분만 후 조혈줄기세포 이식을 위한 제대혈 채취량에 미치는 여러 인자에 관한 임상적 고찰

강석호 ( Suk Ho Kang ),곽인평 ( In Pyung Kwak ),이숙환 ( Sook Hwan Lee ),김수희 ( Soo He Kim ),이경주 ( Kyung Ju Lee ),백진영 ( Jin Young Baek ),김동구 ( Dong Ku Kim ) 대한산부인과학회 2006 Obstetrics & Gynecology Science Vol.49 No.1

한국인 다낭성 난포증후군 환자에서 5,10-Methylenetetrahydrofolate Reductase의 677번 유전자 다형성에 관한 연구

이교원,정유미,이숙환,윤태기,곽인평,윤선웅,최중섭,김계현,한종설,김성도,김남근,차광렬,백광현,이수만,Lee, Kyo-Won,Jeong, Yu-Mi,Lee, Sook-Hwan,Yoon, Tae-Ki,Kwak, In-Pyung,Yoon, Seon-Woong,Choi, Joong-Sub,Kim, Kye-Hyun,Han, Jong-Sul,Kim, Sung-Do,K 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.3

Objective: To investigate the association of genetic background between MTHFR C677T genotype and infertile females with polycystic ovarian syndrome. Materials and Methods: We compared 86 infertile females with polycystic ovarian syndrome (PCOS) with 100 healthy fertile females with one or more offspring. Pyrosequencing analysis for MTHFR C677T variation was performed on polymerase chain reaction (PCR) product of study group. To validate pyrosequencing data of C677T variation for randomly selected 50 samples, we compared the pyrosequencing result with the PCR-RFLP (Restriction Fragment Length Polymorphism) result of MTHFR C677T genotype. Results: The prevalence of the C677T mutant homozygous (TT) was significantly lower (p=0.0085) in females with PCOS (8.14%) than in fertile females (21.00%). MTHFR 677 TT genotype had a decreased risk (3.7-fold) of PCOS compared with wild type (MTHFR 677 CC). Conclusion: Our data support a role for MTHFR mutant homozygous (677 TT) genotype in reducing risk in Korean infertile females with Polycystic ovarian syndrome.

조직별 및 나이에 따른 마이토콘드리아 DNA 결손 (${\Delta}mtDNA^{4977}$)의 축적

정혜진,정형민,조성원,김현아,이경술,권황,최동희,곽인평,윤태기,이숙환,Jeong, Hye-Jin,Chung, Hyung-Min,Cho, Sung-Won,Kim, Hyun-Ah,Lee, Kyung-Sool,Kwon, Hwang,Choi, Dong-Hee,Kwak, In-Pyung,Yoon, Tae-Ki,Lee, Sook-Hwan 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.3

Objectives: Controversial arguments exists on both the case for and against on the accumulation of mitochondrial DNA (mtDNA) deletion in association to tissue and age. The debate continues as to whether this mutation is a major contributor to the phenotypic expression of aging and common degenerative diseases or simply a clinical insignificant epiphenomenon. The objective of this study was to determine whether the accumulation of mtDNA deletion is correlated with age-related and tissue-specific variation. Materials and Methods: One hundred and fifty-seven tissues from blood, ovary, uterine muscle, and abdominal muscle were obtained from patients ranging in age from 31$\sim$60 years. After reviewing the clinical reports, patients with mitochondrial disorder were excluded from this study. The tissues were obtained at gynecological surgeries with the consent of the patient. Total DNA isolated from blood, ovary, uterine muscle, and abdominal muscle was amplified by two rounds of PCR using two pairs of primers corresponding to positions 8225-8247 (sense), 13551-13574 (antisense) for the area around deleted mtDNA and 8421-8440 (sense), 13520-13501 (antisense) for nested PCR product. A statistical analysis was performed by $x^2$-test. Results: About 0% of blood, 94.8% of ovary, 71.4% of uterine muscle, and 86.1% abdominal muscle harbored mtDNA deletion. When we examined the proportion of deleted mtDNA according to age deletion rate was 90% of ovary, 63.6% of uterine muscle, 77.7% of abdominal muscle in thirties and 100% of all tissue in fifties. Conclusion: The findings of this study suggest that the mtDNA deletion is varied in tissue-specific pattern and increases with aging.

한국인 다낭성 난포증후군 환자에서 SHP 유전자 변이 분석

이수만,최흥식,이숙환,한정희,남보현,곽인평,남윤성,김남근,이교원,전혜선,Lee, Su-Man,Choi, Hueng-Sik,Lee, Sook-Hwan,Han, Jung-Hee,Nam, Bo-Hyun,Kwak, In-Pyung,Nam, Yoon-Sung,Kim, Nam-Keun,Lee, Kyo-Won,Jeon, Hye-Sun 대한생식의학회 2001 Clinical and Experimental Reproductive Medicine Vol.28 No.2

Objective: We inversigated Small Heterodimer Partner (SHP) gene mutation in Korean Polycystic Ovarian Syndrome (PCOS) patients. SHP protein regulates the activity of nuclear receptors which regulate the cellular development and differentiation. Recently, the mutation of SHP gene was found in the obesity and diabetes patients in Japanese group, and suggested that its mutation may involved in pathogenic mechanism of PCOS. Methods: This study was performed in 20 PCOS patients and 20 normal women. The DNAs were extracted from the peripheral bloods, and amplified at each exon (1 and 2) of SHP gene by PCR method. Subsequently, each PCR product was digested with the restriction enzyme indicated below for studying restriction fragment length polymorphism (RFLP). After enzyme digestion, the results of RFLP were compared PCOS patients with control women to find any sequence variation. Results: We examined 9 regions of exon 1 with Msp I, Pvu II, Dde I and 3 regions of exon 2 with Pst I, Dde I. There is no heterozygous or homozygous mutation in patients and control women at these restriction sites. Conclusion: The genetic analysis at our restriction sites in the SHP gene did not show any genetic variation in Korean PCOS patients. Our PCR-RFLP analysis was not covered the entire SHP gene (68 bp/1,006 bp), we need to further analysis of the entire SHP gene.

인간 포배란의 유리화동결 융해 후 임신 및 분만에 관한 연구

최동희,정형민,정미경,이숙환,남윤성,박찬,곽인평,윤태기,Choi, Dong-Hee,Chung, Hyung-Min,Chung, Mi-Kyung,Lee, Sook-Hwan,Nam, Yoon-Seung,Park, Chan,Kwak, In-Pyung,Yoon, Tae-Ki 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4

Objective: This study was performed to evaluate whether vitrification method could be used for the cryopreservation of human blastocysts derived from IVF program. Methods: Surplus embryos were obtained from consented IVF patients. Controlled ovarian hyperstimulation was done with midluteal GnRH agonist, gonadotropin and hCG. After oocyte retrieval and insemination, fresh embryo transfer was done at $4{\sim}8$ cell stage. The surplus embryos after ET were cultured in blastocyst medium up to 6 days after oocyte retrieval. Obtained blastocysts were cryopreserved with our vitrification method. Blastocysts were exposed to 1.5 Methylene glycol (EG) in phosphate buffered saline (PBS) for 2.5 minutes, followed by 5.5 M EG plus 1 M sucrose for 20 seconds. Then 1 to 3 blastocysts were mounted on electron microscope (EM) grid and the grid was plunged into liquid nitrogen for storage. For thawing, blastocyst-containing EM grids were sequentially transferred in 1.0 M, 0.5 M, 0.25 M, 0.125 M and 0 M sucrose solution at the intervals of2.5 minutes. And blastocysts were cultured for about 6 hours and only re-expanded blastocysts were transferred to uterus of the patients on 4 to 5 days after ovulation in natural cycle or on 18 to 19 day of artificial cycle. Results: From Oct. 1998 to Jul. 1999, 34 patients were agreed to participate in this study. The mean age and duration of infertility of the patients were 31.6 years and 4.1 years, respectively. Among 34 cycles. replacements could be done in 20 cycles (58.8%). A total 93 blastocysts were thawed and 48 (51.6%) of them survived. Thirty-eight blastocysts, mean 1.9 embryos per patient, were transferred, resulting in 5 clinical pregnancies which consisted of 1 triplet, 2 sets of twins and 2 singleton pregnancies. The pregnancy rate per transfer was 25% and implantation rate was 23.6%. Five patients delivered 7 healthy babies including 2 sets of twins at term. Conclusion: Successful pregnancies and deliveries were established after transfer of vitrified human blastocysts. Vitrification using ethylene glycol as cryoprotectant and electron microscope grid is a rapid and simple method that can be effectively applied for the cryopreservation of human blastocysts.

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.

체외수정 및 배아이식술에서 레이저를 이용한 보조부화술의 효용성에 대한 연구

이정현,한지은,김유신,원형재,조정현,곽인평,엄진희,박은아,최윤정,이동률,윤태기,Lee, Jung-Hyun,Han, Ji-Eun,Kim, You-Shin,Won, Hyung-Jae,Cho, Chung-Hyun,Kwak, In-Pyung,Eum, Jin-Hee,Park, Eun-A,Choi, Yoon-Jung,Lee, Dong-Ryul,Yoon, Tae-Ki 대한생식의학회 2008 Clinical and Experimental Reproductive Medicine Vol.35 No.3

목 적: 보조부화술이 적용되는 좋지 않은 예후를 보여주는 선별된 환자군을 대상으로 산성 용액을 이용한 AHA 방법과 레이저를 이용한 AHL 방법의 효용성을 비교하여, 보다 효과적으로 임신율과 착상률을 높일 수 있는 보조부화술 방법을 찾고자 하였다. 연구방법: 2006년 2월부터 9월까지 체외수정 시술을 시행한 환자 중 보조부화술이 필요한 328주기를 대상으로 산성용액을 이용한 AHA 방법 (180주기)과 ZILOS-tk 레이저를 이용한 AHL 방법 (148주기)으로 나누어 시행하였다. 보조부화술을 시행한 환자군은 환자의 나이가 38세 이상이거나 투명대의 두께가 $18{\mu}m$ 이상, 기저 FSH 농도가 15 mIU/ml이상, 체외수정 시술을 3번 이상 실패한 환자, 이식하는 배아의 상태가 양호하지 않은 환자들로 이상에 적용요인이 있으면 시행하여 무작위로 보조부화술 방법간에 환자들의 임상적 특징과 임신율과 착상률을 분석하였다. 결 과: 전체 보조부화술을 시행한 환자군에 AHL 방법과 AHA 방법간에 임신율 (42.6%, 63/148 vs. 33.3%, 60/180)과 착상률 (17.4%, 82/470 vs. 16.0%, 89/556)에 유의적 차이는 나타나지 않았다. 그러나 나이가 많은 환자군인 Group 1은 임신율 (37.0%, 20/54 vs. 18.7%, 14/75)과 착상률 (14.4%, 23/160 vs. 7.1%, 15/210)이 AHL 방법이 AHA 방법보다 유의적 (p<0.05)으로 높게 나타났다. 전체 환자군이나 선별된 각 군내에 보조부화술 방법간에 환자의 임상적 특징은 유의적 차이가 나타나지 않았다. 3번 이상 체외시술에 실패한 환자군 [Group 2: 43.8% (21/48)과 31.6% (25/79)], 투명대의 두께가 $18{\mu}m$ 이상인 환자군 [Group 3: 43.8% (32/73)과 34.1% (28/82)], 이식한 배아의 질이 양호하지 않은 환자군[Group 4: 25.0% (7/28)과 14.6% (6/41)]에서는 AHL 방법이 임상결과는 좋았으나 유의적 차이는 없었다. 결 론: 레이저를 이용한 AHL 방법이 나이가 많은 환자군과 3번 이상 체외수정 시술에 실패한 환자군에서 AHA방법에 비해 높은 임신율과 착상률을 나타내었다. 결론적으로, AHL을 이용한 보조부화술이 임상적으로 보다 효과적이고 안전한 방법이라고 사료된다. Objective: To evaluate efficacy of assisted hatching by laser (AHL) and acidified Tyrode solution (AHA) in selected groups of IVF-ET patients who have a poor prognosis. Methods: From February 2006 to September 2006, total of 328 IVF-ET cycles with advanced female age (${\geq}38$ years), recurrent implantation failure (${\geq}3$ cycles), thick zona (${\geq}17{\mu}m$), and/or poor quality of embryo were randomly divided into assisted hatching by acidified Tyrode solution (AHA, n=180) and the assisted hatching using the ZILOS-tk laser (AHL, n=148) groups. Clinical outcomes were analyzed and compared between AHA and AHL group based on the patient characteristics. Results: In all AHL and AHA group, there were no significant differences in pregnancy (42.6%, 63/148 vs. 33.3%, 60/180) and implantation rates (17.4%, 82/470 vs. 16.0%, 89/556) However, in advanced female age group (Group 1), pregnancy (37.0%, 20/54 vs. 18.7%, 14/75) and implantation rates (14.4%, 23/160 vs. 7.1%, 15/210) in AHL group were significantly (p<0.05) higher than those of AHA, although there was no difference in patient parameters of both groups. And, the clinical outcome of groups with recurrent implantation failure (Group 2), thick zona pellucida (Group 3) and poor quality embryo (Group 4) were improved in AHL compared to those of AHA: 43.8% (21/48) and 31.6% (25/79) in Group 2, 43.8% (32/73) and 34.1% (28/82) in Group 3, 25.0% (7/28) and 14.6% (6/41) in Group 4, but no significance. Conclusion: The AHL improved the pregnancy and implantation rates in patients with advanced female age and recurrent implantation failure when compared to outcomes achieved from AHA. Therefore, this AHL technique may be a efficient and safe method for patients with poor prognosis.

HAIR - AN 증후군 1 례

이숙환(Sook Hwan Lee),윤태기(Tae Ki Yoon),차광열(Kwang Yul Cha),곽인평(In Pyung Kwak),남윤성(Yoon Sung Nam) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.3

Polycystic ovary disease is a heterogenous endocrinopathy with many interacting causal factors. One potential such factor is chronic hyperinsulinemia. multiple, independent lines of evidence suppart the contention that chronic hyperinsulinemia causes ovarian hyperandragenism. This evidence includes: (1) mutations in the insulin receptor gene that cause severe hyperinsulinemia appear to be associated with ovarian hyperandrogenism, (2) insulin stimulates ovarian thecal and sttomal androgen seaetion in vitro, and (3) in some experimental models, manipulation of circulating insulin concentrations results in changes in circulating androgens. Although the association between hyperinsulinemia and hyperandrogenism remains to be fully explained at the molecular level, chronic hyperinsulinemia appears to be an important cause of hyperandrogenism. We have experienced a case of HAIR AN syndrome showing hyperandrogenism, insulin resistance and acanthosis nigricans in infertile patient. So we report this case with a brief review of literatures.

Y 상염색체의 전좌를 동반한 무정자증 1 례

이숙환(Sook Hwan Lee),윤태기(Tae Ki Yoon),차광열(Kwang Yul Cha),곽인평(In Pyung Kwak),김현주(Hyun Joo Kim),남윤성(Yoon Sung Nam) 대한산부인과학회 1999 Obstetrics & Gynecology Science Vol.42 No.3

Aneuploidy results from nondisjunction in either the meiotic division of the parents or the early cleavage divisions of the affected individuals. The sex chromosomes show a wide range of viable aneuploidy than do the autosomes. The incidence of 47,XXY and 47,XYY children increases with maternal age, as does that of autosomal trisomies, whereas the incidence of 45,X children does not increase with maternal age. In the group of sex chromosome aueuploidies, the 47,XXY and 47,XYY conditions occur with nearly equal hequency at birth. Translocations between X or Y chromosomes and an autosome or between an X chromosome and the Y chromosome cause sterility in human males. It has been assumed that a translocation involving either(or both) of the sex chromosomes would interfere with inactivation of the XY bivalent and thaeby disturb spermatogenesis. We bave experienced a case of Y-autosome translocation in azoospermic patient. So we report this case with a brief review of literatures.

반복적 시험관아기 배아이식 실패 환자에서 정맥내 면역글로블린요법

이숙환(Sook Hwan Lee),윤내영(Nae Young Youn),윤태기(Tae Ki Yoon),곽인평(In Pyung Kwak),이은정(Eun Jung Lee),최동희(Dong Hee Choi),한세열(Se Yul Han) 대한산부인과학회 2000 Obstetrics & Gynecology Science Vol.43 No.1

목적: 반복적인 시험관아기-배아이식 실패 환자에서 정맥내 면역글로블린의 효과를 보고자 하였다. 방법: 총 9예의 반복적인 배아이식 실패 환자에서 새로운 시험관아기 시술시 배아이식 직전에 정맥내면역글로블린을 500mg/kg 투여하였다. 결과: 단 1예에서 착상에 성공하였고, 1예는 화학적임신이었다. 시술중 정맥내면역글로블린의 특이한 부작용은 없었다. 결론: 정맥내면역글로블린요법은 반복적 배아이식 실패환자에서 유용한 방법으로 여겨지지 않으나, 향후 면역요법에 대한 지속적인 연구가 필요할 것으로 사료된다. Objective: The implantation failure after embryo-transfer (ET) is a major continuing problem in in vitro fertilization (IVF). This study was undertaken to determine the effectiveness of intravenous immunoglobulin for treatment of individuals experiencing repeated unexplained in vitro fertilization-embryo transfer (IVF-ET) failure. Methods: A total of nine consecutive infertile patients who failed to become pregnant after previous IVF-ET replacing at least three or more normal developed embryos each were included in our study. During the subsequent new IVF-ET cycle, each women received intravenous immunoglobulin 500㎎/㎏ before the embryo transfer. Results: Only one implantation occurred. There were no remarkable side effects. A specific effect of intravenous immunoglobulin for patients with repeated IVF-ET failure could not be demonstrated. Conclusion: High-dose intravenous immunoglobulin may not be useful for patients with repeated failure of embryo transfer.