BACKGROUND/AIMS: To analyze the effects of preexisting lamivudine (LAM) resistance and applying antiviral treatment (adefovir [ADV] add-on LAM combination treatment) on long-term treatment outcomes, and comparing the clinical outcomes of antiviral-naïve chronic hepatitis B patients receiving entecavir (ETV) monotherapy. METHODS: This study enrolled 73 antiviral-naïve patients who received 0.5-mg ETV as an initial therapy and 54 patients who received ADV add-on LAM combination treatment as a rescue therapy from July 2006 to July 2010. RESULTS: During 24-month treatments, the decreases in serum log10HBV-DNA values (copies/mL) were significantly greater in the antiviral-naïve patients treated with ETV than the patients receiving ADV add-on LAM combination treatment. The biochemical response rates for alanine aminotransferase normalization at 6 months (ETV) and 12 months (ADV add-on LAM) were 90.4% (66/73) and 77.8% (42/54), respectively (P=0.048). A Kaplan-Meier analysis indicated that the rates of serologic response, viral breakthrough, and emergence of genotypic resistance did not differ significantly between the two patient groups. There were also no significant intergroup differences in the rates of disease progression (PD) and new development of hepatocellular carcinoma (HCC). CONCLUSION: The long-term clinical outcomes of antiviral-naïve patients treated with ETV and LAM-resistant patients receiving ADV add-on LAM combination treatment were comparable in terms of the emergence of HCC and disease progression.
Adenine/*analogs & derivatives/pharmacology/therapeutic use ; Adult ; Alanine Transaminase/blood ; Antibodies, Viral/blood ; Antiviral Agents/*therapeutic use ; DNA, Viral/blood ; Disease Progression ; Drug Resistance, Viral/drug effects ; Drug Therapy, Combination ; Female ; Follow-Up Studies ; Genotype ; Guanine/analogs & derivatives/pharmacology/therapeutic use ; Hepatitis B e Antigens/blood ; Hepatitis B virus/drug effects/genetics/isolation & purification ; Hepatitis B, Chronic/*drug therapy ; Humans ; Lamivudine/pharmacology/therapeutic use ; Male ; Middle Aged ; Organophosphonates/pharmacology/*therapeutic use ; Treatment Outcome

BACKGROUND/AIMS: The optimal timing for discontinuing oral antiviral therapy in patients with HBeAg-positive chronic hepatitis B (CHB) is unclear. The aim of our study was to investigate sustained remission after stopping antiviral therapy in patients with HBeAg-positive CHB. METHODS: We analyzed the medical records of 58 patients who were HBeAg-positive and had discontinued antiviral therapy. Antiviral therapy was discontinued after HBeAg seroconversion and HBV DNA negativity for 6-12 months with consolidation therapy. Virologic relapse was defined as an increase in serum HBV DNA >2,000 IU/mL. RESULTS: No difference was observed between the virologic non-relapse and virologic relapse groups in baseline HBV DNA level (p=0.441) or duration of seroconversion (p=0.070). Time-to-undetectable HBV DNA during treatment was shorter in the virologic non-relapse group (29 patients) compared to the relapse group (29 patients) (4.9+/-2.6 vs. 13.2+/-12.7 months; p<0.01). Cumulative relapse rates were 12.7 in month 3, 32.7 in month 6, 47.3 in month 12, and 52.7% in month 18. We determined by multivariate analysis that the consolidation period (> or =18 months, p=0.020) and early virologic response (HBV DNA <20 IU/mL) at six months during antiviral therapy (p=0.017) were significant predictors for sustained remission. CONCLUSIONS: A consolidation period of at least 18 months and early virological response at six months during antiviral therapy were associated with sustained remission in patients with HBeAg-positive CHB after treatment.
Adult ; Aged ; Antiviral Agents/*therapeutic use ; DNA, Viral/analysis ; Female ; Hepatitis B e Antigens/*blood ; Hepatitis B virus/genetics/isolation & purification ; Hepatitis B, Chronic/*drug therapy ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Proportional Hazards Models ; Recurrence ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Withholding Treatment

In the present study, the genomic sequence characteristics of HN-JY40 strains of the hepatitis E virus (HEV) isolated from pigs in Henan Province, China, were analyzed and the evolutionary relationship between HN-JY40 and other sequenced strains examined. The whole genome of HN-JY40 was sequenced and analyzed by reverse transcription-polymerase chain reaction, 3' rapid amplification of cDNA ends (3' RACE) and 5' RACE. Bioinformatic analyses were carried out with Megalign, Expasy, clustal x, and MEGA 4 software. The genome of HN-JY40 was 7 223 bp in size upon removal of polyA sequences. Sizes were 9 bp and 69 bp at 5' and 3' noncoding regions, respectively. The genome of HN-JY40 was predicted to contain three open reading frames (ORFs): ORF1 (5 124 bp) encoding 1 707 amino acids; ORF2 (2 025 bp) encoding 674 amino acids; ORF3 (345 bp) encoding 114 amino acids. Phylogenetic-tree analyses indicated that HN-JY40 is a typical type-IV virus that belongs to a new subgenotype of HEV genotype 4. We sequenced and analyzed the whole genome of HN-JY40. This strategy elicited the genomic characteristics of the HEV isolated from pigs in Henan Province as well as the evolutionary relationships between HN- JY40 and other HEV isolates from pigs. We revealed that the ORF1 of HN-JY40 (153-432 nt) and human HK 104-2004 had high similarity, which offers molecular evidence for uncovering the interspecies transmission of the HEV.
Amino Acid Sequence ; Animals ; Base Sequence ; China ; Cloning, Molecular ; Genome, Viral ; Hepatitis E ; veterinary ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; Sequence Homology, Amino Acid ; Swine ; Swine Diseases ; virology

OBJECTIVETo explore the effect of the cytoplasmic DNA sensor DAI on replication of hepatitis B virus (HBV) and its possible mechanism.

METHODSThe hepatocyte-derived cell line HepG2 was co-transfected with DAI siRNA and the HBV1.3 replicative plasmid PHY106, and the cells were divided into two experimental groups. Six hours later, total RNA was extracted from the first group of cells and expression of IFIT1 and IL-6 were detected by real-time RT-PCR. The second group of cells was incubated for 4 days, after which the cell supernatant was collected and the HBV surface antigen (HBsAg) and envelope antigen (HBeAg) were detected by ELISA. In addition, HBV core particles were extracted and applied to southern blot assay to detect the intracellular HBV replication intermediates (rcDNA, dlDNA and ssDNA). Next, the HepG2 cells were triple transfected with siRNA targeting the type I interferon pathway molecule TBK1 and DAI simultaneously and HBV1.3, after which HBV viral proteins were detected. Two-group comparisons were made using the independent sample t-test, and more-than-2-group comparisons were made using ANOVA.

RESULTSDAI gene expression was down-regulated in response to DAI siRNA transfection. Cells with down-regulated DAI showed inhibited HBV replication (in a dose-dependent manner), accompanied by reduced levels of HBsAg (0.0195+/-0.0050 vs.

CONTROL0.3150+/-0.0200, P less than 0.05, t = 14.77) and HBeAg (0.0140+/-0.0040 vs.

CONTROL0.01235+/-0.0135, P less than 0.05, t = 7.777). No effect of down-regulated DAI was observed for the expression of IFIT1 of IL-6. siRNA-mediated down-regulation of TBK1 and DAI simultaneously led to reduced expression of HBsAg and HBeAg.

CONCLUSIONDown-regulation of DAI gene expression inhibited HBV replication and HBV protein expression, but the underlying mechanism was not related to the type I interferon or NF-kB signaling pathway.

Carrier Proteins ; metabolism ; DNA-Binding Proteins ; genetics ; metabolism ; Down-Regulation ; Gene Expression Regulation ; Hep G2 Cells ; Hepatitis B Surface Antigens ; isolation & purification ; Hepatitis B e Antigens ; isolation & purification ; Hepatitis B virus ; physiology ; Humans ; Interleukin-6 ; metabolism ; NF-kappa B ; metabolism ; Plasmids ; RNA, Small Interfering ; genetics ; Signal Transduction ; Transfection ; Virus Replication

Hepatitis E, caused by hepatitis E virus (HEV) infection, usually leads to an acute clinical course, and is the most common diagnosis among cases of acute viral hepatitis. From 2008, there have been increasing reports of chronic HEV infection in immunocompromised patients such as organ transplant recipients. Without intervention with antiviral treatment, approximately 60% of HEV infections in organ transplant recipients evolve into chronic HEV infections. Of these chronic hepatitis E patients, 10% may develop liver fibrosis and progress to liver cirrhosis. This article reviews chronic HEV infection and treatment in organ transplant recipients.
Animals ; Antiviral Agents ; therapeutic use ; Hepatitis E ; drug therapy ; virology ; Hepatitis E virus ; genetics ; isolation & purification ; physiology ; Hepatitis, Chronic ; drug therapy ; virology ; Humans ; Transplant Recipients ; Transplants ; virology

BACKGROUND: Most mutations in the reverse transcriptase (RT) gene of the hepatitis B virus (HBV) are related to resistance to antiviral agents. Cross-sectional studies on the mutations of this gene are rare. Thus, we analyzed the mutation patterns of RT genes and their biochemical parameters. METHODS: From 2009 to 2012, 301 blood specimens from patients with chronic hepatitis B at Daegu Catholic University Medical Center were retrospectively analyzed for the RT gene sequence of HBV, ALT, hepatitis B e antigen (HBeAg), and HBV DNA. The mutation patterns of the RT gene were compared with the biochemical parameters. RESULTS: Of the 301 patients, 100 (33.2%) had no RT gene mutations. The remaining showed the following mutation patterns: rtM204I/V (50.2%), rtL180M (39.2%), and rtA181T/V (19.6%). Combined mutations were found in 146 cases (48.5%). Of these, the combination of amino acid changes at rt180+rt204 (49.3%) was most frequently detected, followed by rt181+rt236 (11.0%) and rt173+rt180+rt204 (9.6%). In the mutated group, HBV DNA and HBeAg positive rates were significantly higher (P<0.05 for both). Phenotypic analysis showed that lamivudine resistance was most frequently detected (34.6%), followed by adefovir resistance (15.6%). Multidrug resistance was detected in 48 cases (15.9%). The adefovir-resistant group had a higher proportion of cases with HBV loads greater than 2,000 IU/mL. CONCLUSIONS: We found correlations between the mutation status of the RT domain and biochemical parameters such as HBV DNA and HBeAg positive rate. The presence of RT gene mutations could therefore be utilized to predict clinical status.
Adenine/analogs & derivatives/therapeutic use ; Antiviral Agents/therapeutic use ; DNA, Viral/analysis ; Drug Resistance, Multiple, Viral ; Drug Resistance, Viral ; Hepatitis B e Antigens/blood ; Hepatitis B virus/*enzymology/isolation & purification ; Hepatitis B, Chronic/drug therapy ; Hospitals, University ; Humans ; Lamivudine/therapeutic use ; Mutation ; Organophosphonates/therapeutic use ; Phenotype ; RNA-Directed DNA Polymerase/*genetics ; Republic of Korea ; Retrospective Studies

Swine hepatitis E virus (HEV) is widespread throughout pigs in both developing and industrialized countries. This virus is an important zoonotic agent and a public concern worldwide. Infected pigs are asymptomatic, so diagnosing swine HEV relies on detection of the virus or antibodies against the virus. However, several obstacles need to be overcome for effective and practical serological diagnosis. In this study, we developed an enzyme-linked immunosorbent assay (ELISA) that used a purified recombinant capsid protein of swine HEV. The potential clinical use of this assay was evaluated by comparing it with a commercial kit (Genelabs Technologies, Diagnostics, Singapore). Results of the ELISA were highly correlated with those of the commercial kit with a sensitivity of 97% and specificity of 95%. ROC (receiving operator characteristic) analysis of the ELISA data produced a value of 0.987 (95% CI, 0.977~0.998, p < 0.01). The cut-off value for the ELISA was also determined using negative pig sera. In summary, the HEV-specific ELISA developed in the present study appears to be both practical and economical.
Animals ; Antibodies, Anti-Idiotypic/*analysis/blood/genetics ; Capsid Proteins/*genetics/metabolism ; Enzyme-Linked Immunosorbent Assay/*methods/veterinary ; Hepatitis E/diagnosis/immunology/*veterinary/virology ; Hepatitis E virus/genetics/*isolation & purification/metabolism ; Immunoglobulin G/blood/genetics ; ROC Curve ; Recombinant Proteins/genetics/metabolism ; Swine ; Swine Diseases/*diagnosis/immunology/virology

The recent increase in the number of cases of indigenous hepatitis E virus (HEV) infection highlights the importance of identifying the transmission routes for the prevention of such infections. Presented herein is the first case of acute HEV infection after ingesting wild roe deer meat in South Korea. A 43-year-old male presented with abdominal discomfort and jaundice. He had not recently traveled abroad, but had eaten raw roe-deer meat 6-8 weeks before the presentation. On the 7th day of hospitalization the patient was diagnosed with acute viral hepatitis E. Phylogenetic analysis of his serum revealed genotype-4 HEV. This case supports the possibility of zoonotic transmission of HEV because the patient appears to have been infected with genotype-4 HEV after ingesting raw deer meat.
Adult ; Alanine Transaminase/blood ; Animals ; Bilirubin/blood ; Deer/virology ; Genotype ; Hepatitis E/*diagnosis/transmission/virology ; Hepatitis E virus/classification/*genetics/isolation & purification ; Humans ; Male ; Phylogeny ; RNA, Viral/analysis ; Republic of Korea ; Travel

OBJECTIVETo determine the genotype and phylogenetic characteristics of hepatitis E virus (HEV) strains isolated from the human and swine in Anqing City.

METHODSTwenty seven sera from sporadic hepatitis E patients and 400 commercial swine bile samples were collected in Anqing City. According to the collection time, the bile samples were equally divided into 4 groups which were named group A, B, C and D respectively. Nested reverse transcription polymerase chain reaction (RT-PCR) and DNA sequencing technology were performed to obtain the DNA sequences of HEV RNA Open Reading Frame 2 (ORF2) (150 nt) for all the serum and bile samples. The sample sequences and prototype sequences from the GenBank were aligned and their nucleotide sequence identities were calculated. A phylogenetic tree constructed according to the Bayesian inference method was used to analyze the genotype and phylogenetic relationship between the human and swine HEV strains isolated in Anqing City.

RESULTSThe male-to-female sex ratio of the patients was 2.86:1 and the average age was 56.78 years old. Sixteen out of 27 serum (59.26%) samples were HEV RNA positive. Human HEV strains isolated in Anqing City shared 74.75% - 82.99%, 75.26% - 83.64%, 72.77% - 80.57% and 88.03%-91.63% nucleotide sequence identities with prototype I, II, III and IV HEV strains respectively. HEV RNA was detected in 22 out of 400 bile samples (5.5%). The swine HEV detection rates for group A, B, C and D were 7.00%, 3.00%, 9.00% and 3.00% respectively, showing no significant difference among these groups (χ(2) = 5.20, P = 0.16). Swine HEV strains isolated in Anqing City shared 75.24% - 83.42%, 75.93% - 84.19%, 72.86% - 80.64% and 88.15% - 91.79% nucleotide sequence identities with prototype I, II, III and IV HEV strains respectively. Phylogenetic analysis revealed that all the HEV strains isolated from both the human and swine belonged to genotype IV and scattered in evolutionary branches without significant species aggregation.

CONCLUSIONIt's suggested that genotype IV HEV was the dominant genotype among the human and swine in Anqing City and probably transmitted between them in this area.

Aged ; Animals ; Base Sequence ; China ; epidemiology ; Female ; Genotype ; Hepatitis E ; epidemiology ; veterinary ; virology ; Hepatitis E virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Phylogeny ; Swine ; virology ; Swine Diseases ; epidemiology ; virology

OBJECTIVETo analyze antiviral effects of entecavir in patients with hepatitis B virus-related cirrhosis.

METHODS104 patients of hepatitis B virus-related cirrhosis with no previous history of antiviral therapy were treated with entecavir 0.5 mg once daily. 37 patients were taken hepatic histologic examination before and after the treatment.

RESULTSMean reductions of serum HBV DNA was 5.1 log10 96 weeks after the treatment, HBV DNA became undetectable in 98.1% patients, and ALT became normal in 80.7% patients; HBeAg seroconversion occurred in 13.9% of the 72 HBeAg positive patients; 61.5% of these patients were infected with genotype C HBV, and 26.9% were infected with genotype B HBV. The genotype of HBV was not associated with the therapeutical effect. Child-pugh score was associated with the progression of the disease: the proportion of patients with disease progression was highest in Child-Pugh C grade patients and lowest in Child-Pugh A grade patients. The level of the HBV DNA load was positively correlated with Knodell HAI score at the baseline and 96 weeks after the treatment.

CONCLUSIONEntecavir treatment results in suppression of HBV replication and delayed progression of fibrosis in patients with hepatitis B virus-related cirrhosis.

Adult ; Alanine Transaminase ; blood ; Antiviral Agents ; therapeutic use ; DNA, Viral ; blood ; Female ; Genotype ; Guanine ; analogs & derivatives ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; drug effects ; genetics ; isolation & purification ; Hepatitis B, Chronic ; complications ; drug therapy ; virology ; Humans ; Liver Cirrhosis ; drug therapy ; etiology ; virology ; Male ; Middle Aged ; Time Factors ; Treatment Outcome ; Virus Replication ; drug effects