• 한국양식학회지
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• 제11권1호
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• pp.119-124
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• 1998

농어의 난성숙과 배란유도를 위한 C하(21)-스테로이드와 HCG의 효능 비교실험이 in vitro에서 이루어졌다. 분리된 난모세포를 대상으로 17${\alpha}$-hydroxy, 20${\beta}$-dihydroprogesterone(17${\alpha}$20${\beta}$OHP), 17${\alpha}$-hydroxy, 20${\beta}$-dihydroprogesterone(17${\alpha}$20${\beta}$OHP : 5~100ng/ml)와 HCG(5~500IU/ml)를 사용하여 관찰한 난성숙 유도효과, 즉 GVM(germinal vesicle migration)과 GVBD(germinal vesicle breakdown)를 보면, 5ng/ml 농도의 17${\alpha}$20${\beta}$OHP와 17${\alpha}$20${\beta}$OHP를 제외한 모든 실험군에서의 반응은 대조군보다 효과가 있는 것으로 나타났으며, 특히 17${\alpha}$20${\beta}$OHP 50ng/ml 농도에서 가장 높은 성숙 유도효과를 보였다. GVBD 유도효과에 대한 17${\alpha}$20${\beta}$OHP(50ng/ml), HCG(50IU/ml) 그리고 이들 호르몬을 혼합처리한 17${\alpha}$20${\beta}$OHP+HCG에서는 혼합처리한 실험군이 가장 높은 반응을 보여 이들의 상호 보완적 작용을 관찰할 수 있었다. 난소조직을 대상으로 HCG와 17${\alpha}$20${\beta}$OHP를 사용하여 in vitro에서 농어의 배란유도효과를 관찰한 결과 사용한 모든 농도(17${\alpha}$20${\beta}$OHP : 1~1000ng/ml, HCG : 1~500IU/ml)에서 배란반응을 보였다. 특히 저농도 50ng/ml 또는 50IU/ml 이하로 처리하는 것이 더 효율적인 것으로 나타났다. 또한 HCG 처리가 17${\alpha}$20${\beta}$OHP보다 배란 반응에 더 민감하게 작용하는 것으로 생각된다.

• 한국양식학회지
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• 제20권3호
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• pp.160-167
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• 2007

극동산 뱀장어의 자연 수정란을 이용하여 $23{\pm}0.5^{\circ}C$의 여과 해수가 공급되는 실내수조에서 난발생과 pre-leptocephalus 자어발달을 관찰하였다. 친어 관리에 있어서 암컷은 연어 뇌하수체 추출물(20 mg/fish)과 수컷은 HCG (1 IU/g BW)를 매주 복강 주사하여 인위적인 성성숙 및 자연산란을 유도하였다. 수정란은 직경 1.0 mm 정도의 분리 부성란으로, 전형적인 반상분할(discoidal cleavage)을 통해 분할이 이루어졌다. 부화는 $23^{\circ}C$의 여과 해수에서 수정 후 38시간부터 개시되었다. 부화 직후의 자어는 전장 약 3.0 mm, 근절수는 평균 42개였다. 부화자어의 입과 항문은 부화 후 4.5일째에 열렸고, 난황은 부화 후 6.5 일째에 거의 흡수되었다. 자어는 14.5일 동안 생존하였고, 이때 전장 $5.87{\pm}0.25mm$, 근절은 98개였다. 그러나 수정란과 prelaptocephalus 자어발생의 형태학적 특징은 자연수정과 건식법에 의한 인공수정과 큰 차이점은 없었다.

• 한국양식학회지
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• 제21권2호
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• pp.96-101
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• 2008

능성어 암컷의 난모세포 발달에 따른 성숙 및 배란유도를 위하여 제주도 연안에서 채집하여 사육중인 능성어 51마리(전장 $69.1{\pm}1.0cm$, 체중 $5.8{\pm}0.3kg$)를 대상으로 암컷의 생식소 발달을 cannulation으로 조사하였다. 난황구기 단계 이상 난모세포(난경 $300{\sim}500{\mu}m$)를 가진 어미를 대상으로 HCG 500 IU/kg BW로 주사하였다. HCG 처리 후 난경변화는 24시간째에 $300{\sim}700{\mu}m$, 48시간째에는 $800{\mu}m$ 이상이 $91.3{\sim}98.8%(95.1{\pm}3.7%)$이었다. HCG 처리 실험구에서는 실험어 8마리 중 7마리가 배란되었고, 총 배란된 알 2,480 mL이었으며, 이중 부상란은 1,360 mL이었다. 수정률과 부화율은 각각 $56.2{\sim}94.9%(77.8{\pm}2.9%),\;70.0{\sim}97.7%(92.2{\pm}2.1%)$이었다. 이들 결과들은 HCG 500 IU/kg BW 농도로 처리했을 때 난황구기 난모세포를 가진 능성어 암컷의 성숙 및 배란에 효과가 있을 것으로 사료된다.

• Fisheries and Aquatic Sciences
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• 제12권4호
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• pp.293-298
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• 2009

Several studies have reported that nonylphenol (NP) and 2,2', 4,6,6'-pentachlorobiphenyl (PCB104) exhibit estrogenic activity. To investigate the estrogenic potency of NP and PCB104 during oocyte maturation, fully vitellogenic oocytes (0.76 mm diameter in average) of yellow fin goby, Acanthogobius flavimanus, were exposed in vitro to these chemicals at different concentrations (0.1, 1, 10, 100, and 1,000 ng/mL) with the exogenous precursor $17\alpha$-hydroxyprogesterone ($17{\alpha}OHP$) 50 ng/mL in the presence or absence of human chorionic gonadotropin (HCG). The production of testosterone (T), estradiol-$17\beta$ (E2), and $17\alpha,20\beta$-dihydroxy-4-pregnen-3-one ($17\alpha20{\beta}OHP$) in response to NP or PCB104 were measured by radioimmunoassay. Steroid levels were also expressed as E2/T and E2/$17\alpha20{\beta}OHP$ ratios. In the absence of HCG, no significant differences in either NP or PCB104 treatment groups were observed. In the presence of HCG, NP treatment did not show significant differences in the production of T, E2, and $17\alpha20{\beta}OHP$ at any concentrations tested, but E2/T ratios were decreased at concentrations of 0.1, 1, 10, and 1,000 ng/mL compared with the control group. PCB104 decreased E2 production at concentrations of 0.1, 10, and 1000 ng/mL, but did not show significant differences in the production of T and $17\alpha20{\beta}OHP$ at any concentration tested. While E2/T ratios were decreased at PCB104 concentrations of 0.1, 1, 10, and 1,000 ng/mL, E2/$17\alpha20{\beta}OHP$ ratios were also decreased at 0.1, 10, and 1,000 ng/mL compared with the control. Results indicate that both NP and PCB104 appeared to have antiestrogenic effects during this phase.

• Clinical and Experimental Reproductive Medicine
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• 제47권3호
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• pp.213-220
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• 2020

Objective: The aim of this study was to explore the potential adverse effect of spontaneously decreasing serum estradiol (SE) levels on in vitro fertilization (IVF) outcomes. Methods: This retrospective single-subject study analyzed IVF cycles conducted at a hospital IVF unit between 2010 and 2017. Overall, 2,417 cycles were analyzed. Only cycles with spontaneously decreasing SE before human chorionic gonadotropin (hCG) triggering were included. Each patient served as her own control, and subsequent cycles were analyzed for recurrent SE decreases. The main outcome was the number of oocytes retrieved. Results: Cycle characteristics were similar between the study (SE decrease) and control groups, with the exception of the median SE on the day of hCG triggering (899.7 pg/mL; interquartile range [IQR], 193-2,116 pg/mL vs. 1,566.8 pg/mL; IQR, 249-2,970 pg/mL; p< 0.001). The study group, relative to the control group, had significantly fewer total oocytes (5 [IQR, 2-9] vs. 7 [IQR, 3-11]; p= 0.002) and significantly fewer metaphase II (MII) oocytes (3 [IQR, 1-6] vs. 4 [IQR, 2-8]; p= 0.001) retrieved. The study group had fewer cleavage-stage embryos than the control cycles (3 [IQR, 1-6] vs. 4 [IQR, 2-7]; p= 0.012). Compared to cycles with a ≤ 20% SE decrease, cycles with a > 20% decrease had significantly fewer total and MII oocytes retrieved. SE decrease recurred in 12% of patients. Conclusion: A spontaneous decrease in SE levels adversely affected IVF outcomes, with a linear correlation between the percentage decrease and the number of oocytes retrieved. SE decrease can repeat in later cycles.

• 대한핵의학회지
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• 제36권2호
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• pp.133-139
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• 2002

목적: 섬광 근접측정법은 항원 항체 반응 후 결합 분획과 유리분획을 분리하는 과정이 필요없다. 이러한 원리를 검체 내 hCG와 항 ${\alpha}$ hCG 항체간의 항원 항체 반응에 적용하고자 한다. 대상 및 방법: 항 ${\alpha}$ hCG 항체를 biotin과 결합시켜 SPA bead에 부착된 streptavidin과 부착 가능하게 만들었다. 이 측정법은 항 ${\alpha}$ hCG 항체가 부착된 SPA beads에 대해 혈청내 hCG와 표지항원인 $[^{125}I]hCG$간의 경쟁 반응을 기본 원리로 이용하였다. Biotin 표지 항 ${\alpha}$ hCG 항체를 $[^{125}I]hCG\;100{\mu}{\ell}$와 표준용액이나 환자 혈청 $200{\mu}{\ell}$이 들어있는 실온에서 20분 방치하였다. 그리고 streptavidin이 붙은 SPA beads $20{\mu}{\ell}$를 바이알에 넣고 10분 더 방치한다. 환자 혈청의 수치를 표준 응답곡선을 통해 계산하였다. 결과: SPA측정법에 사용되는 방사성 핵종의 방사능 양에 따라 반응용액 속에서 SPA bead와 자유 방사성 핵종에 의한 배후 방사능이 측정값에 영향이 없음을 확인하였다. SPA 방법을 응용한 측정에서 적합한 표준 응답곡선을 얻었고, 실제 환자혈청에서의 hCG 농도를 결정할 수 있었다. 결론: 이 실험을 통해 SPA 방법을 이용한 측정법이 임상진단에 유용하게 사용될 수 있을 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제17권3호
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• pp.317-324
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• 2004

The present study was conducted to investigate the involvement of nitric oxide (NO) in cumulus expansion, oocyte mortality and meiotic maturation of porcine cumulus enclosed oocytes (CEOs) cultured in two different models when gonadotropins, including follicle-stimulating hormone (FSH) and human chorionic gonadotropin (hCG) were presented or not. And the interaction between NO and $\beta$-mercaptoethanol ($\beta$-ME), a free radical scavenger was also investigated. Two models refer to spontaneous maturation model and hypoxanthine (HX) medium model. All the 3,433 eligible CEOs were incubated at $39^{\circ}C$ and the cumulus expansion, oocyte morphology and nuclear phase were evaluated 44 h after incubation. (1) In spontaneous maturation model, NO stimulates the cumulus expansion and $\beta$-ME delayed it. NO doesn't affect the oocyte meiotic resumption but inhibits the oocytes to develop to metaphase II. (2) In HX medium model, NO or $\beta$-ME doesn't affect the expansion in the absence of gonadotropins, but in the presence of gonadotropins, NO or $\beta$-ME inhibits the expansion. In the presence of gonadotropins, NO inhibits the oocyte meiotic resumption and it especially inhibits the oocyte to develop to metaphase II, and $\beta$-ME reverses such inhibitory effects. The cooperation of gonadotropins and $\beta$-ME stimulates the meiotic resumption and especially, promotes the CEOs to develop to metaphase II in both models. Moreover, HX might contribute to the fragility of oocyte zona pellucida and gonadotropins, nitric oxide and $\beta$-ME could alleviate it separately, and cooperatively. It is concluded that NO exerts different functions in two models and $\beta$-ME affected the functions of NO in different models.

• 한국발생생물학회지:발생과생식
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• 제16권1호
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• pp.39-45
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• 2012

To verify the sex steroids which are involved in oocyte maturation of the blacktip grouper, $Epinephelus$ $fasciatus$, we incubated vitellogenic oocytes (0.41 and 0.50 mm in average diameter) in the presence of exogenous steroid precursor ($[^3H]17{\alpha}$-hydroxyprogesterone). Steroids were extracted, separated and identified by thin layer chromatography. The major metabolites produced were androstenedione, estradiol-$17{\beta}$, estrone and progestogens. Progestogen metabolites in the oocytes of 0.50 mm were more abundant than those of 0.41 mm. Also, we investigated the $in$ $vitro$ effects of human chorionic gonadotropin (HCG; 5, 50 and 500 $IU/m{\ell}$), $17{\alpha},20{\beta}$-dihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}P$) and $17{\alpha},20{\beta}$-trihydroxy-4-pregnen-3-one ($17{\alpha}20{\beta}21P$; 5, 50 and 500 $ng/m{\ell}$, respectively) on oocyte maturation. In the oocytes of 0.41 mm, treatment with 50 IU HCG stimulated GVBD ($55.30{\pm}1.20%$) compared with controls ($32.41{\pm}3.13%$, $p$<0.05). In the oocytes of 0.50 mm, treatment of $17{\alpha}20{\beta}P$ (50 and 500 $ng/m{\ell}$) stimulated GVBD ($50.13{\pm}2.52$ and $51.77{\pm}5.91%$, respectively) compared with controls ($36.81{\pm}2.89%$, $p$<0.05). Treatment with 500 IU HCG also stimulated GVBD ($49.59{\pm}5.15%$) compared with controls ($p$<0.05). Taken together, these results suggested that both HCG and $17{\alpha}20{\beta}P$ were effective on in vitro oocyte maturation and $17{\alpha}20{\beta}P$ may act as a maturation inducing hormone in blacktip grouper.

• 한국수정란이식학회지
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• 제27권4호
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• pp.245-252
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• 2012

The purpose of the present study was to investigate the effect of IFN-${\tau}$ on prostaglandin synthesis, cyclooxygenase-2 (COX-2) gene expression in vitro and concentration of progesterone (P4) in endometrial cells. Epithelial and stromal cells cultured in vitro were isolated from bovine endometrium and stimulated with increasing doses of IFN-${\tau}$ (0, 0.02, 0.2 and 2 ug/ml). Human chorionic gonadotropin (hCG, 1.5 IU/ml) was used as a positive control. Prostaglandin $E_2$ and $F_{2{\alpha}}$ levels in the culture media were analyzed by enzyme immunoassays and total RNA was extracted from the cells for RT-PCR. P4 concentrations of blood samples were assayed by chemiluminescent immuno assays system. In epithelial cells, COX-2 gene expression was increased in the presence of IFN-${\tau}$ (p<0.05), but it was not significantly different in all groups of stromal cells except for 2 ug/ml IFN-${\tau}$ group (p<0.05). Although IFN-${\tau}$ did not affect $PGE_2$ and $PGF_{2{\alpha}}$ production in epithelial cells, it decreased $PGE_2$ and $PGF_{2{\alpha}}$ production significantly in stromal cells (p<0.05). In vivo experiment, blood concentration of P4 was significantly increased after addition of IFN-${\tau}$ (1 ug/ml). The results indicate that PG production was mediated by COX-2 expression in stromal cells but it was not affected in epithelial cells and this suggest that treatment of IFN-${\tau}$ could improve the implantation environment of uterine by maintenance of high P4 concentration.

• Clinical and Experimental Reproductive Medicine
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• 제43권2호
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• pp.126-132
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• 2016

Objective: The purpose of this study was to identify useful clinical factors for the identification of patients with polycystic ovary syndrome (PCOS) who would benefit from in vitro maturation (IVM) treatment without exhibiting compromised pregnancy outcomes. Methods: A retrospective cohort study was performed of 186 consecutive patients with PCOS who underwent human chorionic gonadotropin-primed IVM treatment between March 2010 and March 2014. Only the first IVM cycle of each patient was included in this study. A retrospective case-control study was subsequently conducted to compare pregnancy outcomes between IVM and conventional in vitro fertilization (IVF) cycles. Results: Through logistic regression analyses, we arrived at the novel finding that serum $anti-M{\ddot{u}}llerian$ hormone (AMH) levels and the number of fertilized oocytes in IVM were independent predictive factors for live birth with unstandardized coefficients of 0.078 (95% confidence interval [CI], 1.005-1.164; p=0.037) and 0.113 (95% CI, 1.038-1.208; p=0.003), respectively. Furthermore, these two parameters were able to discriminate patients who experienced live births from non-pregnant IVM patients using cut-off levels of 8.5 ng/mL and five fertilized oocytes, respectively. A subsequent retrospective case-control study of patients with PCOS who had serum AMH levels ${\geq}8.5ng/mL$ showed that IVM had pregnancy outcomes comparable to conventional IVF, and that no cases of ovarian hyperstimulation syndrome were observed. Conclusion: Serum AMH levels are a useful factor for predicting pregnancy outcomes in PCOS patients before the beginning of an IVM cycle. IVM may be an alternative to conventional IVF for PCOS patients if the patients are properly selected according to predictive factors such as serum AMH levels.