Transcriptomics and metabolomics were used to investigate the underlying mechanisms of action of LWWL.RESULTS In HepG2.1403F cells,LWWL(0.8 mg/mL)exhibited inhibitory effects on HBV DNA,hepatitis B surface antigen and pregenomic RNA(pgRNA)at rates of 51.36%,24.74%and 50.74%,respectively.The inhibition rates of LWWL(0.8mg/mL)on pgRNA/covalently closed circular DNA in HepG2.1403F,HepG2.2.15 and HepG2.A64 cells were 47.78%,39.51%and 46.74%,respectively.Integration of transcriptomics and metabolomics showed that the anti-HBV effect of LWWL was primarily linked to pathways related to apoptosis(PI3K-AKT,CASP8-CASP3 and P53 pathways).Apoptosis flow analysis revealed that the apoptosis rate in the LWWL-treated group was significantly higher than in the control group(CG)among HBV-replicating cell lines,including HepG2.2.15(2.92%±1.01%vs 6.68%±2.04%,P<0.05),HepG2.A64(4.89%±1.28%vs 8.52%±0.50%,P<0.05)and HepG2.1403F(3.76%±1.40%vs 7.57%±1.35%,P<0.05)(CG vs LWWL-treated group).However,there were no significant differences in apoptosis rates between the non-HBV-replicating HepG2 cells(5.04%±0.74%vs 5.51%±1.57%,P>0.05),L02 cells(5.49%±0.80%vs 5.48%±1.01%,P>0.05)and LX2 cells(6.29%±1.54%vs 6.29%±0.88%,P>0.05).TUNEL staining revealed a significantly higher apoptosis rate in the LWWL-treated group than in the CG in the HBVreplicating mouse model,while no noticeable difference in apoptosis rates between the two groups was observed in the non-HBV-replicating mouse model.CONCLUSION Preliminary results suggest that LWWL exerts a potent inhibitory effect on wild-type and drug-resistant HBV,potentially involving selective regulation of apoptosis.These findings offer novel insights into the anti-HBV activities of LWWL and present a novel mechanism for the development of anti-HBV medications.

Advances in immunotherapy for hepatitis B virus associated hepatocellular carcinoma patients

Hepatitis B virus(HBV)infection plays an important role in the occurrence and development of hepatocellular carcinoma(HCC),and the rate of HBV infection in liver cancer patients in China is as high as 92.05%.Due to long-term exposure to chronic antigens from the gut,the liver needs to maintain a certain level of immune tolerance,both to avoid severe inflammation caused by non-pathogenic antigens and to maintain the possibility of rapid and violent responses to infection and tumors.Therefore,HBV infection interacts with the tumor microenvironment(TME)through a highly complex and intertwined signaling pathway,which results in a special TME in HCC.Due to changes in the TME,tumor cells can evade immune surveillance by inhibiting tumor-specific T cell function through cytotoxic T-lymphocy-associated protein-4(CTLA-4)and programmed cell death 1(PD-1)/programmed cell death ligand 1(PD-L1).Interferons,as a class of immune factors with strong biological activity,can improve the TME of HBV-HCC through various pathways.In recent years,the systematic treatment of HCC has gradually come out of the dilemma.In addition to the continuous emergence of new multi-target anti-vascular tyrosine kinase inhibitor drugs,immune checkpoint inhibitors have opened up a new avenue for the systematic treatment of HCC.At present,immunotherapy based on PD-1/L1 inhibitors has gradually become a new direction of systematic treatment for HCC,and the disease charac-teristics of patients included in global clinical studies are different from those of Chinese patients.Therefore,whether a group of HCC patients with HBV background and poor prognosis in China can also benefit from immunotherapy is an issue of wide concern.This review aims to elucidate the advances of immuno-therapy for HBV related HCC patients with regard to:(1)Immunotherapy based on interferons;(2)Immunotherapy based on PD-1/L1 inhibitors;(3)Immunotherapy based on CTLA4 inhibitors;(4)Adoptive cell transfer;(5)Combination immunotherapy strategy;and(6)Shortcomings of immunotherapy.

Pituitary stalk interruption syndrome complicated with liver cirrhosis:A case report

BACKGROUND Pituitary stalk interruption syndrome(PSIS)is a rare disorder,often characterized by delayed growth and development,short stature,and hypogonadism as the main clinical manifestations.It is not clear whether PSIS can lead to liver cirrhosis.CASE SUMMARY This paper reported a case of liver cirrhosis of unknown origin.The patient was admitted to Beijing Ditan Hospital Affiliated to Capital Medical University in November 2023.The diagnosis of PSIS complicated with liver cirrhosis was established after a series of blood tests and pituitary magnetic resonance imaging examination.CONCLUSION We also reviewed the literature from both domestic and international sources to deepen the clinical understanding of PSIS in conjunction with liver cirrhosis among medical practitioners.

Impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis

Background:Fuzheng Huayu tablet is a traditional Chinese medicine(TCM)used for the treatment of liver fibrosis and cirrhosis.However,whether the combination with Fuzheng Huayu tablet could affect the antiviral efflcacy of nucleos(t)ide remains a concern.The objective of this trial was to explore the impact of Fuzheng Huayu tablet on antiviral effect of entecavir in patients with hepatitis B cirrhosis.Methods:A prospective,randomized control trial was conducted.Patients with compensated hepatitis B cirrhosis were randomly divided into the treatment group(entecavir capsule plus Fuzheng Huayu tablet)and the control group(entecavir capsule plus simulant of Fuzheng Huayu),and followed up for 48 weeks.The dynamic changes of HBV DNA load,the rate of serological conversion of HBeAg,liver function,renal function and liver stiffness measurement(LSM)were monitored.The general clinical data and adverse events were also recorded.Results:There was no significant difference in the rate of virological response and cumulative virological response between the treatment group and the control group(P>0.05).After 48 weeks of treatment,the HBeAg seroconversion rate,biochemical response rate and LSM value were 21.05%and 4.76%(P=0.164),86.96%and 65.96%(P=0.017),9.5 kpa and 10.6 kpa(P=0.827)in the treatment group and the control group,respectively.No serious adverse events related to the study therapy occurred during the trial.Conclusions:The antiviral entecavir combined with Fuzheng Huayu tablet did not affect the antiviral efflcacy of entecavir,but could improve the rate of biochemical response,and had a tendency to improve the rate of serological conversion of HBeAg and liver fibrosis in patients with hepatitis B cirrhosis.Fuzheng Huayu tablet is clinically safe for patients with hepatitis B cirrhosis.

Changes in the Cytokine Profiles of Patients with Chronic Hepatitis B during Antiviral Therapy

Objective To investigate the changes in the cytokine profiles of chronic hepatitis B(CHB)patients undergoing antiviral treatment.Methods Hepatitis B e antigen(HBeAg)-positive patients were treated with Pegylated interferon(PEGIFN)and entecavir(ETV).Clinical biochemistry and cytokines were detected at baseline and every 3 months.Results In all,200 patients completed 48 weeks of treatment,100 in the PEG-IFN group and 100 in the ETV group.During 3–6 months of treatment,compared with baseline,the PEG-IFN group showed a significant decrease in interferon-gamma(IFN-γ),interleukin-17 A(IL-17 A),interleukin-6(IL-6),interleukin-10(IL-10),and transforming growth factor beta(TGF-β)(P<0.001)and a significant increase in interferon-alpha 2(IFN-α2)(P<0.001).In the ETV group,IL-10 and TGF-β1 decreased significantly(P<0.001).After 3 months,the levels of IFN-α2,IL-17 A,and tumor necrosis factor-alpha(TNF-α)in the PEGIFN group were significantly higher than those in the ETV group(P<0.01).The levels of IL-6 and TGF-β3 were significantly lower than those in the ETV group(P<0.01).After 6 months,the levels of IFN-α2,IFN-γ,and TNF-αin the PEG-IFN group were significantly higher than those in the ETV group(P<0.01),while the levels of IL-6 and TGF-β3 were significantly lower than those in the ETV group(P<0.01).Compared with ETV,PEG-IFN had higher HBeAg and HBsAg disappearance rates.Conclusion During antiviral therapy,a change in the cytokine profile occurred;in the aspect of immune control and functional cure,PEG-IFN was significantly better than ETV.

Cytokine Profiles and Virological Marker Monitoring during 48 Weeks Peginterferon Alfa Treatment for HBeAg-Positive Chronic Hepatitis B

Objective This study aimed to investigate whether cytokine profiles and virological markers might add value in monitoring the effects of peginterferon(PEG-IFN)therapy for hepatitis B e-antigen(HBeAg)positive chronic hepatitis B(CHB).Methods HBeAg positive patients with CHB were treated with PEG-IFN for 48 weeks.Clinical biochemical,and HBV serological indexes,as well as cytokines,were detected at baseline and every12 weeks.Results A total of 116 patients with CHB were enrolled in this study;100 patients completed the 48-week treatment and follow-up,of whom 38 achieved serum HBeAg disappearance,25 achieved HBeAg seroconversion,37 showed HBsAg decreases≥1 log10 IU/mL,9 showed HBsAg disappearance,and 8became HBsAb positive.The cytokine levels at baseline and during treatment were similar between the HBeAg disappearance group and non-disappearance group.The disappearance of HBeAg was independently associated with HBeAg levels at weeks 12 and 24,and with the HBeAg decline at week 24(P<0.05).The HBsAg response was independently associated with HBsAg,the HBsAg decline,HBeAg,the HBeAg decline at week 12,and HBsAg at week 24(P<0.05).Conclusion There was no significant correlation between the response to interferon(IFN)and cytokines during PEG-IFN treatment.The changes in virological markers predicted the response to IFN after 48 weeks.

Novel Protein C6ORF120 Promotes Apoptosis through Mitochondria-dependent Pathway in CD4+ T Lymphocytes

Generally,a healthy immune system should be in dynamic balance,which can be maintained by both promoting and resisting inflammation.Lymphocyte apoptosis is indispensable for maintaining homeostasis[1]and participates in the entire process of lymphocyte differentiation,development,maturation,and immune effects.It has been reported that a large amount of lymphocyte apoptosis occurs in lymphoid organs during severe trauma[2].Lymphocytes consist of T and B lymphocytes,among which CD4^(+)T cells were the focus of this study.CD4^(+)T lymphocytes play an important role in the innate immunity.Apoptosis of CD4^(+)T lymphocytes is an important biological process that induces CD4^(+)T cell depletion[3].Numerous studies have shown that CD4^(+)T cell apoptosis participates in many pathological processes of diseases such as HIV infection,cancer,and systemic sclerosis[4].Classical apoptosis is induced by factors that can activate several pathways,including the mitochondrial,endoplasmic reticulum,and death receptor pathways[5].The mitochondrial pathway is mainly activated by the Bcl-2 family[6].The endoplasmic reticulum(ER)pathway is affected by endoplasmic reticulum disorders.Some external factors can trigger the death receptor pathway,such as the binding of TNF-TNFR and the combination of Fas-FasL[7].Considering these pathways,it is feasible to study the specific mechanisms of lymphocyte apoptosis,primarily in CD4^(+)T cells.

Calcitriol attenuates liver fibrosis through hepatitis C virus nonstructural protein 3-transactivated protein 1-mediated TGF β1/Smad3 and NF-κB signaling pathways

BACKGROUND Hepatic fibrosis is a serious condition,and the development of hepatic fibrosis can lead to a series of complications.However,the pathogenesis of hepatic fibrosis remains unclear,and effective therapy options are still lacking.Our group identified hepatitis C virus nonstructural protein 3-transactivated protein 1(NS3TP1) by suppressive subtractive hybridization and bioinformatics analysis,but its role in diseases including hepatic fibrosis remains undefined.Therefore,additional studies on the function of NS3TP1 in hepatic fibrosis are urgently needed to provide new targets for treatment.AIM To elucidate the mechanism of NS3TP1 in hepatic fibrosis and the regulatory effects of calcitriol on NS3TP1.METHODS Twenty-four male C57BL/6 mice were randomized and separated into three groups,comprising the normal,fibrosis,and calcitriol treatment groups,and liver fibrosis was modeled by carbon tetrachloride(CCl4).To evaluate the level of hepatic fibrosis in every group,serological and pathological examinations of the liver were conducted.TGF-β1 was administered to boost the in vitro cultivation of LX-2 cells.NS3TP1,α-smooth muscle actin(α-SMA),collagen I,and collagen Ⅲ in every group were examined using a Western blot and real-time quantitative polymerase chain reaction.The activity of the transforming growth factor beta 1(TGFβ1)/Smad3 and NF-κB signaling pathways in each group of cells transfected with pcDNA-NS3TP1 or siRNA-NS3TP1 was detected.The statistical analysis of the data was performed using the Student’s t test.RESULTS NS3TP1 promoted the activation,proliferation,and differentiation of hepatic stellate cells(HSCs)and enhanced hepatic fibrosis via the TGFβ1/Smad3 and NF-κB signaling pathways,as evidenced by the presence of α-SMA,collagen I,collagen Ⅲ,p-smad3,and p-p65 in LX-2 cells,which were upregulated after NS3TP1 overexpression and downregulated after NS3TP1 interference.The proliferation of HSCs was lowered after NS3TP1 interference and elevated after NS3TP1 overexpression,as shown by the luciferase assay.NS3TP1 inhibited the apoptosis of HSCs.Moreover,both Smad3 and p65 could bind to NS3TP1,and p65 increased the promoter activity of NS3TP1,while NS3TP1 increased the promoter activity of TGFβ1 receptor I,as indicated by coimmunoprecipitation and luciferase assay results.Both in vivo and in vitro,treatment with calcitriol dramatically reduced the expression of NS3TP1.Calcitriol therapy-controlled HSCs activation,proliferation,and differentiation and substantially suppressed CCl4-induced hepatic fibrosis in mice.Furthermore,calcitriol modulated the activities of the above signaling pathways via downregulation of NS3TP1.CONCLUSION Our results suggest that calcitriol may be employed as an adjuvant therapy for hepatic fibrosis and that NS3TP1 is a unique,prospective therapeutic target in hepatic fibrosis.

Difference and clinical value of metabolites in plasma and feces of patients with alcohol-related liver cirrhosis

BACKGROUND Alterations in plasma and intestinal metabolites contribute to the pathogenesis and progression of alcohol-related liver cirrhosis(ALC).AIM To explore the common and different metabolites in the plasma and feces of patients with ALC and evaluate their clinical implications.METHODS According to the inclusion and exclusion criteria,27 patients with ALC and 24 healthy controls(HCs)were selected,and plasma and feces samples were collected.Liver function,blood routine,and other indicators were detected with automatic biochemical and blood routine analyzers.Liquid chromatography-mass spectrometry was used to detect the plasma and feces metabolites of the two groups and the metabolomics of plasma and feces.Also,the correlation between metabolites and clinical features was analyzed.RESULTS More than 300 common metabolites were identified in the plasma and feces of patients with ALC.Pathway analysis showed that these metabolites are enriched in bile acid and amino acid metabolic pathways.Compared to HCs,patients with ALC had a higher level of glycocholic acid(GCA)and taurocholic acid(TCA)in plasma and a lower level of deoxycholic acid(DCA)in the feces,while L-threonine,L-phenylalanine,and L-tyrosine increased simultaneously in plasma and feces.GCA,TCA,L-methionine,L-phenylalanine,and L-tyrosine in plasma were positively correlated with total bilirubin(TBil),prothrombin time(PT),and maddrey discriminant function score(MDF)and negatively correlated with cholinesterase(CHE)and albumin(ALB).The DCA in feces was negatively correlated with TBil,MDF,and PT and positively correlated with CHE and ALB.Moreover,we established a P/S BA ratio of plasma primary bile acid(GCA and TCA)to fecal secondary bile acid(DCA),which was relevant to TBil,PT,and MDF score.CONCLUSION The enrichment of GCA,TCA,L-phenylalanine,L-tyrosine,and L-methionine in the plasma of patients with ALC and the reduction of DCA in feces were related to the severity of ALC.These metabolites may be used as indicators to evaluate the progression of alcohol-related liver cirrhosis.

Clinical analysis of Klebsiella pneumoniae infection in patients with liver cirrhosis in Beijing

BACKGROUND The incidence of Klebsiella pneumoniae(K.pneumoniae)infection in patients with cirrhosis has been increasing over recent years,posing certain difficulties in clinical treatment.AIM To analyze the clinical features of patients with liver cirrhosis and identify the risk factors to help the early diagnosis and treatment of these diseases.METHODS Clinical data and laboratory tests were collected from 72 patients with cirrhosis confirmed by secretion or blood culture of K.pneumoniae infection at Beijing Ditan Hospital,Capital Medical University,between May 2016 and October 2018.Data from hospitalized patients with liver cirrhosis and K.pneumoniae infections,in-cluding age,sex,antimicrobial use,length of stay,site of infection,distribution of pathogenic bacteria,complications,invasive operations,laboratory indicators,treatment,and clinical regression,were extracted and retrospectively analyzed.Clinical data and biochemical values were included in the multivariate logistic regression analysis.RESULTS A total of 52 men and 20 women,with an age range from 29 to 85 years and an average age of 57.7±12.54,were analyzed.The incidence of hospital K.pneu-moniae infection in patients with cirrhosis was approximately 19.44%.The most common the infection site was the bloodstream,followed by the respiratory tract,abdominal cavity,and biliary tract.Risk factors for infection were old age,long hospital stays,gastrointestinal bleeding,and low serum albumin levels,while prophylactic antibiotics were protective factors.The multivariate analysis suggested that other infections,chronic diseases,and invasive procedures were independent factors.CONCLUSION In clinical practice,the length of hospital stays should be shortened as much as possible,invasive operations should be reduced,antibiotics should be rationally used,and the patients’liver function should be timely improved.This is of great significance for reducing the incidence of hospital infection.

Impact of ursodeoxycholic acid therapy in autoimmune liver disease patients with COVID-19 and its clinical prognosis

To explore the impact of ursodeoxycholic acid(UDCA)on severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)infection and clinical outcomes in patients with autoimmune liver disease(AILD).Patients diagnosed with AILD were enrolled and divided into a UDCA group and a non-UDCA group based on whether they received UDCA treatment.Relevant data were collected regarding AILD diagnosis,treatment,biochemical indicators,and imaging examination.The incidence of SARS-CoV-2 infection and the prognosis of AILD patients were observed.A total of 1,138 patients completed follow-up.The usage rate of hormone(P=0.003)and immunosuppressant(P=0.001)used for treating AILD in the non-UDCA group was markedly lower than in the UDCA group.The UDCA usage rate was markedly lower in SARS-CoV-2 infected patients than in uninfected patients(P=0.003).The rate of SARS-CoV-2 infection in the non-UDCA group was significantly higher than in the UDCA group(P=0.018).Logistic regression analysis showed that UDCA use(P=0.003)was correlated to a lower incidence of SARS-CoV-2,while immunosuppressant use(P=0.017)increased the incidence.Recovery time from SARS-CoV-2 infection was markedly longer for those receiving UDCA treatment than those in the non-UDCA group(P=0.018).UDCA is associated with low SARS-CoV-2 incidence in AILD patients,while immunosuppressant increases its incidence instead.Patients receiving UDCA treatment have a longer recovery time after being infected.

Early Serum HBsAg Kinetics as Predictor of HBsAg Loss in Patients with HBeAg-Negative Chronic Hepatitis B after Treatment with Pegylated Interferonα-2a

Hepatitis B surface antigen(HBsAg)loss is an ideal treatment endpoint for patients with chronic hepatitis B(CHB).We investigated the predictive value of on-treatment HBsAg levels for HBsAg loss in hepatitis B e antigen(HBe Ag)-negative CHB patients who received 120-week PEG-IFNα-2a treatment.Serum HBV DNA,HBsAg,and anti-HBs levels were assayed at baseline and every 3 months during the treatment.Of 81 patients,12 achieved HBsAg loss,20 achieved HBsAg\100 IU/mL,and 49 maintained HBs Ag C 100 IU/mL.HBsAg loss rate was only 3.7%at 48 weeks,while it reached to 11.1%and 14.8%after treatment of 96 weeks and 120 weeks.The cutoff HBs Ag levels at 12 weeks predicting HBsAg loss at 96 weeks and 120 weeks of treatment were 400 IU/mL and 750 IU/mL,with AUC 0.725 and 0.722,positive predictive value(PPV)29.41%and 30.56%,and negative predictive value(NPV)93.75%and 97.78%,respectively.The cutoff HBsAg levels at 24 weeks predicting HBsAg loss at 96 weeks and 120 weeks of treatment were 174 IU/m L and 236 IU/mL respectively,with AUC 0.925 and 0.922,PPV 40.0%and 46.15%,and both NPV 100%.The predictive ability of the cutoff HBsAg levels at 24 weeks was better than that at 12 weeks for HBs Ag loss at either 96 or 120 weeks(χ~2=3.880,P=0.049 andχ~2=4.412,P=0.036).These results indicate that extended therapy is critical to HBsAg loss in HBe Ag-negative CHB patients during PEG-IFN treatment,and the HBsAg level at 24 weeks can be used to predict HBsAg loss during tailoring PEG-IFN therapy.

Consolidation treatment needed for sustained HBsAg-negative response induced by interferon-alpha in HBeAg positive chronic hepatitis B patients

Hepatitis B surface antigen(HBsAg)clearance is considered as functional cure in patients with chronic hepatitis B(CHB).This study aimed to assess the durability of HBsAg clearance achieved by interferon-based therapies in patients with CHB who were originally positive for hepatitis B envelope antigen(HBeAg).In this prospective study,HBeAg-positive CHB patients with confirmed HBsAg loss under interferon-based therapies were enrolled within 12 weeks from end of treatment and followed up for 48 weeks.Virological markers,biochemical indicators,and liver imaging examinations were observed every 3–6 months.Sustained functional cure was analysed as primary outcome.Factor associated with sustained HBsAg loss or reversion was also investigated.The rate of HBsAg loss sustainability was 91.8%(212/231).Patients receiving consolidation treatment for 12–24weeks or≥24 weeks had higher rates of sustained HBsAg negativity than those receiving consolidation treatment for<12 weeks(98.3%and 91.2%vs.86.7%,P=0.068),and the former groups had significantly higher anti-HBs levels than the later(P<0.05).The cumulative incidence of HBsAg reversion and HBV DNA reversion was 8.2%and 3.9%,respectively.Consolidation treatment of≥12 weeks[odd ratio(OR)3.318,95%confidence interval(CI)1.077–10.224,P=0.037)was a predictor of sustained functional cure,and HBeAg-positivity at cessation of treatment(OR 12.271,95%CI 1.076–139.919,P=0.043)was a predictor of HBsAg reversion.Interferon-alpha induced functional cure was durable and a consolidation treatment of≥12–24 weeks was needed after HBsAg loss in HBeAg-positive CHB patients.

Upregulation of GLT25D1 in Hepatic Stellate Cells Promotes Liver Fibrosis via the TGF-β1/SMAD3 Pathway In Vivo and In vitro

Background and Aims:Collagenβ(1-O)galactosyltransferase 25 domain 1(GLT25D1)is associated with collagen production and glycosylation,and its knockout in mice results in embryonic death.However,its role in liver fibrosis remains elusive,particularly in hepatic stellate cells(HSCs),the primary collagen-producing cells associated with liver fibrogenesis.Herein,we aimed to elucidate the role of GLT25D1 in HSCs.Methods:Bile duct ligation(BDL)-induced mouse liver fibrosis models,primary mouse HSCs(mHSCs),and transforming growth factor beta 1(TGF-β1)-stimulated LX-2 human hepatic stellate cells were used in in vivo and in vitro studies.Stable LX-2 cell lines with either GLT25D1 overexpression or knockdown were established using lentiviral transfection.RNA-seq was performed to investigate the genomic differences.HPLCMS/MS were used to identify glycosylation sites.Scanning electronic microscopy(SEM)and second-harmonic generation/two-photon excited fluorescence(SHG/TPEF)were used to image collagen fibril morphology.Results:GLT25D1 expression was upregulated in nonparenchymal cells in human cirrhotic liver tissues.Meanwhile,its knockdown attenuated collagen deposition in BDL-induced mouse liver fibrosis and inhibited mHSC activation.GLT25D1 was overexpressed in activated versus quiescence LX-2 cells and regulated in vitro LX-2 cell activation,including proliferation,contraction,and migration.GLT25D1 also significantly increased liver fibrogenic gene and protein expression.GLT25D1 upregulation promoted HSC activation and enhanced collagen expression through the TGF-β1/SMAD signaling pathway.Mass spectrometry showed that GLT25D1 regulated the glycosylation of collagen in HSCs,affecting the diameter of collagen fibers.Conclusions:Collectively,the upregulation of GLT25D1 in HSCs promoted the progression of liver fibrosis by affecting HSCs activation and collagen stability.

Efficacy of Biejiajian Pill on Intestinal Microbiota in Patients with Hepatitis B Cirrhosis/Liver Fibrosis:A Randomized Double-Blind Controlled Trial

Objective:To analyze the efficacy of Biejiajian Pill(BJJP)on intestinal microbiota in patients with hepatitis B cirrhosis/liver fibrosis,and explore its relationship with liver fibrosis.Methods:This was a prospective,randomized double-blind controlled trial.Using the stratified block randomization method,35 patients with hepatitis B liver cirrhosis/liver fibrosis were randomly assigned(1:1)to receive entecavir(0.5 mg/d)combined with BJJP(3 g/time,3 times a day)or placebo(simulator as control,SC group,simulator 3 g/time,3 times a day)for 48 weeks.Blood and stool samples were collected from patients at baseline and week 48 of treatment,respectively.Liver and renal functions as well as hematological indices were detected.Fecal samples were analyzed by 16S rDNA V3-V4 high-throughput sequencing,and intestinal microbiota changes in both groups before and after treatment were compared,and their correlations with liver fibrosis were analyzed.Results:Compared with the SC group,there was no significant difference in liver function,renal function and hematology indices in the BJJP group,however,the improvement rate of liver fibrosis was higher in the BJJP group(94.4%vs.64.7%,P=0.041).Principal coordinate analysis(PCoA)based on weighted Unifrac distance showed significant differences in intestinal microbiota community diversity before and after BJJP treatment(P<0.01 and P=0.003),respectively.After 48 weeks'treatment,the abundance levels of beneficial bacteria(Bifidobacteria,Lactobacillus,Faecalibacterium and Blautia)increased,whereas the abundance levels of potential pathogenic bacteria,including Escherichia coli,Bacteroides,Ruminococcus,Parabacteroides and Prevotella decreased,among which Ruminococcus and Parabacteroides were significantly positively correlated with degree of liver fibrosis(r=0.34,P=0.04;r=0.38,P=0.02),respectively.The microbiota in the SC group did not change significantly throughout the whole process of treatment.Conclusion:BJJP had a certain regulatory effect on intestinal microbiota of patients with hepatitis B cirrhosis/liver fibrosis(ChiCTR1800016801).

Comprehensive investigation of HBV-related hepatocellular carcinoma and choice of anti-HBV therapy

Chronic hepatitis B virus(HBV)infection is a global public health problem,which can cause chronic hepatitis,liver cirrhosis,hepatocellular carcinoma(HCC),and other diseases.Antiviral therapy is the most critical measure to slow down the progression of chronic hepatitis B,prevent or delay cirrhosis,HCC,and other kinds of liver decompensation events.At present,the anti-hepatitis B virus drugs are mainly nucleoside(acid)analogues(NAs)and interferon.Each kind of antiviral drug has different effects on the clinical outcome of hepatitis B patients(such as HCC).In this paper,we discussed the biological characteristics,natural course and prognosis of HBV infection,the mechanism of HBV-related HCC,the effect of different antiviral drugs on patients’outcome,predictive biomarkers and model for HBV clinical outcome,predictors of sustained response and recurrence after withdrawal of antiviral therapy,consideration of expanding therapeutic indications and antiviral therapy,hoping to give a hand to the clinical diagnosis and treatment of HBV.

Application of autoantibody detection in chronic liver disease

Autoantibody(AAb)detection has become one of the standards of diagnosis for autoimmune liver disease(AILD),and some AAbs have become specific biomarkers of AILD.In addition,AAbs can be detected in patients with non-AILDs,such as viral hepatitis and alcoholic liver disease.However,the distribution characteristics and pathogenic mechanisms of AAbs in patients with non-AILD are unclear.This article summarizes the characteristics of AAbs in several common clinical chronic liver diseases(CLDs)and discusses the value of AAb analysis in CLD.