Methotrexte 투여가 생쥐 장관의 Alkaline Phosphatase 활성에 미치는 영향

이여일(YI Lee),박원집(WJ Park),문병갑(BK Moon) 대한산부인과학회 1980 Obstetrics & Gynecology Science Vol.23 No.12

This experiment was performed to study the effects of methotrexate on the mucus secretion and alkaline phosphatase activity of mucosal epithelium in gut. Methotrexate was administered intraperitoneally in a dose of 0.5mg/mouse 48 hours interval. Experimental groups were divided into the single, two times and three times treated ones. Mouse were sacritied 24 hours after final treatment and stains for H&E, PAS and alkaline phosphatase were made in section of jejunum, ileum and colon. The result obtained were as follows. 1. Denudation of surface mucosal epithelium and abnormal dilatation of intestinal glands were prominent in small intestine as compared with those in large intestine, following methotrexate treatments. 2. Reduction of PAS positive materials was shown in the goblet cells and intestinal glands in a proportion to methotrexate treatments, which was more prominent than that of large intestine. 3. Activity of alkaline phosphatase was increased in lamina propria, intestinal glands, and submucosal tissue of jejunum and ileum but decreased in surface mucosal epithelium and exfoliated epithelium in a proportion to methotrexate treatments, which are milder in colonic mucosa than that of small intestine.

경질초음파검사를 이용한 고위험자연유산 환자의 관리에 관한 연구

이여일,장장순 대한산부인과학회 1992 Obstetrics & Gynecology Science Vol.35 No.9

1989년 8월부터 1990년 9월까지 만 1년 2개월동안 자연유산이 일어날 위험이 많아고 인정된 43명의 임산부를 대상으로 임신 첫 3개월동안 경질초음파검사를 이용해서 1주간격으로 추적하여 다음과 같은 성적을 얻었다. 1. 대상환자는 불임치료 후 임산부가 65.1%로 가장 많았고, 절박유산이 37.2%, 반복 또는 습관성유산이 11.6%, 그리고 전번 자연유산이 9.3%의 순이었다. 2. 전체 대상환자의 유산율은 23.3% 이었으며, 불임치료 후 임신시는 17.9%, 절박유산시는 31.2% 그리고 반복 또는 습관성유산과 전번 자연유산은 각각 20%와 25%의 유산율을 나타내었다. 3. 임신유지군에서 임신낭포는 임신 4주, 태아심박동은 임신 5∼6주, 태아 두둔 거리측정은 임신 5∼6주, 난황난은 임신 6∼7주, 양막강, 사지, 제대, 그리고 뇌실은 임신 7∼8주, 그리고 대뇌겸과 태아운동은 임신 8주이후에 관찰되었고, 태아 두둔거리에 따른 임신주수와 임상적 임신주수는 거의 일치된 소견을 보였다. 4. 직경 26mm이상의 임신낭포에 태아심박동이 없던 경우는 결국 모두 유산되었으며, 일단 임신낭포가 나타나면 임신유지군에서 전 예가 2주이내에 태아심박동을 확인할 수 있어, 임신낭포안에 심박동이 없더라도 최소한 2주이상의 추적관리가 필수적이며 또한 태아심박동이 발견되기만 하면 대부분인 97.1%에서 임신이 지속되었다. 5. 임신유지 실패군은 대부분이 배아나 태아의 구조물이나 기능이 없거나, 있더라도 추적 중에 태아 사망 후 소실되었다. 6. 따라서 경질초음파검사를 이용한, 자연유산이 우려된 임신 첫 3개월이내의 임산부에 대한 추적관리는 배아의 구조물과 기능의 변화를 감안하여 일정한 간격으로 연속적으로 시행한다면 효과적이고 효율적으로 수행될 수 있을 것으로 사료된다. Transabdominal sonography has been, for the past two decades, used as an effective diagnostic and research tool in obstetrics. Its use in the first trimester is relatively limited and mostly diagnostic in nature. The introduction of the higher frequency transvaginal transducer probe, with its higher resloution of the images, opens new possibilities to study early gestation. There is a need for techniques which will allow an early diagnosis to be made in high risk patients of early pregnancy failure, preferably with such certainty that a more active line of management can be pursed if so desired. A total of 164 transvaginal scans were performed from 4 to 12 weeks` gestation in the high risk patients of early pregnancy failure. The sequential appearance of 9 embryonal structures or functions were examined: (1) the gestational sac appeared in week 4: (2) the fetal pole with detectable heart beat was first seen in week 5~6: (3) the yolk sac was present from week 6~7:(4) the single unpartitioned ventricle in the brain, the amnionic sac and the body stalk or umbilical cord marked week 7: (5) the falx cerebri and fetal motion appeared during week 8, respectively in continued pregnancy. A sac greater than 26 mm in diameter without fetal heart beat is a poor prognostic sign. Before a live embryo is visualized in the intrauterine gestational sac, no sonographic features were found to be reliable in differentiating viable from nonviable pregnancy (presenting as an empty gestational sac) on a single sonographic examination. Serial vaginal sonography is effective and efficient in the diagnosis and management of early pregnancy failure.

자궁강내 인공수정에 관한 임상적 연구

이여일,임영경 대한산부인과학회 1992 Obstetrics & Gynecology Science Vol.35 No.7

Methods used for artificial insemination with husband's semen ( AIH ) have undergone continuous revision in the hope of increasing the pregnancy rate obtained. We report our methodology using technology developed for in vitro fertilization to prepare sperm for intrauterine insemination. Our overall pregnancy rate is 40%. In this preliminary study several parameters are compared with the pregnancy rates achieved.

Control Mechanisms of Ovarian Follicle Development by Follicle Stimulating Hormone and Pituitary Adenylate Cyclase-activating Polypeptide

이여일,신진옥,김미영,전상영,Lee, Yu-Il,Shin, Jin-Ok,Kim, Mi-Young,Chun, Sang-Young The Korean Society for Reproductive Medicine 2006 Clinical and Experimental Reproductive Medicine Vol.33 No.1

목 적: 본 연구는 흰쥐 난소를 실험모델로 하여 미성숙 전동 난포의 성장에 대한 pituitary adenylate cyclase-activating polypeptide (PACAP)의 영향을 얄아보고자 하였다. 연구방법: 미성숙 전동 난포를 생후 21일된 흰쥐로부터 분리하여 PACAP을 첨가하거나 첨가하지 않은 무혈청 배양액에서 3일 동안 배양하고, 푸로게스테론 호르몬의 생성, 난포의 성장, 과립막세포의 증식 및 유전자의 동태 등을 관찰하였다. 증식의 정도는 thymidine incorporation 방법으로 검색하고 유전자의 변동은 Northern 분석을 이용하였다. 결 과: PACAP으로 처리한 군은 난포의 직경이 75% 증가한 반면 난포자극호르몬인 FSH로 처리한 군은 65% 증가하였고, PACAP 처리는 과립막 세포의 증식을 강화시켰다. FSH와 PACAP 공히 배양된 흰쥐 난포의 과립막 세포와 FSH에 반응하는 세포주인 GFSHR-17에서의 프로게스테론 생성을 촉진시켰고, PACAP이 FSH의 작용을 증진시켜 SF-1과 아로마타제 유전자 발현을 촉진시켰다. 결 론: 본 연구는 PACAP이 과립막증식과 스테로이드합성을 통하여 전동 난포의 성장을 촉진함을 시사하였고, 또한, SF-1, 아로마타제 등에 대한 FSH의 작용을 도와주는 역할을 PACAP이 담당하므로 PACAP은 초기 난포성장에 필요한 난소국소인자임을 유추할 수 있었다. Objective: Pituitary adenylate cyclase-activating polypeptide (PACAP), a novel hypothalamic neuropeptide, has been suggested to play a role in ovarian folliculogenesis. The present study evaluated the effect of PACAP on the growth of preantral follicles. Methods: Preantral follicles were mechanically isolated from ovaries of 21-day-old rats and cultured in groups for 3 days in serum-free medium in the absence or presence of PACAP-38 ($10^{-6}M$). Results: Treatment with PACAP-38 resulted in an increase in follicle diameter by 75% whereas treatment with follicle stimulating hormone (FSH) increased follicle diameter by 65%. PACAP-38 treatment enhanced the granulosa cell proliferation as measured by thymidine incorporation analysis. Furthermore, the production of progesterone by cultured granulosa cells and GFSHR-17 cell line was stimulated by PACAP-38. Interestingly, PACAP enhanced FSH action on stimulation of SF-1 and aromatase gene expression. Conclusion: The present results demonstrate that PACAP stimulated preantral follicle growth by potentiating proliferation and by stimulating steroidogenesis.

Gonadotropin Regulation of Regulator of G Protein Signaling 2 (RGS-2) Expression in the Rat Ovary

이여일,이은숙,김선애,김미영,조문경,전상영,Lee, Yu-Il,Lee, Eun-Suk,Kim, Sun-Ae,Kim, Mi-Young,Cho, Moon-Kyoung,Chun, Sang-Young The Korean Society for Reproductive Medicine 2008 Clinical and Experimental Reproductive Medicine Vol.35 No.2

연구방법: 미성숙 백서 난소의 과배란 유도를 위해 PMSG를 주사하고, 배란을 위해서 hCG를 주입하였다. RGS-2의 유전자 발현양상을 조사하기 위하여는 Northern blot 분석과 in situ hybridization 분석을 시행하였다. 결 과: 미성숙 백서에 성선자극호르몬인 PMSG를 복강내 주사했을 때 RGS-2 mRNA 발현에 영향을 미치지 않음을 Northern blot analysis로 확인할 수 있었으나, hCG를 주입했을 때는 1시간에서 3시간 내에 발현이 증가됨을 알 수 있었다. In situ hybridization으로 살펴본 RGS-2 mRNA의 발현세포는 난포의 크기에 관계없이 난자였으나, hCG로 처리한 후에는 배란 전 난포와 성장중인 난포의 과립막 세포이었다. 그러나, RGS-2 단백의 발현은 hCG 처치와 관계없이 난포막 세포이었다. 상기 생체 실험과 마찬가지로 시험관에서도 배란 전 난포의 과립막 세포에 대한 LH 처리는 RGS-2 유전자 발현을 1시간 내에 촉진하였다. 또한, 성선자극호르몬 분비호르몬 2 길항제도 이러한 LH의 촉진작용을 증진시켰다. 결 론: 본 연구로 배란 전 과립막 세포에서 성선자극호르몬인 LH/hCG와 성선자극호르몬 분비호르몬 길항제에 의해 RGS-2의 발현이 증진되는 양상으로 보아 RGS-2가 배란과정 동안에 Gq protein 신호전달을 조절할 것으로 추정된다. Objective: The purpose of the present study was to examine the hormonal regulation of RGS-2 in the rat ovary. Methods: Immature rats were injected with 10 IU of PMSG to induce multiple growth of preovulatory follicles and 10 IU of hCG to induce ovulation. Northern blot analysis performed for gene expression and in situ hybridization performed for mRNA localization. Results: Northern blot analysis revealed that pregnant mare's serum gonadotropin (PMSG) treatment did not affect RGS-2 mRNA levels. In contrast, human chorionic gonadotropin (hCG) treatment of PMSG-primed rats resulted in an increase in RGS-2 expression within $1{\sim}3\;h$. The major cell-types expressing RGS-2 mRNA were oocytes regardless of follicle size. Interestingly, hCG treatment caused the stimulation of RGS-2 gene expression in granulosa cells of preovulatory and growing follicles. In contrast, cell types expressing RGS-2 protein were theca cells regardless of hCG treatment. Like in vivo, treatment of preovulatory granulosa cells with LH in vitro stimulated RGS-2 levels within 1 h. Interestingly, GnRH antagonist II enhanced the stimulatory action of LH. Conclusion: The present study demonstrates the LH/hCG induction of RGS-2 in preovulatory granulosa cells and suggests a role of RGS-2 in Gq protein signaling pathway during ovulation.

신생아제대혈청이 난자성숙과 난구세포 분산에 미치는 영향

이여일,박현정,권영숙,Lee, Yu-Il,Park, Hyun-Jeong,Kwon, Young-Suk 대한생식의학회 1998 Clinical and Experimental Reproductive Medicine Vol.25 No.1

This study was performed to investigate the stimulating effect on oocyte maturation and cumulus cell expansion in TC199 media by human cord serum (HCS) supplementation. Immature mouse oocyte cumulus complexes (OCCs) were cultured in TC199 media supplemented with bovine serum albumin (BSA), HCS and human chorionic gonadotropin (hCG) instead of luteinizing hormone (LH) respectively, and the expression of cumulus expansion and oocyte maturation were observed. After 4hr and 24hr culture with or without OCCs, media containing 0.4% BSA, 10% HCS and 10 IV hCG respectively were collected and analyzed for changing concentrations of estradiol $(E_2)$, progesterone $(P_4)$, testosterone (T), and $PGF_{2\alpha}$. There were no elevation of $E_2$, T, and $PGF_{2\alpha}$ by OCCs culture, but minute elevation of $P_4$ level by 24hr OCCs culture in hCG supplementation (p=0.048). The stimulating pattern of cumulus expansion of OCCs by HCS and hCG supplementation was similar to our previously report using Ham's F-10 media, however oocyte maturation rates after 24hr OCCs culture in all media were increased by $20\sim30%$ compared to Ham's F-10 media. These results suggest that LH in HCS induce cumulus expansion probably by $P_4$ secretion of OCCs, and TC199 is efficient media for immature mouse oocyte maturation.

체외수정 실패 정자에 대한 전자현미경적 연구

이여일,나재형,이재혁,정상우,Lee, Yu-Il,Na, Jae-Hyung,Lee, Jae-Hyuk,Juhng, Sang-Woo 대한생식의학회 1994 Clinical and Experimental Reproductive Medicine Vol.21 No.2

Failure of in vitro fertilization may occur even though oocyte and semen parameters seem satisfactory. Quantified ultrastructural study of spermatozoa was performed in three cases of failed in vitro fertilization. The results were compared to those of four fertile men. Quantification was achieved by cataloguing cell defects of the spermatozoon heads and mid/principal pieces of the flagella. Using the data from each specimen, the percentages of total cellular abnormalities in the head/mid/principal pieces were established. The percentages of anomalies of the midpiece and of the principal piece were not significantly different between failed cases and controls. The percentage of cell alterations of the head (96-100 vs 75${\pm}$3,4%), the percentage of combined anomalies of the head (80-86 vs 52.5${\pm}$1.9%), and the percentages of nuclear shape deformation (68-86 vs 47.5${\pm}$6.3%), acrosomal defects (86-96 vs 50${\pm}$4.3%), and postacrosomal sheath defects (78-88 vs 44.5${\pm}$7.2%) of the head were significantly different between failed cases and controls. Due to the cost and time involved in processing semen samples for electron microscopy, the widespread application of this technique to all couples presenting for IVF certainly is not warranted. However, in selected instances electron microscopy may play a crucial role in identifying an occult male factor.

정자 세정후 Swim-up 처치가 정자의 운동성에 미치는 영향

이여일,Lee, Yu-Il 대한생식의학회 1991 Clinical and Experimental Reproductive Medicine Vol.18 No.1

Thirty five couples were treated by intrauterine insemination with sperm prepared by a washing and swim-up method. Fifteen women conceived(42.9%). Sperm washing and swim-up was found to significantly improve sperm motility for men of infertile couples and the increment of percent sperm motility after sperm preparation allowed significant differentiation of pregnant and nonpregnant patients in asthenozoospermia(submotile) group (p<0.01). The author suggest that the increment of percent sperm motility after sperm washing and swim-up could be a useful screening tool for in vitro procedure proposed to improve fertility in the intrauterine insemination of asthenozoospermia.

불임여성의 고푸로락틴혈증에 대한 임상적 고찰

이여일 全南大醫大 1986 전남의대학술지 Vol.23 No.1

착상전 생쥐 배아 발달에 대한 Apoptotic Gene의 발현

이여일,이진,김미영,전상영,Lee, Yu-Il,Lee, Jin,Kim, Mi-Young,Chun, Sang-Young 대한생식의학회 2003 Clinical and Experimental Reproductive Medicine Vol.30 No.1

Objective : The aim of this study was to evaluate the influence of three different media on preimplatation embryo development and the expression of Bcl-2, Mcl-1, Bax, and Bok in mouse. Materials and Methods: Two-cell embryos were retrieved from ICR female mice (4 weeks old) at 48 hr after hCG injection and cultured in Ham's F-10, HTF, and G1.2 media. The developmental rate of 2-cell embryos was evaluated from 24 hr to 72 hr after culture. RT-PCR was performed for the detection of Bcl-2, Mcl-1, Bax, and Bok gene expression. Results: The rates of morula and blastocyst in HTF and G1.2 media (88%, 98.1%) were significantly higher than those in Ham's F-10 media (39.6%) at 48 hr. Likewise, the rates of hatching and hatched blastocyst in HTF and G1.2 media (21.9%, 52.9%) were higher than those in Ham's F-10 media (3.5%) at 72 hr. Bcl-2 and Bax mRNAs were highly detected in embryos cultured in Ham's F-10 when compared in embryos cultured in HTF and G1.2. In contrast, the expression of Mcl-1 and Bok was not significantly different. Conclusion: These results show that HTF and G1.2 culture media increase the rate of blastocyst formation and stimulate Bcl-2 and Bax gene expression in mouse preimplantation embryos.