19-nortestosterone(NORA)와 19-norandrostenedione(NT)는 정소내 aromatization과정중 중간 대사물로 검출된다. 본 연구는 이들을 장기간 투여하여 정소 및 부속기관의 무게, 혈청내 testosterone(T)의 농도, 정자형성에 미치는 영향을 조사하였다. NORA, NT 및 TE를 $300/50{\mu}l$농도로 주당 3회씩 12주간 정소에 직접 주사(intratesticular injection, i.t.)하였다. 또한 GnRH antagonist(RS68439)를 처리하여 혈청내 생식소자극호르몬 (GTH)을 감소시킨후 위 호르몬들을 동일방법으로 처리하여 이들의 보상작용을 조사하였다. NORA는 정소무게를 감소시키지 않았으나 GnRH antagonist를 처리하여 감소된 정소무게를 현저하게 보상하였다(P<0.05). NORA, NT, TE는 모두 부속기관의 무게를 증가시켰으며, RS68439가 감소시킨 부속기관의 무게를 현저히 증가시켰다. 그러나 이들은 부정소(epididymis)에는 영향을 주지않았으며 RS68439처리군에서는 보상작용을 나타내었다. 이들의 처리로 혈청내 LH농도는 완전히 감소하였으나 FSH의 농도에는 영향을 나타내지 않았다. 그리고 혈청내 T의 농도를 증가시켰다. NORA는 정자형성과정중 7단계의 spermatid의 수를 현저히 감소시켰다. 위 결과로 보아 NORA는 GnRH antagonist로 FSH의 분비가 억제된 쥐의 FSH분비를 촉진하며, T의 농도를 증가시켜 정자형성과정을 억제하는 것으로 추론된다.

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.

Maternal serum ${\beta}$-specific human chorionic gonadotropin(${\beta}$-hCG) and pregnancy-specific ${\beta}_1$-glycoprotein($SP_1$) levels were determined more than one per week during 11-41 days post embryo transfer(ET) in 21 consecutive pregnancies after in vitro fertilization(IVF), which included 8 normal singleton pregnancies, 3 twin pregnancies, 4 clinical abortions, 1 ectopic pregnancy, and 5 preclinical abortions. The sensitivity of serum ${\beta}$-hCG and $SP_1$ radioimmunoassays was 3mIU/ml and 0.7ng/ml relatively. At the 7th to 8th week of gestation, ultrasonographic confirmation of fetal pole and fetal heartbeat was performed. Both serm ${\beta}$-hCG and $SP_1$ levels showed logarithmic increase, but log[$SP_1$] had more steep rising curve and had wider variation than log[${\beta}$-hCG] in normal singleton pregnancies. In 3 twin pregnancies and one ectopic pregnancy, both serum ${\beta}$-hCG and $SP_1$ levels located within the 95% confidence interval of the mean levels of 8 normal singleton pregnancies(normal range). In 2 clinical abortions which had a fetal pole without heartbeat, serum ${\beta}$-hCG level showed lower limit of the normal range or just below, but all $SP_1$ levels showed within the normal range. In other 2 clinical abortions which were diagnosed as blighted ovum, both serum ${\beta}$-hCG levels from 11 days post-ET and serum $SP_1$ levels from later days compared with ${\beta}$-hCG were below the normal range. In 5 preclinical abortions, serum $SP_1$ levels were within the normal range but serum ${\beta}$-hCG levels were far below the normal range. In conclusion, both serum ${\beta}$-hCG and $SP_1$ levels increased exponentially with similar pattern in normally conceived pregancy after IVF-ET. Both serum ${\beta}$-hCG and $SP_1$ levels could predict outcome of early pregnancy to a certain degree, but log[${\beta}$-hCG] levels had more significant correlation with outcome of pregnancy compared with log[$SP_1$] levels. In addition, ultrasonographic examination of fetal poles and fetal heartbeats gives very important clinical information and prognosis.

The purpose of this study is to improve fertilization rate in IVF-ET program of patients with male infertility used micromanipulation technique, partial zona dissection (PZD) or micro-insemination by sperm transfer (MIST). The results were as follows 1. The fertilization rate of non-micromanipulated oocytes and micromanipulated (PZD) oocytes were 12.5% (n=2) and 42.2% (n=19), respectively, and showed significant differences between two groups (p<0.05). 2. The fertilization rate of micromanipulated (MIST) oocytes was 30% (n=27). 3. The damage rate of Group 1 (PZD) and Group 2 (MIST) were 15.7% (3/19) and 29.6% (8/27), respectively. 4. One pregnancy resulted following replacement of micromanipulated (MIST) embryos in 4 patients.

Clinical effects of intramuscular injection of 250mg testosterone cypionate every 2-3 weeks were investigated in 15 impotent patients with low or low normal serum testosterone level (Hypo-gonadotrophic hypogonadism was excluded). The results were obtained as follows. 1. Among 15 patients, 7(46.7%) showed markedly improved potency, 6(40%) partially improved potency and 2(13.7%) no improvement of potency. There was no correlation between effec-tiveness of testosterone replacement and age, testicular size, serum testosterone level or LH, FSH level. 2. Among 12 patients who had showed improved potency, 8(66.7%) complained of redeveloped decrease in potency during testosterone replacement. In conclusion, testosterone cypionate for treatment of the impotent patients with low or low normal serum testosterone level was effective, but further studies are necessary to investigate cause of redeveloped decrease in potency during testosterone replacement.

항정자항체를 검사할 수 있는 면역분석법개발을 위하여 immunoaffinity chromatography로 분리 정제한 정자표현항원을 microtiter plate에 고정화 시킨것과 효소와 화학발광체로 표지된 2차항체를 사용하여 ELISA법과 CIA법을 개발하고 이 방법의 이용 가능성을 검토하기 위하여 기존 방법인 Kibrick test법으로 임상소견이 다른 남성혈청을 분석 비교하여 다음과 같은 결과를 얻었다. 1. 인간정자 표면항원을 분리하기 위한 immunoaffinity column을 제작키 위하여 인간정자를 토끼에 주사하여 정자에 대한 항혈청을 생산하였으며 이를 Protein A-Sepharose column으로 분리 정제하여 CNBr activated Sepharose-4B에 coupling시켜 immunoaffinity column을 제작하였다. 이 column에 균질화된 정자를 반응시키고 SDS를 넣은 Tris-HCI buffer로 용출시켰을때 60KD정도의 분자량을 갖는 분획을 얻었다. 2. 분리 정제된 human IgG를 microtier plate에 농도를 달리하여 고정화 시키고 ELISA용 Goat anti-human IgG-HRP conjugate와 CIA용 Rabbit anti-human IgG-ABEI-H conjugate와 반응시켜 그 활성도를 측정하였던 바 농도에 따라 반응의 정도가 감소하였다. 3. ELISA법으로 양성혈청의 희석곡선을 작성하였을때 경사가 완만한 것과 급한 것의 두 종류의 경향을 띈 곡선으로 대별되었으며 완만한 경사를 나타내는 것에서 1:160 희석치에서 O.D가가 0.1이하를 음성, 0.1이상 0.2이하를 약양성(weak positive) 그리고 0.2이상을 양성으로 판별하였다. 4. ELISA, CIA 그리고 Kibrick test로 동일시료를 분석 비교하였던바 ELISA와 CIA는 거의 동일한 상관관계를 보였으나 Kibrick test와는 50% 수준만 일치함을 보였다.

We cryopreserved mouse morulae by a simple ultra-rapid method of freezing embryos directly in $LN_2$ after holding 2min in a $LN_2$ vapor, and thawed them in $37^{\circ}C$ water bath. The time requirements for permeation and dehydration by 2.0 M glycerol and 0.2 M sucrose before freezing were studied. When the embryos were equilibrated for 10 min, the optimun post-thaw survival was obtained. Embryos those developed normally to blastocyst after in vitro culture for over 24hrs were regarded as survival ones. Two experiments to assess post-thaw survival following predehydration in various mixtures of glycerol and sucrose were also accomplished. When sucrose was held constant (0.2 M) and glycerol concentration varied (1.5-3.5 M), post-thaw survival was best (78.0%) in 3.0 M glycerol. When glycerol was held constant (3.0M) and sucrose concentration varied (0.0-1.0M), optimun post-thaw survival (78.0%) was found in 0.2 M sucrose.

Development of a solid-phase chemiluminescence immunoassay for the detection of progesterone in serum extract was described. The chemiluminescence immunoassay was establised utilizing anti-progesterone monoclonal antibody that coated on polystyrene tubes and progesterone-ABEI conjugate as tracer. The light yield generated from antibody bound conjugate was counted on clinilumat luminometer by oxidation with microperoxidase and peroxide. The chemiluminescence immunoassay was high specific and accurate and detects as little as 3.9ng/ml of progesterone. The intra-assay CV ranged from 6% to 11.5% and inter-assay CV ranged from 13.6% to 18.7%. This assay system was good correlated with conventional kit radioimmunoassay system (r=0.98).