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ROH, HANG SIK,SONG, HYE MIN,YUN, BO REUM,KANG, HYUN KYUNG,CHOI, KEUM SUK,PARK, YUN JU,KIM, DONG SUB,KIM, SEUNG HEE,MO, IN PIL,AN, BEUM-SOO,AHN, CHI YOUNG Spandidos Publications 2015 MOLECULAR MEDICINE REPORTS Vol.11 No.4
<P>Single radial immunodiffusion (SRID) assay requires a reference antigen and an antibody to the hemagglutinin (HA) of an influenza vaccine. As it takes 2???3 months to develop the reference antigen, vaccine development is delayed in cases of an influenza pandemic. In the present study, the measurement of the HA content of influenza vaccines was assessed using size exclusion high performance liquid chromatography (SE???HPLC) for the rapid development of a pandemic vaccine. When the 2009 H1N1 reference antigen, pandemic 2009 H1N1 vaccine and 2010 seasonal influenza vaccines were analyzed by SE???HPLC, the HA of the reference antigen and vaccines was specifically separated. The presence and specificity of HA were evidenced with immunoprecipitation and ELISA assays. For the influenza vaccines, the chromatogram pattern and retention time of HA were similar among the antigen types (2009 H1N1, 2010 H3N2 and 2010 B). In addition, when SE???HPLC was applied, the ratio of HA chromatogram to peak area revealed a significant correlation with HA concentration for the reference antigen and vaccine. The result of the HA content calculation based on SE???HPLC exhibited 99.91???100% similarity, compared with that of SRID. These findings suggest that the measurement of peak area ratio/HA content using SE???HPLC may be a substitute for SRID and rapidly measure HA content to enable faster development of a vaccine during an influenza pandemic.</P>
원지(Root of Polygala teunifolia Willd)추출물의 급성 경구투여 독성 연구
노항식 ( Hang Sik Roh ),정자영 ( Ja Young Jeong ),석지현 ( Ji Hyun Seok ),하헌용 ( Hun Yong Ha ) 대한한방부인과학회 2013 大韓韓方婦人科學會誌 Vol.26 No.4
In this study, it was carried out to evaluate the acute oral toxicity of Root of Polygala teunifolia Willd. in Sprague-Dawley (SD) rats. Methods: Male and female rats were administered orally with Root of Polygala teunifolia Willd. water extract of 1,000 mg/kg (low dosage group), 2,000 mg/kg (middle dosage group) and 4,000 mg/kg (high dosage group). We daily observed number of deaths, clinical signs and gross findings for 7 days. After 7 days, we measured body and organs weight. Also we analyzed hematological changes. Results: No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology and serum biochemistry. But we found out histopathological changes in liver fat tissues of female. In addition, there were no significant changes of gross body and individual organs weight. Conclusions: These results suggest that water soluble extract of Root of Polygala teunifolia Willd. has not acute oral toxicity and oral LD50 value was over 4,000 mg/kg in SD rats.
Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine
Kang, Hyunkyung,Roh, Hang Sik,Song, Hyemin,Lee, Kwangmoon,Chung, Seung-Tae,Ban, Sang-ja,Mo, In Pil,An, Beum-Soo,Ahn, Chi-Young Korean Society of ToxicologyKorea Environmental Mu 2016 Toxicological Research Vol.32 No.4
The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.
Evaluation of Haemagglutinin Content by RP-HPLC to Generate Pandemic Influenza Vaccine
Hyunkyung Kang,Hang Sik Roh,Hyemin Song,Kwangmoon Lee,Seung-Tae Chung,Sang-ja Ban,In Pil Mo,Beum-Soo An,Chi-Young Ahn 한국독성학회 2016 Toxicological Research Vol.32 No.4
The potency of influenza vaccine is determined based on its hemagglutinin (HA) content. In general, single radial immunodiffusion (SRID) assay has been utilized as the standard method to measure HA content. However, preparation of reagents for SRID such as antigen and antibody takes approximately 2~3 months, which causes delays in the development of influenza vaccine. Therefore, quantification of HA content by other alternative methods is required. In this study, we measured HA contents of H1N1 antigen and H1N1 influenza vaccine by reverse phase-high performance liquid chromatography (RP-HPLC) methods. The presence of HA1 and HA2 was investigated by silver staining and Western blot assay. In addition, accuracy and repeatability of HA measurement by RP-HPLC were evaluated. Comparison of HA concentration by SRID and RP-HPLC revealed a precise correlation between the two methods. Our results suggest that RP-HPLC assay can replace SRID in the event of a pandemic flu outbreak for rapid vaccine development.
Expression Profile of Cathepsin B in the Fat Body of Bombyx mori during Metamorphosis
Bo Yeon Kim,Kwang Sik Lee,Young Moo Choo,Hyung Joo Yoon,Pil Don Kang,Soo Dong Woo,Hung Dae Sohn,Jong Yul Roh,Yeon Ho Je,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05
Background: Proteolytic enzymes are involved in insect molting and metamorphosis and play a vital role in the programmed cell death of obsolete organs. Here we show the expression profile of cathepsin B in the fat body of the silkworm Bombyx mori during development. We also compared the expression profile of B. mori cathepsins B (BmCatB) and D (BmCatD) in the fat body during the larval-pupal transformation of B. mori in the BmCatB or BmCatD RNA interference (RNAi) process. Results: BmCatB is ecdysone-induced and expressed in the fat body of B. mori during the molting, and the larval-pupal and pupal-adult transformations, and its expression leads to programmed cell death. In particular, BmCatB is highly expressed in the fat body of B. mori during the larval-pupal transformation and BmCatB RNAi treatment resulted in the arrest of the larval-pupal transformation. RNAi-treated BmCatB knock-down sustained the expression of BmCatD during the larval-pupal transformation. On the other hand, BmCatD RNAi up-regulated the expression of BmCatB in the fat body of final instar larvae. Conclusion: Based on these results, we conclude that BmCatB is involved in the programmed cell death of the fat body during B. mori metamorphosis and that BmCatB and BmCatD contribute collaboratively to B. mori metamorphosis
Young Moo Choo,Kwang Sik Lee,Hyung Joo Yoon,Bo Yeon Kim,Mi Ri Sohn,Jong Yul Roh,Yeon Ho Je,Nam Jung Kim,Iksoo Kim,Soo Dong Woo,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2010 한국응용곤충학회 학술대회논문집 Vol.2010 No.10
Bee venom contains a variety of peptides and enzymes, including serine proteases. While the presence of serine proteases in bee venom has been demonstrated, the role of these proteins in bee venom has not been elucidated. Furthermore, there is currently no information available regarding the melanization response or the fibrin(ogen)olytic activity of bee venom serine protease, and the molecular mechanism of its action remains unknown. Here we show that bee venom serine protease (Bi-VSP) is a multifunctional enzyme. In insects, Bi-VSP acts as an arthropod prophenoloxidase (proPO)-activating factor (PPAF), thereby triggering the phenoloxidase (PO) cascade. Bi-VSP injected through the stinger induces a lethal melanization response in target insects by modulating the innate immune response. In mammals, Bi-VSP acts similarly to snake venom serine protease, which exhibits fibrin(ogen)olytic activity. Bi-VSP activates prothrombin and directly degrades fibrinogen into fibrin degradation products, defining roles forBi-VSP as a prothrombin activator, a thrombin-like protease, and a plasmin-like protease. These findings provide a novel view of the mechanism of bee venom in which the bee venom serine protease kills target insects via a melanization strategy and exhibits fibrin(ogen)olytic activity.
택사(Alismatis Rhizoma) 추출물의 반복 경구투여 독성 연구
노항식,석지현,정자영,이종권,김태성,최혜경,하헌용,Roh, Hang-Sik,Seok, Ji-Hyun,Jeong, Ja-Young,Lee, Jong-Kwon,Kim, Tae-Sung,Choi, Hye-Kyung,Ha, Hun-Yong 대한한방안이비인후피부과학회 2014 한방안이비인후피부과학회지 Vol.27 No.1
Objectives : This study was carried out to evaluate the repeated dose oral toxicity of Alismatis Rhizoma in Sprague-Dawley(SD) rats. Methods : Male and female rats were administered orally with Alismatis Rhizoma water extract of 500 mg/kg(low dosage group), 1,000 mg/kg(middle dosage group) and 2,000 mg/kg(high dosage group). We daily observed number of deaths, clinical signs and gross findings for 14 days(twice a day). After 14 days, we measured body and organs weight. Also we analyzed hematological changes. Results : No dead SD rats and no clinical signs were found during the experiment period. Also other specific changes were not found between control and treated groups in hematology and serum biochemistry. In addition no significant changes of gross body and individual organs weight. Conclusions : These results suggest that water soluble extract of Alismatis Rhizoma has not repeated dose oral toxicity and oral LD50 value was over 2,000 mg/kg in SD rats. As a result, we can determine Alismatis Rhizoma is a relatively safe substance.