Construction of Stably Transformed Bm 5 Cells by Using Autographa californica Nuclear Polyhedrosis Virus IE 0 Gene

Cho, Eun Sook,Jin, Byung Rae,Sohn, Hung Dae,Choi, Kwang Ho,Kim, Soung Ryul,Kang, Seok Woo,Yun, Eun Young,Kim, Sang Hyun,Kim, Keun Young,Je, Yeon Ho,Kang, Seok Kwon 한국잠사학회 1998 한국잠사곤충학회지 Vol.40 No.2

To construct transfurmed Bm5 cells, Autographa californica nuclear polyhedrosis virus(AcNPV) IE1 gene, an immediate early viral gene was firstly used in this study. AcNPV IE1 gene, which shares on 95.3% nucleotide sequence homology with Bombyx mori nuclear polyhedrosis virus (BmNPV) IE1 gene, was isolated and cloned into pBluescript. Neomycin gene from pKO-neo was inserted under the control of the IE1 promoter to yield pAcIE1-neo. The plasmid pAcIE1-neo was transfected into Bm5 or Sf9 cells, and neomycin-resistant cells were selected in TC100 medium containing 10% fetal bovine serum (FBS) and 1 mg/ml G4l8 for two weeks. Individual clones were picked and each was amplified for further characterization. The genomic DNA from neomycin-resistant cells was isolated and characterized by PCR using AcNPV IEI gene-specific primers and by Southern blot analysis using neomycin gene probe. We concluded that AcNPV IE1 gene was functional in B. mori-derived Bm5 cells as well as Spodoptera fugjprrda-derived Sf9 cells to produce stably-transformed insect cells

Production of the Eggs with Abnormal Shape from the Domestic Silkworm , Bombyx mori , Infected with Autographa californica Nuclear Polyhedrosis Virus

(Hung Dae Sohn),(Byung Rae Jin),(Sang Mong Lee),(Nam Sook Park),(Hye Jin Park),(Eun Young Yun),(Seok Woo Kang),(Keun Young Kim) 한국잠사학회 2000 International Journal of Industrial Entomology Vol.1 No.2

Basic, Research : Role of 5-hydroxytryptamine Receptor 2A Antagonist in Hepatic Fibrosis

( Dae Won Jun ),( Waqar Khalid Saeed ),( Tae Yeob Kim ),( Joo Hyun Sohn ),( Kang Nyeong Lee ),( Hang Lak Lee ),( Oh Young Lee ),( Byung Chul Yoon ),( Ho Soon Choi ) 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background: The aim of this study was to determine whether 5-HT2A receptor antagonists affect the activation or apoptosis of HSCs in vitro and/or in vivo. Methods: For the in vitro experiments, the viability, apoptosis and wound healing ability of LX-2 cells were examined after treatment with various 5-HT2A receptor antagonists. Levels of HSC activation markers (procollagen type I, α-SMA, TGF-β and Smad 2/3) were measured. For in vivo experiments, rats were divided into three groups: control group, cirrhosis, 5-HT2A antagonist group. Results: 5-HT2A receptors expression was essentially absent in inactive LX-2 cells but was induced in activated LX-2 cells. Expression of the 5-HT2A and 5-HT2B receptors was significantly decreased by sarpogrelate, with a somewhat greater effect on the 5-HT2A receptor. Similar results were obtained with primary hepatic stellate cells. There was a time and dose dependent decrease in sarpogrelate-treated cell proliferation compared to untreated cells (P<0.05). Ketanserin and ritanserin also had anti-proliferative effects. Ketanserin and sarpogrelate significantly increased HSC apoptosis, with the effect strongest for ketanserin in TUNEL assay. The expression of α-SMA was decreased by sarpogrelate in a dose dependent manner. LX-2 cell migration was significantly suppressed in sarpogrelate or ketanserin- treated cultures and wound healing was delayed compared to cultures treated with only PDGF. There was less severe periportal and septal fibrosis in the rats in the treatment group, but the difference was not statistically significant in Masson`s trichrome stain. But expression of α-SMA was lower in the treatment group than in the cirrhotic group (61.0±7.2% vs 136.7±11.7%, P<0.05), and the treatment group expression was lower than in the disease group (58.9±0.8% vs 118.2±18.2%, P<0.05). Conclusions: 5-HT2A receptor antagonists can reduce inflammation and the activation of HSCs in this cirrhotic model.

Plenary Session 1 : PS-1-1 ; Effects of Lactobacillus paracasei on kupffer cell polarization in non-alcoholic steatohepatitis model

( Won Sohn ),( Dae Won Jun ),( Kang Nyeong Lee ),( Hang Lak Lee ),( Tae Yeob Kim ),( Joo Hyun Sohn ),( Oh Young Lee ),( Byung Chul Yoon ),( Ho Soon Choi ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.-

Background: Recently, gut microbiota has been received attention in pathogenesis of non-alcoholic fatty liver disease (NAFLD). The protective effects of Lactobacillus paracasei on NAFLD and possible mechanism were investigated. Methods: Forty male C57BL/6 mice were randomized into 4 groups for 12 weeks: control, NASH model (high fat+10% fructose diet), Lactobacillus paracasei and Lactobacillus plantarum groups. Hepatic fat deposition, inflammation, serum ALT, AST, and bilirubin were tested. Kupffer cell polarization was evaluated by flow cytometry using M1 and M2 marker. Intestinal permeability was measured by urinary Cr-EDTA amount. Results: Mean body weight of L. paracasei and L. plantarum group were lower than NASH model (38 g vs. 43g, p<0.05). Compared with NASH model mice, serum ALT and AST were significantly decreased in both L. paracasei group and L. plantarum group (ALT: 135.5 U/L vs. 92.9 and 53.7U/L, p<0.05). Intestinal permeability was decreased in both L. paracasei and L. planatrum group compared to NASH model (p<0.05).While hepatic fat deposition and lobular inflammation was significantly decreased in both L. paracasei and L. plantarum groups. Kupffer cell infiltration was significantly low in only L. paracasei group (p<0.05). M2 macrophage population increased in L. paracasei compared to NASH group (62.1% vs 50.2%, p<0.05). But Kupffer cell polarization of L. plantarum group did not showed differences compare to NASH group. Conclusions: Probiotics attenuate hepatic fat deposition and decrease ALT and AST levels in NASH model. L. paracasei, but not L. plantarum prevented steatohepatitis via modulation of Kupffer cell polarization.

Plenary Session 1 : PS-1-1 ; Effects of Lactobacillus paracasei on kupffer cell polarization in non-alcoholic steatohepatitis model

( Won Sohn ),( Dae Won Jun ),( Kang Nyeong Lee ),( Hang Lak Lee ),( Tae Yeob Kim ),( Joo Hyun Sohn ),( Oh Young Lee ),( Byung Chul Yoon ),( Ho Soon Choi ) 대한간학회 2012 춘·추계 학술대회 (KASL) Vol.2012 No.1

Background: Recently, gut microbiota has been received attention in pathogenesis of non-alcoholic fatty liver disease (NAFLD). The protective effects of Lactobacillus paracasei on NAFLD and possible mechanism were investigated. Methods: Forty male C57BL/6 mice were randomized into 4 groups for 12 weeks: control, NASH model (high fat+10% fructose diet), Lactobacillus paracasei and Lactobacillus plantarum groups. Hepatic fat deposition, inflammation, serum ALT, AST, and bilirubin were tested. Kupffer cell polarization was evaluated by flow cytometry using M1 and M2 marker. Intestinal permeability was measured by urinary Cr-EDTA amount. Results: Mean body weight of L. paracasei and L. plantarum group were lower than NASH model (38 g vs. 43g, p<0.05). Compared with NASH model mice, serum ALT and AST were significantly decreased in both L. paracasei group and L. plantarum group (ALT: 135.5 U/L vs. 92.9 and 53.7U/L, p<0.05). Intestinal permeability was decreased in both L. paracasei and L. planatrum group compared to NASH model (p<0.05).While hepatic fat deposition and lobular inflammation was significantly decreased in both L. paracasei and L. plantarum groups. Kupffer cell infiltration was significantly low in only L. paracasei group (p<0.05). M2 macrophage population increased in L. paracasei compared to NASH group (62.1% vs 50.2%, p<0.05). But Kupffer cell polarization of L. plantarum group did not showed differences compare to NASH group. Conclusions: Probiotics attenuate hepatic fat deposition and decrease ALT and AST levels in NASH model. L. paracasei, but not L. plantarum prevented steatohepatitis via modulation of Kupffer cell polarization.

대장암 환자에서 선암종과 선종성 용종의 Leptin 발현

손원 ( Won Sohn ),전대원 ( Dae Won Jun ),이오영 ( Oh Young Lee ),이학현 ( Hak Hyun Lee ),백유흠 ( Yoo Hum Baek ),이강녕 ( Kang Yeoung Lee ),이상표 ( Sang Pyo Lee ),이항락 ( Hang Lak Lee ),윤병철 ( Byung Chul Yoon ),최호순 ( Ho So 대한내과학회 2007 대한내과학회지 Vol.72 No.4

목적: 지방세포에서 분비하는 호르몬인 leptin은 비만에 중요한 역할을 할 뿐 아니라 위장관암을 포함한 일부 악성 종양에서 성장 인자로써 작용할 수 있다. 이러한 배경하에 본 연구에서는 대장암에서 leptin 발현과 임상병리학적 특성 및 질환과의 상관성을 알아보고자 하였다. 대장암으로 진단받고 장절제 수술을 시행한 환자 중 장절제 표본에 정상 대장 점막, 선종성 용종, 선암종을 동시에 동반하고 있는 환자를 선별하여 면역조직화학적으로 leptin 발현을 조사함으로써 대장 선종-암종 연속성에 따른 발현 차이를 분석하였다. 방법: 대장암으로 진단받고 장절제 수술을 시행한 환자 중 장절제 표본에 정상 대장 점막, 선종성 용종을 동시에 동반하고 있는 24명의 환자를 대상으로 선택하여 분석하였다. 24개의 표본에서 각각 정상 대장 점막, 선종성 용종, 선암종 조직에서의 leptin 발현을 면역조직염색으로 알아보았다. 결과는 반정량적으로 판독하였으며, 염색 강도는 음성인 경우를 0, 양성이면서 약한 경우를 1+, 중등도인 경우를 2+, 강한 경우를 3+으로 하였다. 결과: Leptin 발현은 정상 대장 점막에서 4.2% (1명/24명)인 반면에 선종성 용종에서 33.3% (8명/24명), 선암종에서 50.0% (12명/24명)로 선종성 용종에서의 leptin 발현이 정상 대장 점막에서보다 더 높았으며(p<0.05), 선암종에서도 정상 대장 점막보다 더 높았다(p<0.05). 그러나 선종성 용종과 선암종 군간에 leptin 발현은 통계학적으로 유의한 차이가 없었다(p>0.05). 대장암 조직에서의 Leptin 발현 유무와 나이, 성별, 체질량 지수(BMI: Body Mass Index), 암병기, 임파선 전이 여부등과는 상관관계를 보이지 않았으나 종양의 크기는 leptin 발현 음성인 군이 3.9±1.4 cm, leptin 발현 양성인 군이 5.6±2.2 cm으로 leptin 발현 양성인 군이 음성인 군에 비하여 종양 크기가 더 컸다(p<0.05). 결론: 본 연구에서는 동일 환자에서 선종성 용종이나 선암종에서 정상 대장 점막 보다 leptin 발현이 통계학적으로 의의있게 높았고, leptin 발현 양성인 군이 음성인 군보다 대장암의 크기가 더 컸다는 결과로 보아 leptin이 대장에서 선종성 용종과 선암종의 발생에 중요한 역할을 할 것으로 사료된다. 그러나 선종성 용종과 선암종에서의 leptin 발현은 차이가 없었기에 leptin과 선종성 용종에서 선암종으로의 진행과의 상관성은 찾지 못하였다. 따라서 추후 좀더 많은 연구가 필요할 것으로 생각된다. Background: Though leptin, the adipocytes-derived hormone, plays an important role in obesity, it can act as a growth factor for several cancers including gastrointestinal malignancies. Based on this background, we investigated whether leptin expression correlated with the clinicopathological characteristics or disease outcome in patients with colon cancer. We immunohistochemically analyzed the expression of leptin in a colon adenoma-carcinoma sequence in the normal colon mucosa, an adenomatous polyp and adenocarcinoma tissue, from a surgical resection for each patient. Methods: We collected samples from 24 patients with a colorectal adenocarcinoma that was removed in either a total colectomy or hemicolectomy, and who presented with an adenomoatous polyp and an adenocarcinoma in the same surgical specimen. Leptin expression was assessed using immunohistochemical methods and was evaluated by grading the staining intensity as 0, +1, +2, +3. Results: Whereas leptin expression was observed in 4.2% (1/24) of the normal colon mucosa, adenomatous polyps and adenocarcinomas showed 33.3% (8/24) and 50.0% (12/24) expression of leptin, respectively (p<0.05), suggesting that leptin expression in the adenomatous polyps and adenocarcinomas was higher than in the normal colon mucosa (p<0.05). There was no significant difference in leptin expression between the adenomatous polyps and adenocarcinomas, statistically. There was no relationship between leptin expression and patients age, sex, BMI (body mass index), cancer stage, and lymph node metastasis. However, the tumor size in the positive leptin expression group was larger than in the negative leptin expression group (5.6±2.2 cm vs 3.9±1.4 cm; p<0.05). Conclusions: Since leptin expression in adenomatous polyps and adenocarcinomas was higher than in the normal colon mucosa and leptin expression significantly correlated with the tumor size, leptin might play a role in the development of an adenomatous polyp and an adenocarcinoma in the colon. However, leptin does not contribute to the progression of colon adenoma, and further evaluation studies will be required. (Korean J Med 72:352-359, 2007)

Molecular Cloning and Characterization of a Ferritin Heavy Chain Subunit of the Cricket Teleogryllus emma

Mi Ri Sohn,Nam Jung Kim,Hung Dae Sohn,Byung Rae Jin 한국응용곤충학회 2009 한국응용곤충학회 학술대회논문집 Vol.2009 No.05

We describe here the cloning and characterization of a cDNA encoding the ferritin heavy chain homologue (TeFerHCH) from the cricket Teleogryllus emma. The TeFerHCH gene spans 1,009 bp and consisted of four introns and five exons coding for 217 amino acids residues. The TeFerHCH subunit contained the conserved motifs for the ferroxidase center typical of vertebrate ferritin heavy chains and the iron-responsive element (IRE) sequence with a predicted stem-loop structure was present in the 5'-untranslated region (UTR) of TeFerHCH mRNA. TeFerHCH was grouped with the S type (HCH) in a phylogenetic tree. The TeFerHCH cDNA was expressed as approximately 27 kDa polypeptide in baculovirus-infected insect Sf9 cells. Northern blot analysis revealed that TeFerHCH exhibited ubiquitous expression and was upregulated by wounding and iron overload in the fatbody, suggesting a functional role for TeFerHCH in iron metabolism.

LC : Randomized Clinical Trial: Effects of Multi-Species Probiotics on Small Intestinal Bacterial Overgrowth in Patients with Chronic Liver Disease: A Placebo Controlled Study

( Dong Shin Kwak ),( Dae Won Jun ),( Jae Gu Seo ),( Won Seok Chung ),( Soon-eun Park ),( Tae Yeob Kim ),( Joo Hyun Sohn ),( Kang Nyeong Lee ),( Waqar Khalid Saeed ),( Hang Lak Lee ),( Oh Young Lee ),( 대한간학회 2013 춘·추계 학술대회 (KASL) Vol.2013 No.1

Background: This study was conducted to investigate the effi - cacy of probiotics administration in alleviating SIBO and intestinal permeability in chronic liver disease. Methods: Fifty three patients with chronic liver disease were randomized to receive either probiotics or placebo. After 4 weeks, the changes in SIBO, intestinal permeability and clinical symptoms were examined and compared. The lactulose-based hydrogen breath test was conducted to identify SIBO. The intestinal permeability was assessed by comparing the absorbability of lactulose and mannitol. The changes in digestive symptoms, composition of fecal bacteria, and liver function were also compared after four weeks of administration. Results: The positive rate of SIBO was 26% in chronic liver disease patients. After four weeks later, in the probiotics group, 24% of patients showed improvement of SIBO, but in placebo group, there was no patient whose SIBO was improved and 16% showed aggravation of SIBO (P<0.05). The treatment group showed significant increase in the level of fecal B. lactis, L. rhamnosus, and L. acidophilus (P<0.05). Although probiotics also contained B. bifidum, B. longum, and S. thermophiles, the changes in the levels of B. bifidum, B. longum, and S. thermophilus were not significant. By contrast, there were no significant changes in the levels of the ingested bacteria in the placebo group. About half of treatment and 31.3% of placebo group reported improvement in intestinal permeability, the difference between two groups was statistically insignificant (P=0.248). Improvement in abdominal pain of the probiotics and placebo group was 3.17±1.6 and 1.95±2.4 respectively (P=0.056) while improvement in gastrointestinal symptoms of the probiotics and placebo group was 3.35±1.7 and 2.05±2.3 respectively, suggesting that digestive symptoms of the probiotics group improved compared to placebo (P=0.047). Conclusions: 4 weeks probiotics administration in chronic liver disease patients was effective in alleviating SIBO and clini- cal symptoms but ineffective in improving intestinal permeability and liver function.

Molecular Cloning of a cDNA Encoding the Putative Family of the Larval Midgut Serine Protease from the Firefly, Pyrocoelia rufa

Lee Kwang-Sik,Sohn Hung-Dae,Lee Sang-Mong,Jin Byung-Rae 한국응용곤충학회 2002 한국응용곤충학회 학술대회논문집 Vol.2002 No.1

Molecular Cloning of a cDNA Encoding a Putative Family of the Antioxidant Protein, Peroxiredoxin, from the Firefly, Pyrocoelia rufa

Lee Kwang-Sik,Sohn Hung-Dae,Lee Sang-Mong,Jin Byung-Rae 한국응용곤충학회 2002 한국응용곤충학회 학술대회논문집 Vol.2002 No.1