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착상전 배아의 분리된 할구에서 중기염색체 상을 획득하기 위한 효율적인 방법의 개발에 대한 연구: 미세소관 형성 저해제의 효과
임천규,민동미,이형송,김진영,궁미경,강인수,전진현,Lim, Chun-Kyu,Min, Dong-Mi,Lee, Hyoung-Song,Kim, Jin-Young,Koong, Mi-Kyoung,Kang, Inn-Soo,Jun, Jin-Hyun 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.2
Objectives: The development of an useful method for obtaining metaphase chromosomes from a biopsied blastomere would allow differentiation between embryos with balanced and normal chromosome complements in the preimplantation genetic diagnosis for chromosomal translocations. This study was performed to evaluate the effects of microtubule depolymerizing agents (MTDAs) on the blastomeres of mouse and human preimplantation embryos, and to establish an effective method for obtaining metaphase chromosomes of biopsied blastomeres in human early embryos. Materials and Methods: Early embryos (2-4 cell stage) from superovulated mice (ICR strain) were collected and treated with single or mixture MTDAs, such as vinblastine, nocodazole and colcemid. After the treatment of MTDAs for 16 hours, the metaphase aquisition (MA) rates were evaluated by the observation of chromosome status with bis-benzimide or DAPI staining. The optimal condition from the above experiment was applied to human embryos, which were developed from abnormal fertilization (3-pronuclei). Fluorescence in-situ hybridization (FISH) with whole chromosome probes was conducted on the human metaphase chromosomes by the MTDAs. Results: In mouse embryos, the effective concentrations of each MTDAs for obtaining metaphase chromosomes were $1.0{\mu}M$ of vinblastine (20.3%), $5.0{\mu}M$ of nocodazole (28.1%) and $1.0{\mu}M$ colcemid (55.6%), respectively. The highest MA rate (91.2%) in the mouse embryos was obtained by a mixture of vinblastine ($1.0{\mu}M$) and nocodazole ($1.0{\mu}M$). In the human embryos, the metaphase chromosomes of blastomeres were obtained in 44 of 113 blastomeres (38.9%) by treatment of the mixture of vinblastine and nocodazole. FISH signals of the metaphase chromosomes were successfully observed in human individual blastomeres. Conclusions: The treatment of a mixture MTDAs for obtaining metaphase chromosomes was an efficient method, and the MA rate was above 90% in the mouse embryos. However, only a relatively small proportions of the blastomeres yielded metaphase chromosomes by the MTDAs in the human embryos. The inconsistent effects of MTDAs may be related to the variation of different species and the poor developmental potency of abnormally fertilized human embryos. We should develop more reliable and efficient methods for obtaining the metaphase chromosomes in the biopsied blastomeres of human preimplantation embryos.
형광직접보합법을 이용한 착상전 유전진단 기법의 최적화와 경험 축적에 의한 임신율의 향상
임천규,민동미,이형송,변혜경,박소연,류현미,김진영,궁미경,강인수,전진현,Lim, Chun-Kyu,Min, Dong-Mi,Lee, Hyoung-Song,Byun, Hye-Kyung,Park, So-Yeon,Ryu, Hyun-Mee,Kim, Jin-Young,Koong, Mi-Kyoung,Kang, Inn-Soo,Jun, Jin-Hyun 대한생식의학회 2004 Clinical and Experimental Reproductive Medicine Vol.31 No.1
Objectives: This study was performed to evaluate the laboratory system for successful PGD using fluorescence in situ hybridization (FISH) and the clinical outcome of PGD cycles in five years experiences. Methods: A total of 181 PGD-FISH cycles of 106 couples were performed, and diagnosed chromosome normality in the preimplantation embryos. The laboratory and clinical data were classified by the following optimization steps, and statistically analyzed. Phase I: Blastomere biopsy with two kinds of pipettes, removal of cytoplasmic proteins without treatment of pepsin and culture of biopsied embryos with single medium; Phase II: Blatomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with single medium; Phase III: Blastomere biopsy with single pipette, removal of cytoplasmic proteins with pepsin and culture of biopsied embryos with sequential media. Results: A total of 3, 209 oocytes were collected, and 83.8% (2, 212/2, 640) of fertilization rate was obtained by ICSI procedure. The successful blastomere biopsies were accomplished in 98.6% (2, 043/2, 071) of embryos, and the successful diagnosis rate of FISH was 94.7% (1, 935/ 2, 043) of blastomeres from overall data. Embryo transfers with normal embryos were conducted in 93.9% (170/181) of started cycles. There was no difference in the successful rate of biopsy and diagnosis among Phase I, II and III. However, the pregnancy rate per embryo transfer of Phase III (38.8%, 26/67) was significantly (p<0.05) higher than those of Phase I (13.9%, 5/36) and Phase II (14.9%, 10/67). Conclusions: The laboratory optimization and experience for the PGD with FISH procedure can increase the pregnancy rate to 38.8% in the human IVF-ET program. Our facility of PGD with FISH provides the great possibility to get a normal pregnancy for the concerned couples by chromosomal aberrations.
김설호,박수호,이천지 한국품질경영학회 2019 한국품질경영학회 학술대회 Vol.2019 No.-
승강기 최적 운행 문제는 잘 알려진 NP문제 중 하나입니다. 우리 팀은 강화학습을 사용해 승강기의 최적 이동경로를 구하기까지 필요한 Computation의 양을 추정해보았습니다. 다른 Heuristic한 알고리즘이 고려하지 못하는 Case를 모두 고려할 수 있다는 점에서 강화학습을 선택하였고, Partially Observable MDP라, 알고리즘을 세우기 어려운 상황에서도 Reasonable 한 결과를 낼 수 있을 것이라 기대했습니다. 환경을 구성 한 변수는 승강기 수, 각 층별 최대 대기 인원, 승강기 최대 탑승 인원, 건물의 최고층 위 4가지이며, 1층에서는 무작위 층으로 올라가고, 나머지 층에서는 1층으로 내려오는 것으로 상황을 가정하였으며 각 층별 대기 인원수만 알 수 있었습니다. 상황 평가 기준은 인당 대기시간 총 합의 최소화이었으며, 200 episode동안 평균이 1step도 줄지 않았을 때 수렴했다고 보았습니다. 연구를 진행하며 일어날 수 있는 문제로는 Combinatorial space of trajectories문제, Local minimum문제, POMDP 문제가 있었고, 이를 해결하기 위해 Safe Reinforcement, Distributional Reinforcement, Hierarchy Reinforcement를 사용하였습니다. 실험 결과, Computation양은 승강기 최대 탑승 인원, 각 층별 최대 대기 인원과는 상관관계가 거의 없고, 승강기수와 건물의 최고층과 관계가 있다는 결과가 도출되었습니다. 한계점으로는 우리가 찾았던 수렴점이 Global Minimum이 아닌, Local Minimum일 가능성을 배제 할 수 없다는 것이며, 컴퓨터 성능의 한계로 모든 경우의 수를 고려하지 못하여 구체적인 상관관계를 구하지 못했다는 점입니다.
소 체내포 핵이식에 의한 핵-세포질 상호작용에 관한 연구
정희태,최종엽,박춘근,김정익,민동미 韓國受精卵移植學會 2000 한국동물생명공학회지 Vol.15 No.1
This study was conducted to investigate the effects of quiescent treatment of donor cells and activation treatment time of recipient cytoplasm on nuclear remodeling and in vitro development of somatic cell-cloned bovine embryos. Serum starved, confluent and nonquiescent cycling adult skin cells were teansferred into enucleated oocytes. Nuclear transfer oocytes were activated at 30 min, 1 and 2 hrs after electrofusion. Some nuclear transfer embryos(23% to 35%) extruded a polar body, which was not affected by quiescent treatment of donor cells and activiation time of recipient cytoplasm. About 68% of nuclear transfer embryos fused with a serum starved cells has a chromatin clump, but which was not different from embryos fused with confluent(51%) and nonquiescent(47%) cells. The proportion of embryos with a single chromatin clump was sightly increased when nuclear transfer embryos were activated within 30 min after fusion(69%) compared to those were activated at 1 and 2 hrs after fusion, but there was not significantly different. Development rates to the blastocyst stage were 8.6% and 15.9% when serum starved and confluent cells were transferred, which were higher than that of control group. Developmental rate to the blastocyst stage was higher in embryos were activated within 30 min after fusion (17.3%) compared to those of embryos were activated at 1 and 2 hrs after fusion (P<0.05). From the present result, it is suggested that quiescent treatment of donor cells and activation time of recipient cytoplasm can affect the in vitro development. Quiescent plasm activation within 30 min after fusion could increase the number of embryos with a normal chromation structure, which results in increased in vitro development.
이형송 ( Lee Hyeong Song ),최혜원 ( Choe Hye Won ),임천규 ( Im Cheon Gyu ),민동미 ( Min Dong Mi ),변혜경 ( Byeon Hye Gyeong ),김진영 ( Kim Jin Yeong ),궁미경 ( Gung Mi Gyeong ),유한욱 ( Yu Han Ug ),김수찬 ( Kim Su Chan ),전진현 ( 대한산부인과학회 2004 Obstetrics & Gynecology Science Vol.47 No.4
목적: 착상전 유전진단 (preimplantation genetic diagnosis: PGD)은 체외수정에서 획득한 수정란으로부터 할구를 분리한 후 염색체 또는 유전자 검사를 통해 정상적인 배아만을 선별적으로 자궁에 이식함으로써 유전적으로 이상이 있는 환아를 출산할 확률이 높은 부부들에서 환아의 임신과 출산을 예방할 수 있다. 본 연구에서는 체외수정을 통해 얻어진 배아에서 하나 혹은 두개의 할구를 분리하고, duplex nested PCR 방법을 사용 Objective: Preimplantation genetic diagnosis (PGD) is reserved for couples with a risk of transmitting a serious and incurable disease, and hence avoids the undesirable therapeutic abortion. Herein, we report the result of PGD to carriers at risk of trans
유전질환 및 염색체 이상의 예방을 위한 착상전 유전진단의 결과
김진영,임천규,송인옥,유근재,양광문,한국선,허걸,송지홍,전진현,민동미,박소연,전종영,궁미경,강인수,Kim, Jin-Yeong,Lim, Chun-Kyu,Song, In-Ok,Yoo, Keun-Jai,Yang, Kwang-Moon,Han, Kuk-Sun,Hur, Kuol,Song, Ji-Hong,Jun, Jin-Hyun,Min, Dong-Mi,Park, So- 대한생식의학회 2002 Clinical and Experimental Reproductive Medicine Vol.29 No.4
Objective s: Chromosome aneuploidy is associated with recurrent abortion and congenital anomaly and genetic diseases occur repeatedly in the specific families. Preimplantation genetic diagnosis (PGD) can prevent aneuploidy or genetic disease by selecting normal embryos before implantation and is an alternative to prenatal diagnosis. The aim of this study is to assess the outcome of PGD cycles by using FISH or PCR, and to determine the clinical usefulness and values in patients with risk of chromosomal aneuploidy or genetic disease. Materials and Methods: From 1995 to Apr. 2001, a total of 108 PGD cycles in 65 patients with poor reproductive outcome were analyzed. The indications of PGD were translocation (n=49), inversion (n=2), aneuploidy screening (n=7), Duchenne muscular dystrophy (n=5) and spinal muscular atrophy (n=2). PGD was applied due to the history of recurrent abortion, previous birth of affected child or risk of aneuploidy related to sex chromosome aneuploidy or old age. Blastomere biopsy was performed in 6$\sim$10 cell stage embryo after IVF with ICSI. In the single blastomere, chromosome aneuploidy was diagnosed by using FISH and PCR was performed for the diagnosis of exon deletion in DMD or SMA. Results: The FISH or PCR amplification was successful in 94.3% of biopsied blastomeres. The rate of transferable balanced emb ryos was 24.0% in the chromosome translocation and inversion, 57.1% for the DMD and SMA, and 28.8% for the aneuploidy screening. Overall hCG positive rate per transfer was 17.8% (18/101) and clinical pregnancy rate was 13.9% (14/101) (11 term pregnancy, 3 abortion, and 4 biochemical pregnancy). The clinical pregnancy rate of translocation and inversion was 12.9% (11/85) and abortion rate was 27.3% (3/11). In the DMD and SMA, the clinical pregnancy rate was 33.3% (3/9) and all delivered at term. The PGD results were confirmed by amniocentesis and were correct. When the embryos developed to compaction or morula, the pregnancy rate was higher (32%) than that of the cases without compaction (7.2%, p<0.01). Conclusions: PGD by using FISH or PCR is useful to get n ormal pregnancy by reducing spontaneous abortion associated with chromosome aneuploidy in the patients with structural chromosome aberration or risk of aneuploidy and can prevent genetic disease prior to implantation.