Kim, Ha-Dong;Choi, Yoon-Chul;Lee, Bum-Yong;Junn, Eun-Sung;Ahn, Jeong-Keun;Kang, Chang-Won;Park, In-Won
BMB Reports
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v.28
no.6
/
pp.509-514
/
1995
A putative secondary structure of the mRNA for the human hepatitis B virus (HBV) X gene is proposed based on not only chemical and enzymatic determination of its single- and double-stranded regions but also selection by the computer program MFOLD for energy minimum conformation under the constraints that the experimentally determined nucleotides were forced or prohibited to base pair. An RNA of 536 nucleotides including the 461-nucleotide HBV X mRNA sequence was synthesized in vitro by the phage T7 RNA polymerase transcription. The thermally renatured transcripts were subjected to chemical modifications with dimethylsulfate and kethoxal and enzymatic hydrolysis with single strand-specific RNase T1 and double strand-specific RNase V1, separately. The sites of modification and cleavage were detected by reverse transcriptase extension of 4 different primers. Many nucleotides could be assigned with high confidence, twenty in double-stranded and thirty-seven in Single-stranded regions. These nucleotides were forced and prohibited, respectively, to base pair in running the recursive RNA folding program MFOLD. The results suggest that 6 different regions (5 within X mRNA) of 14~23 nucleotides are Single-stranded. This putative structure provides a good working model and suggests potential target sites for antisense and ribozyme inhibitors and hybridization probes for the HBV X mRNA.
Hyeokjun Yun;Bo Kyeung Jung;In Soo Rheem;Kap No Lee;Jae Kyung Kim
International Journal of Advanced Culture Technology
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v.11
no.2
/
pp.276-283
/
2023
Purpose: We aimed to determine the differences in the levels of serum thyroid hormone (free T4 [FT4]) and thyroid stimulating hormone [TSH]) as biomarkers for hepatitis B virus (HBV) infection status, with respect to age and sex. Methods: We retrospectively analyzed serum samples from 200 patients who underwent HBV testing from August 2022 to September 2022. Serum samples were collected from patients suspected of having HBV infection who visited this hospital. Thyroid hormone levels were measured, and patients were grouped according to age and sex. Results: Differences in TSH and FT4 levels in the serum of patients in the HBV-positive and -negative groups were not significant. Among the HBV-positive patients in the younger age group (<60 years), TSH and FT4 levels were 1.78 ± 0.09 µIU/mL (normal: 0.4-5.0 µIU/mL) and 1.24 ± 0.02 ng/mL (normal: 0.8-1.9 ng/mL), respectively, whereas among the HBV-positive patients in the older age group (≥60 years), TSH and FT4 levels were 2.22 ± 0.17 µIU/mL and 1.24 ± 0.07 ng/mL, respectively. Conclusions: The presence of HBV did not markedly affect serum thyroid hormone levels. Our findings shed light on the conflicting evidence on the association between thyroid hormone levels and HBV infection. We, Hyeokjun Yun and Bo Kyeung Jung are co-first authors which made substantial contribution equally to the conception and designed of this work. Jae Kyung Kim, In soo Rheem and Kap No Lee made significant contributions to the acquisition and analysis of the data.
Juhee Son;Mi-Jeong Kim;Ji Su Lee;Ji Young Kim;Eunyoung Chun;Ki-Young Lee
IMMUNE NETWORK
/
v.21
no.5
/
pp.37.1-37.17
/
2021
Hepatitis B virus X (HBx) protein has been reported as a key protein regulating the pathogenesis of HBV-induced hepatocellular carcinoma (HCC). Recent evidence has shown that HBx is implicated in the activation of autophagy in hepatic cells. Nevertheless, the precise molecular and cellular mechanism by which HBx induces autophagy is still controversial. Herein, we investigated the molecular and cellular mechanism by which HBx is involved in the TRAF6-BECN1-Bcl-2 signaling for the regulation of autophagy in response to TLR4 stimulation, therefore influencing the HCC progression. HBx interacts with BECN1 (Beclin 1) and inhibits the association of the BECN1-Bcl-2 complex, which is known to prevent the assembly of the pre-autophagosomal structure. Furthermore, HBx enhances the interaction between VPS34 and TRAF6-BECN1 complex, increases the ubiquitination of BECN1, and subsequently enhances autophagy induction in response to LPS stimulation. To verify the functional role of HBx in liver cancer progression, we utilized different HCC cell lines, HepG2, SK-Hep-1, and SNU-761. HBx-expressing HepG2 cells exhibited enhanced cell migration, invasion, and cell mobility in response to LPS stimulation compared to those of control HepG2 cells. These results were consistently observed in HBx-expressed SK-Hep-1 and HBx-expressed SNU-761 cells. Taken together, our findings suggest that HBx positively regulates the induction of autophagy through the inhibition of the BECN1-Bcl-2 complex and enhancement of the TRAF6-BECN1-VPS34 complex, leading to enhance liver cancer migration and invasion.
The familial environment may also play an important role in the epidemiology of HCV infection through vertical and horizontal transmission by infected household members. However, it is still controversial whether familial clustering of HCV occurs. We experienced a case of familial clustering of hepatitis C virus. A 10-year old girl presented with nausea, vomiting and anorexia for a month was diagnosed as hepatitis C. Her mother, grandmother, a maternal aunt and her daughter had contracted with HCV. Her laboratory findings showed AST/ALT 63/122 IU/L, positive anti-HCV Ab and HCV RNA ($3.54{\times}10^5copies/mL$). Pathologic findings of the liver biopsy revealed chronic hepatitis with minimal lobular activity, mild porto-periportal activity and mild portal fibrosis. After treatment with interferon-${\alpha}$ 2b for 6 months, the clinical symptoms and laboratory findings were normalized.
Choi, Young Hee;Lee, Chang Ho;Ko, Myong Suk;Han, Hyun Joo;Kim, Sang Geon
Toxicological Research
/
v.33
no.4
/
pp.343-350
/
2017
Lamivudine belongs to the set of antiviral agents effective against hepatitis B virus infection. Given case reports on liver injuries after certain antiviral agent treatments, this study examined the effects of lamivudine on alanine aminotransferase (ALT) and total bilirubin (TB) using a medical system database. A total of 1,321 patients taking lamivudine alone or with others were evaluated using laboratory hits in an electronic medical system at Seoul National University Hospital from 2005 through 2011. The patients were grouped according to prior ALT results: G#1, ALT < 40 IU/L; G#2, 40 IU/L ${\leq}$ ALT < 120 IU/L; G#3, 120 IU/L ${\leq}$ ALT < 240 IU/L; and G#4, ALT ${\geq}$ 240 IU/L. In G#1 and G#2 patients, lamivudine or adefovir treatment decreased ALT and TB compared to prior values. In G#3 and G#4 patients with three times the upper limit of normal (ULN) ${\leq}$ ALT < 15 times the ULN, both ALT and TB were decreased after treatment with lamivudine alone, or adefovir following lamivudine therapy, indicating that lamivudine therapy ameliorated liver functions. However, in G#4 patients who experienced severely advanced hepatitis (ALT ${\geq}$ 15 times the ULN, or ${\geq}$ 600 IU/L), lamivudine augmented TBmax ($6.3{\rightarrow}13.3mg/dL$) despite a slight improvement in ALT ($839{\rightarrow}783IU/L$), indicative of exacerbation of bilirubinemia. Patients who used adefovir after lamivudine also showed a high incidence of hyperbilirubinemia when they experienced severely advanced hepatitis. Treatment with adefovir alone did not show the effect. In conclusion, lamivudine may increase the risk of hyperbilirubinemia in patients with severely advanced hepatitis, implying that caution should be exercised when using lamivudine therapy in certain patient populations.
The nonstructural protein 5B (NS5B) of hepatitis C virus (HCV) is the viral RNA-dependent RNA polymerase (RdRp), which is the essential catalytic enzyme for the viral replication and is an appealing target for the development of new therapeutic agents against HCV infection. A small amount of serum from a single patient with hepatitis C was used to get the genome of a Korean HCV isolate. Sequence analysis of NS5B 1701 nucleotides showed the genotype of a Korean isolate to be subtype 1b. The soluble recombinant HCV NS5B polymerase lacking the C-terminal 24 amino acids was expressed and purified to homogeneity. With the highly purified NS5B protein, we established in vitro systems for RdRp activity to identify potential polymerase inhibitors. The rhodanine family compounds were found to be potent and specific inhibitors of NS5B from high throughput screening (HTS) assay utilizing the scintillation proximity assay (SPA) system. The binding mode of an inhibitor was analyzed by measuring various kinetic parameters. Lineweaver-Burk plots of the inhibitor suggested it binds not to the active site of NS5B polymerase, but to an allosteric site of the enzyme. The activity of NS5B in in vitro polymerase reactions with homopolymeric RNA requires interaction with multiple substrates that include a template/primer and ribonucleotide triphosphate. Steady-state kinetic parameter, such as Km, was determined for the ribonucleotide triphosphate. One of compounds found interacts directly with the viral polymerase and inhibits RNA synthesis in a manner noncompetitively with respect to UTP. Furthermore, we also investigated the ability of the compound to inhibit NS5B-directed viral RNA replication using the Huh7 cell-based HCV replicon system. The investigation is potentially very useful for the utility of such compounds as anti-hepatitic agents.
Purpose: The purpose of this study is to examine the impacts of infection control education on dental hygienists' perceptions for hepatitis B and their behaviors. Materials and methods: Study participants were chosen by random selection among dental hygienists working in Seoul, Korea. A total of 150 questionnaires were sent out for the survey from April 2013 to May 2013, of which 140-excluding incomplete responses-were used for the study. Chi-square tests and t-tests were used (SPSS 19.0), and post-hoc analysis was performed as well. The maximum significance level was 0.05. Results: Average 1.53 times infection control education was taken, but dental hygienists' perceptions for hepatitis B and their behaviors did not show statistical differences whether the education was done or not. Practices to prevent infection showed differences depending on categories, especially disinfection and sterilization were well performed by educated group. The most common reasons for not taking the education and noncompliance with infection prevention guidelines are lack of time and opportunity due to busy schedule. Conclusion: 1. The more highly educated, the greater number of patients per day, and the greater size of hospitals, the better infection control education was conducted. 2. Although hepatitis B is one of the most common chronic liver diseases in Korea, no significant correlation between perceptions of hepatitis B and infection control education was found. 3. Dental hygienists who received infection control education performed more efficient practices for protection against infections than those who did not.
Purpose : This study was conducted to assess the current(2003-2005) prevalence of anti-HBs and immunologic memory for Hepatitis B vaccine in children from the central area of Korea. Methods : Subjects were chosen from children and adolescents who received tests for hepatitis B surface antigen(HBsAg) and anti-HBs at Dankook University Hospital from March 2003 to May 2005. Among these, antibodies to hepatitis B core antigen(IgG anti-HBc) were checked. A single booster vaccination was performed on children whose anti-HBs titers were under 10 mIU/mL. One month after booster vaccination we rechecked the anti-HBs titer. Results : A total of 3,277 subjects were tested for HBsAg/anti-HBs, and 1,913(58.4 percent) of them were positive for anti-HBs. Of these, 29 subjects(0.9 percent) were positive for HBsAg. Positive results for anti-HBs by age were 78.6 percent for 6-12 months of age, 62.7 percent for 1-3 years of age, 51.9 percent for 4-6 years of age, 49.5 percent for 7-12 years of age, 63.4 percent for 13-15 years of age and 72.2 percent for 16-18 years of age. The 80 subjects who were tested negative for HBsAg/anti-HBs received a single booster vaccine, 71 subjects were tested positive for antibodies. IgG anti-HBc titer was checked for 169 of the subjects, 5 subjects were positive. Conclusion : In our study, a significant anamnestic response was observed in 88.8 percent of children. This is believed to be a result of the relatively long immunologic memory effect of the hepatitis B vaccination in children from the central area of Korea.
Purpose: Lamivudine is known to be effective for the treatment of chronic hepatitis B in adults. However, data on lamivudine therapy in pediatrics is limited. The aim of this study was to evaluate the efficacy and durability of lamivudine therapy for chronic hepatitis B in Korean children. Methods: A total of 44 children (27 males and 17 females, ages 6 months to 14.8 years, mean age 6.7 years) with chronic hepatitis B who received lamivudine (3 mg/kg/day, max 100 mg) for at least 12 months were enrolled. We evaluated the serum AST, ALT and serological HBV markers (HBsAg and anti-HBs, HBeAg and anti HBe, and HBV DNA) periodically. Predictive three year cumulative seroconversion rates were obtained using the Kaplan-Meier method. Results: Twenty one (48%) of 44 children achieved seroconversion of HBeAg by three years, while 23 (42%) children did not. HBV DNA was cleared in 34 (77%) children and the serum ALT levels were normalized in 41 children (93%). The three year cumulative seroconversion rates were 60% for HBeAg, and the clearance rates were 76% for HBV DNA. Eighteen children who discontinued lamivudine after HBeAg seroconversion maintained the therapeutic response for three years (treatment duration 13~58 months mean 24 months). Viral breakthrough developed in 12 children (27%) during the therapy and the YMDD mutation was documented in 11 children (25%). The mean duration for the development of a mutation was 22.7 months. Loss of HBsAg occurred in 6 children (14%). The pretreatment ALT levels were higher in responders; however, the differences were not statistically significant (p>0.05). Conclusion: The results of this study showed that lamivudine treatment had a favorable effect and durable therapeutic response in children with chronic hepatitis B. Long term follow-up and alternative therapy are warranted for those patients who do not respond to this treatment.
Choi, Yujung;Bae, Kil Seoung;Kim, Ki Hwan;Koh, Dae Kyun;Kim, Jong-Hyun
Pediatric Infection and Vaccine
/
v.25
no.2
/
pp.72-81
/
2018
Purpose: This prospective study aimed to investigate the therapeutic efficacy of lamivudine in children with chronic hepatitis B virus (HBV) infection. Methods: During July 2003 through October 2015, children with chronic hepatitis B who visited our institution were included in this study. Fifty-five patients, who received first-line treatment of lamivudine (3 mg/kg, 100 mg maximum) for over three months, were enrolled. After initiating lamivudine, alanine aminotransferase (ALT), HBV-DNA, and HBV markers were followed up at 1 month, 3 months, and every 3 months, thereafter. The treatment endpoint was determined as 1) normalization of ALT, 2) HBeAg seroconversion, and 3) anti-HBe positivity for twelve consecutive months. Results: Thirty-one male (56.4%) and 24 female (43.6%) patients were included. The mean age at treatment initiation was 8.1 years. The mean duration of treatment was 23.4 months. ALT normalization was found in 98.2% (54 of 55). Anti-HBe seroconversion was found in 70.6% (36/51). Loss of HBsAg was found in 10.9% (6/55). All biochemical responses occurred under age seven. The rate of virologic response (defined as HBV-DNA <2,000 IU/mL) at six months after treatment initiation was 78.7% (37/47). At twelve months after reaching treatment endpoint, 87.2% (34/39) maintained their virologic response. Resistance to lamivudine was found in 16.4% (9/55). Conclusions: Lamivudine treatment in Korean pediatric patients with chronic hepatitis B showed better outcomes compared with other studies that implemented similar protocols in foreign populations. Further studies are needed to investigate the efficacy of newly recommended antiviral drugs on the Korean pediatric population.
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