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김남근,정형민,이유진,남윤성,최동희,손태종,이숙환,고정재,차광열,Kim, Nam-Keun,Chung, Hyung-Min,Lee, Eu-Gene,Nam, Yoon-Sung,Choi, Dong-Hee,Sohn, Tae-Jong,Lee, Sook-Hwan,Ko, Jung-Jae,Cha, Kwang-Yul The Korean Society for Reproductive Medicine 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.2
연구목적: 한국여성의 다낭성 난포증후군 환자에서 황체형성 호르몬 exon 2 유전자의 변이를 탐색하여 이들 변이와 질환과의 관련성 여부를 밝히고자 하였다. 연구재료 및 방법: 21명의 다낭성 난포증후군 환자를 대상으로 황체형성 호르몬 exon 2(Trp8Arg;TGG to CGG and Ile15Thr; ATC to ACC)의 변이를 탐색하였다. 혈액에서 Genomic DNA를 추출하여 PCR로 증폭한 후 RFLP 방법으로 변이형을 구분하였다. 결과: 황체형성 호르몬 exon 2의 변이형이 다낭성 난포증후군 환자에서 28.6%로 이미 조사된 바 있는 대조군의 16.7% 보다 약간 높게 나타났으나 통계적으로 유의한 차이는 없었다(p>0.05). 결론: 황체형성 호르몬 exon 2의 변이가 한국인의 다낭성 난포증후군 발병과 관련이 있는지를 밝히기 위해 더 많은 개체에 대한 연구가 요구된다.
생쥐 Preantral 난포의 체외배양: FSH의 종류와 농도 및 초자화 냉동보존의 영향
이숙현,신창숙,정형민,고정재,차광렬,이경아,Lee, Sook-Hyun,Shin, Chang-Sook,Chung, Hyung-Min,Ko, Jung-Jae,Cha, Kwang-Yul,Lee, Kyung-Ah 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4
Objectives: 1) To compare the efficacy of urofollitropin (Follimon) to that of recombinant human FSH (rhFSH) on the growth and maturation of mouse early preantral follicles in vitro, and 2) effect of vitrification on the growth and maturation of preantral follicles and oocytes . Methods: Isolated early preantra1 follicles (100-130 ${\mu}m$ diameter) were cultured for 12 days in 20 ${\mu}l$ ${\alpha}$-MEM media drop under the mineral oil. Follimon or rhFSH was added to the culture medium at various concentrations (0, 10, 100, and 1000 mIU/ml). Results: With Follimon, the dose of 10 mIU/ml showed the best follicle survival, growth, and MIl rate of oocyte than the other concentrations. Whereas the optimal dose of rhFSH was 100 mIU/ml. Despite the different optimal doses, the efficacy of two different FSHs on the follicle growth and maturation was similar. Isolated mouse preantral follicles were cryopreserved by vitrification and cultured in vitro for 12 days with 100 mIU/ml rhFSH. Despite the decreased follicular survival rate after thawing, the follicular growth and maturation rate of its oocyte were comparable to those of the fresh follicle. Conclusion: Results from the present study revealed that 1) the optimal doses of Follimon and rhFSH for in-vitro culture of mouse follicles are different, and 2) the frozen-thawed follicles develop normally after vitrification.
CMSX-10 단결정 초내열합금에서 일방향 응고 공정변수에 따른 응고조직 변화도
구지호 ( Ji Ho Gu ),정형민 ( Hyung Min Jung ),이정석 ( Jeong Seok Lee ),윤혜영 ( Hye Young Yoon ),서성문 ( Seong Moon Seo ),조창용 ( Chang Yong Jo ),이재현 ( Je Hyun Lee ) 대한금속재료학회(구 대한금속학회) 2013 대한금속·재료학회지 Vol.51 No.3
Directional solidification experiments were carried out on a single crystal Ni base superalloy CMSX-10 over a wide range of growth velocity and temperature gradient conditions. Under the imposed temperature gradients, the primary dendrite arm spacings decreased proportionally with the growth velocity. Microstructural characteristics at the solid/liquid interface were found to depend strongly on the growth velocities and temperature gradients. Based on the experimental observations, a growth velocity versus temperature gradient map was selected for planar to cellular and cellular to dendritic transitions during solidification. As a result, the microstructure selection map for the alloy was established by comparing it to theoretical models. Good agreement was obtained from the results for planarcellular and cellular-dendritic transitions. The columnar-equiaxed transition, however, was calculated using Hunt`s model for castings. The results are discussed in terms of the combined effects of constitutional undercooling and nucleation undercooling ahead of the growing dendrite tips.
자궁내막증과 다낭성 난포증후군 불임환자에서 $LH{\beta}$ Exon 3 (Gly102Ser) 유전자의 돌연변이 분석
김남근,이유진,조민순,남윤성,정형민,정기화,오유경,고정재,차광열,Kim, Nam-Keun,Lee, Eu-Gene,Cho, Min-Soon,Nam, Yoon-Sung,Chung, Hyung-Min,Chung, Ki-Wha,Oh, Yu-Kyoung,Ko, Jung-Jae,Cha, Kwang-Yul The Korean Society for Reproductive Medicine 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3
연구목적: 본 연구는 자궁내막증과 다낭성 난포증후군 불임환자들을 대상으로 $LH{\beta}$ exon 3 (Gly102Ser) 유전자의 돌연변이를 탐색하고자 시도하였다. 연구재료 및 방법: 그 대상으로 26명의 자궁내막증 환자와 52명의 다낭성 난포증후군 환자 그리고, 50명의 출산 경험이 있는 건강한 여성을 대조군으로 사용하였다. 이들을 대상으로 한 돌연변이 탐색은 PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) 방법으로 수행되었다. 결과 : 그 결과 자궁내막증과 다낭성 난포증후군 환자 및 출산 경험이 있는 건강한 여성에서 그 변이형이 나타나지 않았다. 결론: 따라서, 자궁내막증과 다낭성 난포증후군 불임환자의 $LH{\beta}$ exon 3 돌연변이형은 중국인 집단에만 존재할 가능성이 높으며, 더 많은 불임환자들을 대상으로 한 연구가 요구된다.
착상전 유전진단을 이용하여 성공적으로 분만한 반복자연유산 1례
남윤성,이숙환,엄기붕,이은정,정형민,차광열,Nam, Yoon-Sung,Lee, Sook-Hwan,Oum, Ki-Bung,Lee, Eun-Jung,Chung, Hyung-Min,Cha, Kwang-Yul 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.3
Objective: To report the successful delivery in a patient of recurrent spontaneous abortion caused by chromosomal abnormality. Material and Method: Case report. Results: Twelve oocytes were obtained by in vitro fertilization. Of eleven oocytes fertilized, two embryos turned out to be normal by using fluorescent in situ hybridization on blastomere biopsy. The patient succeeded in pregnancy and the result of amniocentesis was found to be normal. She delivered the healthy female baby by cesarean section. Conclusions: The successful delivery is possible in recurrent spontaneous abortion related with reciprocal translocation by using preimplantation genetic diagnosis.
수정에 실패한 인간 난자에 있어서의 염색체의 수의 이상
손원영,이경아,박상희,한세열,윤태기,정형민,곽인평,차광열,Son, Weon-Young,Lee, Kyung-Ah,Park, Sang-Hee,Han, Sei-Yul,Yoon, Tae-Ki,Jung, Hyung-Min,Kwak, In-Pyung,Cha, Kwang-Yul 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2
Many oocytes fail to fertilize and cleave in vitro and many embryos transferred back to uterus fail to implant or maintain implantation. Chromosomal abnormalities in the male and female gametes may contribute to this loss. The higher incidence of meiotic chromosomal abnormalities bas been found in oocytes than in sperm. The wide range of incidence of chromosomal abnormalities in unfertilized oocytes has been reported in human IVF program (26-63%). However, factors affecting chromosomal abnormalities are not well understood. The present study has been conducted to investigate effects of the method for ovarian hyperstimulation, women's age, and the number of oocytes retrieved per patients on the incidence of numerical chromosomal abnormalities. Five hundred eighty four unfertilized metaphase II oocytes were subjected to chromosomal analysis. Included unfertilized oocytes were from 220 patients (mean $age=32.7{\pm}3.0$) and three hundred thirty oocytes were legible for analysis. Two hundred fourty five oocytes out of 330 (73.3%) were normal, while 38 (11.5%) were hyperploidy, 35 (10.6%) were hypoploidy, and 12 (3.6%) were diploidy. Significant difference in chromosomal abnormalities was not found between two patient groups stimulated by follicular stimulating hormone/human menopausal gonadotrophin (FSH/HMG) (25.9%) and gonadotrophin-releasing hormone agonist/follicular stimulating hormone/human menopausal gonadotrophin (GnRHa/FSH/HMG) (28%). There was a tendency of increasing chromosomal abnormalities in unfertilized oocytes from older patients (<30 yrs: 20.3%, 30-34yrs: 26.9%, >34 yrs: 35.3%). The number of oocytes retrieved per patient had no effect the incidence of chromosomal abnormalities (1-5: 31. 4%, 6-10: 29.8%, 11-15: 28.6%, > 15: 16.5%). These results from the present study suggest that the chromosomal abnormalities observed in the unfertilized oocytes has not affected by the stimulation methods, patient's age, and the number of oocytes retrieved per patients.
hESC로부터 유래된 혈관내피전구세포를 위한 최적의 배양조건 확립
김주미 ( Ju Mi Kim ),문성환 ( Sung Hwan Moon ),이민지 ( Min Ji Lee ),오인록 ( In Rok Oh ),신정민 ( Jeong Min Shin ),박순정 ( Soon Jung Park ),정선화 ( Sun Hwa Chung ),김문규 ( Mun Kyou Kim ),이경일 ( Kyung Il Lee ),정형민 ( Hyung 한국조직공학과 재생의학회 2008 조직공학과 재생의학 Vol.5 No.3
Human embryonic stem cell(hESC) can be very valuable source for the cell therapy due to their pluripotency when they differentiated into specialized cell types. Conventionally, differentiated cell population were obtained from spontaneously formed embryoid body(EB) which could contain various cell types. In this study, we described the optimization of culture condition for endothelial precursor cells derived from human embryonic stem cells(hESC-EPCs). First, we compared the expression of endothelial precursor cell markers such as CD133, CD34 and KDR in day 7 hEBs and isolated CD133 and KDR double positive population as hESC-EPCs. Second, we compared the culture medium and ECM coating to optimize culture condition for hESC-EPCs. As the result, we could confirm EGM-2 and collagen coating were optimized condition to culture of hESC-EPCs. After conducting optimized culture condition for hESC-EPCs, we examined the maintenance of characteristics of hESC-EPCs cultured in optimized culture condition as vascular cells. Finally, we could confirm the characteristics of hESC-EPCs cultured in optimized condition as vascular cells were maintained in developed optimized culture condition. Furthermore, the optimized culture condition developed in this study will be useful to culture of other vascular lineage cells derived from hESCs.
발효, 세포배양, 생물공정 세포 크기 차이를 이용한 유세포 분석을 통한 인간배아줄기세포 유래 기능성 혈관세포의 확립
이지혜 ( Ji Hye Lee ),김주미 ( Ju Mi Kim ),정형민 ( Hyung Min Chung ),채정일 ( Jung Il Chae ) 한국미생물생명공학회 2011 한국미생물·생명공학회지 Vol.39 No.4
Human embryonic stem cells have been highlighted as a valuable cellular source in the regenerative medicine field, due to their pluripotency. However, there is the challenge of the establishment of specific functional cell type forms of undifferentiated human embryonic stem cells (hESC). To establish and purify functional cell types from hESCs, we differentiated undifferentiated hESCs into vascular lineage cells and sorted the specific cell population from the whole cell population, depending on their cell volume, and compared them with the non-sorted cell population. We observed that about 10% of the PECAM positive population existed in the VEGF induced differentiating human embryoid body (hEB), and differentiated hEBs were made into single cells for cell transplantation. After making single cells, we performed cell sorting using a fluorescence-activated cell sorter (FACs), according to their cell volume on the basis of FSC region gating, and compared their therapeutic capacity with the non-sorted cell population through cell transplantation into hindlimb ischemic disease model mice. 4 Weeks after cell transplantation, the recovery rate of blood perfusion reached 54% and 17% in the FSC regions of sorted cells- and non-sorted cells, respectively. This result suggests that derivation of a functional cell population from hESCs can be performed through cell sorting on the basis of cell volume after preliminary differentiation induction. This approach may then greatly contribute to overcoming the limitations of marker sorting.
Conservation of the lethal giant larvae Tumor Suppressor Gene in Bos taurus coreanae
Baek, Kwang-Hyun,Kim, Yong-Soo,Chung, Hyung-Min,Ko, Jung Jae,Cha, Kwang Yul 한국유전학회 2002 Genes & Genomics Vol.24 No.2
We cloned a homologue of lethal giant larvae (lgl) tumor suppressor oncogene, bgl-l, in Bostaurus coreanae by RT-PCR using primers designed based on the conserved sequences for the lgl family members. The sequence analysis revealed that it encodes a 1,036 amino acid polypeptide containing a domain characteristic of WD-40 proteins. The amino acid sequence of bgl-1 showed a homology of 98.3%, 87.3%, 30.5%, 16.7%, 12.3% and 10.6% identity to that of mouse, human, fly, C. elegans, yeast SOP1 and SOP2, respectively. The complementation analysis revealed that the bovine bgl-1 partially restored Na^+ tolerance in the absence of yeast lgl homologues, suggesting that the bgl-l is a bovine homologue of the lgl family and is conserved for cellular functions.