[PA-0012] CRISPR/Cas9-multiple guide RNAs construction and case study for targeting the FAD2 and FATB multi-genes in soybean

Won-Nyeong Kim(Won-Nyeong Kim),Hyun Uk Kim(Hyun Uk Kim) 한국육종학회 2022 한국육종학회 공동학술발표집 Vol.2022 No.-

FAD2 and FATB family gene editing by multi sgRNA-Cas9 system to produce high oleic/ low saturated fatty acid soybean and discovery of transcription factor to enhance oil production

Won Nyeong Kim,Hyun Uk Kim 한국육종학회 2023 한국육종학회 공동학술발표집 Vol.2023 No.-

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.

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.

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.

The Differences in Prevalence and Sociodemographic Characteristics of Irritable Bowel Syndrome According to Rome 2 and Rome 3

( Dong Won Park ),( Oh Young Lee ),( Sung Gon Shim ),( Dae Won Jun ),( Kang Nyeong Lee ),( Hye Young Kim ),( Hang Lak Lee ),( Byung Chul Yoon ),( Ho Soon Choi ) 대한소화기기능성질환·운동학회 2010 Journal of Neurogastroenterology and Motility (JNM Vol.16 No.2

Background/Aims: Irritable bowel syndrome (IBS) is one of the most frequently observed disorders by primary care and practitioners. The aim of this study was to estimate the prevalence of IBS using the Rome II and III criteria in the general Korean population and also to compare sociodemographic differences between subjects diagnosed by these criteria. Methods: Telephone interview surveys were performed with a total of 1,009 individuals in Korea, 15 years of age or older. The questionnaire, based on the Rome II and III criteria, was validated. Results: Among the 1,009 subjects, the prevalence of IBS was 8.0% under the Rome II criteria (81 subjects; 6.4%, male; 9.6%, female), and 9.0% (91 subjects; 7.0%, male; 11.0%, female) under the Rome III criteria. The accordance rate of Rome II and III was 73.5%. Both groups showed highest frequency in the age of 30s (13.9% vs. 15.3% respectively). Female subjects showed a higher prevalence than male subjects under Rome III (91 subjects; 11.0% in female, 7.0% in male; p < 0.05), but not under Rome II criteria. Many patients older than 50 years were added when analyzed under the Rome III criteria, but not under the Rome II criteria (p = 0.017). Conclusions: The Rome III criteria were Less restrictive and showed good agreement with the Rome II criteria. The prevalence of IBS was increased in young women.(J Neurogastroenterol Motil 2010;16:186-193)

가로방향척수염으로 발현된 Churg-Strauss 증후군

조용원 ( Yong Won Cho ),장혁원 ( Hyuk Won Chang ),채진녕 ( Jin Nyeong Chae ),김상현 ( Sang Hyon Kim ) 대한류마티스학회 2009 대한류마티스학회지 Vol.16 No.4

Churg-Strauss syndrome (CSS), also known as allergic granulomatous angiitis, is a rare disorder characterized by the presence of asthma, eosinophilia and small-to-medium-sized vessels vasculitis. The peripheral nervous system is frequently involved in CSS, but central nervous system (CNS) involvement is rare. Furthermore, transverse myelitis (TM) as a presenting symptom in patients with CSS is extremely rare. We report here on a 60-year-old female who presented peripheral eosinophilia, lung eosinophilic infiltration, lung vasculitis, and TM. She was diagnosed as CSS based on clinical manifestation, pathologic findings, and the use of magnetic resonance imaging (MRI).

한 대학병원에서 급성 폐색전증으로 진단된 환자들의 임상적 특성 및 예후

채진녕 ( Jin Nyeong Chae ),최원일 ( Won Il Choi ),박지혜 ( Jie Hae Park ),조병학 ( Byung Hak Rho ),김재범 ( Jae Bum Kim ) 대한결핵 및 호흡기학회 2010 Tuberculosis and Respiratory Diseases Vol.68 No.3

Background: Pulmonary embolism (PE) is a common clinical problem in the West that is associated with substantial morbidity and mortality. The diagnostic modality has been changed since 2001. This study retrospectively reviewed the PE mortality with the aim of identifying the risk factors associated with mortality since the multidetector computed tomography (MDCT) was introduced. Methods: We analyzed 105 patients with acute PE proven by multidetector CT or ventilation perfusion scan. The primary outcome measure was the all-cause mortality at 3 months. The prognostic effect of the baseline factors on survival was assessed by multivariate analysis. Results: The main risk factors were prolonged immobilization, stroke, cancer and obesity. Forty nine percent of patients had 3 or more risk factors. The overall mortality at 3 months was 18.1%. Multivariate analysis revealed low diastolic blood pressure and the existence of cancer to be independent factors significantly associated with mortality. Forty two PE patients were examined for the coagulation inhibitors. Four of these patients had a protein C deficiency (9.5%), and 11 had a protein S deficiency (26%). Conclusion: PE is an important clinical problem with a high mortality rate. Close monitoring may be necessary in patients with the risk factors.

Original Article : Lipoic Acid Prevents the Changes of Intracellular Lipid Partitioning by Free Fatty Acid

( Dong Chan Kim ),( Dae Won Jun ),( Eun Chul Jang ),( Sang Heum Kim ),( Eun Kyung Kim ),( Sang Pyo Lee ),( Kang Nyeong Lee ),( Hang Lak Lee ),( Oh Young Lee ),( Byung Chul Yoon ),( Ho Soon Choi ) 대한간학회 2013 Gut and Liver Vol.7 No.2

Background/Aims: It is suggested that the hepatic lipid composition is more important than lipid quantity in the pathogenesis of non-alcoholic steatohepatitis. We examined whether lipoic acid (LA) could alter intrahepatic lipid composition and free cholesterol distribution. Methods: HepG2 cells were cultured with palmitic acid (PA) with and without LA. Apoptosis, changes of the mitochondrial structure, intracellular lipid partitioning, and reactive oxygen species (ROS) activity were measured. Results: Free fatty acid (FA) increased apoptosis, and LA co-treatment prevented this lipotoxicity (apoptosis in controls vs PA vs PA+LA, 0.5% vs 19.5% vs 1.6%, p<0.05). LA also restored the intracellular mitochondrial DNA copy number (553±33.8 copies vs 291±14.55 copies vs 421±21.05 copies, p<0.05) and reversed the morphological changes induced by PA. In addition, ROS was increased in response to PA and was decreased in response to LA co-treatment (41,382 relative fluorescence unit [RFU] vs 43,646 RFU vs 41,935 RFU, p<0.05). LA co-treatment increased the monounsaturated and polyunsaturated FA concentrations and decreased the total saturated FA fraction. It also prevented the movement of intracellular free cholesterol from the cell membrane to the cytoplasm. Conclusions: LA opposes free FA-generated lipotoxicity by altering the intracellular lipid composition and free cholesterol distribution. (Gut Liver 2013;7:221-227)

한국 토종오리의 개체 식별 및 품종 구분을 위한 Microsatellite 마커 탐색

서동원(Dong Won Seo),술타나(Hasina Sultana),최누리(Nu Ri Choi),김연수(Yeon Su Kim),진실(Shil Jin),허강녕(Kang Nyeong Heo),진선덕(Seon Deok Jin),이준헌(Jun Heon Lee) 한국가금학회 2015 韓國家禽學會誌 Vol.42 No.1

& & 최근, 한국의 소비자들이 건강에 대한 관심이 증가하면서 단일불포화 지방산이 풍부해 건강에 긍정적인 영향을 줄 수 있는 오리고기의 수요가 급격하게 증가하고 있다. 하지만 대부분의 종 오리는 수입에 의존하고 있는 실정이기 때문에, 토종오리의 개발 및 보급이 필요한 실정이며, 이는 종자주권의 확립 및 농가 소득 증대에도 매우 필요한 일이라 할 수 있다. 따라서 본 연구에서는 24개의 microsatellite 마커를 확보하였으며, 이들 마커의 대립유전자수는 1~16개, 이형접합도는 0∼0.865, 다형성은 0∼0.841로 확인되었다. 이들 마커를 이용하여 임의 집단에서 동일개체 출현빈도를 계산한 결과는 임의 집단 1.64×10<SUP>-16</SUP>, 전형매 집단 2.60×10<SUP>-7</SUP>, 반형매 집단 1.30×10<SUP>-12</SUP>으로 높은 개체식별률과 친자확인도를 확인할 수 있었다. 하지만 이들 마커를 이용한 계통분석 결과, 토종오리와 실용오리 집단을 정확하게 구분하기에는 어려운 것으로 확인되었다. 따라서 추가연구를 통해 토종오리의 순종화 및 더 정확한 토종오리와 실용오리 집단 구분이 가능한 마커 개발이 필요할 것으로 사료된다. & & Recently, duck meat consumption has been rapidly increased because consumers recognized duck meat for healthy food. In relation to this, Korean duck industry need to develop Korean native duck (KND) breed for both conservation perspective and self-sufficient of the breeding stocks. In this study, 24 microsatellite (MS) markers were investigated for classification of KND and commercial duck (CD) breeds in the Korean market. Using these MS markers, the calculated number of alleles (K), expected heterozygosity (He) values and polymorphic information contents (PIC) were 1∼16, 0∼0.865 and 0∼0.841, respectively. Also, the expected probability of identical values in random individuals (PI), random sib (PIsib) and random half-sib (PIhalf-sib) were estimated as 1.64×10<SUP>-16</SUP>, 2.60×10<SUP>-7</SUP> and 1.30×10<SUP>-12</SUP>, respectively. The results indicated that the expected probabilities of identity powers were enough for the individual identification. However, KND and CD breeds were not fully discriminated well using the 24 MS markers, which may CD and KND has shared same origin or crossbred. Therefore, further studies will be ultimately needed for developing a genetically pure line of KND breed even though the DNA markers used. Finally, these results will provide useful information for individual traceability system in ducks.