Asymptomatic Chronic Hepatitis Research Articles

INCREASED TNF-? mRNA EXPRESSION IN PBMC CELLS INFECTED WITH HEPATITIS B VIRUS EXPOSED TO PURPLE SWEET POTATO EXTRACT (Ipomoea batatas L.)

Hepatitis B Virus infection is a serious health problem throughout the world that can cause death. Purple sweet potatoes contain quite high levels of anthocyanins. Anthocyanins have been widely proven to act as immunostimulators by influencing levels of the cytokine TNF-?. This study aims to prove that purple sweet potato extract has an effect on TNF-? mRNA expression in PBMC cell cultures infected with the hepatitis B virus. The research sample was a PBMC cell culture taken from an asymptomatic chronic Hepatitis B patient. The samples from the normal saline control group were 18 wells and the treatment group with purple sweet potato extract with an anthocyanin content of 0.265 mg/100g was 18 wells. Purple sweet potato extract is made using the maceration technique. PBMCs were isolated from blood using Ficoll-Paque. PBMC cells were cultured in 24 well plates with complete RPMI 1640 medium. Examination of TNF-? mRNA expression using Relative 1-step qRT-PCR quantification. The results showed the mean difference in TNF-? mRNA expression between the treatment and control groups: 20.08 fg/µL (p<0.05), there was a significant increase in TNF-? mRNA expression. The results of this study can be concluded that administration of purple sweet potato extract in vitro can increase TNF-? mRNA expression in PBMC cultures infected with hepatitis B virus.

A Prospective Study on Fetomaternal Outcomes in Asymptomatic Chronic Hepatitis B Pregnant Women in a Tertiary Level Hospital

Introduction: Hepatitis B surface Antigen (HBsAGg) prevalence among pregnant women in India is between 0.9-3.1%. The most important factor in determining the prevalence is transmission from asymptomatic chronic Hepatitis B Virus (HBV) mother to newborn. This study was done to observe antiviral treatment and fetomaternal outcome in asymptomatic HBV mothers. Aim: To study fetomaternal outcomes in asymptomatic chronic hepatitis B pregnant women. Materials and Methods: This prospective cohort study enrolled 125 HBsAg seropositive singleton pregnancy over a period of 18 months from October 2020 to March 2022 at Department of Obstetrics and Gynaecology, VMMC and Safdarjung Hospital, New Delhi, India. Chronic Hepatitis B (CHB) was diagnosed when HBsAg, HBeAg or HBV DNA was present and IgM antiHBc was absent. Tenofovir was started in consultation with the Gastroenterologist in women with HBeAg positivity or with high HBV DNA titre ≥200,000 IU/mL or Alanine Transaminase (ALT)> two times the Upper Normal Limit (UNL). All women were followed with Liver Function Test (LFT) till delivery and six weeks postpartum and their fetomaternal outcome were noted. The p-value <0.05 was taken as significant. Descriptive statistics was analysed with Statistical Package for the Social Sciences (SPSS) version 17.0 software. Results: Tenofovir was started in 26 (20.8%) women. LFT flare was seen in 15.15% (15/99) women who were not on treatment and 0% in women on treatment. In women without treatment, log HBV DNA level was significantly increased while it was reduced in the women who received tenofovir and the reduction was significant (p-value <0.05). HBeAg was positive in 61.5% women on treatment and 0% in untreated women. No significant association was found in maternal outcomes i.e., Gestational Diabetes Mellitus (GDM), Antepartum Haemorrhage (APH), Preeclampsia, Premature Rupture Of Membrane (PROM), Preterm labour, Postpartum Haemorrhage (PPH) and High Dependency Unit (HDU) stay, in treated and untreated women. Foetal outcomes such as birth weight, APGAR score, Neonatal Intensive Care Unit (NICU) admission, prematurity, Foetal Growth Restriction (FGR) and neonatal jaundice also showed no significant association between antiviral treated and untreated women. Conclusion: There was no significant association of antiviral treatment with maternal and foetal outcome. Tenofovir is safe and reduces the LFT flare in CHB mothers. HBV DNA levels were reduced in treated women which may have reduced the incidence of Mother To Child Transmission (MTCT), which was not studied in the present study. HBeAg seropositivity and ALT >2 times the ULN can replace the need of HBV DNA titres for initiation of antiviral therapy in India.

Health state utilities of patients with hepatitis B and C and hepatitis-related conditions in Japan

Health state utilities are global measurements of quality of life and have been used to evaluate health outcomes for the cost-utility analysis. This study aimed to estimate the health state utilities of patients with hepatitis B (HB), hepatitis C (HC), and hepatitis-related diseases in Japan. We distributed a self-administered questionnaire, including the EuroQol 5-Dimension 5-Level (EQ-5D-5L), to 9,952 outpatients with several clinical conditions caused by HB or HC virus infection (such as asymptomatic chronic hepatitis, chronic hepatitis, compensated cirrhosis, and decompensated cirrhosis) and estimated the condition-specific utilities of patients with HB or HC. In patients with more severe conditions (patients with acute hepatitis, fulminant hepatitis, and hepatocellular carcinoma and patients undergoing post-liver transplantation), the utilities of these severe conditions were estimated by three hepatitis experts using the EQ-5D-5L. The means of the utilities for acute hepatitis, fulminant hepatitis, asymptomatic chronic hepatitis, chronic hepatitis, compensated cirrhosis, compensated cirrhosis, hepatocellular carcinoma stage I/II, hepatocellular carcinoma stage III/IV, and post-liver transplantation were 0.529, − 0.111, 0.904, 0.868, 0.845, 0.722, 0,675, 0,428, and 0.651 and 0.876, 0.821, 0.737, 0.671, 0.675, 0.428, and 0.651 in HB and HC, respectively. To the best of our knowledge, this is the first study that comprehensively assessed the health state utilities of patients with HB, HC and hepatitis-related conditions from a nationwide survey in Japan using the EQ-5D-5L.

Ultrasound markers in early diagnosis of asymptomatic chronic hepatitis. A prospective study.

Nowadays, chronic viral hepatitis represents one of the most common pathological entities with over 400 million cases worldwide, with high diagnosis and treatment cost, severe complications in late stages and an annual death-rate around 1 million. The purpose of this study was to identify noninvasive ultrasound markers in order to establish an early diagnosis in asymptomatic chronic hepatitis. We enrolled in a prospective study 111 patients diagnosed with asymptomatic chronic hepatitis: 53 patients with hepatitis B (group A) and 58 patients with hepatitis C (group B). All patients underwent ultrasound exam; we evaluated the dimensions of the left hepatic lobe, right hepatic lobe, caudate lobe, portal vein, liver stiffness, presence of hepatic hilar lymph nodes, number and dimensions of the adenopathies. Increased liver size was found in over 16% of patients in group A and over 20% of patients in group B. We registered an enlarged portal vein in 3.77% of patients in group A and in 3.45% of patients in group B. Adenopathies were found in 64.16% of group A and 82.76% of group B. Using long adenopathic axis as marker, we obtained a NPV over 0.9, PPV of 0.5 in the detection of chronic hepatitis B and a PPV of 0.9 in the detection of chronic hepatitis C in asymptomatic patients, a specificity of 0.98 for both chronic hepatitis B and C, a sensitivity of 0.81 for group B and 0.64 for group A. The presence of hilar liver adenopathies represents an important marker in detecting asymptomatic chronic viral hepatitis.

The effect of Hepatitis D co-infection on the immunologic and molecular profile of Hepatitis B in asymptomatic Chronic Hepatitis B patients in southwest Nigeria

ABSTRACTIntroduction: Hepatitis D infection causes severe form of viral hepatitis in humans and only affects those with hepatitis B either as a co-infection or superinfection. The aim of this study was to determine the prevalence of Hepatitis D and its effect on the immunologic and molecular profile of Hepatitis B among asymptomatic Chronic Hepatitis B patients in Abeokuta.Methodology: A cross-sectional study of 99 chronic HBV patient who met the inclusion criteria. All the patients were tested for HBsAg, anti HCV, HDV antigen, anti HDV, HBsAg quantification, and HBV DNA quantification. Associations were tested for and P value less than 0.05 was considered significant.Results: The participants included 53 (58%) male and 38 (42%) females with ages ranging from 18 to 69 (means 39 ± 11) years. Ten (11%) participants were positive for HDV-Ag while 1 (1.1%) was positive for anti-HDV. Five (5.5%) were positive for HIV 1 &2 while 1 (1.1%) was positive for anti HCV. HBV DNA quantification ranged from 15 to 17,000,000 IU/ml while HBsAg quantification ranged from 0.25 to45,520 IU/ml. There was no statistically significant relationship between HDV-Ag and age (p = .51), sex (p = .73), HBV DNA (p = .8) and HBsAg quantification (p = 1).Conclusion: The prevalence of HDV-Ag among asymptomatic treatment naïve chronic hepatitis B patients in Abeokuta was 11% and there was no significant difference in the levels of HBV DNA and HBsAg among those with or without hepatitis D.

Asymptomatic chronic hepatitis c with F0-1 fibrosis: Wait and monitor

The short-term progression of asymptomatic chronic hepatitis C patients with mild fibrosis isexcellent. Non-invasive test is easy and use to monitor the progression of fibrosis. Interferon regimen isthe treatment for HCV patients, currently direct-acting antiviral drug (DDA) regimens is more effectiveand shorter duration. Many DDA regimens can have sustained viral response (SVR) rates of morethan 90% and safety outcomes. Furthermore, all-oral interferon free regimens are not cost-effectivefor patients with mild fibrosis disease. The HCV treatment should be deferred for a few years and theprogression of fibrosis is regularly monitored. This option is safe until new treatment, which ischeaper, shorter, more effective and more convenient, become available.
 
 Keywords: symptomatic, direct-acting antiviral drug, fibrosis progression, cost

Asymptomatic chronic hepatitis C with F0-1 fibrosis: Treat now

Patients with chronic hepatitis C with no or minimal liver fibrosis (F0-F1 Fibrosis) were 50% of the entire group of patients with chronic hepatitis C. Although this group of patients might have to defer treatment due to treatment prioritization strategy, as the excellent performance of regimens with few side-effects, if in the future they would have access the treatment, the number of patient with chronic hepatitis C would decrease dramatically. After following up with treatment-naive patients for five years, 60% of them had the fibrosis progression. Patients with minimal fibrosis had possibility of advanced fibrosis at 30%. Findings indicated that the infection in the old aged patient, infection for long period, chronic hepatic inflammation, steatosis, insulin resistance and alcohol drinking affected the rapid fibrosis progression. Having treatment from F0-F1 stage was count for the long-term benefit as it reduced the development of advanced fibrosis for 15% and the survival rate at 15 years better than the non - treatment group, since it minimized the chance of cirrhosis and the extrahepatic mortality.After viral eradication, it would minimize health service and following up expenses on chronic patients.Symptom conditions such as fatique, impaired cognitive function, obsessive-compulsive, depression,and anxiety were improved that affected the better quality of life. The expenses and longer quality-adjustlife year of the treatment from minimal or no fibrotic stage should be taken into account comparingwith the treatment from significant fibrotic stage according to the health economic evaluation.Early treatment in patients with minimal or no fibrotic stage who had factors affecting the rapidfibrosis progression would result in cost-effectiveness.
 
 Keywords: chronic hepatitis C, fibrosis progression, cost-effectiveness, quality-adjust life years, increment cost effectiveness ratio

Hepatitis B virus pre-S/S variants in liver diseases.

Chronic hepatitis B is a global health problem. The clinical outcomes of chronic hepatitis B infection include asymptomatic carrier state, chronic hepatitis (CH), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). Because of the spontaneous error rate inherent to viral reverse transcriptase, the hepatitis B virus (HBV) genome evolves during the course of infection under the antiviral pressure of host immunity. The clinical significance of pre-S/S variants has become increasingly recognized in patients with chronic HBV infection. Pre-S/S variants are often identified in hepatitis B carriers with CH, LC, and HCC, which suggests that these naturally occurring pre-S/S variants may contribute to the development of progressive liver damage and hepatocarcinogenesis. This paper reviews the function of the pre-S/S region along with recent findings related to the role of pre-S/S variants in liver diseases. According to the mutation type, five pre-S/S variants have been identified: pre-S deletion, pre-S point mutation, pre-S1 splice variant, C-terminus S point mutation, and pre-S/S nonsense mutation. Their associations with HBV genotype and the possible pathogenesis of pre-S/S variants are discussed. Different pre-S/S variants cause liver diseases through different mechanisms. Most cause the intracellular retention of HBV envelope proteins and induction of endoplasmic reticulum stress, which results in liver diseases. Pre-S/S variants should be routinely determined in HBV carriers to help identify individuals who may be at a high risk of less favorable liver disease progression. Additional investigations are required to explore the molecular mechanisms of the pre-S/S variants involved in the pathogenesis of each stage of liver disease.

Multiple Eruptive Myxoid Dermatofibroma in a Male with Chronic Hepatitis B Infection

We report the first published case of multiple eruptive myxoid dermatofibroma (MEMDF) in a male with chronic hepatitis B infection presenting with eruptive lesions showing marked deposits of dermal mucin. Alcian blue and immunohistochemistry confirmed the diagnosis of myxoid dermatofibroma. Further work-up showed asymptomatic chronic hepatitis B infection without cirrhosis. This case highlights an extremely rare histologic variant and the importance of screening for altered immunity in patients with eruptive dermatofibromas.

Future complications of chronic hepatitis C in a low-risk area: projections from the hepatitis c study in Northern Norway

BackgroundHepatitis C (HCV) infection causes an asymptomatic chronic hepatitis in most affected individuals, which often remains undetected until cirrhosis and cirrhosis-related complications occur. Screening of high-risk subjects in Northern Norway has revealed a relatively low prevalence in the general population (0.24%). Despite this, late complications of HCV infection are increasing. Our object was to estimate the future prevalence and complications of chronic HCV infection in the period 2013–2050 in a low-risk area.MethodsWe have entered available data into a prognostic Markov model to project future complications to HCV infection.ResultsThe model extrapolates the prevalence in the present cohort of HCV-infected individuals, and assumes a stable low incidence in the projection period. We predict an almost three-fold increase in the incidence of cirrhosis (68 per 100,000), of decompensated cirrhosis (21 per 100,000) and of hepatocellular carcinoma (4 per 100,000) by 2050, as well as a six-fold increase in the cumulated number of deaths from HCV-related liver disease (170 per 100,000 inhabitants). All estimates are made assuming an unchanged treatment coverage of approximately 15%. The estimated numbers can be reduced by approximately 50% for cirrhosis, and by approximately one third for the other endpoints if treatment coverage is raised to 50%.ConclusionThese projections from a low-prevalence area indicate a substantial rise in HCV-related morbidity and mortality in the coming years. The global HCV epidemic is of great concern and increased treatment coverage is necessary to reduce the burden of the disease.

Expression Profiling of Cellular MicroRNA in Asymptomatic HBsAg Carriers and Chronic Hepatitis B Patients.

Background MicroRNAs (miRNAs) may serve as potential molecular markers to predict liver injury resulting from chronic hepatitis B (CHB). In the present study, we want to study the expression profile and clinical significance of miRNAs at different stages of CHB virus infection. Methods Using miRNA microarray, we investigated the global expression profiles of cellular miRNA in asymptomatic hepatitis B antigen carriers (ASCs) and CHB patients, compared with healthy controls (HCs). Results We identified 79 and 203 differentially expressed miRNAs in the peripheral blood mononuclear cells of ASCs and CHB patients compared to HCs, respectively. Some of these miRNAs were common to ASCs and CHB patients, but another set of miRNAs that showed differential expression between ASCs and CHB patients was also identified. Gene ontology and pathway enrichment analysis showed that the target genes of the identified miRNAs played a role in important biological functions, such as learning or memory, cell-cell adherens junction, ion channel inhibitor activity, TGF-beta signaling pathway, and p53 signaling pathway. Conclusion We identified some significant differentially expressed miRNA in different phases of HBV infection, which might serve as biomarkers or therapeutic targets in the future.

Chronic viral hepatitis C in pediatric age group; assessment of viral activity and hepatic fibrosis by 1H magnetic resonance spectroscopy and diffusion weighted imaging in asymptomatic patient

BackgroundChronic hepatitis C is the most common cause of chronic liver disease and cirrhosis. Egypt is the highest affected country with a prevalence of 22%. In children, seroprevalence of HCV is 0.2% in children less than 11years of age and 0.4% in children equal and more than 11years of age. Aim of the workThe purpose of this study was to assess the value of 1H MRS and DW-MRI as noninvasive tool in evaluation of activity and fibrosis of hepatic parenchyma in asymptomatic children with chronic hepatitis C. Subjects and methodsAcross-section study was conducted over a period of two years, included thirty children of asymptomatic chronic hepatitis C virus infection (mean age±SD 14.1±2.8years) and twenty healthy children as controls were included. Abdominal ultrasonography, percutaneous liver biopsy, MRS and DW-MRI were done to all cases. ResultsThe results showed that HCV infection was more common in our males (83.3%). The results of METAVIR grades showed 29 cases (99.9%) had activity while 17 cases (56.4%) had fibrosis. The results of MRS and DW-MRI showed significant differences between cases and controls and positive correlations between of the results of 1H MRS with the results of liver biopsy (METAVIR grades and METAVIR stages). ConclusionEarly diagnosis of asymptomatic chronic hepatitis C is essential to prevent or delay end stage chronic parenchymal liver disease. 1H MRS may be a potential noninvasive helpful diagnostic tool in the assessment of staging and fibrosis of asymptomatic chronic hepatitis C. The increase in metabolites were correlated with histopathological changes. DW-MRI can be considered as an effective predictor in the assessment of activity in chronic hepatitis C.

G1896A Precore Mutation and Association With HBeAg Status, Genotype and Clinical Status in Patients With Chronic Hepatitis B.

Background:Precore stop codon (G1896A) mutation is one of the commonest mutations found in patients with chronic hepatitis B. However, over the years, this mutation was not reported much in Malaysia.Objectives:We therefore investigated the presence of G1896A mutation in Malaysian population and its association with HBeAg status, clinical stage, hepatitis B virus (HBV) genotype and e-seroconversion rate.Patients and Methods:Serum samples from 93 patients confirmed as hepatitis B carriers were collected for molecular assay. The whole genome of HBV was amplified by polymerase chain reaction and directly sequenced. The precore and basal core promoter regions were analyzed for presence of mutations.Results:The most commonly observed mutation in the precore region was C1858T with 64.5% prevalence. The precore mutation of interest (G1896A) was identified in 25.8% of isolates. The basal core promoter mutations detected were A1762T-G1764A (26.9%), C1653T (8.6%), A1752G (10.8%) and C1766T (2.2%). No significant association was observed between G1896A mutation and HBeAg-negativity. Nonetheless, G1896A was highly prevalent among HBV genotype B. Clinical association revealed that subjects with G1896A mutations were mainly detected in asymptomatic chronic hepatitis B (58.3%) and liver cirrhosis (41.7%). One subject was diagnosed with fulminant hepatitis (4.2%) and 8.3% had hepatocellular carcinoma (HCC).Conclusions:Our data suggested an intermediate prevalence of G1896A mutation among Malaysian hepatitis B carriers. The stop codon mutation has a significant association with genotype B and patients with chronic hepatitis B and liver cirrhosis.

Clinical characteristics of chronic hepatitis C in children with natural course of the disease

Aim. Identification of the clinical features of chronic hepatitis C in children with the natural course of the disease. Methods. Examined were 168 children with chronic hepatitis C in the age of 0-17 years. Features of chronic hepatitis C was determined by the manifestation of infection, duration of disease, route of infection, objective data, the results of clinical and laboratory, instrumental and morphological studies of the dynamics for the period 2000-2013. Results. Depending on the variant of the infection, all children were divided into two groups: those with asymptomatic (121 patients, 72%) and manifest (47 patients, 28%) forms. Asymptomatic chronic hepatitis C in children characterized by the absence of complaints, normal values of alanine aminotransferase (36.4% of cases) and the minimal viral load (38.8% of cases). The clinical picture of chronic hepatitis C in children with manifest course characterized by the presence of clinical symptoms of low intensity as astenovegetatic (55.3%) and dyspeptic (55.3%) syndromes and extrahepatic manifestations (22.3%). According to the elastometry in more than half of the children (58.2%) liver fibrosis was identified. In 3 children in early childhood (up to 3 years) there was the elimination of the virus. Conclusion. Chronic hepatitis C in children in 72% of cases is asymptomatic; manifest form of chronic hepatitis C differs from asymptomatic form by a faster rate of disease progression.

Correlation between the promoter basal core and precore mutations and HBsAg quantification in French blood donors infected with hepatitis B virus.

Hepatitis B virus (HBV) basal core promoter (BCP) and precore (PC) mutations, HBV viral load and HBV surface antigen (HBsAg) quantitation were screened to assess correlations between these HBV markers in asymptomatic chronic hepatitis B carriers in France. From January 2006 to July 2007, 200 sera were collected from patients who were discovered to be HBsAg-positive when they volunteered to give blood. Direct sequencing of precore/core gene was used to detect A1762T/G1764A mutations in the BCP and G1896A in the PC region. HBV viral load and HBsAg were quantified with two commercials assays. The prevalence of the BCP and PC mixed/mutants were 37% and 60% respectively (P = 0.0001). HBV DNA level and HBsAg titer were significantly lower in subjects harboring the mixed/mutant PC virus compared to those infected by the wild phenotype. No significant difference was observed in HBV viral loads of blood donors infected by wild or mixed/mutant BCP viruses. Mutant or mixed PC virus was associated with male gender, HBeAb-positive status and HBV/D and HBV/E genotypes. BCP mutations were associated with age, and both HBV/A-HBV/E genotypes.The genetic properties of HBV in this cohort showed that most of the blood donors had a negative HBeAg serological status and harbored the PC mutant phenotype in combination with low levels of both HBV DNA and HBsAg. As the study was conducted in healthy subjects who could be considered as asmptomatic carriers, these results suggest a possible protective effect of the G1896A mutation against severe liver lesions.

Aqueous Flare and Choroidal Thickness in Patients with Chronic Hepatitis C Virus Infection: A Pilot Study

To investigate the status of the blood-aqueous barrier and to evaluate the subfoveal choroidal thickness (SCT) in patients with asymptomatic untreated chronic hepatitis C virus (HCV) infection without any anterior or posterior ocular involvement and to search for possible correlations. Observational case-control study. A total of 80 eyes of 20 HCV-positive patients (male-to-female ratio, 12:8; mean age, 46.9±7.23 years) and 20 healthy controls (male-to-female ratio, 10:10; mean age, 48.2±8.71 years) were examined. Participants underwent a complete ophthalmologic examination. Aqueous flare was quantified objectively by using the noninvasive laser flare cell meter FC-500 (Kowa Company Ltd, Tokyo, Japan), whereas SCT was evaluated by using enhanced depth imaging optical coherence tomography (Spectralis OCT; Heidelberg Engineering GmbH, Heidelberg, Germany). A Wilcoxon rank-sum test was performed to compare ocular findings between HCV patients and controls, and correlations were assessed by using the Spearman rank test. Retinal and choroidal thickness and anterior chamber inflammation of HCV patients and healthy controls. Patients with HCV showed significantly higher aqueous flare values (8.37±2.25 photon counts/ms vs. 4.56±1.45 photon counts/ms; P<0.0001) and a significantly increased SCT (362.7±46.5 μm vs. 320.25±32.82 μm; P<0.0001) than healthy controls. Moreover, subjects with liver fibrosis had higher flare values than those with no significant hepatic fibrosis (9.62±1.99 photon counts/ms vs. 6.97±2.19 photon counts/ms; P = 0.0003) and thicker choroids (379.15±44.75 μm vs. 346.3±43.27 μm; P = 0.024). Statistical analysis revealed that there was a positive correlation between aqueous flare values and SCT in HCV patients (r = 0.69; P<0.0001) and between flare and the degree of liver fibrosis (r = 0.67; P = 0.0001). This study showed that impairment of the blood-aqueous barrier and thickened choroids are features of asymptomatic HCV patients, and that choroidal thickness increases as the degree of subclinical inflammation of the anterior chamber increases. Patients with significant liver fibrosis have the highest flare values and the thickest choroids.

Association Between a Polymorphism in Cannabinoid Receptor 2 and Severe Necroinflammation in Patients With Chronic Hepatitis C

The cannabinoid receptor 2 (CB2) has been implicated in liver disease. The single-nucleotide polymorphism rs35761398 in cannabinoid receptor 2 gene (CNR2), which encodes the CB2, substitutes glutamine (Q) 63 with arginine (R), and reduces the function of the gene product. We investigated the effects of CNR2 rs35761398 in patients with hepatitis C virus (HCV) infection. We studied 169 consecutive patients with asymptomatic chronic hepatitis (tested positive for anti-HCV and HCV RNA) at 2 liver units in southern Italy. First, liver biopsy samples were collected from July 2009 through December 2011. All patients were naive to antiviral therapy; CNR2 genotype was determined by polymerase chain reaction analysis. Patients with the CB2-63 QQ variant had higher serum levels of aminotransferase than those with the CB2-63 QR or RR variants; they also had higher histologic activity index (HAI) scores (8.6 ± 3.8) than patients without the CB2-63 RR variant (5.3 ± 3.6; P < .005) or those with the CB2-63 QR variant (5.8 ± 3.3; P < .001). Patients with the different variants of CNR2 did not differ in fibrosis stage or steatosis score. Moderate or severe chronic hepatitis (HAI score, >8) was identified more frequently (55.5%) in patients with the CB2-63 QQ variant than in those with the 63 QR (20%; P < .005) or RR variants (17.4%; P < .005). In logistic regression analysis, the CB2-63 QQ variant and fibrosis score were independent predictors of moderate or severe chronic hepatitis (HAI score, >8; P < .0001). The CB2-63 QQ variant of CNR2 is associated with more severe inflammation and hepatocellular necrosis in patients with HCV infection.

Quantification and genotyping of hepatitis B virus in a single reaction based on dual molecular beacon real-time PCR

Objective To develop a new method for simultaneous quantifying and genotyping of HBV in a single reaction based on dual molecular beacon real-time PCR. Methods Genotype B and C recombinant plasmids were constructed as the standards and genotype-specific primers and molecular beacons were designed for each genotype.The molecular beacons of genotype B and C were labeled with FAM and Hex respectively.In this way,a simultaneous qualification and genotyping method for HBV DNA in a single real-time PCR reaction system was developed.Firstly,10-fold gradient dilution of genotype B and C standard plasmids (103-1011 kIU/L) were utilized to evaluate the linear ranges and sensitivity of this approach.The clinical specificity was tested with twenty different serum specimens (5 cases with hepatitis C virus,5 cases with herpes simplex virus and 5 cases with human papilloma virus as well as 5 healthy volunteers) ; the reproducibility was assessed by intra-assay and inter-assay coefficient of variation (CV) of cycle threshold (Ct) value through 10 repeated detections within a batch and between batches of the B,C standard plasmids (108,106and 104kIU/L).Then the accuracy of qualifying and genotyping of the self-built method was evaluated by a parallel examination with 132 HBV infected patients by use of two commercial kits as the references.Finally,these HBVpositive patients were divided into 4 groups:asymptomatic carrier (n=21),chronic hepatitis (n=77),liver cirrhosis (n=25) and hepatocellular carcinoma (n=9) to investigate the relationship of genotypes,stages of disease progression and HBV DNA load. Results A simultaneous qualification and genotyping assay was successfully built and its genotyping sensitivity was 103kIU/L and the linear range was 103-1011k IU/L.The intra-assay CV of B genotyping was 1.51% to 1.80% and the interassay CV was 2.11% to 3.03%,while the intra-assay CV of C genotyping was 1.79% to 1.95% and the inter-assay CV was 2.53% to 2.91%.The results of non HBV infected cases and healthy volunteers showed negative.In the test of 132 HBV infected patients,the general coincident rate of genotyping results comparing our assay and HBV DNA genotyping kit was 90.9% (120/132,Kappa=0.832,P<0.05).The HBV DNA quatitive results between the assay[5.07 (3.89-6.33)]and HBV DNA quatitive kit[5.19 (4.15-6.32) lg kIU/L] were well correlative (R2=0.8477,P<0.05).69 genotype B cases,51 genotype C cases and 12 B/C mixed-genotype cases were detected by dual molecular beacon real-time PCR method and their HBV DNA load were 4.54 (3.83-6.17),5.53 (4.02-6.55),4.58 (3.68-4.98) lg kIU/L respectively.Where the patients with genotype C had higher DNA load than the patients with other two genotypes (Z=-2.195 and-2.162,P<0.05).The HBV DNA load of asymptomatic group,chronic hepatitis group,liver cirrhosis group and hepatocellular carcinoma group were 7.02 (6.35-7.84),4.94 (4.16-6.25),4.37 (3.50-5.17) and 3.45 (3.25-4.92) lg kIU/L, respectively.Among them,the asymptomatic group was significantly higher than those of other three groups (Z=-4.244,-4.568 and-3.489,P<0.001) and DNA load comparing with the chronic hepatitis group,liver cirrhosis group and hepatocellular carcinoma group also showed statistically different (Z=-2.894 and-2.413,P<0.05).However,compared with the liver cirrhosis group and hepatocellular carcinoma group there was no significant difference (Z=-0.995,P=0.335). Conclusion A dual molecular beacon real-time PCR assay which can simultaneously quantifying and genotyping HBV DNA with highly accuracy,sensitive and specificity is successfully developed.(Chin J Lab Med,2013,36:333-338) Key words: Hepatitis B virus; DNA; viral; Genotype; Polymerase chain reaction

Low prevalence of hepatitis B virus pre‐s deletion mutation in Indonesia

The molecular epidemiological study of hepatitis B virus (HBV) in Indonesia is still limited. This study was aimed to identify the prevalence of HBV pre-S deletion/insertion mutations, and to assess the association of pre-S deletion mutation with liver disease progression in Indonesia. Pre-S mutations were identified by direct sequencing. Of the 265 subjects, 32 samples (12.1%) harbored pre-S deletion/insertion mutations. The prevalence of those pre-S mutations was 2.7% (2/75), 12.9% (8/62), 16.7% (11/66), and 17.7% (11/62) in asymptomatic carrier, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma groups, respectively. Statistical analysis showed significant difference among them (P = 0.024). In HBV genotype B (HBV/B), pre-S1, pre-S1/S2, and pre-S2 deletion mutations were detected respectively in 3 (17.6%), 4 (23.5%), and 9 (52.9%) of 17 samples. On the other hand, in HBV/C, 12 of 15 samples (80.0%) showed a pre-S2 deletion mutation, and only 2 samples (13.3%) demonstrated a pre-S1/S2 deletion mutation. These results suggest that in HBV/B deletion mutation tends to occur in pre-S1 or pre-S1/S2 region, while in HBV/C the deletion mutation usually occurs in the pre-S2 region. Analysis of complete genome of four viruses confirmed that 3 isolates were classified into HBV/B3, and 1 isolate was HBV/C1. However, SimPlot and BootScan analyses showed that isolate 08.10.002 was an intragenotypic recombinant between HBV/B3 and HBV/B4. As conclusion, the prevalence of HBV pre-S mutations was relatively low in Indonesian patients compared to those from Taiwan, Japan, and other Asian countries. There was a weak association between pre-S deletion mutation and progressive liver disease.

Asymptomatic chronic hepatitis B virus infection does not increase the risk of diabetes mellitus: A ten‐year observation

Chronic hepatitis C virus infection has been known to increase the risk of diabetes. Whether this association holds true for chronic hepatitis B virus (HBV) infection remains unclear. We thus conducted this study to investigate the influence of asymptomatic chronic HBV infection on the incidence of diabetes in a longitudinal cohort. A total of 1233 adults who received health examinations in 1997-1998 and in 2000-2001 were enrolled. Among them, 483 subjects who received a third health examination in 2006-2008 were further sampled. The prevalence and incidence of diabetes between asymptomatic HBV carriers and non-HBV controls were compared using the chi(2)-test and logistic regression. In 1997-1998, the prevalence rates of diabetes (9.49 and 12.0%) and glucose intolerance (28.5 and 25.4%) in HBV carriers and non-HBV controls were comparable (P > 0.05). There was no significant correlation between asymptomatic HBV infection and the presence of diabetes in subjects examined in 1997-1998, 2000-2001, or 2006-2008 when adjusted for age, gender, and body mass index (P > 0.05). In 296 non-diabetic subjects during 1997-1998, the ten-year incidence of diabetes/glucose intolerance was similar between HBV carriers and non-HBV controls (40.0 and 38.7%, P > 0.05). Moreover, no significant correlation was found between asymptomatic HBV infection and the incidence of diabetes/glucose intolerance in 2006-2008 (P = 0.775). Compared to non-HBV controls, subjects with asymptomatic chronic HBV infection do not have an increased risk of diabetes, and thus HBV itself is not pro-diabetic.