국산 Fluorescence in Situ Hybridization 시스템을 이용한 다양한 검체에서의 염색체 분석

문신용,방명걸,오선경,류범용,황도영,정병준,최진,손철,장준근,김종원,김석현,최영민,Moon, Shin-Yong,Pang, Myung-Geol,Oh, Sun-Kyung,Ryu, Buom-Yong,Hwang, Do-Yeong,Jung, Byeong-Jun,Choe, Jin,Sohn, Cherl,Chang, Jun-Keun,Kim, Jong-Won,Kim, Se 대한생식의학회 1997 Clinical and Experimental Reproductive Medicine Vol.24 No.3

Fluorescence in situ hybridization (FISH) techniques allow the enumeration of chromosome abnormalities and from a great potential for many clinical applications. In order to produce quantitative and reproducible results, expensive tools such as a cooled CCD camera and a computer software are required. We have developed a Chromosome Image Processing System (Chips) using FISH that allows the detection and mapping of the genetic aberrations. The aim of our study, therefore, is to evaluate the capabilities of our original system using a black-and-white video camera. As a model system, three repetitive DNA probes (D18Z1, DXZ1, and DYZ3) were hybridized to variety different clinical samples such as human metaphase spreads and interphase nuclei obtained from uncultured peripheral blood lymphocytes, uncultured amniocytes, and germ cells. The visualization of the FISH signals was performed using our system for image acquisition and pseudocoloring. FISH images were obtained by combining images from each of probes and DAPI counterstain captured separately. Using our original system, the aberrations of single or multiple chromosomes in a single hybridization experiment using chromosomes and interphase nuclei from a variety of cell types, including lymphocytes, amniocytes, sperm, and biopsied blastomeres, were enabled to evaluate. There were no differences in the image quality in accordance with FISH method, fluorochrome types, or different clinical samples. Always bright signals were detected using our system. Our system also yielded constant results. Our Chips would permit a level of performance of FISH analysis on metaphase chromosomes and interphase nuclei with unparalleled capabilities. Thus, it would be useful for clinical purposes.

과배란유도에 의한 난소과자극증후군 발생 고위험군에 있어서 알부민 정맥투여요법의 효과에 관한 연구

문신용,노재숙,이경순,서창석,김석현,최영민,신창재,김정구,이진용,장윤석,Moon, Shin-Yong,Roh, Jae-Sook,Lee, Kyung-Soon,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

Ovarian hyperstimulation syndrome(OHSS) is one of the well-known complication of controlled ovarian hyperstimulation(COH). Though there have been numerous measures to prevent the occurrence of OHS, it has not been completely preventable until now. The fluid shift from the intravascular space to the third space is due to decreased oncotic pressure of the serum. The objective of this study was to evaluate if IV administration of 20% albumin in those patients with OHSS risk can make prevention of severe OHSS. We retrospectively analysed 70 patients undergoing IVF-ET who had serum peak estradiol($E_2$) level of >2,500 pg/ml and/or the number of oocytes retrieved over 20. The treatment group(n=39) received albumin while the control group(n=31) did not. After 40 grams of human albumin diluted in 1,000 ml of 0.9% sodium chloride solution, the treatment group received half of the fluid during oocyte retrieval, the remainder in the recovery suite. The results were as follows; There were significant differences in the levels of serum peak $E_2$ and number of oocytes retrieved between the two groups(p<0.05). However, there were no significant differences in the incidence of OHSS and pregnancy rate or multifetal pregnancy rate. In conclusion, administration of albumin to OHSS risk patients did not reduce the rate of OHSS in IVF-ET. However, if we consider the fact that there were differences in the level of peak serum $E_2$ and oocyte numbers, further prospective study may be needed.

과배란유도시 난포기 LH의 생물학적 활성이 성숙난자의 수정율 및 난할율에 미치는 영향에 관한 연구

문신용,김석현,황태영,신창재,이진용,장윤석,Moon, Shin-Yong,Kim, Seok-Hyun,Hwang, Tae-Young,Shin, Chang-Jae,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.1

To investigate the factors that affect the fertilization and cleavage rates of mature oocytes, 44 patients undergoing controlled ovarian hyperstimulation(COH) with FSH/hMG/hCG regimen for IVF - ET were analyzed. During follicular phase, serum LH levels were measured by radioimmunoassay and bioassay. Based on the mean follicular immunoactive LH(i-LH) and bioactive LH(b-LH) levels, patients were divided into 3 groups, respectively. There were no significant differences in basal serum FSH levels on menstrual cycle day 3, serum estradiol($E_2$) and progesterone ($P_4$) levels on the day of hCG administration, and the numbers of follicles aspirated and oocytes retrieved among groups. In relation to the mean follicular i-LH levels, the fertilization and cleavage rates of mature oocytes did not show a significant difference among groups. However, in groups with higher mean follicular b-LH levels, the fertilization and cleavage rates were reduced significantly. During late follicular phase, day-to-day variance in b-LH levels was not significant, but there was a significant difference among groups. There was no significant correlation between serum P. and b-LH levels. These data suggest that the fertilization and cleavage rates of mature oocytes are adversely affected by the raised mean follicular b-LH levels, and monitoring of serum b-LH levels is more useful in COH when compared with i-LH. It appears that the reduced rates are not due to the attenuated endogenous LH surge.

인간 배아의 동결보존에 관한 연구

문신용,김정훈,김석현,최영민,신창재,김정구,이진용,장윤석,Moon, Shin-Yong,Kim, Chung-Hoon,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.2

Controlled ovarian hyperstimulation(COH) for in vitro fertilization and embryo transfer(IVFET) often results in the production of more embryos than can be efficaciously transferred at one time. However, embryo cryopreservation provides a mechanism by which additional embryos can be stored for later thawing and transfer. From November, 1990 to October, 1992, we completed 42 transfer cycles of cryopreserved pronucleus(PN) l-cell embryos using the fixed protocol of hormonal replacement therapy in a physiological manner regardless of individual ovarian function. Artificial endometrial stimulation was performed with only exogenous estradiol and progesterone(E-P) in 36 transfer cycles (Group I) and with gonadotropin-releasing hormone agonist(GnRHa) and exogenous estradiol and progesterone(GEEP) in 6 transfer cycles(Group II ). The results were as follows. 1. The Survival rate of total cryopreserved-thawed embryos was 64.9%(198/305): 64.9% (172/265) in Group I and 65.0% (26/40) in Group II. 2. Total 168 embryos were transferred with an average of 4.7 per ET in Group I and total 26 embryos were transferred with an average of 4.3 per ET in Group II. 3. The pregnancy rate(PR) per cryopreserved-thawed ET and the implantation rate was 33.3 %(14/42) and 6.7%(13/194), respectively. The PRs per cryopreserved-thawed ET were 30.6% (11/36) in Group I and 50.0% (3/6) in Group II without significant difference. 4. The take home baby rate was 11.1%(4/36) in Group I and 33.3% (2/6) in Group II.

한국인(韓國人) Turner 증후군(症候群)의 세포유전학적(細胞遺傳學的) 연구(硏究)

문신용,정영훈,차상헌,박태동,이진용,장윤석,송정자,최규완,Moon, Shin-Yong,Jung, Young-Hoon,Cha, Sang-Heon,Park, Tae-Dong,Lee, Jin-Yong,Chang, Yoon-Seok,Song, Jeong-Ja,Choe, Gyu-Wan 대한생식의학회 1985 Clinical and Experimental Reproductive Medicine Vol.12 No.1

The purpose of this study is to investigate .the cytogenetic characteristics of the Turner's syndrome in Korea. For this study selected were eighty-two patients with Turner's syndrome, who were diagnosed by the chromosomal analyses of the cultured peripheral blood lymphocytes, at the Cytogenetic Laboratory, Institute of Reproductive Medicine and Population, College of Medicine, Seoul National University, for the period of 11 years and 6 months from May 1971 through November 1982. Among the 82 patients with Turner's syndrome, 21 (25.6%) had 45,X karyotype, 57 (69.4%) had mosaicism of 45,X/46,XX (39), 45, X/46, XX/47, XXX (9), 45, X/47, XXX (5), 45, X/46, XY (4). Remaining 4 patients had 46, $XX_{p-}$, 46, $XX_{q-}$and 46,X,i($X_q$), respectively.

체외수정시술을 위한 과배란유도시 GnRH Agonist(Lupron)와 성선자극호르몬 복합 투여의 효용성에 관한 연구

문신용,이진용,장윤석,Moon, Shin-Yong,Lee, Jin-Yong,Chang, Yoon-Seok 대한생식의학회 1990 Clinical and Experimental Reproductive Medicine Vol.17 No.1

In 105 patients with the past history of poor response to the previous controlled ovarian hyperstimulation(COH) due to poor follicular growth or premature LH surge, the effectiveness of pituitary suppression with gonadotropin-releasing hormone agonist(GnRH agonist) in IVF/GIFT program was evaluated in 112 cycles of COH using a combination regimen of Leuprolide acetate (Lupron TAP Pharmaceuticals, USA) and FSH/hMG or pure FSH from May to December, 1989 at SNUH. Starting on day 21 of the menstrual cycle(MCD #21, Day 1), Lupron (1.0mg/day, subcutaneous) was administered once a day till next MCD #3(suppression phase). After the confirmation of pituitary suppression, ovarian follicular growth was stimulated with FSH/hMG or pure FSH from MCD #3(Day + 1), and Lupron was continued with hMG or FSH until hCG administration (D 0) (stimulation phase). After suppression phase, serum E2 level decreased from 183.7${\pm}$95.1(Day 1) to 17.4${\pm}$12.3pg/ml (Day +1), and serum progesterone level from 19.17${\pm}$8.67 to 0.12${\pm}$0.05ng/ml. But there was no decresas in serum LH and FSH levels; LH from 12.74${\pm}$6.21 to 15.49${\pm}$4.93mIU/ml,FSH from 7.60${\pm}$3.84 to 8.58${\pm}$3.15 rnlU/ml. There was no occurrence of premature LH surge during COH. Eleven cycles(9.8%) were cancelled due to poor follicular growth during stimulation phase, and 3 cycles (3.0%) failed in the transvaginal oocytes fretrieval. Serum E2 level was 1366.8${\pm}$642.4 on D 0 and 1492.3${\pm}$906.9pg/ml on D+1. 7.00${\pm}$3.32 follicles(FD${\geq}$12mm) were observed on D 0, and 6.11${\pm}$4.15 oocytes were retrieved, with the oocyte retrieval rate per follicle of 95.0%. 3.59${\pm}$2.57 oocytes were fertilized and cleaved with the oocyte cleavage rate of 55.7%. In 83 IVF patients, 4.08${\pm}$2.39 embryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET 2.39 mebryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET of 19.3%. In 6 GIFT patients, 7.83${\pm}$3.31 oocytes were retrieved and transferred with maximum number of 6, but no pregnancy was obtained. When compared with the previous 108 cycles of COH using FSH/hMG or pure FSH regimen, the cancellation rate during COH was significantly decreased, and all the parameters of the outcome of COH including the pregnancy rate were increased. These data suggest that GnRH agonist therapy for pituitary suppression is an effective adjunct to the current gonadotropin regimens for COH in IVF/GIFT and can increase the probability of oocytes retrieval and pregnancy, especially in the previous poor responders.

Klinefelter 증후군 및 Y염색체 이상으로 진단된 환자들의 염색체 결과 및 임상 양상

문신용,최석태,고희정,오선경,한정호,서창석,김석현,최영민,김정구,이진용,Moon, Shin-Yong,Choi, Seok-Tae,Ko, Hee-Jung,Oh, Sun-Kyung,Han, Jung-Ho,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Lee, Jin-Yong 대한생식의학회 1999 Clinical and Experimental Reproductive Medicine Vol.26 No.1

Klinefelter's syndrome is a very important disease in gynecologic endocrinologic fields, because the patients with this karyotype complain of infertility, azospermia and ambiguous genitalia. Y chromosome is an important chromosome which determine genetic sex and the structure of gonad and genitalia. In this study, to elucidate the cytogenetic characteristics and clinical features of Klinefelter's syndrome and Y chromosomal abnormalities in Korea, we studied 303 cases of Klinefelter's syndrome and 11 cases of Y chromosomal abnormalities which were diagnosed by chromosomal analysis at the Cytogenetic Laboratory, Institute of Reproductive Medicine and Population, Seoul National University for 12 years from January 1984 to December 1996. The results of this study showed as follows: 1. In a total of 9275 cases, there were 303 cases (3.3%) of Klinefelter's syndromes, 11 cases (0.1%) of Y chromosomal abnormalities. 2. In 102 cases of patients showed typical clinical features of Klinefelter's syndrome, 101 cases (99%) of them were diagnosed to Klinefelter's syndrome in karyotyping. 3. In 303 cases of Klinefelter's syndrome, there were 277 cases (91.4%) of 47,XXY complement, 16 cases (5.3%) of mosaicism, 2 cases (0.7%) of 48,XXXY, 5 cases (1.7%) of 48,XXYY and 3 cases (1.0%) of 49,XXXXY. 4. In 303 cases of Klinefelter's syndrome, 284 cases (93.7%) of them were diagnosed after puberty and only 19 cases (6.3%) of them were diagnosed before puberty. 5. In 303 cases of Klinefelter's syndrome, there were 146 cases (48.2%) of patients with infertility-associated chief complaints, 101 cases (33.3%) of patients with typical clinical features of Klinefelter's syndrome, 22 cases (7.3%) of patients with ambiguous genitalia. 6. In patients with Klinefelter's syndrome, 48,XXYY and 49,XXXXY had serious symptoms such as mental retardation, developmental delay, Down syndrome-like features, congenital anomalies, but 48,XXYY and other mosaicisms had infertility-associated symptoms or ambiguous genitalia. 7. The 8 cases of polysomy Y (XYY complement) showed several serious symptoms such as Down syndrome-like features, mental retardation, fragile X syndrome-like feature, congenital anomalies, ambiguous genitalia which could be detected before puberty.

자궁내막증과 동반된 불임환자의 보조생식술을 이용한 치료

문신용,이경순,노재숙,서창석,김석현,최영민,신창재,김정구,이진용,장윤석,Moon, Shin-Yong,Lee, Kyung-Soon,Roh, Jae-Sook,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Lee, Jin-Young,Chang, Yoon-Seok 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

Though the endometriosis is not always related with infertility, endometriosis causes infertility in some patients. There are many treatment modalities of infertile patients who have endometriosis. In recent years, Assisted Reproductive Technology(ART) have been widely accepted as being a useful tool for the treatment of infertile endometriotic patients. The objective of this study was to evaluate the outcome of ART in infertile endometriotic patients who have been carried out IVF-ET from Jan, 1992 to Dec, 1994 and to compare the results between COH/IUI and IVF-ET in the patients with endometriosis stage I. Tubal disease only patients were grouped(308 patient, 956 cycles) as a control. Endometriosis group was subdivided into 4 groups according to American Fertility Society classification; endometriosis stage I (45 patients, 61 cycles), stage II (26 patients, 39 cycles), stage III (26 pateitns, 37 cycles), stage IV (33 patients, 50 cycles). The outcomes of IVF-ET in endometriosis patients were as follows; The oocyte recovery rates were significantly lower in stage III, IV endometriosis. In case of stage III endometriosis, the fertilization rate was significantly lower than other stages of endometriosis. Clinical pregnancy rates per cycle were not different between the tubal group(22%) and the endometriosis group(25%). According to endometriosis stage, the implantation rate and clinical pregnancy rate were significantly lower in stage IV (5.6%, 16%) compared with other stages (I; 10.0%, 26%, II;9.8%, 31%, III;12.6%, 32%). It suggests that some factor like autoantibodies may inhibit implantation of embryos in stage IV endometriosis. To evaluate the possibility that simply increasing the number of gametes at the site of fertilization might account for pregnancies attributed to IVF-ET, the authors retrospectively analyzed the outcome of couples undergoing IUI during hMG cycles and CC cycles between 1992 and 1994 in the women with endometriosis stage 1. In case of stage I endometriosis, though the COH/IUI group showed lower FSH level and lesser age profile than IVF-ET group, IUI group has resulted in lower pregnancy rates(19.2%) compared with the IVF-ET group(26.2%). In conclusion, endometriotic infertile patients can get comparable pregnancy rates with the tubal factor infertility patients during IVF-ET program. Moreover even in stage I endometriosis, IVF-ET may be an more effective treatment modality than COH/IUI.

인간 정자의 생식력 평가에 있어 첨제반응율과 햄스터 난자 침투 분석법의 비교연구

문신용,류범용,오선경,서창석,김석현,최영민,신창재,김정구,장윤석,이진용,Moon, Shin-Yong,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Shin, Chang-Jae,Kim, Jung-Gu,Chang, Yoon-Seok,Lee, Jin-Yong 대한생식의학회 1995 Clinical and Experimental Reproductive Medicine Vol.22 No.2

This study was designed to determine the relationship between sperm acrosome reaction following ionophore challenge(ARIC) and hamster ovum sperm penetration assay(SPA) as assessment of fertilizing capacity of male. ARIC test and SPA were performed in 23 fertile and 19 subfertile men. The results were as follows; Sperm concentration was significantly higher in fertile group compared with subfertile group: $114.6{\pm}64.40$ vs $61.3{\pm}46.50{\times}10^6/ml$. However, there were no significantly differences in seminal volume, motility and motility index, respectively. There was a significantly correlation between spontaneous and induced AR in fertile and subfertile group, respectively. ARIC value was significantly higher in fertile group, compared with subfertile group: $12.0{\pm}5.57%$ vs $2.6{\pm}4.96%$. Both Penetration rate(PR) and Penetration index(PI) were significantly higher in fertile group, compared with subfertile group: $97.4{\pm}7.40%$ vs $64.9{\pm}36$. 20% and $5.4{\pm}2.88$ vs $1.5{\pm}1.47$, respectively. The Positive predictive value(PPV), Negative predictive value(NPV), sensitivity and specificity of ARIC test (cut-off: 8.5) and SPA(PI cut-off : 3.0) in predicting fertility were 95.0%, 81.8%, 82.6%, 94.7% and 95.2%, 85.7%, 87.0% and 94.7%, respectively. There was no significantly difference in predicting fertility between ARIC test and SPA. In conclusion, ARIC test was shown to have a predictive value for fertilizing capacity comparable to that of the hamster ovum sperm penetration assay. Therefore, ARIC test may be a simple and cost-effective addition to existing semenology instead of SPA.

생쥐배아의 부화와 탈각에 미치는 Pronase의 영향

문신용,최성미,김희선,류범용,오선경,서창석,김석현,최영민,김정구,최규홍,이진용,Moon, Shin-Yong,Choi, Sung-Mi,Kim, Hee-Sun,Ryu, Buom-Yong,Oh, Sun-Kyung,Suh, Chang-Suk,Kim, Seok-Hyun,Choi, Young-Min,Kim, Jung-Gu,Choi, Kyu-Hong,Lee, Jin-Y 대한생식의학회 2000 Clinical and Experimental Reproductive Medicine Vol.27 No.4

Objective: Hatching of the blastocyst from the zona pellucida (ZP) is a key event in mammalian implantation. In vivo, two factors have been identified as possible mediators of hatching: lysis of the ZP by substances elaborated either from the embryo or female reproductive tract and pressure exerted on the zona by expansion of the blastocyst. Two methods of zona manipulation were already in use to enhance the ability of embryos to hatch: mechanical PZD and chemical ZD by acidic Tyrode's solution. But several controversies of each method have been reported. The purpose of this study was to investigate the effect of pronase for mouse embryo hatching. Methods: Mouse embryos were obtained following ovulation induction of $F_1$ animals. Fresh and cryo-thawed morula embryos were exposed to 0.5, 1.0, 2.0, 5.0 ${\mu}g/ml$ pronase in Ham's F10 for 72 hrs. Main outcome measures were the rates of partial hatching and completely hatched blastocysts, and cell number of it. Results: In fresh and cryo-thawed group, the rates of completely hatched blastocyst were significantly higher in 5 ${\mu}g/ml$ pronase treatment group than control group. There was no difference in completely hatched blastocyst total cell number between pronase treatment group and control group. This suggest that pronase treatment did not harmful in mouse embryo development. In pronase treatment group, zona pellucida were thinner than control group. Conclusion: The addition of pronase to culture media may accelerate the hatching of embryo. So, enzymatic treatment of the zona may provide a valuable and effective assisted hatching technique for human in-vitro fertilization-embryo transfer.