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α-1,3-Galactosyltransferase Knock Out(GalT KO) 돼지유래 골수 중간엽 줄기세포의 특성 규명
옥선아,오건봉,황성수,임석기,김영임,박진기 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.3
A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal α -1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from α-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% CO2 incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers (CD45-, 29+, 90+ and 105+) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited CD45-, 29+, 90+ and 105+ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at 1x103 cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs (15.0 ± 0.4 μm) had a little larger cell size than Wild BM-MSCs (13.5 ± 0.3 μm). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.
김말애,임석옥,김민정 경희대학교 체육과학연구소 1997 體育學論文集 Vol.25 No.-
The purpose of this study is to investigate the body composition of female by part of dance major in university. For this purpose a total of 32 female students, attending university K in Seoul participated in the study. The subjects were selected randomly 24 female students majored in dance in university(Korean dance: 8, Modern dance: 8, Ballet: 8), and 8 regular female students, who did not have any dance experience. The results of the study were as follows; 1.Skinfolds thickness 1)In factors of triceps showed in Control group 17.71±7.97 mm, Korean dance group 12.98±3.14mm, Modern dance group 11.85±3.48 mm, Ballet group 11.11 ±3.48 mm. Among these group there was significantly(p<.05). 2)In factors of subscapular showed in Control group 11.26±4.59 mm, Modern dance group 9.33± 2.29 mm, Korean dance group 9.13± 1.52 mm, Ballet group 8.74±2.53 mm. Among these group there was no significantly. 3)In factors of abdomen showed in Control group 12.29±5.24 mm, Modern dance group 7.75±2.94mn, Korean dance group 8.58 ± 3.36 mm, Ballet group 6.97± 2.27 mm. Among these group there was significantly (p<.05). 4)In factors of thigh showed in Control group 21.51±8.22 mm, Korean dance group 76.30± 1.92 mm, Modern dance group 15.45±3.08 mm, Ballet group 14.45±3.94 mal. Among these group there was significan11y(P<.05). 2.Body fat 1)In factors of body density showed in Ballet group 1.0633 ±0.0072 g/ml. Modern dance group 1.0615±0.0059 g/ml. Korean dance group 1.0602±0.0031 g/ml, Control groupp 1.0512±0.0144 g/ml Among these group there was significantly(P<.05). 2)In factors of %fat showed in Control group 20.59±6.11 %, Korean dance group 76.82± 1.24 %, Modern dance group 16.34±2.41 %, Ballet group 15.61 ±2.95 %, 킬song these group there was significantly ( p<.05). 3)In factors of fat showed in Control group 10.87±5.62 kg, Korean dance group 8.45± 1.21 kg. Modern dance group 8.14± 1.49 kg, Ballet group 7.76± 1.81 kg. Among these group there was no significantly. 4)In factors of LBM showed in Ballet group 41.67±4.04 kg, Korean dance group 41.61±3.14 kg, Modern dance group 41.51 ±2.47 kg, Control group 39.76±3.36 kg. Among these group there was no significantly.
이병철,오선옥,임석병,유창식,김진천 대한대장항문학회 2017 Annals of Coloproctolgy Vol.33 No.2
Purpose: Although the standard treatment for patients with locally advanced rectal cancer managed by preoperative chemoradiotherapy (CRT) is a radical resection, local excisions are used in highly-selective cases. Recently, transanal minimally-invasive surgery (TAMIS) has emerged as a feasible technique for local excision of midrectal lesions. We assess the feasibility of using TAMIS to treat patients with locally advanced rectal cancer who showed good response to CRT. Methods: From October 2010 to June 2013, 35 consecutive patients with rectal cancer managed by using preoperative CRT underwent TAMIS. After a single-incision laparoscopic surgery port had been introduced into the anal canal, a fullthickness local excision with conventional laparoscopic instruments was performed. We retrospectively reviewed a prospectively collected database of these cases. Results: Of the 35 patients analyzed, 18 showed pathologic complete responses and 17 had residual lesions (2 ypTis, 4 ypT1, 9 ypT2, and 2 ypT3); 34 (97.1%) showed clear deep, lateral margins. The median distance of lesions from the anal verge was 5 cm. All procedures were completed laparoscopically, and the median operating time was 84 minutes. No intraoperative events or morbidities were seen in any of the patients, except one with wound dehiscence, who was treated conservatively. The median postoperative hospital stay and follow-up period were 4 days and 36 months, respectively. During the study period, no patients died, but 5 (14.3%) experienced recurrence, including one recurrence at the TAMIS site. Conclusion: TAMIS seems to be a feasible, safe modality for treating patients with locally advanced rectal cancer who show good response to preoperative CRT.
α1,3-Galactosyltransferase 유전자 좌위에서 Membrane Cofactor Protein을 효과적으로 발현하는 자성 돼지 섬유아세포의 생산
오건봉,김벨라,황성수,옥선아,임석기,박진기 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.3
Xenotransplantation of pig organs into primates results in fatal damage, referred as hyperacute rejection (HAR), and acute humoral xenograft rejection (AHXR), to the organ graft mediated by antibodies pre-existing and newly-producing in primates against their cognate pig antigens. Functional ablation of α1,3-galactosyltransferase (Gal-T KO) of pig which is an enzyme involved in synthesis of Gala1-3Galb1-4GlcNAc-R antigen is essentially required to prevent HAR. Moreover, additional genetic modification under Gal-T KO background for enforced expression of human complement regulatory proteins which can inhibits complement activation is known to effectively imped HAR and AHXR. In this study, we constructed a membrane cofactor protein (MCP) expression cassette under control of human EF1α promoter. This cassette was inserted between homologous recombination regions corresponding to Gal-T locus. Subsequently this vector was introduced into ear skin fibroblasts of female pig by nucleofection. We were able to obtained 40 clones by neomycin selection and 4 clones among them were identified as clones targeted into Gal-T locus of MCP expression cassette by long-range PCR. Real time RT-PCR was shown to down-regulation of Gal-T expression. From these results, we demonstrated human EF1α promoter could induce efficient expression of MCP on cell surface of fibroblasts of female pig.
Hh-Ag1.5 처리가 돼지 체외수정란의 발육 및 세포사멸에 미치는 영향
권대진,여재훈,노원근,곽태욱,오건봉,옥선아,임석기,박진기,황성수 韓國受精卵移植學會 2013 한국동물생명공학회지 Vol.28 No.3
The present study was performed to investigate the effect of Hh-Ag1.5, a small-molecule chemical agonist of SMOothened receptor, on the in vitro maturation and development of in vitro fertilized (IVF) embryos in pigs. Oocytes or fertilized embryos were cultured in a maturation or embryo culture medium supplemented with 0 (control), 25, 50 or 100 nM of Hh-Ag1.5, respectively. Although the maturation rate were not different among treatment groups, the blastocyst formation rate in the group treated with 25 nM Hh-Ag1.5 was significantly increased compared to other groups (P<0.05). While the highest dose of Hh-Ag1.5 (100 nM) did negatively affect to the embryo development and cell number in blastocysts compared to other groups (P<0.05), the apoptotic cell index in blastocysts was significantly lower in 25 and 50 nM groups than in control and 100 nM groups (P<0.05). The mRNA expression of the proapoptotic gene Bax and the ratio of Bax/Bcl-XL decreased in among treatment groups compared to control (P<0.05). The embryo quality related genes, Tert and Zfp42, were significantly decreased in 50 and 100 nM groups compared with control and 25 nM groups (P<0.05). In conclusion, the addition of 25 nM Hh-Ag1.5 to in vitro maturation and culture medium can enhance the developmental potential as well as quality of IVF embryos in pig.