Efficacy of 104-week Telbivudine-based optimization strategy in patients with HBeAg-negative chronic hepatitis B virus infections.

BACKGROUND: Evaluate the safety and efficacy of 104-week regimen of Telbivudine(LdT)-based optimization strategy for Chinese patients who have chronic hepatits B(CHB) with HBeAg-negative. METHODS: This multi-center, open-label, prospective study enrolled 108 HBeAg-negative CHB patients who received LdT (600 mg/day) for 24 weeks, Adefovir (ADV) was added if HBV DNA remained detectable at week 24, otherwise LdT was maintained to use until 104 weeks. HBV DNA, alanine amino transferase (ALT), hepatitis B surface antigen(HBsAg), creatinine kinase(CK), and estimated glomerular filtration rate (eGFR) were measured, safety was assessed. RESULTS: Eighty-eight patients (81%) had HBV-DNA undetectable at 24 weeks and maintained to receive LdT monotherapy until 104 weeks, whereas the other 20 patients had HBV-DNA detectable and ADV was used in combination. For all patients, 72% of patients reached ALT normalization at 24 weeks, which increased to 80% at 52 weeks and 104 weeks, respectively.. 81% of total patients had undetectable HBV-DNA at 24 weeks, 92% at 52 weeks, and 94% at 104 weeks. The HBsAg titre declined steadily from baseline to 104 weeks (3.62 vs. 2.98 log10 IU/mL, p < 0.05), and the eGFR increased steadily from baseline to 104 weeks (92.9 vs. 104.4 mL/min/1.73 m2, p < 0.05). Although 79 patients (73%) had at least one time of elevated CK, most of these patients had CK elevated in Grade 1/2. CONCLUSIONS: LdT was well tolerated and effective, and 94% of patients achieved virological suppression after 104 weeks. TRIAL REGISTRATION: This study was registered in clinicaltrials.gov on January 31, 2012 and the ID No. was NCT01521975 .

Myeloid-derived suppressor cells expansion is closely associated with disease severity and progression in HBV-related acute-on-chronic liver failure.

Dysregulation of the host immune responses induced by host hepatitis B virus (HBV) interactions has been observed in acute-on-chronic liver failure (ACLF). Myeloid-derived suppressor cells (MDSCs), well known for their immunomodulatory properties, can suppress T-cell function by regulating the expression of CD3 ζ chain in cancer and autoimmune/infectious diseases while rarely have been studied in ACLF. In this study, MDSCs, CD4+ /CD8+ T cells, and CD3 ζ chain were analyzed by flow cytometry in peripheral blood mononuclear cells obtained from HBV-related ACLF patients, chronic hepatitis B (CHB) patients and healthy controls. ACLF patients were followed up for dynamic detection of MDSCs and observation of outcomes after treatment. Interestingly, peripheral CD14+ CD33+ CD11b+ HLA-DR-/low MDSCs from ACLF patients were significantly increased compared to those from CHB patients and healthy controls. CD4+ /CD8+ T cell frequency and CD3 ζ chain expression in T cells were decreased in ACLF patients compared to those of healthy controls and were negatively correlated with matched MDSC frequency. Meanwhile, the frequency of MDSCs was closely correlated with biochemical parameters that are relevant for liver injury rather than virological parameters. Moreover, a lower level of MDSCs was correlated with a better short-term prognosis (within 4 weeks but not at 8 weeks), and MDSCs remained high in ACLF patients whose conditions worsened within a 4-week follow-up period after treatment. These results suggest that MDSCs are closely involved in cell-mediated immunity in HBV-related ACLF and that peripheral MDSC expansion is closely associated with disease severity and progression in HBV-related ACLF, which may serve as a predictor of short-term prognosis.

Association of frailty with activity levels and sedentary behaviours in patients with hepatitis B cirrhosis: A cross-sectional study.

BACKGROUND AND AIMS: Research on the association between activity levels and sedentary behaviour with frailty in patients affected by hepatitis B cirrhosis is sparse. This study aimed to explore the association of frailty with activity levels and sedentary behaviours in patients with hepatitis B cirrhosis. DESIGN: This cross-sectional study followed the STROBE checklist. METHODS: This study was conducted in Guangzhou, China, between August 2021 and October 2022. The frailty condition of patients with hepatitis B cirrhosis was assessed using the liver frailty index (LFI). Their physical activity levels and sedentary time were assessed using the International Questionnaire of Physical Activity. Pearson correlation and binary logistic regression were used to analyse the data. RESULTS: Among the 503 patients with hepatitis B cirrhosis in the final analysis, 107 (21.3%) were identified as frail. Frailty was negatively correlated with walking time (r = -0.174, p < 0.001), moderate-intensity activity time (r = -0.185, p < 0.001), high-intensity activity time (r = -0.243, p < 0.001) and total activity time (r = -0.256, p < 0.001). Patients with insufficient activity (<150 min/week) and sedentary behaviour (≥420 min/day) were found to have 2.829 times higher risk of frailty than those with sufficient activity (≥150 min/week) and no sedentary behaviour (<420 min/day) (95% CI: 1.380, 5.799). CONCLUSION: Patients with hepatitis B cirrhosis who exhibited frailty demonstrated limited physical activity and engaged in sedentary behaviours. NO PATIENT OR PUBLIC CONTRIBUTION: Patients with hepatitis B cirrhosis contributed their data to the study.

Short-term entecavir versus lamivudine therapy for HBeAg-negative patients with acute-on-chronic hepatitis B liver failure.

BACKGROUND: Selection of drugs for antiviral therapy of patients with hepatitis B virus (HBV)-related acute-on-chronic liver failure (ACLF) remains difficult. This study was undertaken to evaluate the short-term efficacy of entecavir versus lamivudine on hepatitis B e antigen (HBeAg)-negative patients with ACLF. METHODS: The data of 182 HBeAg-negative patients with ACLF were retrospectively collected from patient profiles of the hospital. In these patients, 93 HBeAg-negative patients with ACLF were treated orally with 0.5 mg of entecavir and 89 were treated orally with 100 mg of lamivudine every day. The gender and age were matched between the two groups. Biochemical items, the model for end-stage liver disease (MELD) score, and HBV DNA level were matched at baseline between the two groups and monitored during treatment. The 3-month mortalities of the two groups were compared. RESULTS: No significant differences were found in biochemical items, MELD score, and HBV DNA level at baseline (P>0.05). HBV DNA level decreased within 3 months in both groups (P<0.05), regardless of the pretreatment MELD score. In patients with the same range of pretreatment MELD scores, treatment duration, posttreatment HBV DNA levels, percentage of HBV DNA level <2.7 lg copies/mL, biochemical items, MELD scores and 3-month mortality were similar in the two groups (all P>0.05). Pretreatment MELD score was not related to posttreatment HBV DNA levels (P>0.05), but related to a 3-month mortality in both groups (both P<0.001). CONCLUSIONS: In HBeAg-negative patients with ACLF, the short-term efficacy of entecavir versus lamivudine was similar. The degree of pretreatment liver failure significantly affected the outcome of treatment.

Reduction in Intrahepatic cccDNA and Integration of HBV in Chronic Hepatitis B Patients with a Functional Cure.

Background and Aims: Functional cure (FC) is characterized by the clearance of the hepatitis B surface antigen from the serum of patients with chronic hepatitis B (CHB). However, the level of intrahepatic covalently closed circular DNA (cccDNA) and hepatitis B virus (HBV) integration remains unclear. We conducted this study to determine them and reveal their value in the treatment of CHB. Methods: There were two sessions to elucidate the changes in intrahepatic cccDNA and HBV integration after antiviral therapy. In the first session, 116 patients were enrolled and divided into FC, non-functional cure (NFC), and CHB groups, including 48 patients with functionally cured CHB, 27 with CHB without functional cure after antiviral treatment, and 41 with treatment-naïve CHB. Patients were tested for both intrahepatic cccDNA and other viral markers. All patients in the FC group were followed up for at least 24 weeks to observe relapse. In the second session, another ten patients were included for in-depth whole-genome sequencing to analyze HBV integration. Results: Thirteen patients in the FC group were negative for intrahepatic cccDNA. Intrahepatic cccDNA was much higher in the CHB group compared with the FC group. Seven patients had HBsAg seroreversion, including two with virological relapse. Integration of HBV was detected in one (33.3%) functionally cured patients and in seven (100%) with CHB. 28.0% of the HBV breakpoints were assigned in the 1,500 nt to 1,900 nt range of the HBV genome. Conclusions: After achieving an FC, the rate of intrahepatic cccDNA and HBV integration was significantly reduced in patients with CHB. For those patients who cleared intrahepatic cccDNA, the chances of developing virological relapse were even lower.

Clinical and Genetic Characteristics of Alagille Syndrome in Adults.

Background and Aims: Alagille syndrome (AGS) is an autosomal dominant multisystem disorder caused by mutations in the JAG1 and NOTCH2 genes. AGS has been rarely reported in adult patients, mainly because its characteristics in adults are subtle. The study aimed to improve the understanding of adult AGS by a descriptive case series. Methods: Eight adults diagnosed with AGS at our hospital between June 2016 and June 2019 were included in the study. Clinical data, biochemical results, imaging results, liver histopathology, and genetic testing were analyzed. Results: Three female and five male patients with a median age of 24.5 years at the time of diagnosis were included in the analysis. The clinical manifestations were adult-onset (62.5%, 5/8), cholestasis (50%, 4/8), butterfly vertebrae (62.5%, 5/8), systolic murmurs (12.5%, 1/8), typical facies (12.5%, 1/8), posterior embryotoxon, and renal abnormalities (0/8). Genetic sequencing showed that all patients had mutations, with four occurring in the JAG1 gene and four in the NOTCH2 gene. Six were substitution mutations, one was a deletion mutation, and one was a splicing mutation. Five had been previously reported; but the others, one JAG1 mutation and two NOTCH2 mutations were unique and are reported here for the first time. Conclusions: The clinical manifestations highlighted by the current diagnostic criteria for most adults with AGS are atypical. Those who do not meet the criteria but are highly suspicious of having AGS need further evaluation, especially genetic testing.

Hepatitis B virus X protein induces p16 gene promoter methylation through upregulation of DNA methylation transferases DNMT1 and DNMT3A.

BACKGROUND: As a tumor suppressor, p16 can competitively block the cyclin D1-CDK4/6 complex to arrest the cell cycle in the G1 phase. Lack of p16 gene expression can lead to infinite cell proliferation and malignant transformation. OBJECTIVES: To determine whether the hepatitis B virus X protein (HBx) regulates the methylation and expression of the p16 gene. MATERIAL AND METHODS: We constructed a eukaryotic expression vector carrying the HBx gene and green fluorescent protein (GFP), and transfected it into HepG2 cells to build cell lines stably expressing GFP and GFP-HBx. The p16 protein level and p16 gene methylation were measured in these cell lines. We further detected the mRNA expression of DNA methyltransferases (DNMTs) 1, 3A and 3B. The DNMT1, DNMT3A and DNMT3B gene promoter sequences were inserted into a reporter vector (pGL3-Basic) to build recombinant vectors, which were then transiently transfected into different cell lines. After 48 h, the luciferase activity was measured. RESULTS: The level of p16 protein in GFP-HBx/HepG2 cells was significantly lower than in HepG2 and GFP/HepG2 cells. The CpG methylation was present in the p16 gene promoter region of GFP-HBx/HepG2 cells. The DNMT1 and DNMT3A mRNA levels in GFP-HBx/HepG2 cells were significantly elevated compared to that in the HepG2 cells (p = 0.0495). The luciferase activity in GFP-HBx/HepG2 cells transfected with the pGL3-DNMT1/3A pro plasmid was significantly higher than in HepG2 and GFP/HepG2 cells (both p < 0.05). CONCLUSIONS: The HBx can induce p16 gene promoter methylation and inhibit the expression of p16 in HepG2 cells. This occurs due to HBx activation of DNMT1 and DNMT3A promoters and the induction of p16 promoter methylation, which downregulates the expression of p16 protein.

[Dynamics of serum HBV DNA levels during the terminal phases of acute-on-chronic hepatitis B liver failure with different HBeAg status].

OBJECTIVE: To investigate the dynamics and clinical significance of serum hepatitis B virus (HBV) DNA levels during the terminal phase of acute-on-chronic liver failure (ACLF) with different hepatitis B e antigen (HBeAg) status. METHODS: One-hundred-and-seven patients with terminal ACLF were tested for HBeAg status by electrochemiluminescence immunoassay and serum HBV DNA levels by real-time PCR at three chronological time ranges, representing increasing severity of disease phases prior to death (day 0): 29-56 d, 15-28 d, and 0-14 d. RESULTS: In the 37 HBeAg(+) patients, HBV DNA levels at above-mentioned phases were 6.10+/-1.63, 5.61+/-1.50, and 5.29+/-1.96 log10 copies/mL. In the 70 anti-HBe(+) patients, HBV DNA levels were 4.63+/-1.82, 5.81+/-1.78, and 4.93+/-1.73 log10 copies/mL. Phase to phase comparisons revealed that the HBV DNA level in the HBeAg(+) group was significantly higher than that in the anti-HBe(+) group at 29-56 d (P less than 0.05), and that 15-28 d and 0-14 d were not significantly different (P more than 0.05). Intragroup comparisons of phases revealed no significant differences in the HBeAg(+) group (P more than 0.05), but a significant difference between 15-28 d and 0-14 d (P less than 0.05) for the anti-HBe(+) group. CONCLUSION: Serum levels of HBV DNA in patients with HBeAg positivity are higher than those in patients with anti-HBe positivity as the disease phase of ACLF nears fatality. Following the deterioration to liver failure, the HBV DNA load in HBeAg(+) patients remains stable while that in anti-HBe(+) patients decreases.

The relationship between serum exosome HBV-miR-3 and current virological markers and its dynamics in chronic hepatitis B patients on antiviral treatment.

Background: Hepatitis B virus (HBV) can encode microRNA-HBV-miR-3, which can be detected in both HBV-infected cell lines and peripheral blood exosomes of chronic hepatitis B (CHB) patients. This study was conducted to further evaluate its relationship with the current viral markers and their dynamics during antiviral therapy. Methods: We used Stem-loop real time quantitative PCR (RT-qPCR) to quantify HBV-miR-3 in the serum exosomes of CHB patients by extracting exosomes using the Supbio exosome separation kit and designing primers and Taqman probes specific for HBV-miR-3. We conducted a cross-sectional study and two cohort studies. In the cross-sectional study, 48 treatment-naive (TN) CHB patients were enrolled. In the nucleoside analogues (NAs) cohort study, 20 hepatitis B e antigen (HBeAg) negative CHB patients with negative HBV DNA on NA therapy were followed up for 96 weeks. In the NAs + pegylated interferon (Peg-IFN) cohort study, 40 patients with hepatitis B surface antigen (HBsAg) <1,500 IU/mL, negative HBV DNA, and HBeAg after NAs treatment were enrolled and were switched to Peg-IFN therapy for 48 weeks. HBV-miR-3 titers and other viral markers were detected at different time points. Results: HBV-miR-3 only existed in CHB patients with a concentration of 6.41±3.55 log10 copies/mL. HBV-miR-3 was positively correlated with HBV DNA, pregenomic RNA (pgRNA), and HBsAg. In the NAs cohort, HBsAg, pgRNA, and HBV-miR-3 levels showed little fluctuation during the 96 weeks of NA treatment (P>0.05). In the NAs + PEG-IFN cohort, HBsAg, pgRNA, and HBV-miR-3 levels declined significantly during the 48 weeks of sequential therapy (P<0.05). Conclusions: HBV-miR-3 was positively correlated with HBV DNA, pgRNA, and particularly HBsAg in TN CHB patients. Peg-IFN following NA therapy had a positive impact on HBsAg, pgRNA, and HBV-miR-3 decrease.

The relationship between the clearance of HBsAg and the remodeling of B cell subsets in CHB patients treated with Peg-IFN-α.

BACKGROUND: The seroconversion of the hepatitis B antigen is the ideal outcome for long-acting interferon-pegylated interferon-α (Peg-IFN-α) treatment among patients with chronic hepatitis B (CHB). B-cell response plays an important role in the process of hepatitis B antigen clearance, but the specific mechanism by which B-cell improve hepatitis B virus (HBV) is still unclear. METHODS: A total of 103 CHB patients participated in this study. The patients received 24 weeks of Peg-IFN-α treatment. Flow cytometry was used to detect B-cell surface markers' cluster of differentiation cluster of differentiation CD19, CD24, and CD27 in the peripheral blood mononuclear cells (PBMCs) of CHB patients before and after 24 weeks of Peg-IFN-α treatment. RESULTS: After 24 weeks of Peg-IFN-α treatment, the content of memory B cells (CD19+CD27+) and effector B cells (CD19+CD38+) increased significantly. Further analysis showed that the clearance of the hepatitis B antigen was correlated with the change value, ΔT, of plasma cells before and after treatment. The B-cell subsets (CD19+CD24+; CD19+CD40+; CD19+CD40+; CD19+CD80+), was also tested and the results showed that CD19+CD24+ and CD19+CD80+ content also increased significantly after treatment. CONCLUSIONS: After Peg-IFN-α treatment, the B-cell subsets of CHB patients are remodeled. Thus, Peg-IFN-α treatment appears to play an important role in the remodeling of B cell subsets and the clearance of HBV antigens. The results of this study provide a theoretical basis and guidance for the clinical treatment of CHB.

The clinical value of serum hepatic parenchyma cell volume-normalized hepatitis B surface antigen levels in hepatitis B e antigen -positive and -negative chronic hepatitis B patients.

BACKGROUND: While serum hepatitis B surface antigens (HBsAg) play an important role in the diagnosis and assessment of treatment results of hepatitis B virus (HBV) infections, it remains unclear whether HBsAg levels normalized to hepatic parenchymal cell volume (HPCV) is a superior indicator of disease state. This study compared the absolute and HPCV-normalized serum HBsAg levels in hepatitis B e antigen (HBeAg)-positive and HBeAg-negative patients with chronic hepatitis B (CHB). METHODS: Patients admitted to our institution with CHB were retrospectively included and categorized into the HBeAg-positive and HBeAg-negative groups. HPCV was calculated based on pathological examination of liver biopsy specimens and theory of sphere geometry. The difference between HBsAg levels and HBsAg normalized to HPCV, and also correlation between HBsAg levels and liver inflammation and fibrosis was analyzed. RESULTS: Absolute HBsAg levels (P=0.004), but not HPCV-normalized HBsAg levels (P=0.071) were significantly higher in HBeAg-positive patients compared to HBeAg-negative patients. In HBeAg-positive CHB patients, absolute HBsAg levels were positively correlated with liver inflammation grade (R=0.285, P=0.001) and hepatic fibrosis stage (R=0.351, P<0.001), as were HPCV-normalized HBsAg levels (R=0.640 and 0.742, both, P<0.001). However, in HBeAg-negative CHB patients, only HPCV-normalized HBsAg level were correlated with liver inflammation grade and hepatic fibrosis stage (R=0.640 and 0.785, both, P<0.001). CONCLUSIONS: HPCV-normalized serum HBsAg levels, rather than absolute HBsAg levels, were positively correlated with liver inflammation grade and hepatic fibrosis stage in both HBeAg-positive and HBeAg-negative CHB patients. Thus, HPCV-normalized HBsAg levels may more accurately reflect the pathological progress of CHB patients compared to absolute HBsAg levels.

Efficacy and safety of tenofovir alafenamide fumarate for preventing mother-to-child transmission of hepatitis B virus: a national cohort study.

BACKGROUND: Data on tenofovir alafenamide fumarate (TAF) for preventing mother-to-child transmission of hepatitis B virus (HBV) are lacking. AIMS: To investigate the efficacy and safety of TAF therapy for preventing hepatitis B mother-to-child transmission. METHODS: Mothers with chronic HBV infection, positive for hepatitis B e-antigen and with HBV DNA >200 000 IU/mL received TAF for preventing mother-to-child transmission were enrolled retrospectively from multiple centres with data collection on mother-infant dyads up to postpartum week 24-28. Primary measurements were the mother-to-child transmission rate and infants' malformation rate. Secondary assessments included maternal HBV DNA reduction at delivery, and maternal or infant adverse events during follow up. RESULTS: Among 71 mothers enrolled, the mean (±SD) age was 30.3 (±2.2) years. TAF was initiated during the second or third trimester and continued to delivery with a mean (±SD) duration of 12.8 (±4.0) weeks. At delivery, 85.9% (61/71) of the mothers achieved HBV DNA <200 000 IU/L. Seventy-three infants (two sets of twins) were born from mothers treated with TAF and none had congenital defects or malformations. All infants received HBV immunoglobulin and vaccine at birth with additional HBV vaccinations at one and six months. At age 24-28 weeks, all infants had negative hepatitis B surface antigen and undetectable levels of HBV DNA (<100 IU/mL). Body weight, height, and head circumferences were comparable to national standards for physical development. No severe adverse effects were reported in either mothers or infants. CONCLUSIONS: TAF for highly viraemic mothers effectively prevented mother-to-child transmission of hepatitis B. There were no safety concerns for either mothers or infants with 24-28 weeks of follow up.

Effect of healthcare insurance policy on the quality of life of chronic hepatitis C patients receiving interferon α-2a plus ribavirin therapy.

The aim of this study was to evaluate the effect of pegylated interferon α-2a plus ribavirin therapy on the quality of life (QOL) of chronic hepatitis C patients when this treatment was paid for by healthcare insurance. The QOL questionnaire (GQOLI-74) was used to assess patient QOL. A total of 42 cases received 1-year pegylated interferon α-2a plus ribavirin treatment paid for by Guangzhou Medical Insurance (group A), and 30 cases received treatment self-subsidized by the patients themselves (group B). Another 30 patients did not receive interferon therapy (group C). All groups completed the evaluation twice; prior to interferon treatment (T0) and at the end of treatment (T1). There was no statistically significant difference among the three groups (P>0.05). At T1, patients in group A had higher scores for each questionnaire dimension and a higher total score than those of group C (P<0.05). Patients in group B also had higher scores than those of group C (P<0.05), except for material well-being (P=0.305). Compared with group B, patients in group A had higher scores for mental function, material well-being and a higher total score (P<0.05). Patients in group A had higher scores for each dimension and a higher total score at T1 than at T0 (P=0.05), while patients in group B had higher scores for physical function, social function and a higher total score at T1 than at T0 (P=0.05). Pegylated interferon α-2a plus ribavirin treatment is able to improve the QOL of chronic hepatitis C patients. Patients whose treatment was financed by medical insurance exhibited increased improvement in QOL compared to those who paid for their own treatment.