• Animal cells and systems
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• v.5 no.3
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• pp.195-198
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• 2001

Hepatitis B virus (HBV) polymerase, which possesses the activities of terminal binding, DNA polymerase, reverse transcriptase and RNaseH, has been shown to accomplish viral DNA replication through a pregenomic intermediate. Because the HBV polymerase has not been purified, the expression of HBV polymerase was examined in an E. coli expression system that is under the regulation of arabinose operon. The expressed individual domain containing terminal binding protein, polymerase, or RNaseH turned out to be insoluble. The activities of those domains were not able to be recovered by denaturation and renaturation using urea or guanidine-HCI. The expressed reverse transcriptase containing the polymerase and RNaseH domains became extensively degraded, whereas the proteolysis was reduced in a Ion- mutant. These results indicate that Lon protease proteolyzes the HBV reverse transcriptase expressed in E. coli.

• Journal of the Korean Chemical Society
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• v.51 no.1
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• pp.36-42
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• 2007

Hepatitis B is a serious public health problem leading to chronic infection and liver cancer. Quantitation of circulating hepatitis B virus (HBV) is important for monitoring disease progression and for assessing the response to antiviral therapy. In this study, by using Real-Time PCR and novel Micro-PCR assay method, we measured HBV concentration in the clinical sample. A total of 120 serum samples from patients with HBV infection collected was in Dankook university hospital to compare the detection limit, sensitivity, specificity and reproducibility of the two assay methods. These findings of this study suggest that Micro-PCR and Real-Time PCR assay methods are comparable to each other in there detection limit, sensitivity, and reproducibility for HBV DNA quantitation. However, Micro-PCR assay is more efficient than Real-Time PCR method, because Real-Time PCR is not so time - consuming, technically easy and need to reagent of a small quantity. It will be useful for rapid and reliable clinical diagnosis of HBV in many countries.

• IMMUNE NETWORK
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• v.3 no.1
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• pp.23-28
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• 2003

Background: Intracellular antibody specific to hepatitis B virus X protein (HBx) might be useful for studying the role of HBx in hepatocellular carcinogenesis and HBV replication. Methods: With variable region genes for H7 monoclonal anti-HBx Ab, we constructed a vector for bacterial expression of single chain Ab (scFv) and a vector for eukaryotic cell expression of it. The expression of H7 scFv and its binding activity against HBx was examined by immunoblotting and immunofluorescence microscopy. Results: H7 scFv expressed in bacterial cells retained reactivity to HBx. We demonstrated its intracytoplasmic expression in CosM6 eukaryotic cells. Conclusion: This is the first study showing the expression of intracellular anti-HBx Ab in eukaryotic cells. H7 scFv may be a good tool to study the function of HBx in HBV infection.

• BMB Reports
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• v.29 no.2
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• pp.175-179
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• 1996

Viral surface proteins are known to play an essential role in attachment of the virus particle to the host cell membrane. In case of the hepatitis B virus (HBV) several reports have described potential receptors on the target cell side, but no definite receptor protein has been isolated yet. As for the viral side, it has been suggested that the preS region of the envelope protein, especially the preS1 region, is involved in binding of HBV to the host cell. In this study, preS1 region was recombinantly expressed in the form of a maltose binding protein (MBP) fusion protein and used to identify and visualize the expression of putative HBV receptor(s) on the host cell. Using laser scanned confocal microscopy and by FACS analysis, MBP-preS1 proteins were shown to bind to the human hepatoma cell line HepG2 in a receptor-ligand specific manner. The binding kinetic of MBP-preS1 to its cellular receptor was shown to be temperature and time dependent. In cells permeabilized with Triton X-100 and treated with the fusion protein, a specific staining of the nuclear membrane could be observed. To determine the precise location of the receptor binding site within the preS1 region, several short overlapping peptides from this region were synthesized and used in a competition assay. In this way the receptor binding epitope in preS1 was revealed to be amino acid residues 27 to 51, which is in agreement with previous reports. These results confirm the significance of the preS1 region in virus attachment in general, and suggest an internalization pathway mediated by direct attachment of the viral particle to the target cell membrane.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.23 no.4
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• pp.311-318
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• 2020

Chronic hepatitis B viral (HBV) infection remains a major health threat, especially in high-prevalence areas. Most infants infected by mother-to-infant HBV transmission become chronic carriers. In Taiwan, many important preventive interventions have been implemented to block the perinatal transmission of HBV in the past 35 years. The first nationwide universal HBV vaccination program was launched in Taiwan in July 1984. The three-dose HBV vaccine completion rate reached 98.1% in 2018. The prevalence of Hepatitis B surface antigen (HBsAg) decreased from 9.8% in pre-vaccinated period in 1984 to 0.5% in the vaccinated cohort in 2014. The incidence of hepatocellular carcinoma in children aged 6-9 years significantly declined from 0.52 to 0.13 per 100,000 children born before and after 1984, respectively. Furthermore, we have performed a maternal HBV screening program during pregnancy since 1984, with the screening rate peaked at 93% in 2012. The HBsAg- and HBeAg-seropositive rate in pregnant women declined from 13.4% and 6.4% in 1984-1985 to 5.9% and 1.0% in 2016, respectively. To closely control perinatal HBV infection, we have administered hepatitis B immunoglobulin immediately after birth and checked the serum level of HBsAg and anti-HBs in high-risk babies born to HBsAg-seropositive mothers, irrespective of their HBeAg status, since July 2019. We have also adopted short-term antiviral treatments such as tenofovir 300 mg daily in the third trimester for highly viremic mothers and reduced the perinatal infection rates from 10.7 to 1.5%. Through all these efforts, we expect to meet the global goal of eliminating HBV infection by 2030.

• Journal of Microbiology
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• v.45 no.6
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• pp.528-533
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• 2007

In a previous study we generated an anti-Hepatitis B Virus (HBV) preS1 humanized antibody (HzKR127) that showed in vivo HBV-neutralizing activity in chimpanzees. However, the antigen-binding affinity of the humanized antibody may not be sufficient for clinical use and thus affinity maturation is required for better therapeutic efficacy. In this study, phage display technique was employed to increase the affinity of HzKR127. All six amino acid residues (Glu95-Tyr96-Asp97-Glu98-Ala99-Tyr100) in the heavy (H) chain complementary-determining region 3 (HCDR3) of HzKR127 were randomized and phage-displayed single chain Fv (scFv) library was constructed. After three rounds of panning, 12 different clones exhibiting higher antigen-binding activity than the wild type ScFv were selected and their antigen-binding specificity for the preS1 confirmed. Subsequently, five ScFv clones were converted to whole IgG and subjected to affinity determination. The results showed that two clones (B3 and A19) exhibited an approximately 6 fold higher affinities than that of HzKR127. The affinity-matured humanized antibodies may be useful in anti-HBV immunotherapy.

• Asian Pacific Journal of Cancer Prevention
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• v.16 no.1
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• pp.35-39
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• 2015

Background: Hepatocellular carcinoma (HCC) is one of the most frequent cancers in South East Asian countries including Cambodia, where prevalence of chronic carriers of hepatitis B and C virus (HBV and HCV) is reported to be very high. We reviewed HCC cases admitted to a cancer hospital in Phnom Penh, which is the only one hospital for cancer treatment and care in Cambodia during the study period. Materials and Methods: Information was collected from medical records of 281 cases (210 males and 71 females) diagnosed as primary HCC from 2006 to 2011. Results: The subjects were 7-81 years old with a median age of 53 years. Hypochondriac pain was the most common complained symptom (74%). One third of the cases presented with jaundice. Nearly half had ascites at their first visit. One third had liver cirrhosis. Nearly three fourths of the cases presented with tumor sized more than 50 mm in diameter, and in almost all cases (97.4%) the size was more than 20 mm. Among 209 subjects tested, hepatitis virus carriers were 75.6%; 46.4% for HBV only, 21.5% for HCV only, and 7.7% for both viral infections. Median age of patients with HBV was about ten years younger than those with HCV. Conclusions: This study revealed the characteristics of HCC cases in Cambodia, although there were several limitations. Most HCC cases were infected with HBV and/or HCV, and diagnosed at late stages with complications. This implicated that public health intervention to prevent HBV and HCV infection is of high priority.

• Clinical and Experimental Pediatrics
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• v.50 no.9
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• pp.823-834
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• 2007

Interferon (IFN) alpha has been the first line therapy of chronic hepatitis B in children, but HBeAg seroconversion occurred in 26% of treated children compared to 11% of controls in multinational randomized controlled study. Recently, lamivudine was shown to be a potent inhibitor of Hepatitis B virus (HBV) reproduction both in HBeAg positive and in HBeAg negative (the pre-core mutant form) chronic hepatitis in randomized studies worldwide. Lamivudine therapy led to considerable improvement in the seroconversion rate of HBeAg in children with chronic hepatitis B, though long-term therapy resulted in the expansion of lamivudine-resistant mutant viruses. Combination therapy with lamivudine plus alpha-IFN does not seem to improve HBe Ag seroconversion. Above all, the most effective way to prevent hepatitis B is universal HBV vaccination.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.10 no.2
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• pp.166-172
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• 2007

Purpose: It has recently been reported that de novo HBV infection following liver transplantation is caused by grafts from HBcAb positive donors, and this phenomenon has been observed in one third of the liver transplant patients in our center. Therefore, we investigated the presence of HBV virus DNA in liver tissues obtained from HBcAb positive donors to determine the mechanism by which de novo HBV infection occurs. Methods: This study was conducted on 6 patients that were HBsAg negative, HBsAb positive, and HBcAb positive who were donors for liver transplantation between November 1997 and November 1998 at Asan Medical Center. We isolated DNA from a portion of liver biopsy tissues that were obtained during the operation, and then identified the surface and core region of HBV DNA using nested PCR. In addition, four children who received liver grafts from these donors were monitored to determine if they became afflicted with non-HBV related diseases while receiving prophylaxis consisting of short-term HBIG treatment and long-term treatment with an antiviral agent. Results: The surface antigen region was identified in all 6 donors and the core antigen region was observed in 4 of the 6 donors. However, no episodes of de novo HBV infection with prophylaxis were observed. Conclusion: The results of this study support the results of previous studies, which indicated that HBV infection may be the main cause of de novo HBV infection in patients that receive HBsAb positive and HBcAb positive donor grafts.

• Korean Journal of Clinical Laboratory Science
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• v.43 no.2
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• pp.57-67
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• 2011

In this study, we investigated the correlation between the administration of various antiviral agents and the alternation of specific biomarkers induced by the hepatitis B virus (HBV). Eligible subjects diagnosed with chronic hepatitis B were prescribed with antiviral drugs at the Gastroenterology Internal Medicine Department of E University Hospital in Daejeon between May 2004 and September 2009. Lamivudine was prescribed to 66 out of 100 patients. Of the 12 patients, 6 (50.0%) showed a change from being HBe-antigen-positive to being HBe-antigen-negative. Of the 39 patients, 23 (59.0%) showed higher than 40 IU/L alanine aminotransferase (ALT). Of the 65 patients, 41 (63.1%) showed HBV DNA decrease of 1 log, and were prescribed with Lamivudine. Adefovir was prescribed to 3 out of 100 patients. Of the 12 patients, 1 (8.3%) showed a change from being HBe-antigen-positive to being HBe-antigen-negative, and was prescribed with Adefovir. Entecavir was prescribed to 19 (19.0%) out of 100 patients. Of the 12 patients, 3 (25.0%) showed a change from being HBe-antigen-positive to being HBe-antigen-negative. Of the 12 patients, 3 (125.8%) showed higher than 40 IU/L ALT. Of the 65 patients, 14 (21.5%) showed HBV DNA decrease of 1 log, and were prescribed with Entecavir. Clavudine was prescribed to 7 out of 100 patients. Of the 12 patients, 1 (8.3%) showed a change from positive HBe antigen to negative HBe antigen. Of the 39 patients, 5 (12.8%) showed higher than 40 IU/L ALT. Of the 65 patients, 6 (9.2%) showed HBV decrease of 1 log, and were prescribed with Clavudine. These results do not show a statistically significant correlation between drugs and biomarkers. Data on combination therapy using Lamivudine and Adefovir show no statistically significant difference between drugs and biomarkers. Medications for periodic inspection was not correlated to HBe-antigen-negative conversion, ALT, and HBV DNA. HBV DNA was significantly reduced in patients with high levels of AST(aspartic acid aminotransferase) and ALT before treatment. In addition, the decrease of HBV DNA after 12 months of treatment was less frequently observed in patients treated with Lamivudine compared with other drugs. This result is associated with Lamivudine resistance. Although the association of drugs with diagnostic markers and the correct choice of treatment is difficult to determine, these results may be useful for further research on diagnosis and treatment of the hepatitis B virus.