• Archives of Pharmacal Research
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• v.29 no.11
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• pp.1042-1048
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• 2006

Plasmid DNA vaccines encoding the hepatitis B virus (HBV) surface and hepatitis C virus (HCV) envelope antigens, respectively, were constructed, and attempt were made to find the possibility of a divalent vaccine against HBV and HCV. The expression of each plasmid in Cos-1 cells was confirmed using immunocytochemistry. To measure the induced immune response by these plasmids in vivo, female BALB/c mice were immunized intramuscularly with $100\;{\mu}g$ of either both or just one of the plasmids. Anti-HBV and HCV-specific antibodies and related cytokines were evaluated to investigate the generation of both humoral and cellular immune responses. As a result, specific anti-HBV and anti-HCV serum antibodies from mice immunized with these plasmids were observed using immunoblot. The levels of IL-2 and RANTES showing a $Th_{1}$ immune response were significantly increased, but there was no change in the level of IL-4 ($Th_{1}$ immune response) in any of the immunized groups. Compared with each plasmid DNA vaccine, the combined vaccine elicited similar immune responses in both humoral and cell-mediated immunities. These results suggest that the combined DNA vaccine can induce not only comparable immunity experimentally without antigenic interference, but also humoral and $Th_{1}$ dominant cellular immune responses. Therefore, they could serve as candidates for a simultaneous bivalent vaccine against HBV and HCV infections.

• Childhood Kidney Diseases
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• v.10 no.2
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• pp.162-173
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• 2006

Purpose : To report the decreasing indicence of HBV(Hepatitis B virus)-associated membranous nephropathy in children after HBV vaccination and to elucidate the clinical course and treatment strategies of IMN(Idiopathic membranous nephropathy). Methods : We retrospectively reviewed the clinico-pathological findings of HBV-MN and IMN patients who underwent a renal biopsy from 1986 to 2005. We compared the HBV-MN and the IMN groups and the remission and the non-remission groups of patients with IMN. Results : Among 24 cases of MN patients, HBV-MN comprised 6 cases(25%) and IMN 18 cases(75%). Clinical masnifestations were nephrotic syndrome(3 cases, 50%), nephritic syndrome(1 case, 16.7%), asymptomatic(2 cases, 33.4%) in the HBV-MN group, asymptomatic(10 cases, 55.5%), nephrotic syndrome(5 cases, 27.8%), and gross hematuria(3 cases, 16.7%) in the IMN groups. From 1996 to 2000, there were 2 cases(28%) of HBV-MN and 5 cases(72%) of IMN. After 2001 all 10 cases were IMN. In the HBV-MN group, 4 cases(66.7%) received interferon and 1 cases received methylprednisolone pulse therapy. In the IMN group, 16 cases(88.9%) received methylprednisolone, 8 cases(44.4%) were in complete remission, 2 cases(11.1%) were in partial remission, 2 cases(11.1%) were in chronic renal failure, and 5 cases(27.8%) were lost to follow-up with sustained proteinuria, 1 case(5.6%) continued to have frequent relapse of nephrotic syndrome without renal insufficiency. In the comparison between remission and non-remission groups, nephrotic range proteinuria and hypertension were more significantly common in the non-remission group(P<0.05). Conclusion : With HBV vaccination, HBV-MN has decreased markedly. IMN is a rare glomerular disease in children. Because the prognosis for patients with nephrotic range proteinuria is poor this group needs more aggressive treatment.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.242.1-242.1
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• 2003

The nucleoside analogue, L-FMUS was synthesized from L-FMAU which has been shown to have significant antiviral activity against hepatitis B virus (HBV). It was prepared by two steps. First, 5'hydroxyl of L-FMAU was substituted by thioacetyl group using diisopropylazodicarboxylate(DIAD), Triphenyl phosphine(PPh3) and thioacetic acid in anhydrous THF. (omitted)

• Journal of Korean society of Dental Hygiene
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• v.1 no.2
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• pp.181-192
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• 2001

The purpose of this was to evaluate the level of knowledge about Hepatitis B and AIDS among dental hygiene students at six dental hygiene educational programs in nationwide, compare the findings to those of Song's study. 1703 dental hygiene students surveyed by mail and answered a questionnaire, using the questionnaire used developed by DiClemente el al., on Hepatitis B vaccination, antibody production, experience of unintentional needle stick injuries, experience of surgical operation and blood transfusion, personal risk factors for HBV and HIV, management of HBV and HIV postexposure management et al. The data indicated that dental hygiene students (79.0%) in this study had more HIV vaccination than dental hygiene(74.3%) in Song's study. As for recognition of antibody production after Hepatitis B vaccination, only 34.6% of respondents answered they knew having antibody production and 95% of respondents knew not having antibody production. Over one-half of the respondents (55%) answered they didn't even recognize having antibody production. 52.3 percent of dental hygiene students experienced unintentional needle stick injuries. The findings on the knowledge about Hepatitis B and AIDS were that dental hygienists (76.5 points) received higher mean score than those of dental hygiene students (71.0 points) and that the distribution of right answer rate also showed wider range in dental hygiene students (27.7~97.5%) than those of dental hygienist (41.2~99.5%). There were no statistical differences among dental hygiene education programs. Finding of this study support that the curriculum of dental hygiene program should include instruction on sources and methods of transmission of infectious diseases, risk of virus transmission in the workplace and principles of infection control. Furthermore, infection-control practices should be utilized routinely and their application to dental personnel, management of HBV and HIV postexposure management. Furthermore, for preventing the further spread of infectious diseases (HBV, HIV and AIDS${\cdots}$) caused by bloodborne viruses, imposing an obligation rather than recommendation on Hepatitis B vaccination to all dental personnel and routinely utilizing infection-control guidelines for all dental patients in dental practices (include dental educational programs) should be accomplished by coordination the government agency.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.9
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• pp.5489-5494
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• 2013

In the current study we aimed to show the common YMDD motif mutations in viral polymerase gene in chronic hepatitis B patients during lamivudine and adefovir therapy. Forty-one serum samples obtained from chronic hepatitis B patients (24 male, 17 female; age range: 34-68 years) were included in the study. HBV-DNA was extracted from the peripheral blood of the patients using an extraction kit (Invisorb, Instant Spin DNA/RNA Virus Mini Kit, Germany). A line probe assay and direct sequencing analyses (INNO-LIPA HBV DR v2; INNOGENETICS N.V, Ghent, Belgium) were applied to determine target mutations of the viral polymerase gene in positive HBV-DNA samples. A total of 41 mutations located in 21 different codons were detected in the current results. In 17 (41.5%) patients various point mutations were detected leading to lamivudin, adefovir and/or combined drug resistance. Wild polymerase gene profiles were detected in 24 (58.5%) HBV positive patients of the current cohort. Eight of the 17 samples (19.5%) having rtM204V/I/A missense transition and/or transversion point mutations and resistance to lamivudin. Six of the the mutated samples (14.6%) having rtL180M missense transversion mutation and resistance to combined adefovir and lamivudin. Three of the mutated samples (7.5%) having rtG215H by the double base substituation and resistance to adefovir. Three of the mutated samples (7.5%) having codon rtL181W due to the missense transversion point mutations and showed resistance to combined adefovir and lamivudin. Unreported novel point mutations were detected in the different codons of polymerase gene region in the current HBV positive cohort fromTurkish population. The current results provide evidence that rtL180M and rtM204V/I/A mutations of HBV-DNA may be associated with a poor antiviral response and HBV chronicity during conventional therapy in Turkish patients.

• IMMUNE NETWORK
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• v.4 no.1
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• pp.16-22
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• 2004

Background: To develop a novel treatment strategy for hepatitis B virus infection, a major cause of liver chirosis and cancer, we aimed to make human monoclonal antibodies inhibiting RNase H activity of P protein playing in important role in HBV replication. In this regard, phage display technology was employed and demonstrated as an efficient cloning method for human monoclonal antibody. So this study analysed the usability of human monoclonal antibody as protein based gene therapy. Methods: RNase H of HBV was expressed as fusion protein with maltose binding protein and purified with amylose resin column. Single chain Fv (scFv) phage antibody library was constructed by PCR cloning using total RNAs of PBMC from 50 healthy volunteers. Binders to RNase H were selected with BIAcore 2000 from the constructed library, and purified as soluble antibody fragment. The affinity and sequences of selected antibody fragments were analyzed with BIAcore and ABI automatic sequencer, respectively. And finally RNase H activity inhibiting assay was carried out. Results: Recombinant RNase H expressed in E. coli exhibited an proper enzyme activity. Naive library of $4.46{\times}10^9cfu$ was screened by BIAcore 2000. Two clones, RN41 and RN56, showed affinity of $4.5{\times}10^{-7}M$ and $1.9{\times}10^{-7}M$, respectively. But RNase H inhibiting activity of RN41 was higher than that of RN56. Conclusion: We cloned human monoclonal antibodies inhibiting RNase H activity of P protein of HBV. These antibodies can be expected to be a good candidate for protein-based antiviral therapy by preventing a replication of HBV if they can be expressed intracellularly in HBV-infected hepatocytes.

• Pediatric Infection and Vaccine
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• v.11 no.1
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• pp.73-80
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• 2004

Purpose : The serial clinical findings, biochemical results, and serological hepatitis B virus(HBV) markers in Korean children with chronic HBV infection were analyzed to determine the relationships among these factors. Methods : Ninety children have been chosen from those who have visited to the Department of Pediatrics at St. Vincent's Hospital in The Catholic University of Korea from July 1st, 1995 to June 30th, 2000. The sample patients were followed up for over six months. HBV markers and liver function tests were all performed. Results : All children were asymptomatic at presentation. Eighty-three percent of the children had a history of chronic HBV infection in their families. Eighty-one percent were HBeAg positive, 16% were anti-HBe positive, while 3% were all HBeAg and anti-HBe negative. The prevalence of HBeAg among three age groups : 0~5; 6~10; and 11~15 year-old was 90%, 96% and 61% respectively. The prevalence of HBeAg in less than 10 year-old group was significantly higher than 11~15 year-old group(P=0.001). Serum ALT levels were within 40 IU/L in 64% children, 41~80 IU/L in 17%, 81~200 IU/L in 10%, and beyond 201 IU/L in 9%. The percentage of abnormality of ALT levels in HBeAg positive patients was significantly higher than that of HBeAg negative(P=0.036). Eleven of the 73 HBeAg positive children lost their HBeAg and seroconverted to anti-HBe. In these cases, all had transient elevations in ALT levels before HBeAg seroconversions. The annual rates of spontaneous seroconversion of HBeAg and HBsAg were 9.7% and 0.6%, respectively. Conclusion : Recognition of the dynamics of these changes in viral markers and biochemical findings is needed in the selection and evaluation of therapeutic regimens, establishment of treatment, and calling for controlled trials with adequate follow-up. The hepatitis B carrier state may be asymptomatic in children however, continued surveillance of carriers is important to determine the individual adverse prognostic factors of chronic HBV infections.

• Proceedings of the Korean Society of Veterinary Pathology Conference
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• 2002.11a
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• pp.145-145
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• 2002

Chronic viral infection has been reported to cause a range of hepatic lesion, including hepatic fibrosis, cirrhosis and hepatocellular carcinoma (HCC) in a wide variety of animal species. Woodchucks (Marmota monax) chronically infected with woodchuck hepatitis virus (WHV) develop similar progressive hepatic inflammatory and neoplastic lesions that are remarkably similar to those associated with HBV infection in humans. Twenty two-month-old offspring from woodchucks (Marmota monax) experimentally infected with woodchuck hepatitis virus, were purchased. One randomly chosen animal was autopsied. The liver exhibits marked cirrhotic changes characteristic of the pre-transformation phase of WHV. We believe that this may represent a new suitable and cost-effective model for the disease processes associated with hepadnaviruses in a number of other species, most notably Hepatitis B virus infection in man.

• Journal of Microbiology and Biotechnology
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• v.17 no.4
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• pp.701-704
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• 2007

This report describes the full-length sequences of 2HBV clones from a hepatocellular carcinoma (HCC) patient, one with preC mutation (1896A) and the other without preC mutation. The high level of discrepancy in mutation frequency between these 2 strains was observed in the Core (C) region among 4 ORFs. These data support previous results that Korean HBV strains, belonging to genotype C2, are prone to mutations. It is possible that the mutations (BCP and preC mutations) associated with the HBeAg defective production might contribute to the diversity of mutations related to HBV persistence, playing an important role in hepatocarcinogenesis in this patient.

• The Journal of Korean Medicine
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• v.20 no.3 s.39
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• pp.94-104
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• 1999

Objectives: Hepatoma is a very serious disease in Korea and worldwide. Hepatitis B virus (HBV) has proved the most significant cause of hepatoma. We carried out this study to investigate the effect of Injinchunggan-tang (Yinchenqinggan -tang) on inhibiting cell proliferation and DNA synthesis in HepG2.2.15 cell lines and on inhibiting phosphorilation of oncogene (MAP kinase) in NIT /3T3-HEx cells. Methods: First we confinned the Hepatitis B virus producing ability of HepG2.2.15 cells. To investigate the anti-cancer effect of Injinchunggan-tang (Yinchenqinggan-tang), we did the NTS/PMS assay, [3H]-thymidine incorporation assay and transfection of pcDNA-X. We also measured the gene expression through western blotting. Results: Injinchunggan-tang (Yinchenqing gan tang) showed the suppressing effect of HepG2.2.l5 increase in the MTS/PMS assay and the inhibiting effect of DNA synthesis of HepG2.2.15 in the [3H] thymidine incorporation assay. Injinchunggan-tang (Yinchenqinggan-tang) also showed the inhibiting phosphorilation effect of MAP kinase in HBV -X genes. Conclusions: From the above Injinchunggan-tang (Yinchenqinggan-tang) is thought to have an anti-cancer effect on the hepatoma from HBV. It is suggested that further studies on this prescription would give us a better medicine with an anti-cancer effect.