Hepatitis Monthly
Year:2015 | Volume:15 | Issue:8|Papers Count:6

Background: Viral load measurements are commonly used to monitor HCV infection in patients with chronic diseases or determining the number of HCV-genomes in serum samples of patients after sustained virological response. However, in some patients, HCV viral load in serum samples is too low to be detected by PCR, especially after treatment.Objectives: The aim of this study was to develop a highly specific, sensitive, and reproducible in-house quantitative PCR using specific primers and probe cited in highly conservative region of HCV genome that allows simultaneous detection of HCV genotypes 1 - 4.Materials and Methods: In this study, three sets of primer pairs and a TaqMan probe for amplification and detection of selected region within 5’-non-coding (5’NCR) of four HCV genotypes were used. Using plasmid containing 5’NCR region of HCV, standard curve, threshold, and threshold cycle (CT) values were determined. Real-time and nested PCR were performed on HCV genotypes 1 - 4 extracted from plasma and peripheral blood mononuclear cells (PBMCs) samples collected from patients with chronic HCV infection.Results: The lower limit detection of this in-house HCV real-time RT-PCR was determined as 100 RNA copies/mL. Inter- and intra-assay coefficient of variation (CV) of this in-house HCV real-time RT-PCR ranged from 0.9% to 1.8% and 1.76% to 3.94%, respectively. The viral load of the genotyped samples ranged from 2.0 ´ 106 ± 0.31 to 2.7 ´ 105 ± 0.46 copies/mL in serum samples and 5 ´ 102 ± 0.36 to 4.0 ´ 103 ± 0.51 copies/106 cells/mL of PBMCs.Conclusions: The quite sensitive in-house TaqMan real time RT-PCR assay was able to detect and quantify all four main HCV genotypes prevailing around all geographical regions of Iran.

Introduction: Gastrointestinal basidiobolomycosis is an emerging infection, with fewer than 80 cases reported in the English literature.Case Presentation: Also, a few cases of gastrointestinal basidiobolomycosis, accompanied by liver involvement as part of a disseminated disease, have been reported.Conclusions: This is the first case report of an isolated liver involvement of this fungal infection in a 2-year-old girl, who presented with a liver mass resembling a hepatic abscess.

Dear EditorIt was with great interest that we read the article entitled “Management of liver hemangioma using trans-catheter arterial embolization” by Firouznia et al. By way of this article, we can comment on liver hemangiomas and their embolization.

Dear EditorWe have read with great interest the meta-analysis by Yang et al. The reported objective was to evaluate the relation between cytokeratin-18 (CK-18) levels and hepatitis pathogenesis. Eight case-control studies were included in the meta-analysis according to the selection criteria adopted. High serum CK-18 levels were found in patients with hepatitis compared to controls. Moreover, increased serum CK-18 levels were related to fibrosis progression irrespective of ethnicity (Asians, Africans, and Caucasians) and liver disease etiology (chronic hepatitis C, chronic hepatitis B, and nonalcoholic steatohepatitis [NASH]). The authors concluded that serum CK-18 could be adopted as a diagnostic marker of liver injury and as a predictor of hepatitis progression.

Backgrounds: Serum hepatitis B surface antigen (HBsAg) levels are associated with fibrosis in patients with chronic hepatitis B (CHB) infection.Objectives: The aim of our study was to evaluate serum HBsAg level as a biomarker for compensated cirrhosis in hepatitis B e antigen (HBeAg) positive CHB patients.Patients and Methods: Two-hundred and one HBeAg-positive Chinese CHB patients with or without cirrhosis were enrolled in this retrospective study. Cirrhosis was diagnosed based on liver biopsy. Furthermore, patients with decompensated cirrhosis were excluded. A statistical analysis was performed regarding the association between serum HBsAg level and compensated cirrhosis.Results: Patients with compensated cirrhosis had a significantly lower mean serum HBsAg level compared to those without cirrhosis (3.27 Log10 IU/mL VS 4.17 Log10 IU/mL, P < 0.001). Furthermore, examining the correlation with compensated cirrhosis revealed that lower level of serum HBsAg was a significant factor in multivariate analysis. The area under the receiver operating characteristics curve of serum HBsAg was 0.856 for compensated cirrhosis. A positive predictive value of 66.2% and negative predictive value of 90.7% were obtained with a cut-off value of < 3.60 Log10 IU/mL (4000 IU/mL) of serum HBsAg. Moreover, the rate of compensated cirrhosis increased to 75.0% after combining with APRI > 2.Conclusions: In HBeAg positive CHB patients, low serum HBsAg level is a useful predictor of compensated cirrhosis.

Background: Macrophages play critical roles in innate immune response in the liver. Whether macrophages participate in liver innate immunity against HCV replication is poorly understood Objectives: The aim of this study was to investigate the role of macrophages in liver innate immunity against HCV replication.Materials and Methods: Freshly isolated monocytes were purified from peripheral blood of healthy adult donors. Macrophages refer to 7-day-cultured monocytes in vitro. A hepatoma cell line (Huh7) was infected with HCV JFH-1 to generate in vitro HCV infectious system. RT-PCR was used to determine HCV RNA and mRNA levels of genes expression. ELISA was used to measure the protein level of interferon-a (IFN-a) and western blot was used to determine protein expression level of Toll-like receptor 3 (TLR3).Results: HCV dsRNA induced the expression of type I IFN (IFN-a/b) in monocyte-derived macrophages. HCV dsRNA also induced the expression of TLR3 and IFN regulatory factor-7 (IRF-7), the key regulators of the IFN signaling pathway. When HCV JFH-1-infected Huh7 cells were co-cultured with macrophages activated with HCV dsRNA or incubated in media conditioned with supernatant (SN) from HCV dsRNA-activated macrophages, HCV replication was significantly suppressed. This macrophage SN action on HCV inhibition was mediated through type I IFN, which was evidenced by the observation that antibody to type I IFN receptor could neutralize the macrophages-mediated anti-HCV effect. The role of type I IFN in macrophages-mediated anti-HCV activity is further supported by the observation that HCV dsRNA-activated macrophages SN treatment induced the expression of several IFN-stimulated genes (ISGs), ISG15, ISG56, OAS-1, OAS-2, MxA and Viperin in HCV-infected Huh7 cells.Conclusions: Macrophages may play an important role in liver innate immunity against HCV replication through a type I IFN-dependent mechanism.