• Clinical and Experimental Reproductive Medicine
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• 제36권4호
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• pp.265-274
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• 2009

목 적: 미세리보핵산 (microRNA, miR)은 전사 후 (post-transcriptional) 단계에서 목표 유전자 (target gene)의 발현을 억제하여 세포의 발달과 성장에 중요한 역할을 하는 것으로 알려져 있다. 그러나 난포의 성장과정 중의 miR 발현 양상에 대해서는 잘 알려져 있지 않다. 따라서 존 연구는 생쥐 난포의 체외배양 후 생식샘자극호르몬 (gonadotropin)과 사람융모성생식샘자극호르몬 (hCG) 첨가에 따른 난자와 난구세포에서의 miR 발현 양상을 살펴보고자 수행되었다. 연구방법: 생후 12일된 생쥐 (C57BL6)의 난소 적출 후, 전동 난포 (preantral follicle)를 분리하여 무작위로 $20\;{\mu}L$ 점적의 배양액만 있는 군 (control group), 재조합 난포자극호르몬을 첨가한 군 (FSH group), 재조합 황체형성호르몬을 첨가한 군 (LH group), FSH와 LH를 같이 첨가한 군 (FSH+LH group)으로 나누어 배양하였다. 난포가 충분히 성장하였을 때, 다시 hCG를 첨가한 군 (hCG (+) group)과 첨가하지 않은 군 (hCG (-) group)으로 나누어 hCG (-) group에서 난자와 난구세포를 각각 분리하여 RNA를 추출하였다. 36시간 후, 배란된 난자 난구세포 복합체 (cumulus oocyte complex, COC)에서 난자와 난구세포를 각각 분리하여 RNA를 추출하고, mmu--miR-16, -miR-27a, -miR-126, -miR-721 등의 miR에 대한 primer를 이용하여 실시간 중합연쇄반응을 시행하였다. 결 과: 배란율과 MII 난자 생성율은 다른 군들에 비해 FSH+LH군에서 유의하게 높았다. 각 군내의 난자와 난구세포 사이에서도 miR 발현 양상의 차이가 관찰되었다. 또한, 난자와 난구세포에서의 miR 발현 양상은 각 군간 차이가 있었으며, hCG (+)군과 hCG (-)군간에도 차이를 나타냈다. 결 론: 생쥐 난포의 체외배양 중 난자 및 난구세포에서의 miR 발현 양상은 gonadotropin의 종류 및 난자의 성숙도에 따라 다르다. 이러한 결과는 표적 유전자의 발현에 대한 추후 연구를 통한 확인이 필요할 것으로 사료된다.

• 대한수의학회지
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• 제16권1호
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• pp.77-96
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• 1976

In order to investigate the effects and acting mechanism on ovaries, thyroid glands and hypophyses of rabbits in short term administration of sulfadimethoxine (SDM) as medical dose, a total of 90 virgin albino rabbits (mean body weight, 1,362g) were selected at random and alloted to two groups. Rabbits in one group served as controls and the others were administered SDM of 50 mg/kg/day for 5 weeks, and then reared without medication for 4 weeks. Pathological changes of the three organs were observed each week for 9 weeks and the results obtained were summarized as follows: 1. Mean body weights of both groups manifested slow increasing tendency but mean hypophysis weights fluctuated throughout the experimental term. Mean ovary weights of experiments were decreased significantly from the 3rd to 6th week but mean thyroid weights of experiments were increased significantly from the 1st to 6th week compared with those of controls. 2. Many ovarian follicles of each developing stage showed follicular atresia accompanying atrophy or necrosis of oocytes and of disintegrated follicular cells. Theca interna cells and sudanophilic interstitial cells showed atrophy and diminished sudanophilic granules and also liquor folliculi were diminished. These changes icreased from the 1st week, remaining so for 5 weeks and returned to normal status in the 8th or 9th week. 3. The thyroid gland showed a typical hyperplastic goiter. Hypertrophic and hyperplastic epithelia follicular manifested cuboidal or columnar form showing tiny or small vacuoles in cytoplasm. The follicles showed atrophy and decreasing colloidal materials. Necrotic and regenerative changes were also present. The interfollicular vessels showed congestion and hemorrhage. These changes increased from the 1st week, remaining so for 5 weeks and returned to normal status in the 9th week. 4. The rates of differential cell counts of hypophyses revealed increase of basophils (gonadotrophs and thyrotrophs) and decrease of chromophobes. Basophils which had diminished granules stainable with HE, PAS and AF revealed hypertrophy, hyperplasia, and increasing of tiny or small vacuoles in cytoplasm. These changes increased from the 1st week, remaining so for 5 weeks and returned to normal status in the 8th or 9th week. As summarized above histologically, administration of SDM led thyrotrophs and gonadotrophs of pituitary glands to hyperactivity but revealed retrogressive and compensatory changes with functional disturbance in ovaries and thyroid glands. These changes were transitional and attributed to direct actions of the drugs on the ovaries and thyroid glands.

• Clinical and Experimental Reproductive Medicine
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• 제15권1호
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• pp.17-24
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• 1988

Follicular fluid(FF) prostaglandin $E_2$(PG$E_2$) and $PGF_{2{\alpha}}$ levels were compared in 3 groups of spontaneously ovulatory women undergoing ovulation induction with clomiphene citrate (CC) alone or with human menopausal gonadotropin(hMG) (14 patients), hMG(9 patients), or pure FSH/hMG(11 patients) for the purpose of in vitro fertilization. FF volume aspirated did not differ significantly according to the maturity of the oocyte. According to hyperstimulation regimens, the volume of FF from which preovulatory occytes were obtained was significantly less in the hMG-treated group than in the other groups. In follicles of preovulatory oocytes, FF PG$E_2$ values were significantly lower in the FSH treated group than in the Cc.treated or hMG-treated group, and FF $PGF_{2{\alpha}}$ values were significantly higher in the hMG-treated group than in the CC-treated or FSH-treated group. In follicles of immature or atretic oocytes, there was no significant difference in FF PG$E_2$ and PG$F_{2{\alpha}}$ concentrations of the similar morphology of the oocyte according to hyperstimulation regimens. In all cycles, FF PG$E_2$ and PG$F_{2{\alpha}}$ values of preovulatory oocytes were not significantly different from those of immature oocytes, but those of atretic oocytes were relatively lower than those of fertilizable oocytes and it was statistically signifincant in PG$E_2$ values of CC-treated group. In all treatment groups, FF PG$E_2$ and PG$F_{2{\alpha}}$ levels did not show and close relationship with the success of fertilization in vitro and of pregnancy after embryo transfer. Above results suggested that FF PG$E_2$ and PG$F_{2{\alpha}}$ be involved in oocyte maturation and ovulation, but their relationship with the success of in vitro fertilization was not found.

• 한국발생생물학회지:발생과생식
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• 제2권1호
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• pp.1-8
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• 1998

본 연구에서는 자연세포사 (apoptosis)를 유발시키는 것으로 알려진 ceramide를 배양중인 생쥐 과립세포에 처리한 뒤 형광염색, in-situ 3'-end labeling(ISEL), 그리고 flow cytometry 기법을 이용하여, 자연세포사 및 세포주기에 미치는 ceramide의 영향을 조사하였다. Ceramide를 처리하지 않은 대조군에 비하여, ceramide를 처리한 실험군에서 세로의 생존율은 농도에 비레하여 유의하게 감소하였다. 또한 acridine orange에 의한 형광염색 결과, 자연세포사의 양상을 보이는 핵을 갖는 세포의 수가 ceramide의 농도가 증가함에 따라 현저하게 증가되었다. 또한 ISEL을 실시해 본 결과, 자연세포사가 ceramide의 처리농도가 증가됨에 따라 점차적으로 증가되었다. 한편, ceramide를 처리한 과립세포의 세포주기 분석을 위한 flow cytometry 결과도 자연세포사가 일어난 $A_{0}$기에 있는 세포들의 비율이 대조군에 비하여 농도 의존적으로 증가하였으며, $G_{0}$/$G_{1}$ 기에 있는 세포들이 비율은 현저하게 감소됨을 관찰할 수 있었다. 위의 결과로 보아 ceramide는 생쥐 과립세포의 $G_{0}$/$G_{1}$ 기에 특이저긍로 자\ulcorner하여 자연세포사를 유발하며, 난포의 폐쇄시 과립세포의 자연세포사를 유발할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제15권12호
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• pp.1695-1701
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• 2002

The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.

• Clinical and Experimental Reproductive Medicine
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• 제37권3호
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• pp.239-244
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• 2010

목 적: 본 후향적 연구는 성선자극호르몬분비호르몬 작용제 (gonadotropin-releasing hormone [GnRH] agonist)와 길항제 (GnRH antagonist) 치료를 받은 불량반응군의 결과를 비교, 분석하고자 하였다. 연구방법: 총 172회의 체외수정시술 주기에서 GnRH agonist 또는 antagonist protocol로 과배란유도를 시행받고 채취된 난자의 수가 5개 이하인 불량반응군을 대상군으로 하였다. 난포 및 채취된 난자의 수, 수정률 등의 결과를 두 군 간에 비교하였다. 결 과: GnRH agonist long protocol과 antagonist protocol 두 군 간에 난포 및 난자의 수와 수정률은 차이를 보이지 아니하였다. 반면, 과배란유도 제7/8일의 혈중 $E_2$ 농도는 GnRH antagonist군에서 더 높았던 반면, 사용한 평균 성선자극호르몬의 용량은 유의하게 적고 과배란유도 기간은 짧은 것을 확인할 수 있었다 (각각 p<0.01). 결 론: 불량 반응군에서 GnRH agonist long protocol에 비하여 GnRH antagonist protocol의 경우 노력이 상대적으로 적게 필요한 반면 비슷한 임상적 결과를 고려할 때, GnRH antagonist protocol이 상대적으로 우수한 것으로 생각된다.

• Clinical and Experimental Reproductive Medicine
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• 제17권2호
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• pp.173-184
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• 1990

At infertility clinic, department of Obstetrics and Gynegology, Kyung Hee Medical Hospital, 80 patients who underwent IVF-ET from January to July, 1989 were evaluated for the prediction of endogenous LH Surge and its effects on outcome of controlled ovarian hyperstimulation (COH) were compared among LH Surge group without hCG given (N=18), with hCG given (N=5), and no-LH Surge group with hCG given (N=57). LH Surge were occurred in 23(28.7%) out of 80 patients studied. Serum E2 levels on Day-1, Day 0, Day+1, were no significant different among three groups. When basal serum LH/FSH ratio is above 1.0, the possibility of endogenous LH Surge is much higher (56.3% in LH Surge group without hCG given). Serum P4 levels on Day 0 were significantly increased in LH Surge group without hCG given. Cycles which serum P4 level is higher than l.0ng/ml were 70.6% of LH Surge group without hCG given. But there was no significant interrelationship between endogenous LH Surge and serum P4 rising rate as an efficient predictor of the occurrence of endogenous LH Surge in COH for IVF. There was no significant differences in number of follicles, follicular size on Day-1, Day 0, Day+1, and number of oocyte collected per cycle. The oocyte fertilization rate of No-LH surge group with hCG given was significantly higher than LH Surge group without hCG given. There was no significant difference in oocyte cleavage rate among three groups.

• Clinical and Experimental Reproductive Medicine
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• 제38권3호
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• pp.164-167
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• 2011

Objective: There are limited data regarding the significance of elevated serum CA-125 level during IUI cycles, even though it is used widely during the initial evaluation of infertile patients. The aim of this study was to investigate the prognostic value of serum CA-125 levels during IUI cycles. Methods: Among the patients with controlled ovarian stimulation and IUI cycles at Seoul National University Hospital from Jan 2005 through Dec 2009, 92 cases with no identified endometriotic lesion, ovarian tumor, salpingeal lesion, or uterine myoma were selected. To compare the clinical characteristics between the pregnancy group and the non-pregnancy group, the Mann-Whitney U test and Fisher's exact test were used. Results: The overall pregnancy rate was 18.5% (17/92). The pregnancy group showed a higher number of follicles 16 mm in diameter ($p$=0.036), endometrial thickness ($p$ <0.001), ampules of gonadotropin ($p$=0.009), and higher body mass index ($p$=0.022) than the non-pregnancy group. No significant difference was observed in the serum CA-125 level or the proportion of patients with CA-125 exceeding 17 IU/mL between the two groups. Conclusion: The prognostic value of serum CA-125 level among infertile patients with IUI cycles is considered limited.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.233-240
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• 1994

This study was conducted to compare the endocrine milieu, and pregnancy rates in In Vitro Fertilization and Embryo Transfer(IVF-ET) program employing combined with gonadotropin releasing hormone agonist(GnRH-a) and pergonal(LH 75lU+FSH 75lU) when either human chorionic gonadotropin(HCG) or progesterone were used for luteal phase support. A total number of 40 IVF-ET treatment cycles were prospectively studied. Ovarian hyperstimulation method was modified ultrashort protocol using GnRH-a. All patients started Decapeptyl at menstrual cycle day # 2, and HMG was started at # 3 days. When leading follicle was ${\geqq}$18mm or at least two follicles were ${\geqq}$14mm in diameter, HCG 10000lU intramuscularly was injected. After 36 hours HCG administration, oocytes were retrieved as usual guided by transvaginal ultrasound. Embryo were transfered 36-48 hours later. The patient's cycles were prospectively randomized to receive HCG(20cycles) or Progesterone (20cycles) for luteal support. The progesterone group received 25mg 1M starting from the day of ET. The HCG group received 1500IU 1M. on days 0, +2, +5 after ET. Estadiol($E_2$) and Progesterone($P_4$) were measured on the day of oocyte aspiration, ET day, and every 6 days thereafter. Results were follows as; 1. Estradiol, progesterone and LH levels on the day of HCG trigger, retrieved oocytes and number of transfered embryo were not significantly different in both groups. 2. On the day of aspiration and embryo transfered day, $E_2$, $P_4$ level were significantly higher in progesterone group than HCG group(p<0.01). 3. $E_2$, $P_4$ level on 6 days after ET were significantly higher in progesterone group than HCG group(p<0.01). But, $P_4/E_2$ ratio was not different in both groups. 4. $E_2$, $P_4$ level 12 days after ET were decreased abruptly in both groups and higher hormonal level appeared in HCG group(P<0.01). 5. The total pregnancy rate in the HCG group was 40% (8/20) and in the progesterone group 15%(3/20). 6. Comparing the pregnant and nonpregnant cases progesterone group was not different the hormonal status. In HCG group, pregnant cases appeared in higher $P_4$, $P_4/E_2$ ratio than nonpregnanct cases(P<0.01).

• Asian-Australasian Journal of Animal Sciences
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• 제28권2호
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• pp.171-179
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• 2015

The present study was aimed at investigating the adverse effects of dietary zearalenone (ZEA) (1.1 to 3.2 mg/kg diet) on serum hormones, morphologic and apoptotic measurements of genital organs in post-weaning gilts. A total of twenty gilts ($Landrace{\times}Yorkshire{\times}Duroc$) weaned at 21 d with an average body weight of $10.36{\pm}1.21kg$ were used in the study. Gilts were fed a basal diet with an addition of 0, 1.1, 2.0, or 3.2 mg/kg purified ZEA for 18 d ad libitum. Results showed that 3.2 mg/kg ZEA challenged gilts decreased (p<0.05) the serum levels of luteinizing hormone, however, serum levels of prolactin in gilts fed the diet containing 2.0 mg/kg ZEA or more were increased (p<0.05) compared to those in the control. Linear effects on all tested serum hormones except progesterone were observed as dietary ZEA levels increased (p<0.05). Gilts fed ZEA-contaminated diet showed increase (p<0.05) in genital organs size, hyperplasia of submucosal smooth muscles in the corpus uteri in a dose-dependent manner. However, the decreased numbers of follicles in the cortex and apoptotic cells in the ovarian were observed in gilts treated with ZEA in a dose-dependent manner. Degeneration and structural abnormalities of genital organs tissues were also observed in the gilts fed diet containing 1.1 mg/kg ZEA or more. Results suggested that dietary ZEA at 1.1 to 3.2 mg/kg can induce endocrine disturbance and damage genital organs in post-weaning gilts.