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지보근,KideokJin,Jang-HeeHahn,Hyung-GeunSong,HansooLee 생화학분자생물학회 2003 Experimental and molecular medicine Vol.35 No.1
To investigate the functional role of KAI1/CD82, a metastasis suppressor for human prostate cancer, in the regulation of homotypic cel adhesion, we transfected KAI1 cDNA into DU 145 human pros-tate cancer cels and established stable transfec-tant clones with high KAI1/CD82 expression. The KAI1 transfectant cels exhibited significantly in-creased homotypic cell agregation in comparison of the KAI1 transfectants was further enhanced upon exposure to anti-CD82 antibody, suggesting that KAI1/CD82 may be involved in the intracellular signaling for the cel adhesion. Among several signal pathway inhibitors tested, PP1, an inhibitor of Src family kinases, significantly supressed ho-motypic aggregation of the KAI1 transfectant cels. Ligation of KAI1/CD82 with anti-CD82 antibody in-creased endogenous Src kinase activity of the KAI1 transfectant cels. When diferent types of src ex-KAI1-transfected DU 145 cels, kinase-negative mu-tant src transfectant cels exhibited much lower homotypic aggregation than the mock cells trans-fected with an empty vector. Moreover, homotypic aggregation of the mutant src transfectant cels was not enhanced by KAI1/CD82 ligation with anti- CD82 antibody. These results suggest that Src mediates the intracellular signaling pathway of KAI1/CD82 for the induction of homotypic adhe-sion of human prostate cancer cels.
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요추부 추간판 탈출 환자들에서 저산소 유도 인자-1а와 세포사멸
한창환 ( Chang Whan Han ),권순용 ( Soon Yong Kwon ),조윤경 ( Yun Kyoung Cho ),지보근 ( Bo Keun Jee ),김영율 ( Young Yul Kim ),김순희 ( Soon Hee Kim ),강길선 ( Gil Son Khang ) 한국조직공학과 재생의학회 2004 조직공학과 재생의학 Vol.1 No.2
To analyze the expression of a hypoxia inducible factor (HIF)-1а and apoptosis in herniated lumbar discs. 12 human intervertebral discs (IVD) from surgery for lumber disc herniation (LDH) were stained for HIF-1а immunohistochemically and apoptosis through TUNEL method. The patients were on average 35.9 years old. The cases were divided into non-contained (NC: 6 cases) and contained (C: 6 cases) groups. For the control group, IVD from 2 cases of adolescent idiopathic scoliosis (AIS) and 2 of bursting fracture were used. Expression of HIF-1а was visualized in every case, with an average of 65.2±9.5% of the disc cells in the NC group, 33.5±3.6% in the C group, and 13.4±9.7% in the control. Apoptosis occurred in 77.3±7.3% of the cells in the NC group, 45.8±5.5% of C group, and 26.8±8.4% of the control. HIF-1а and apoptosis expressions were both observed more frequently in the LDH groups, and the NC group showed greater expression.(P<0.001) In correlation analysis, the relationship between the degree of HIF-1а expression and apoptosis showed statistically significant correlation. HIF-1а and apoptosis occur physiologically in human IVD. Their expression was the greatest among the NC group.
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양성자 자기공명분광법을 이용한 우울증 동물모델에서의 항우울제 약물 효능 평가
김상영,최치봉,이성호,우동철,윤성익,홍관수,이현승,정재준,지보근,홍승탁,김휘율,최보영,Kim, Sang-Young,Choi, Chi-Bong,Lee, Sung-Ho,Woo, Dong-Cheol,Yoon, Seong-Ik,Hong, Kwan-Soo,Lee, Hyun-Sung,Cheong, Chae-Joon,Jee, Bo-Keun,Hong, Sung-Tak,K 한국의학물리학회 2008 의학물리 Vol.19 No.2
본 기초연구에서는 강제수영검사(forced swimming test, FST)에 의해 우울증이 유발된 쥐의 해마(hippocampus) 부위에서 뇌 대사물질(brain metabolites)중에 choline 신호의 변화를 관찰하였다. 본 연구의 목적은 우울증이 유발된 쥐에 항우울제인 desipramine-HCl을 투여하여 정상 쥐와 약물을 투여한 쥐 사이의 뇌 대사물질의 차이를 자기공명분광법(magnetic resonance spectroscopy: MRS)을 통하여 알아보는 것이다. 실험대상으로는 6주령의 SD rat을 사용하였으며 우울증을 유발시키고 항우울제의 약물평가를 위한 모델링 방법으로 강제수영검사를 시행하였다. 자기공명분광법을 시행하기 위한 localization 방법으로는 PRESS 펄스시퀀스를 사용하였다(TR: 2,500 ms, TE: 144 ms, averaging: 512, complex data point: 2048, scan time: 25 min). 정상 쥐 그룹과 우울증을 유발시킨 뒤 항우울제를 투여한 쥐 그룹 모두 양쪽 해마부위에서의 NAA/Cr, Cho/Cr 비율(ratio)은 차이가 나지 않았다. 또한 왼쪽 해마에서 정상 쥐와 항우울제를 투여한 쥐 사이에서 NAA/Cr, Cho/Cr 비율도 차이가 나지 않았고, 오른쪽 해마부위에서도 통계적으로 유의한 차이가 없었다. 본 연구를 통하여 anti-immobility효과를 나타내는 항우울제를 투여함으로써 choline 신호가 정상 쥐와 비슷하게 회복되는 것을 관찰하였다. 이는 앞으로의 항우울제 약물 평가에 있어 자기공명분광법을 도입함으로써 기존의 주관적인 판단을 배제할 수 없었던 행동변화분석만으로 항우울제의 효과를 평가하는 방법에 비해 객관적이고 정량적인 분석을 할 수 있다는 점에서 아주 유용한 방법이라 사료된다. In this study, we observed the alteration of choline signal intensity in hippocampus region of the depressive rat model induced by forced swimming test (FST). The purpose of this study was to evaluate the antidepressant efficacy in the depressive animal model using MR spectroscopy. Fourteen experimentally naive male Sprague-Dawley rats weighting $160{\sim}180\;g$ were used as subjects. Drug injection group was exposed to the FST except for control group. The drugs were administered subcutaneously (SC) in a volume equivalent to 2ml/kg. And three injections were administered 23, 5, and 1h before beginning the given test. 1H MR spectra were obtained with use of a point resolved spectroscopy (PRESS) localization sequence performed according to the following parameters: repetition time, 2500 ms; echo time, 144 ms; 512 average; 2048 complex data points; voxel dimensions, $1.5{\times}2.5{\times}2.5\;mm^3$ ; acquisition time, 25min. There were no differences in NAA/Cr and Cho/Cr ratio between the right and the left hippocampus both normal control rats and antidepressant-injected rats. Also, no differences were observed in NAA/Cr and Cho/Cr ratio between the normal control rats and the antidepressant-injected rats both the right and the left hippocampus. In this study, we found the recovery of choline signals in the depressive animal model similar to normal control groups as injecting desipramine-HCl which was antidepressant causing anti-immobility effects. Thus, we demonstrated that MR spectroscopy was able to aid in evaluating the antidepressant effect of desipramine-HCl.
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일산화질소에 의해 유도된 연골 세포 사멸에서의 항세포사멸유전자 Bcl-XL의 방어 효과
한창환 ( Chang Whan Han ),신윤학 ( Yun Hak Shin ),권순용 ( Soon Yong Kwon ),조윤경 ( Yun Kyoung Cho ),지보근 ( Bo Keun Jee ),김영율 ( Young Yul Kim ),김순희 ( Soon Hee Kim ),박기숙 ( Ki Sook Park ),강길선 ( Gil Son Khang ) 한국조직공학과 재생의학회 2005 조직공학과 재생의학 Vol.2 No.2
To evaluate whether transfection of human chondrocytes with an anti-apoptotic gene, Bcl-XL, can prevent nitric oxide (NO)-induced chondrocyte apoptosis. We stably transduced early passage chondrocytes with an anti- apoptotic gene Bcl-XL and retrovirus. Bcl-XL transducants were compared with chondrocytes transduced in parallel with a Lac-Z gene. After the cells were treated with 500 microM of SNP, the process of apoptosis was assessed by trypan blue exclusion, enzyme-linked immunosorbent assay (ELISA), and flow cytometry. The functional aspect of the chondrocyte was measured by proteoglycan synthesis assay. Chondrocytes were cultivated in NO, the chondrocytes that were either not transfected or transfected with Lac-Z exhibited a decrease in viable cell number in comparison to the Bcl-XL transfected chondrocytes. The NO significantly increased fragmentation of DNA in chondrocytes that were either not transfected or transfected with Lac-Z, and this increase in DNA fragmentation was significantly greater than in the chondrocytes transfected with Bcl-XL. Flow cytometric result showed that apoptotic chondrocytes were significantly higher in the transfected and Lac-Z transfected chondrocytes than Bcl-XL transfected chondrocytes. Chondrocytes transfected with Bcl-XL were protected from NO-induced impairment of proteoglycan synthesis response to NO. From the present study, we have demonstrate that NO-induced chondrocytes death involve mechanisms subjected to regulation by an anti-apoptotic protein, Bcl-XL. Therefore, Bcl-XL gene therapy has the potential to protect apoptosis in human chondrocyte.
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