Randomised controlled trial: effect of metformin add-on therapy on functional cure in entecavir-treated patients with chronic hepatitis B

Introduction and ObjectivesHepatitis B surface antigen (HBsAg) clearance, indicating functional cure or resolved chronic hepatitis B (CHB), remains difficult to achieve via nucleos(t)ide analogue monotherapy. We investigated whether metformin add-on therapy could help achieve this goal in entecavir-treated patients with hepatitis B e antigen (HBeAg)-negative CHB.Patients and MethodsPatients with HBeAg-negative CHB who met eligibility criteria (entecavir treatment for > 12 months, HBsAg < 1000 IU/mL) were randomly assigned (1:1) to receive 24 weeks of either metformin (1000 mg, oral, once a day) or placebo (oral, once a day) add-on therapy. The group allocation was blinded for both patients and investigators. Efficacy and safety analyses were based on the intention-to-treat set. The primary outcome, serum HBsAg level (IU/mL) at weeks 24 and 36, was analysed using mixed models.ResultsSixty eligible patients were randomly assigned to the metformin (n = 29) and placebo (n = 31) groups. There was no substantial between-group difference in the HBsAg level at week 24 (adjusted mean difference 0.05, 95% confidence interval -0.04 to 0.13, p = 0.278) or week 36 (0.06, -0.03 to 0.15, p = 0.187), and no significant effect of group-by-time interaction on the HBsAg level throughout the trial (p = 0.814). The occurrence of total adverse events between the two groups was comparable (9 [31.0%] of 29 vs. 5 [16.1%] of 31, p = 0.227) and no patient experienced serious adverse events during the study.ConclusionAlthough it was safe, metformin add-on therapy did not accelerate HBsAg clearance in entecavir-treated patients with HBeAg-negative CHB.     

Long-term evaluation of recombinant interferon α2b in the treatment of patients with hepatitis B e antigen-negative chronic hepatitis B in Taiwan

The effect of interferon (IFN) on hepatitis B e antigen (HBeAg)-negative chronic hepatitis B (CHB) has not been fully investigated in Chinese patients. We enrolled 58 HBeAg-negative CHB Chinese patients with hepatitis B viremia in Taiwan to evaluate the response to IFN. 30 patients received recombinant IFN 5 million units 3 times weekly for 6–10 months, and 28 patients who refused IFN treatment served as controls. Rates of virological response and biochemical response were higher in the treated group at the end of treatment (57% vs 18%, P =0.006, and 73% vs 29%, P =0.002, respectively). Both effects were superior in the treated group at 6 months after IFN withdrawal (virological: 30% vs 7%, P =0.06; biochemical: 47% vs 7%, P =0.002). Improvement of liver histological activities with persistently biochemical response was found in 65% of the treated patients. After a mean of 32 months' follow-up, virological response was rarely maintained (17% vs 4%, P =0.228) but biochemical response was better in the treated group (27% vs 4%, P =0.039). None of the treated patients but five controls developed severe complications of CHB during the follow-up period. A larger total IFN dosage or a younger age (≤ 40 years) were associated with 'sustained' virological response. Younger age and higher baseline alanine transaminase values (≥ 120 Ul^–1) were related to 'sustained' biochemical response.     

HBV-RNA,Quantitative HBsAg,Levels of HBV in Peripheral Lymphocytes and HBV Mutation Profiles in Chronic Hepatitis B

A comprehensive characterization of chronic HBV (CHB) patients is required to guide therapeutic decisions. The cumulative impact of classical and novel biomarkers on the clinical categorization of these patients has not been rigorously assessed. We determined plasma HBV-RNA and HBsAg levels, HBV in peripheral lymphocytes (PBMCs) and HBV mutation profiles in CHB patients. Patient demographics (n = 139) and classical HBV biomarkers were determined during a clinical routine. HBV-RNA in plasma and HBV-DNA in PBMCs were determined by RT-PCR. HBsAg levels were determined using Architect. In samples with HBV-DNA viral load >1000 IU/mL, genotype mutations in precore (PC), basal core promoter (BCP), HBsAg and Pol regions were determined by sequencing. Most patients (n = 126) were HBeAg-negative (HBeAgNeg) with significantly lower levels of HBV-RNA, HBV-DNA and HBsAg compared to HBeAg-positive (HBeAgPos) patients (p < 0.05). HBV genotype D prevailed (61/68), and >95% had BCP/PC mutations. Escape mutations were identified in 22.6% (13/63). HBeAgNeg patients with low levels of HBsAg (log IU ≤ 3) were older and were characterized by undetectable plasma HBV-DNA and undetectable HBV-RNA but not undetectable HBV-DNA in PBMCs compared to those with high HBsAg levels. In >50% of the studied HBeAgNeg patients (66/126), the quantitation of HBsAg and HBV-RNA may impact clinical decisions. In conclusion, the combined assessment of classical and novel serum biomarkers, especially in HBeAgNeg patients, which is the largest group of CHB patients in many regions, may assist in clinical decisions. Prospective studies are required to determine the real-time additive clinical advantage of these biomarkers.     

ALT正常的HBV慢性携带者肝组织病理结果分析

了解ALT正常的HBV慢性携带者（ASC）的肝组织病理改变状况,探讨其临床意义及其与HBVDNA、HBeAg的关系。对32例ALT正常的HBV慢性携带者行快速经皮肝穿刺术取肝组织,研究肝组织炎症活动度及纤维化程度分级分期;ELISA法检测血清HBVM,PCR法检测血清HBVDNA。结果没有真正的健康ASC（病理状态为G0S0）,32例ASC中,G1S0有15例,G1S1有13例,G2S1有4例;肝组织学的炎症活动及纤维化改变程度HBVDNA阳性组明显重于HBVDNA阴性组,男、女之间、HBeAg定性检测及HBVDNA水平的比较,无明显差异。     

Clinical presentation and genotype of hepatitis delta in Karachi

AIM: To assess the clinical presentation and genotypes of delta hepatitis in local population. METHODS: In this prospective study, 39 consecutive patients who were positive for HBsAg and hepatitis D virus (HDV) antibody were included. The patients were divided in two groups on the basis of presence or absence of HDV RNA and a comparative study was done. Genotype of HDV was determined in PCR positive patients. RESULTS: Overall there is male dominance, in which 34 patients out of 39 (87.2%) were male. Twenty (51%) patients were from the adjacent areas of three provinces; Sindh, Punjab and Balochistan indicating the higher prevalence of delta hepatitis in this mid region of Pakistan. Patients of all age groups were affected with delta hepatitis (median 31.5 years, range 12-75). HDV RNA was detectable in 23 patients (59%). All the HDV strains belonged to genotypeⅠ. HBV DNA was detectable only in 3 cases who were also HBeAg and HDV RNA positive. Patients with detectable HDV RNA were younger than patients with undetectable RNA; mean age 29.7 ± 12.8 years vs 36.8 ± 15.2. There were no statistically significant differences in the clinical presentation and routine biochemical profile of patients with detectable or undetectable HDV RNA. Clinical cirrhosis was present in 19 (49%) patients; 12 with detectable RNA and 7 with undetectable HDV RNA (P = 0.748). Decompensated disease was seen in eight patients; five and three respectively from each group. Four patients with undetectable RNA and two patients with detectable RNA had normal ALT and ultrasound abdomen. CONCLUSION: HDV may infect at any age, usuallyyoung adult males. GenotypeⅠ is prevalent. With time some of the patients become HDV RNA negative or asymptomatic carrier. Most of the patients have suppressed HBV DNA replication. Significant numbers of patients have cirrhosis.     

Natural course following the onset of cirrhosis in patients with chronic hepatitis B: a long-term follow-up study

Purpose To elucidate the long-term natural course following the onset of cirrhosis in patients with chronic hepatitis B. Methods Ninety-three patients with chronic hepatitis B who had developed cirrhosis during regular follow-up were included in this long-term follow-up study. At the time of cirrhosis detection, 30% of the patients were seropositive for hepatitis B e antigen (HBeAg) and 73% had a HBV-DNA level >10⁴ copies/ml. Follow-up studies included liver biochemistry, viral markers, α-fetoprotein and ultrasonography every 3–6 months. Results During a mean follow-up period of 102 ± 60 (12–246; median 97) months, 32 patients (34.4%) experienced 55 episodes of hepatitis flare (7.0%/year), 15 (53.6%) of 28 HBeAg-positive patients seroconverted to anti-HBe (6.3%/yr) and 12 (12.9%) lost HBsAg (1.5%/year). Overall disease progression was observed in 25 (26.9%, 3.2%/year) patients: 12 (12.9%, 1.5%/year) hepatic decompensation, 21 (22.6%, 2.7%/year) hepatocellular carcinoma and 11 (11.8%, 1.4%/year) died. Multivariate analysis showed that age at onset of cirrhosis (P = 0.015) and persistent HBeAg seropositivity (P = 0.019) were the independent factors for overall disease progression. Conclusions These results suggest that patients with older age at onset of cirrhosis and persistent HBeAg seropositivity following the onset of cirrhosis were independent factors for the disease progression in the first 10-year after the development of cirrhosis in patients with chronic hepatitis B. This work was supported by grants from National Science Council, Taiwan (NSC95-2314-B-182A-032) and the Prosperous Foundation, Taipei, Taiwan.     

Mapping of the hepatitis B virus attachment site by use of infection-inhibiting preS1 lipopeptides and tupaia hepatocytes

BACKGROUND & AIMS: Studies on the early steps in the life cycle of hepatitis B virus have been hampered by the lack of readily available target cells. In this study, we mapped a defined virus attachment site to primary hepatocytes that is essential for infection. METHODS: We used purified virus particles from human carrier plasma as an inoculum and primary cultures of tupaia hepatocytes as susceptible target cells and studied the inhibitory effect of amino-terminally acylated preS1-derived lipopeptides on infection interference. RESULTS: Infectivity of virus could be blocked efficiently in this system by amino-terminally acylated peptides containing amino acids 2-18 from the preS1 domain. The addition of amino acids 28-48 enhanced the inhibitory capacity, whereas amino acids 49-78 did not contribute to inhibition. Myristoylated preS1 peptides 2-48 bound strongly to tupaia hepatocytes but not to nonhepatic cells or rodent hepatocytes and thereby inhibited infection even at concentrations of 1 nmol/L completely. Particles consisting only of the small hepatitis B surface protein-the active component of current hepatitis B vaccines-did not bind at all to tupaia hepatocytes, but the addition of the preS1 domain to the particles allowed binding. CONCLUSIONS: The preS1 sequence 2-48 mediates attachment of the virus to its target cells, whereas the small surface protein seems to be involved in other steps. These findings indicate that the current subunit hepatitis B vaccines may be improved by the addition of distinct preS1 epitopes. Moreover, preS1 lipopeptides are promising candidates for specific antiviral therapy against hepatitis B infections.     

乙型肝炎患者e抗原转换与外周血γδT和Tc17的关系

目的探讨乙型病毒性肝炎患者e抗原(HBeAg)转换与外周血γδT、白细胞介素(IL)-17、γ干扰素(IFN-γ)等的相关性及临床意义。方法随机选取2017年10月至2018年4月在该院就诊的乙型肝炎患者90例作为研究组,选取同期30例健康人作为对照组。然后在研究组内根据HBeAg表达与否进一步分为3个亚组:HBeAg+/Ab-组(n=30)、HBeAg+/Ab+组(n=30)、HBeAg-/Ab+组(n=30)。运用流式细胞仪检测受试者外周血中的γδT细胞、IL-17+γδT细胞、IFN-γ+γδT细胞、IL-17+CD8细胞在外周血单个核细胞(PBMCs)中的比例,对比上述指标在研究组及对照组间、以及乙型肝炎患者HBeAg/Ab不同表达组内的差异。进一步分析乙型肝炎患者HBeAg/Ab不同表达组及不同血清丙氨酸氨基转移酶(ALT)水平下,血清内HBV-DNA水平的差异。结果研究组IFN-γ+γδT细胞、γδT细胞水平、IL-17+CD8细胞要明显高于对照组(P<0.05)。研究组中,HBeAg+/Ab-组和HBeAg+/Ab+组患者PBMCs内总γδT细胞水平要显著高于HBeAg-/Ab+组(P<0.05);HBeAg+/Ab+组乙型肝炎患者IL-17+CD8细胞明显高于HBeAg+/Ab-组和HBeAg-/Ab b组(P<0.05)。HBeAg+/Ab-组慢性乙型肝炎患者血清内HBV-DNA水平要明显高于HBeAg+/Ab+组和HBeAg-/Ab+组(P<0.05)。结论γδT细胞在HBeAg/Ab血清转换中可能发挥一定作用。IL-17+CD8细胞在慢性乙型肝炎良性转归中起重要作用,对于抗病毒治疗的效果可能具有预测意义。     

HBeAg阴性慢性乙型肝炎

HBeAg阴性慢性乙型肝炎(简称e CHB),一般认为是HBV慢性感染自然病程中的一个时期[1],但也不排除一开始即感染了此种类型的HBV所致。据现有的文献报道[2],此种类型的肝炎进展快,易发展至肝硬化、肝癌等终末期肝病,治疗困难。现就e CHB的一些相关问题作一综述。1流行情况e CHB在世界各地区的流行情况有很大差异。据报道,地中海地区如意大利、希腊等国,e CHB占全部慢性乙型肝炎的33%,亚太地区如韩国、马来西亚、我国台湾省等地区占15%～20%,美国和北欧占14%。我国也是HBeAg阴性慢性乙型肝炎的高发区,且在香港占69%[2,3]。2诊断标准到…     

Prevalence of hepatitis B virus precore stop codon mutations in chronically infected children

AIM: To find out whether there is a significant difference in the prevalence of the precore stop codon mutation between HBeAg positive and anti-HBe positive children. METHODS: We investigated a large pediatric population of 155 European children (mean age 10.9 years) with chronic hepatitis B by PCR and direct sequencing. Ninety were HBeAg positive and 65 had seroconversion to anti-HBe. Additionally genotyping was performed. RESULTS: Seventy-four (48%) of the sequenced HBV strains were attributed to genotype D and 81 (52%) to genotype A. In the group of 90 HBeAg positive patients, 2 (2.2%) 1896-G-to-A transitions leading to precore stop codon mutation were found, and in the group of 65 anti-HBe positive children, 5 (7.7%) were identified harbouring HBeAg-minus mutants. The difference was not statistically significant (P=0.13). CONCLUSIONS: HBeAg minus variants as predominant viral HB strains play a minor role in the course of chronic hepatitis B in European children.