Background: Nucleic acid testing (NAT) has been widely used for transfusion - transmitted infection screening at blood banks all over the world to reduce window period, yet the assay has not been implemented in Vietnam. Objective: Using and evaluating cost - effectiveness of NAT in screening HIV, HCV, HBV in blood \r\n', u'donors. Subjects and methods: The study was carried out on 9392 blood donors at National Institute of Hematology and Blood Transfusion from Jan to May 2007 who were HIV, HCV, HBV negative with ELISA. Plasma from donors was pooled (pool size of 8) and tested with UItrio Procleix HIV - 1, HCV, HBV (Chiton). Results: These 9392 plasma samples were pooled into 1174 pool samples to perform NAT. Among 1174 pooled samples, there was only 1 case with negative ELISA - Reactive NAT. The sample was determined as response with probe HCV. From there, one of eight pool samples was identified responding to probe HCV and it was more likely to have been missed in the window period when screened by ELISA.Conclusion: The sample should be further tested with HCV qualitative and quantitative testing to confirm the status of infection. \r\n', u'\r\n', u'
HIV ; Hepatitis B virus/drug effects ; Hepacivirus ; Blood Donors

No abstract available.
Antiviral Agents/therapeutic use ; Hepacivirus/drug effects/physiology ; Hepatitis C/*drug therapy ; Humans

OBJECTIVETo review the updated research on direct antiviral agents (DAAs)-including regimens for hepatitis C virus (HCV), and focus on "difficult-to-treat" HCV-infected patients.

DATA SOURCESThe literature concerning DAAs and hepatitis C cited in this review was collected from PubMed and Google Scholar databases published in English up to July 2013.

STUDY SELECTIONData from published articles regarding HCV and DAAs in clinical trials and in clinical use were identified and reviewed.

RESULTSIt was recognized that some "difficult-to-treat" patients would still exist, even though stronger treatments using such as DAAs, including telaprevir and boceprevir, which lead to higher sustained virological response rates, are available. Such patients include those with advanced fibrosis/cirrhosis, elderly persons, children, HCV-human immunodeficiency virus co-infected patients, HCV-infected recipients, and so on.

CONCLUSIONSCertain "difficult-to-treat" patients would still exist, even though stronger treatment is available. Although evidence from clinical trials is still lacking, interferon-sparing regimens could have stronger effects for eradicating HCV in such cases.

Antiviral Agents ; pharmacology ; therapeutic use ; Hepacivirus ; drug effects ; pathogenicity ; Hepatitis C, Chronic ; drug therapy ; Humans

Antiviral Agents ; administration & dosage ; Drug Therapy, Combination ; Hepacivirus ; drug effects ; genetics ; Hepatitis C, Chronic ; drug therapy ; Humans ; Interferon-alpha ; administration & dosage ; adverse effects ; Ribavirin ; administration & dosage ; Treatment Outcome

OBJECTIVETo investigate the impact of interferon-stimulated exonuclease 20 kDa (ISG20) on replication of genotype 2a hepatitis C virus (HCV) subgenomic replicon RNA and infectivity of the cell culture-derived HCV strain JFH1 to determine the potential of exogenously expressed ISG20 as an anti-viral therapy of chronic hepatitis C.

METHODSPlasma vectors containing wild-type (WT) ISG20 or a catalytically-inactive mutant ISG20m were transiently transfected into Huh7, Huh7.5 and HEK293 cells, and the replication of a monocistronic subgenomic JFH1 RNA replicon, SGRm-JFH1BlaRL, was measured. Huh7.5 cells stably expressing ISG20, ISG20m, or the control vector were established by transducing replication incompetent pCX4-Bsr-myc retroviruses encoding WT ISG20, D94G mutant ISG20, or the empty vector, respectively, and selecting with 5 mug/mL of blasticidin for approximately three weeks. The stable Huh7.5 cells were then transfected with HCV replicon RNA and infected with cell culture-derived HCV to investigate inhibition capacity of ISG20 against HCV.

RESULTSHuh7.5-ISG20, Huh7.5-ISG20m, and Huh7.5-Bsr controls cells stably expressing ISG20, ISG20m, or the control vector, respectively, were constructed successfully; the ectopically expressed ISG20 and ISG20m were distributed in both nucleus and cytoplasm, as detected by immuno uorescence. SGRm-JFH1BlaRL replicated efficiently and with similar kinetics in the Huh7.5-Bsr and Huh7.5-ISG20m cells, with expression levels plateauing at 48-96 h post-transfection. In contrast, at all time points examined, SGRm-JFH1BlaRL replication was 9.1% to 16.7% in the Huh7.5-ISG20 cells. The Huh7, Huh7.5 and HEK293 cells transiently expressing ISG20 also showed 16.7% to 25.0% of HCV replication that the respective controls. In addition, the amount of infectious progeny JFH1 virus released in culture supernatants was 9.1% to 12.5% from the Huh7.5-ISG20 cells than from the Huh7.5-Bsr and Huh7.5-ISG20m cells at 48-72 h post-infection, and the latter two cultures produced similar JFH1 virus yields. Finally, the expression of HCV core protein was also lower in the Huh7.5-ISG20 cells, as detected by immunoblot analysis.

CONCLUSIONExogenous expression of ISG20, either in a transient or stable manner, suppresses not only replication of genotype 2a HCV RNA replicons but also JFH1 virus propagation in cultured hepatocytes. The exonuclease activity of ISG20 is required for its antiviral activities against HCV.

Antiviral Agents ; pharmacology ; Cell Line ; Genome, Viral ; HEK293 Cells ; Hepacivirus ; genetics ; Humans ; RNA, Viral ; genetics ; Replicon ; Virus Replication ; drug effects

Adult ; Blood Donors ; Hepacivirus ; drug effects ; genetics ; Hepatitis C, Chronic ; virology ; Humans ; Interferons ; pharmacology ; Middle Aged ; Viral Load

OBJECTIVETo study the inhibition effect of HCV NS5A on p53 protein transactivity and its possible mechanism.

METHODSLuciferase reporter gene system was used for the study of p53 transactivity on p21 promoter and electrophorectic mobility-shift assay (EMSA) was applied to observe whether HCV NS5A could suppress the binding ability of p53 protein to its specific DNA sequence.

RESULTSEndogenous p53 protein could stimulate p21 promoter activity, and the relative luciferase activity increased significantly (3.49 x 10(5) vs 0.60 x 10(5), t = 5.92, P<0.01). Exogenous p53 protein also up-regulated p21 promoter driving luciferase expression, comparing to the control group (0.47 x 10(5)), the relative luciferase activity increased (5.63 x 10(5)) obviously (t = 10.12, P<0.01). HCV NS5A protein inhibited both endogenous and exogenous p53 transactivity on p21 promoter in a dose-dependent manner (F > or = 20.71, P<0.01). In the experiment of EMSA, p53 could bind to its specific DNA sequence, but when co-transfected with HCV NS5A expressing vector, the p53 binding affinity to its DNA decreased.

CONCLUSIONHCV NS5A can inhibit p53 protein transactivity on p21 promoter through its inhibiting of p53 binding ability to the specific DNA sequence.

Hepacivirus ; genetics ; Humans ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Tumor Suppressor Protein p53 ; drug effects ; genetics ; metabolism ; physiology ; Viral Core Proteins ; genetics ; Viral Nonstructural Proteins ; genetics ; pharmacology

OBJECTIVETo investigate the influence of miR-122 on IFN-α treatment for HCV infection.

METHODSHuh7.5.1 cells infected with HCV were treated with miR-122 mimics (20 nmol/L, 100 nmol/L, 400 nmol/L) and/or IFN-α (1000 IU/ml). The relative expression of HCV RNA was detected by real-time polymerase chain reaction (PCR). Huh7.5.1 cells were treated with different amounts of HCV (107 copies, 106 copies and 105 copies) and/or IFN-α (1000 IU/ml).

RESULTSIFN-α suppressed the replication of HCV in a time-dependent manner, resulting in a ≊ 83% reduction of HCV at 48 h. MiR-122 mimics facilitated replication of HCV RNA in a dose-dependent manner (P<0.05). The antiviral effect of IFN-α was inverted to levels of miR-122 mimics (20 nmol/L, 100 nmol/L, 400 nmol/L), (73.3% ± 3.5% compared with 84% ± 4.5%, P>0.05; 64.67% ± 5.5% compared with 84% ± 4.5%, P>0.05; 56.33% ± 5.1% compared with 84% ± 4.5%, P<0.05). The antiviral effect of IFN-α was inverted to HCV load (105 copies group compared with 107 copies group, P<0.05).

CONCLUSIONMiR-122 facilitates replication of HCV RNA in the cell culture system; and the expression of miR-122 may partly counteract the anti-HCV effect of IFN-α.

Antiviral Agents ; pharmacology ; Cell Line, Tumor ; Hepacivirus ; drug effects ; genetics ; physiology ; Humans ; Interferon-alpha ; pharmacology ; MicroRNAs ; genetics ; RNA, Viral ; genetics ; Transfection ; Virus Replication ; drug effects ; genetics

OBJECTIVETo investigate the inhibitive effects of small interfering RNA (siRNA) on hepatitis C virus (HCV) replication in cells infected by HCV in vitro.

METHODSThe HCV RNA transcripts prepared by pFL-JC1 were transfected into Huh-7.5.1 cells. Na ve Huh-7.5.1 cells were incubated with the supernatants of transfected cells and the expression of HCV core protein in infected cells was detected by indirect immunofluorescence. The infected cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA for 24 h, 48 h and 72 h, respectively. The normal Huh-7.5.1 cells were transfected with 4, 40 and 200 nmol/L of NS5B siRNA. Group of blank, lipofectamine 2000, unrelated siRNA and IFNα-2b (1000 IU/ml) served as controls. The HCV RNA and PKR mRNA levels were examined by quantitative RT-PCR.

RESULTSThe HCV core protein in HCV infected cells was detected. Compared with control groups, the HCV RNA levels in infected cells significantly decreased when transfected with 40 and 200 nmol/L of siRNA for 24 h; 4, 40 and 200 nmol/L of siRNA for 48 h and 72 h (P<0.05). The HCV RNA levels in infected cells treated with IFNα-2b (1000 IU/ml) for 24 h, 48 h and 72 h were significantly lower than those in control groups (P<0.05 or P<0.01). The PKR mRNA levels in Huh-7.5.1 cells transfected with siRNA of three concentrations did not have significant difference, as compared with control groups (P>0.05).

CONCLUSIONsiRNA against HCV NS5B region can effectively inhibit HCV replication in HCV infected cells, but can not activate the dsRNA-dependent protein kinase (PKR).

Cell Line, Tumor ; Hepacivirus ; drug effects ; genetics ; physiology ; Humans ; RNA, Small Interfering ; pharmacology ; Transfection ; Viral Nonstructural Proteins ; genetics ; Virus Replication ; drug effects

Hepatitis c virus (HCV) infection has become one of the global public health problem,while there is no vaccine to prevent HCV infection, the so-called "cocktail" therapy that use a combination of drugs targeting multiple steps in the HCV infection cycle could achieve better curative effect. the process of HCV entering into host cell is the important step of drug intervention, in which HCV envelope protein El and E2, Host cell factors including Heparan sulfate(HS), CD81, scavenger receptor class B type I (SR-BI), Occludin (OCLD), Claudin (CLDN), low densitity lipoprotein receptor (LDLR), dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN), Liver/lymph node specific ICAM-3-grabbing integrin(L-SIGN), trans- ferrin receptor 1 (TfR1) and so on play a important role. The virus and the host factors can be used as targets of hcv entry inhibitors many studies have shown that as novel and promising compounds, HCV entry inhibitors combinating with other drugs can be more effective in the treatment of HCV, this paper have re- viewed targets and inhibitors of HCV enterring into host cell since 1990s.
Animals ; Antiviral Agents ; pharmacology ; Hepacivirus ; drug effects ; physiology ; Hepatitis C ; genetics ; metabolism ; virology ; Humans ; Receptors, Virus ; genetics ; metabolism ; Viral Envelope Proteins ; genetics ; metabolism ; Virus Internalization ; drug effects