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이상길 ( Sang Kil Lee ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),장재영 ( Jae Young Jang ),남기덕 ( Ki Deok Nam ),김남훈 ( Nam Hoon Kim ),주광로 ( Kwang Ro Joo ),동석호 ( Seok Ho Dong ),김병호 ( Byung Ho Kim ),장영운 ( Youn 대한소화기학회 2005 대한소화기학회지 Vol.45 No.5
Background/Aims: Saccharomyces boulardii (S. boulardii) has been reported to be beneficial in the treatment of inflammatory bowel disease, however, little is known about its mechanism of action. Peroxisome proliferator- activated receptor-γ (PPAR-γ) is re
TNBS 유도 대장염에서의 Microarray 분석과 Saccharomyces boulardii의 역할에 관한 연구
이상길 ( Sang Kil Lee ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ) 대한소화기학회 2010 대한소화기학회지 Vol.55 No.1
목적: Saccharomyces boulardii (S. boulardii)는 이미 여러 종류의 설사병에 사용되고 있으며 작용 기전이 정확히 알려져 있지는 않지만 염증성 장질환에서도 치료 효과가 보고되고 있다. 이에 저자들은 S. boulardii의 염증성 장질환에서의 효과와 작용 기전을 알아보기 위하여 2,4,6-trinitrobencene sulfonic acid (TNBS)로 유발한 대장염 동물모델을 이용하여 조직학적 분석과 microarray를 하였다. 대상 및 방법: BALB/c 흰쥐에게 경구 식이용 관을 이용하여 항문으로 TNBS (2mg)와 50% 에탄올 혼합물(0.1 mL)을 주입하였다. S. boulardii는 10 mg을 하루에 두 번 4일 동안 경구 투여시켰다. Microarray는 마크로젠의 DNA chip 기술 매뉴얼에 따라서 시행되었다. 결과: TNBS로 유발한 장염에서 S. boulardii의 처리는 조직학적 손상과 사망률을 감소시켰다(p<0.05). 총 33,013개의 유전자 중에서 TNBS와 S. boulardii에 의해서 유의한 수준인 2배 이상으로 변화한 것이 330개 있었다. 이 중에서 TNBS에 의해서 과발현된 유전자의 과발현폭을 S. boulardii가 감소시킨 것이 193개였다. 이들 유전자를 기능에 따라 분류하면 염증반응과 면역반응에 관여하는 유전자들이 많았다. 결론: S. boulardii의 항염증 효과가 염증반응과 면역 및 방어 반응에 관여하는 유전자의 발현변화를 통해서 나타날 가능성이 높고, 이에 대한 추가 연구가 필요하다. Background/Aims: Saccharomyces boulardii has been reported to be beneficial in the treatment of inflammatory bowel disease. The aim of this work was to evaluate the effect of S. boulardii in a mice model of 2,4,6-trinitrobencene sulfonic acid (TNBS) induced colitis and analyze the expression of genes in S. boulardii treated mice by microarray. Methods: BALB/c mice received TNBS or TNBS and S. boulardii treatment for 4 days. Microarray was performed on total mRNA form colon, and histologic evaluation was also performed. Results: In mice treated with S. boulardii, the histological appearance and mortality rate were significantly restored compared with rats receiving only TNBS. Among 330 genes which were altered by both S. boulardii and TNBS (>2 folds), 193 genes were down-regulated by S. boulardii in microarray. Most of genes which were down-regulated by S. bouardii were functionally classified as inflammatory and immune response related genes. Conclusions: S. boulardii may reduce colonic inflammation along with regulation of inflammatory and immune responsive genes in TNBS-induced colitis. (Korean J Gastroenterol 2010;55:33-45)
한국인 대장 선종성용종 환자에서의 MYH 유전자 변이에 대한 연구
김한수 ( Hansoo Kim ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),주광로 ( Gwang Ro Joo ),동석호 ( Seok Ho Dong ),김병호 ( Byung Ho Kim ),장영운 ( Young Woon Chang ),이정일 ( Jung Il Lee ),장린 ( Rin Chang ) 대한장연구학회 2005 Intestinal Research Vol.3 No.1
Background/Aims: Recently, germ-line mutation in the base-excision-repair gene MYH was identified to cause a novel autosomal recessive form of familial adenomatous polyposis (FAP). Interestingly, a striking evidence for MYH mutations within different ethnic groups has been demonstrated. We have screened 30 patients with multiple adenomatous polyps for MYH mutations to assess its prevalence and ethnic specificity in Korea. Methods: Thirty patients with multiple adenomatous polyps were examined for MYH mutations. Twenty-one men and 9 women presented at a median age of 62.3 years. The mean number of adenomas per patient was 10.0. Sixteen exonic regions and its intronic sequences were amplified by PCR and subjected to SSCP and DNA sequencing analyses. Results: None of the patients was identified to carry any truncating or sequence alterations in MYH. Our screening for the mutational regions, which were recognized from Caucasian patients or affected Indian families, also failed to detect sequence substitutions. Conclusions: Mutation in MYH may be rarely involved in the pathogenesis of multiple sporadic colorectal adenomas in Korea, although large-scale analysis will be required to clarify the presence of specific MYH variants in a subset of patients and its role for the predisposition of multiple colorectal adenomas in Korea. (Intest Res 2005;3:27-32)
대장암에서 관찰되는 Caveolin 의 낮은 발현과 프로모터 CpG 과메틸화에 대한 연구
김남훈 ( Nam Hoon Kim ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),문영수 ( Young Soo Moon ) 대한장연구학회 2007 Intestinal Research Vol.5 No.1
Background/Aims: Abnormal reduction of caveolins has been found in many human cancers while its overexpression also correlates with increased metastatic progression of some tumors. To elucidate the possible implication of caveolin abnormality in human colon tumorigenesis, the expression and mutational status of caveolins was explored. Methods: We investigated 11 human colon cancer cell lines, 49 primary carcinoma tissues, and its matched normal colonic tissues. Both mRNA and protein levels of caveolins (cav-1, cav-2) were evaluated by quantitative RT-PCR and immunoblotting. Effect of cav-1 expression on tumor growth was tested using cell counting and colony formation assay. Cav-1 expression was restored in nonexpressing cells, whereas cav-1 expression was inhibited by siRNA-mediated knockdown in expressing cells. Methylation status of 38 CpG sites was evaluated by bisulfite DNA sequencing. Results: Low expression of cav-1 transcript was found in 54.5% of cancer cell lines, whereas 45.5% of those showed strong expression. Expression level of cav-1 protein was very low in majority of cancer cell lines except two cell lines. Approximately 47% and 10% of primary carcinomas exhibited significant reduction and elevation in cav-1 expression, respectively. Cav-2 expression also showed down- and up-regulation in 28% and 3% of primary tumors, respectively. Cav-1 transcript was re-expressed in nonexpressing cells by 5-aza-dC treatment. Restoration of cav-1 inhibited growth of cav-1-negative cells and reduced phospho-Erk level, whereas ectopic overexpression of cav-1 further stimulated cav-1-expressing cells and activated p53 and p21. Conclusions: Caveolin undergoes epigenetic silencing in a considerable proportion of human colon cancers by aberrant promoter CpG hypermethylation. Also, cav-1 acts two opposite functions as a growth suppressor or growth stimulator in colon cancers. (Intest Res 2007;5:60-72)
대장암에서 종양억제 유전자와 p53의 상위조절인자로서의 hSRBC 연구
김완중 ( Wan Jung Kim ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),김진오 ( Jin Oh Kim ),조주영 ( Joo Young Cho ),심찬섭 ( Chan Sup Shim ) 대한장연구학회 2007 Intestinal Research Vol.5 No.2
Background/Aims: hSRBC [human Serum deprivation response (sdr)-Related gene product that Binds to c-kinase] was identified using PKCδ or BRCA1 as a probe and located at 11p15.5-p15.4 region. Expression of hSRBC protein was also decreased in a number of breast, lung, and ovarian cancer cell lines, suggesting that hSRBC might be a putative tumor suppressor gene. Methods: The expression status of hSRBC was analyzed in 50 primary colon tumors and their adjacent 50 normal tissues, and 20 colon cancer cell lines. Transcript and protein expression of hSRBC was studied by quantitative RT-PCR and Western blot, respectively. siRNA-mediated knockdown of hSRBC expression was utilized to investigate its association with p53. Results: The mRNA expression of hSRBC was decreased in 60% (12/20) of colon cancer cell lines and 44% (22/50) of patient``s colon cancer tissues. Expression of hSRBC mRNA was significantly decreased in tumors compared to non-cancerous cells, while genomic level of hSRBC was not decreased in tumors. hSRBC expression was increased by 5-aza-2’-deoxycytidine treatment and hypermethylation of CpG sites was strongly associated with decreased expression. Ectopic transfection of hSRBC suppressed RKO cell count and hSRBC knockdown by siRNA augmented HCT116 cell numbers. Flow cytometry showed G1 arrest and apoptosis of colon cancer cells by restoration of hSRBC expression in RKO cells. Both basal and etoposide-mediated p53 expression was decreased when hSRBC expression was knockdowned with siRNA. Conclusions: hSRBC expression is frequently decreased by promoter CpG site hypermethylation. hSRBC down-regulates p53 expression in G1 phase and might be a novel upstream regulator of p53. (Intest Res 2007;5:131-143)
대장암에서 관찰되는 XAF1 과메틸화에 의한 유전자외 불활성 연구
장재영 ( Jae Young Jang ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),이길연 ( Kil Yeon Lee ),남기덕 ( Ki Deuk Nam ),김남훈 ( Nam Hoon Kim ),이상길 ( Sang Kil Lee ),주광로 ( Kwang Ro Joo ),동석호 ( Seok Ho Dong ),김병호 ( Byun 대한소화기학회 2005 대한소화기학회지 Vol.45 No.4
Background/Aims: X-linked inhibitor of apoptosis (XIAP) is the most potent member of the IAP family that exerts antiapoptotic effects. Recently, XIAP-associated factor 1 (XAF1) and two mitochondrial proteins, Smac/DIABLO and HtrA2, have been identified to
한국인 염증성 장질환 환자에서의 Natural Resistance-ssociated Macrophage Protein 1 유전자 촉진자의 다형성
이병욱 ( Byoung Wook Lee ),조재호 ( Jai Ho Cho ),이길연 ( Gil Yeon Lee ),김효종 ( Hyo Jong Kim ),지성길 ( Sung Gil Chi ),정용희 ( Yong Hee Jeong ),한요셉 ( Yo Seb Han ),동석호 ( Seok Ho Dong ),김병호 ( Byung Ho Kim ),장영운 ( Youn 대한소화기학회 2003 대한소화기학회지 Vol.41 No.6
Background/Aims: Natural resistance-associated macrophage protein 1 (NRAMP1) gene controls the immune response to intracellular microbial pathogens in human. To identify the association between the NRAMP1 gene and susceptibility of inflammatory bowel disease (IBD) in Korean population, we examined three predominant alleles (allele 2, 3, and 7) that have been known to influence the transcriptional activity of the NRAMP1 gene. Methods: Genomic DNA was extracted from blood lymphocytes by standard method in 70 patients with Crohn`s disease (CD), 72 patients with ulcerative colitis (UC), and 52 healthy controls. To detect sequence polymorphisms in the NRAMP1 gene promoter, we performed nonisotopic genomic PCR-SSCP analysis. PCR products with different SSCP patterns were subjected to DNA sequencing analysis to verify sequence polymorphisms. Results: The allele and carrier frequencies of allele 2 and 3 were similar in both patient and healthy control group. Although the statistical significance did not exist, the allele and carrier frequencies of allele 7 had higher tendency in patients with CD (10.7%, 21.4%) and UC (11.1%, 22.2%) than in the healthy control group (4.8%, 9.6%). Conclusions: Our findings suggest that the specific promoter polymorphism of the NRAMP1 gene may influence susceptibility to IBD in the Korean population. Further studies with a group of subjects are required to clarify our observations. (Korean J Gastroenterol 2003;41:465-472)
이지은 ( Ji Eun Lee ),이선경 ( Seon Kyung Lee ),지성길 ( Sung Gil Chi ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.12
목적 : 침윤성 자궁 경부암 조직에서 신생혈관생성 및 VEGF, TSP-1의 발현 정도를 조사하여 자궁 경부암의 진행 과정에 있어 이들의 상호 관계 및 예후인자로서의 가치를 평가하고자 하였다. 연구 방법 : 자궁경부암 37예와 7예의 정상 자궁경부 조직 절편을 채취하여 VEGF 아단위와 TSP-1의 mRNA 정량분석을 하였다. 미세혈관수는 면역조직화학 염색을 통해 검사하였다. 결과 : 정상 조직과 자궁경부암 조직 모두에서 VEGF와 TSP-1이 발현되었으며, 정상조직에서 발현된 VEGF와 TSP-1의 mRNA값의 평균값보다 2배 이상 발현된 경우를 과발현 (over expression), 1/2 이하의 값을 가지는 경우는 저발현 (low expression)으로 정의하였다. VEGF121은 56.8%에서, VEGF165는 40.5%에서 과발현을, TSP-1은 35.1%에서 저발현을 보였으며 정상 조직에서는 과발현이나 저발현이 나타나지 않았다. 미세혈관수는 VEGF의 과발현과 유의한 상관관계를 보였으나 (p<0.05), TSP-1의 저발현과는 상관관계가 없었다. VEGF의 과발현과 TSP-1의 저발현이 동반되어 나타난 그룹에서의 미세혈관수도 의미 있게 증가하였다 (p<0.05). 결론 : VEGF와 TSP-1의 발현에 따라 미세혈관의 수가 의미 있게 변화하는 것으로 보아 침윤성 자궁경부암에서 VEGF 과다 발현과 TSP-1의 저발현이 함께 나타나는 경우 미세혈관 형성을 촉진시키는 것으로 사료된다. Objective: Vascular Endothelial Growth Factor (VEGF) is a potent stimulator of angiogenesis in solid tumors. Thrombospondin-1 (TSP-1) has inhibitory role in cancer cell proliferation and metastasis. To analyze the correlation with expression of VEGF and TSP-1 including microvessel density (MVD), the levels of VEGF/TSP-1 mRNA expression and microvessel count (MVC) were estimated in patients with invasive cervical carcinomas. Methods: From 1996 to 1999, 37 carcinomas and 7 normal cervical tissues were collected, frozen and stored at -70℃ until used. The levels of VEGF and TSP-1 mRNAs were determined by quantitative RT-PCR. MVD was assessed by immunostaining for factor VIII-related antigen. The results are expressed as the largest number of microvessels present within a single ×40 field, and counted at x 100 field. Results: Quantitative RT-PCR analysis demonstrated abnormally increased VEGF mRNA expression levels (>0.66) in 14 (37.8%) of 37 cervical carcinomas comparing to control groups (mean: 0.32±0.09) and abnormally low TSP-1 mRNA expression levels (<0.72) in 13 (35.1%) of 37 cervical carcinomas comparing to control groups (mean: 0.51±0.07). MVC was higher in tumors showing decreased expression of TSP-1 (but not statistically) (p<0.18) and overexpression of VEGF (p<0.05). When VEGF overexpression was accompanied with reduced TSP-1 expression, the microvessel density showed significantly increased pattern (p<0.05). Conclusion: Our study demonstrates that reduced expression of TSP-1 mRNAs and overexpression of VEGF mRNAs may be an important contributing factor in cervical carcinomas. Moreover, the inversed correlation of VEGF and TSP-1 mRNA expression can be an evidence of angiogenic role in cervical carcinomas.