• Title/Summary/Keyword: viral activity

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Construction of a Transgenic Tobacco Expressing a Polydnaviral Cystatin (폴리드나바이러스 유래 시스타틴 유전자 발현 형질전환 담배 제작)

  • Kim, Yeongtae;Kim, Eunsung;Park, Youngjin;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.54 no.1
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    • pp.7-15
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    • 2015
  • CpBV (Cotesia plutellae bracovirus) is a polydnavirus and encodes a cystatin (CpBV-CST1) gene. Its overexpression suppresses insect immunity and alters insect developmental processes. This study aimed to construct a genetically modified (GM) tobacco to further explore the physiological function of the viral cystatin and to apply to control insect pests. To this end, the transgenic tobacco lines were screened in expression of the target gene and assessed in insecticidal activity. A recombinant vector (pBI121-CST) was prepared and used to transform a bacterium, Agrobacterium tumefasciens. The transformed bacteria were used to generate transgenic tobacco lines, which were induced to grow callus and resulted in about 92% of shoot regeneration. The regenerated plants were screened by PCR analysis to confirm the insertion of the target gene in the plant genome. In addition, the expression of the target gene was assessed in the regenerated plants by quantitative real-time PCR (qRT-PCR). The qRT-PCR analysis showed that the transgenic line plant expressed the target gene about 17 times more than the control tobacco, indicating a stable insertion and expression of the target gene in the transgenic tobacco line. The insecticidal activity was then analyzed using the screened transgenic tobacco lines against the teneral 1st instar larvae of the oriental tobacco budworm, Helicoverpa assulta. Though there was a variation in the insecticidal efficacy among transgenic lines, T9 and T12 lines exhibited more than 95% mortality at 7 days after feeding treatment. These results suggest that CpBV-CST1 is a useful genetic resource to be used to generate GM crop against insect pests.

Characterization of compounds and quantitative analysis of oleuropein in commercial olive leaf extracts (상업용 올리브 잎 추출물의 화합물 특성과 이들의 oleuropein 함량 비교분석)

  • Park, Mi Hyeon;Kim, Doo-Young;Arbianto, Alfan Danny;Kim, Jung-Hee;Lee, Seong Mi;Ryu, Hyung Won;Oh, Sei-Ryang
    • Journal of Applied Biological Chemistry
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    • v.64 no.2
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    • pp.113-119
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    • 2021
  • Olive (Olea europaea L.) leaves, a raw material for health functional foods and cosmetics have abundant polyphenols including oleuropein (major bioactive compound) with various biological activities: antioxidant, antibacterial, antiviral, anticancer activity, and inhibit platelet activation. Oleuropein has been reported as skin protectant, antioxidant, anti-ageing, anti-cancer, anti-inflammation, anti-atherogenic, anti-viral, and anti-microbial activity. Despite oleuropein is the important compound in olive leaves, there is still no quantitative approach to reveal oleuropein content in commercial products. Therefore, a validated method of analysis has to develop for oleuropein. In this study, the components and oleuropein content in 10 types of products were analyzed using a developed method with ultra-performance liquid chromatography to quadrupole time-of-flight mass spectrometry, charge of aerosol detector, and photodiode array. The total of 18 compounds including iridoids (1, 3, 4, 14, and 16-18), coumarin (2), phenylethanoids (5, 9, and 11), flavonoids (6-8, 10, 12, and 13), lignan (15), were tentatively identified in the leaves extract based high resolution mass spectrometry data, and the content of oleuropein in each product was almost identical between two detection methods. The oleuropein in three commercial product (A, G, H) was contained more over the suggested content, and it of five products (B, E, H, I, J) were analyzed within 5-10% error range. However, the two products (C, D) were found far lower than suggested contents. This study provides that analytical results of oleuropein could be a potential information for the quality control of leaf extract for a manufactured functional food.

Efficacy of Three Antiviral Agents and Resistant Cultivars on Tomato Yellow Leaf Curl Virus in Tomato (토마토황화잎말림바이러스병에 대한 저항성 품종과 항바이러스 활성 물질 3종의 효과 검증)

  • Kwon, Yongnam;Cha, Byeongjin;Kim, Mikyeong
    • Research in Plant Disease
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    • v.28 no.2
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    • pp.82-91
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    • 2022
  • Recently, several in vitro studies have reported antiviral activity of agents of systemic acquired resistance against plant virus infection, but the approach has not been applied in a wide range of agricultural fields. The objective of this study was to evaluate the inhibitory effect of the exogenous application of salicylic acid (SA), chitosan (CH), or eugenol (EG) in tomato yellow leaf curl virus (TYLCV) infection of greenhouse-grown tomato plants. In vitro, the initial time of symptom was observed in TYLCV-infected plants (VP) of the resistant cultivar 'Superdotaerang' at 12 days post inoculation (dpi) after application of antiviral agents. At 32 dpi, the disease rate of TYLCV in the CHT+VP (0.1% chitosan and virus infected control) treated plants was 87.5%, lower than that of the other treatment. However, the virus content in the CHT+VP treated plants was higher than those of the other treatments, and SA, EG, and CH did not show significant effect on plant height or shoot and root fresh weight. Our results from summer-cultivated greenhouse-grown tomatoes show that none of the tested agents had an inhibitory activity on viral infection or yield of tomato 'Dotaerangsola'cultivar. In contrast, all treated 'TY Giants' cultivars that possessed TYLCV resistance genes Ty-1 and Ty-3a did not show typical symptoms and the virus content was remarkably lower than those in the TYLCV treated plants in 'Superdotaerang'. The results of this research indicated that the planting of resistant tomato cultivars was effective method instead of using SA, EG, and CH (known as resistance-inducing factors for control) of TYLCV in the field.

Anti-tumor Effect of a Combination of Hongyoung Ethanol Extract and Cisplatinin YD-10B Oral Cancer Cells (YD-10B 구강암세포에서 홍영 에탄올 추출물과 시스플라틴 병용에 의한 항암 효과)

  • Eun-Jung Kim;Sung-Hee Hwang;Sangwook Park
    • Journal of Life Science
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    • v.33 no.6
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    • pp.498-505
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    • 2023
  • Solanum tuberosum Linnaeus cv Hongyoung, which represents red potato, was developed in Korea. Hongyoung is known to have anti-oxidant, anti-inflammatory, anti-viral, and anti-tumor properties, but no research has been conducted on the growth inhibition and apoptosis effects of hongyoung in YD-10B oral cancer cells. In this study, the combined treatment of hongyoung ethanol extract (HEE) and cisplatin were examined to determine its ability to inhibit cancer cell growth, induce apoptosis, and inhibit matrix metalloproteinases (MMP)-2 and MMP-9 cancer metastasis. The cell viability was investigated using a 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H- tetrazolium monosodium salt (MTS) assay, and the ability to induce apoptosis was analyzed using an FACS analyzer. The mRNA expression and protein activity of MMP-2 and MMP-9 were measured via RT-PCR and zymography. The YD-10B oral cancer cells showed an increase in growth inhibition as the concentration of HEE increased. The combination of 200 µM cisplatin and 500 ㎍/ml HEE reduced the growth of the YD-10B oral cancer cells by more than 50% compared to cisplatin alone. When phorbol 12-myristate 13-acetate (PMA)-treated YD-10B oral cancer cells were co-treated with 200 µM cisplatin and 500 ㎍/ml HEE, both the mRNA expression and protein activity of the MMP-2 and MMP-9 decreased. In addition, the percentage of the sub-G1 phase, which indicates apoptosis ability, more than doubled when treated in combination with 200 µM cisplatin and 500 ㎍/ml HEE than when cisplatin alone was used. The results of this study therefore suggest the possibility of using a combination of HEE and cisplatin in the development of effective drugs to treat oral cancer.

Chronic HBV Infection in Children: The histopathologic classification and its correlation with clinical findings (소아의 만성 B형 간염: 새로운 병리조직학적 분류와 임상 소견의 상관 분석)

  • Lee, Seon-Young;Ko, Jae-Sung;Kim, Chong-Jai;Jang, Ja-June;Seo, Jeong-Kee
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.1 no.1
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    • pp.56-78
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    • 1998
  • Objective: Chronic hepatitis B infection (CHB) occurs in 6% to 10% of population in Korea. In ethinic communities where prevalence of chronic infection is high such as Korea, transmission of hepatitis B infection is either vertical (ie, by perinatal infection) or by close family contact (usually from mothers or siblings) during the first 5 years of life. The development of chronic hepatitis B infection is increasingly more common the earlier a person is exposed to the virus, particularly in fetal and neonatal life. And it progress to cirrhosis and hepatocellular carcinoma, especially in severe liver damage and perinatal infection. Histopathology of CHB is important when evaluating the final outcomes. A numerical scoring system which is a semiquantitatively assessed objective reproducible classification of chronic viral hepatitis, is a valuable tool for statistical analysis when predicting the outcome and evaluating antiviral and other therapies. In this study, a numerical scoring system (Ludwig system) was applied and compared with the conventional histological classification of De Groute. And the comparative analysis of cinical findings, family history, serology, and liver function test by histopathological findings in chronic hepatitis B of children was done. Methods: Ninety nine patients [mean age=9 years (range=17 months to 16 years)] with clinical, biochemical, serological and histological patterns of chronic HBV infection included in this study. Five of these children had hepatocelluar carcinoma. They were 83 male and 16 female children. They all underwent liver biopsies and histologic evaluation was performed by one pathologist. The biopsy specimens were classified, according to the standard criteria of De Groute as follows: normal, chronic lobular hepatitis (CLH), chronic persistent hepatitis (CPH), mild to severe chronic active hepatitis (CAH), or active cirrhosis, inactive cirrhosis, hepatocellular carcinoma (HCC). And the biopsy specimens were also assessed and scored semiquantitatively by the numerical scoring Ludwig system. Serum HBsAg, anti-HBs, HBeAg, anti-HBe, anti-HBc (IgG, IgM), and HDV were measured by radioimunoassays. Results: Male predominated in a proportion of 5.2:1 for all patients. Of 99 patients, 2 cases had normal, 2 cases had CLH, 22 cases had CPH, 40 cases had mild CAH, 19 cases had moderate CAH, 1 case had severe CAH, 7 cases had active cirrhosis, 1 case had inactive cirrhosis, and 5 cases had HCC. The mean age, sex distribution, symptoms, signs, and family history did not differ statistically among the different histologic groups. The numerical scoring system was correlated well with the conventional histological classification. The histological activity evaluated by both the conventional classification and the scoring system was more severe as the levels of serum aminotransferases were higher. In contrast, the levels of serum aminotransferases were not useful for predicting the degree of histologic activity because of its wide range overlapping. When the histological activity was more severe and especially the cirrhosis more progressing, the prothrombin time was more prolonged. The histological severity was inversely related with the duration of seroconversion of HBeAg. Conclusions: The histological activity could not be accurately predicted by clinical and biochemical findings, but by the proper histological classification of the numerical scoring system for the biopsy specimen. The numerical scoring system was correlated well with the conventional histological classification, and it seems to be a valuable tool for the statistical analysis when predicting the outcome and evaluating effects of antiviral and other therapies in chronic hepatitis B in children.

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Detection of Serum Hepatitis B Virus DNA According to HBV Markers in Chronic Hepatitis B Liver Disease (만성 B형 간질환에서 간염 B virus 표식자 발현에 따른 DNA의 검출)

  • Lee, Dong-Jun;Choi, Jin-Su;Kim, Joon-Hwan;Lee, Heon-Ju
    • Journal of Yeungnam Medical Science
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    • v.14 no.1
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    • pp.155-167
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    • 1997
  • The identification of serum HBV DNA is very important for the assessment of the disease activity in persistent infection, for the evaluation of the infectivity of an individuals blood. The dot blot, however, has limited sensitivity and sometimes inconsistent with other serological markers and clinical settings. Using the most important recent advance in molecular biology, the polymerase chain reaction(PCR), specific DNA sequences can be amplified more than a million-fold in a few hours and with this technique the detection of the extreme low level of DNA is possible. This study was to determine sensitivity of the PCR for the detection of serum HBV DNA in comparison with dot blot analysis and to investigate the serum HBV DNA status and clinical significance of PCR in patients with chronic HBsAg positive liver disease. The subjects of this study were 17 patients with asymptomatic HBsAg carriers(9 HBeAg positive patients, 8 anti-HBe positive patients), 91 chronic hepatitis B(50 HBeAg positive patients, 41 anti-HBe positive patients), 57 liver cirrhosis(21 HBeAg positive patients, 36 anti-HBe positive patients), 27 hepatocellular carcinoma(10 HBeAg positive patients, 17 anti-HBe positive patients). The results were summerized as following; The detection rates of HBV DNA by dot blot, PCR were 58.9%, 72.2% in HBeAg positive patients, 34.3%, 53.9% in anti-HBe positive patients. The detection rates of HBV DNA by PCR in HBeAg negative patients were 25.0% in asymptomatic HBsAg carriers, 61.0% in chronic hepatitis B, 52.8% in liver cirrhosis, 52.9% in hepatocellular carcinoma. The positive rate for HBV DNA is a significant difference between HBeAg positive and negative asymptomatic HBsAg carriers, but not significantly difference in other groups. In conclusions, this study confirmed that the PCR is much more sensitive than the dot blot analysis in detecting the HBV DNA in the sera of patients with chronic liver disease. The presence of HBV DNA in the serum was detected by PCR with higher sensitivity and it suggested that active viral replication is still going on in most patients with chronic HBsAg positive liver disease irrespective of HBeAg/anti-HBe status, and PCR may be used as a prognostic factor in asymptomatic HBsAg carriers.

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Extract from Prunus mume Sieb. et Zucc. Fruit Prevents LPS-induced Homotypic Aggregation of Monocytic THP-1 Cells via Suppression of Nitric Oxide Production and NF-κB Activation (매실 추출물의 산화질소 생성과 NF-κB 활성 조절을 통한 LPS유도성 THP-1 세포 동형성 응집의 억제 효과)

  • Lee, Hye-Rim;Park, Youngsook;Kim, Hyun Jeong;Lee, Aram;Choi, Jihea;Pyee, Jaeho;Park, Heonyong;Kim, Jongmin
    • Journal of Life Science
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    • v.25 no.7
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    • pp.801-809
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    • 2015
  • Homotypic cell adhesion (homotypic aggregation) in activated monocytes plays a central role in physiological and pathological processes including inflammatory responses, differentiation and migration. The extract of the Prunus mume Sieb. et Zucc. fruit (Maesil) has potential benefits to human health; such as anti-viral, anti-microbial, and anti-cancer activities. Indeed, Maesil extract may modulate inflammatory responses via interference with homotypic aggregation in monocytes. In the present study, the molecular mechanisms underpinning the therapeutic efficacy of Maesil extract in inflammatory diseases were investigated. It was found that Maesil extract inhibited homotypic aggregation in lipopolysaccharide (LPS)-activated monocytes. This was mediated by reduction of nitric oxide (NO) production, partly via inhibition of inducible nitric oxide synthase (iNOS) expression in LPS-activated THP-1 cells. It was confirmed that NO inhibition is a key mechanism in Maesil induced blockade of monocyte aggregation through identification of reversal of this inhibitory effect by the NO-producing agent S-nitroso-N-acetyl penicillamine (SNAP). In addition, Maesil extract significantly attenuated LPS-induced IκB-α phosphorylation and NF-κB translocation into the nucleus. In conclusion, Maesil extract exerts anti-inflammatory effects via inhibition of homotypic aggregation of LPS-activated monocytes through mechanisms involving the suppression of NO production and NF-κB activity, suggesting Maesil extract as a potential therapeutic candidate for the prevention and treatment of chronic inflammatory diseases.

Inhibitory Effect on Infection of Plant Viruses by Filtrate Powder from Culture Broth of Acinetobacter sp. and Its Mode of Action (Acinetobacter sp. 배양여과액 분말제제의 식물바이러스에 대한 감염억제 효과 및 작용)

  • Kim Mi-Soon;Jung Min-Young;Kim Yun-Sung;Jang Cheol;Hwang In-Cheon;Ryu Ki-Hyun;Choi Jang-Kyung
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.91-98
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    • 2006
  • A filtrate powder, designated as KNF2022, produced from culture broth of Acinetobacter sp. KTB3 was tested for their inhibitory effects on Pepper mild mottle virus (PMMoV) infection to Nicotiana glutinosa or N. tabacum cv. Xanthi nc. When 1/100 dilution with distilled water was treated to the plants and PMMoV was inoculated, the inhibition was estimated to be 94.3 and 95.6%, respectively. The same concentrations of KNF2022 inhibited infections of Pepper mottle virus (PepMoV) and Cucumber mosaic virus (CMV) on Chenopodium amaranticolor by 97.1 and 92.5%, respectively. Duration of inhibitory activity of the filtrate powder from Acinetobacter sp. culture broth against PMMoV infection on N. glutinosa was maintained for 2 days at 80% inhibition level, however, the inhibitory effect was diminished from 4 days after treatment to 50% levels. To evaluate inhibitory effects on systemic host plants of the antiviral agent, symptom developments of PMMoV, PepMoV and CMV on KNF2022-treated pepper plants were investigated. Delayed symptom developments until 10 days after inoculation (DAI) were observed for all the three viruses when the viruses were inoculated individually, and these delayed symptom development effects were maintained until 30 DAI in case of PepMoV. Moreover, PepMoV was not detected by RT-PCR and ELISA until 30 DAI. These delayed symptom development effects were diminished in all combinations of three virus co-inoculations due to synergism of three viruses on symptom developments. Inhibitory effect of KNF2022 was verified under electron microscopic examinations using purified virus preparations. Particles of PMMoV and PepMoV were observed on specimens from 5 min after KNF2022 treatment, and the particle sizes were reached in the range of 200-250 nm and 400-600 nm, respectively. Furthermore, the viral particles were destructed and particle sizes were reached in the range of 100-150 nm and 300-500 nm, respectively, on 60 min after treatments. Reduction of local lesion numbers on N. tabacum cv. Xanthi nc and C. amaranticolor were accompanied with reduction of virus particle sizes. In the case of CMV destructed particle numbers were also increased according to incubation period after KNF2022 treatment and local lesions on C. amaranticolor were reduced.

Analysis of Physical Status on COVID-19: Based on Impacts of Physical Activity (COVID-19에 대한 운동중재효과 분석)

  • Kim, Kwi-Baek;Kwak, Yi Sub
    • Journal of Life Science
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    • v.31 no.6
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    • pp.603-608
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    • 2021
  • The purpose of this perspective research is to discuss the potential role of exercise-interventions in COVID-19, terms of prevention and prognosis in the periods of the COVID-19 vaccine. SARCO-CoV-2. COVID-19 was detected as a new virus causing severe cardiovascular and respiratory complications. It emerged as a global public health emergency and national pandemic. It caused more than 1 million deaths in the first 6 months of the pandemic and resulted in huge social and economic fluctuations internationally. Unprecedented stressful situations, such as COVID-19 blue and COVID-19 red impact on many health problems. In healthy individuals, COVID-19 infection may induced no symptoms (i.e., asymptomatic), whereas others may experience flu-like symptoms, such as ARDS, pneumonia, and death. Poor health status, such as obesity and cardiovascular and respiratory complications, are high risk factors for COVID-19 prevention, occurrence, and prognosis. Several COVID-19 vaccines are currently in human trials. However, the efficacy and safety of COVID-19 vaccines, including potential side effects, such as anaphylaxis (a life-threatening allergic reaction) and rare blood clots, still need to be investigated. On the basis of direct and indirect evidence, it seems that regular and moderate physical exercise can be recommended as a nonpharmacological, efficient, and safe way to cope with COVID-19. Physical inactivity and metabolic abnormalities are directly associated with reduced immune responses, including reduced innate, CMI, and AMI responses. Due to prolonged viral shedding, quarantine in inactive, obese and disease people should likely be longer than physical active people. Multicomponent and systemic exercise should be considered for the obese, disease, and elderly people. More mechanism research is needed in this area.

Therapeutic Efficacy of Prednisolone Withdrawal Followed by Recombinant ${\alpha}$ Interferon in Children with Chronic Hepatitis B (소아 만성 B형 간염 환자에서 스테로이드 이탈 요법 후 인터페론 병용 투여의 치료 효과)

  • Ryu, Na-Eun;Kim, Byung-Ju;Ma, Jae-Sook;Hwang, Tai-Ju
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.2 no.2
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    • pp.169-177
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    • 1999
  • Purpose: To evaluate the efficacy of interferon alpha therapy with or without prednisolone in children with chronic hepatitis B. Methods: Twenty-eight children (22 boys, 6 girls, mean age 130 months) had seropositive results for HBsAg, HBeAg and HBV DNA; 11 had chronic persistent hepatitis and 17 had chronic active hepatitis. The patients were divided into two groups depending upon their inflammatory activity on liver biopsy, pretreatment serum ALT levels and HBV DNA levels. Fourteen children (group 1: chronic active hepatitis, ALT ${\geq}$ 100 IU/L and HBV DNA ${\leq}$ 100 pg/$300\;{\mu}L$) received interferon alpha 2a 5 $MU/m^2$ of body surface three times weekly for 6 months. Fourteen children (group 2: chronic persistent hepatitis or chronic active hepatitis with ALT < 100 IU/L or HBV DNA > 100 pg/$300\;{\mu}L$) received prednisolone in decreasing daily doses of 60 mg/$m^2$, 40 mg/$m^2$, and 20 mg/$m^2$, each for 2 weeks, followed after 2 weeks by interferon alpha 2a on the same schedule. At the end of therapy, 3 end points were analyzed: HBeAg seroconversion, serum ALT normalization rate and clearance of serum HBV DNA. Results: At the end of treatment, HBe antigen-to antibody seroconversion was higher but not more significant in group 1 than group 2 (71.4% vs. 50.0%). Only one patient in group 2 who lost HBeAg, also cleared HBsAg. ALT normalization was similar in both groups (64.3% in group 1 vs. 55.6% in group 2). Clearance of serum HBV DNA was observed in 78.6% of patients in group 1 and 64.3% in group 2, but no significant differences. Complete response was similarly achieved in both groups (57.1% in group 1 vs. 50.0% in group 2). Interferon alpha therapy with prednisolone priming was well tolerated and all children finished therapy. Conclusion: The combined therapy with prednisolone followed by interferon alpha may be safe and effective in inducing a serological and biochemical remission of the disease in approximately 50% of children with chronic hepatitis B and with a high level of viral replication and less active disease. However, a controlled study should be performed to confirm these results.

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