• 대한생식의학회:학술대회논문집
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• 2007.11a
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• pp.95-95
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• 2007

• Journal of Life Science
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• v.12 no.3
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• pp.317-324
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• 2002

We investigated the compositional changes of glycoconjugates (GCs) and expression of estrogen receptor (ER)-$\alpha$, c-fos and c-jun in the vagina of normal and ovariectomized rats by histochemical and immunohistochemical methods. The mucinous transformation of the superficial layer that occurred from late diestrus to proestrus was accompanied with extensive enrichment of GCs. According to the cyclic changes of the vagina, distinct reactivity patterns such as SBA affinity in the diestrus and Con A affinity in the diestrus and estrus phase was observed. However, weak staining for GCs was detected in the atrophied vaginal epithelium of ovariectomized rats. ER-$\alpha$ immunoreaction was mainly demonstrated in the basal layer of epithelium and estrus cycle-related variation in the number of ER-$\alpha$ immunoreaction were not pronounced. But the stromal cells showing ER-$\alpha$ immunoreaction were abundantly observed from diestrus to estrus phase. The most numerous c-fos immunoreactive cells were observed in the basal and intermediate layer of epithelium and stromal fells from the proestrus to estrus phase and c-jun in the basal layer of epithelium during estrus phase. The c-jun immunoreaction of stromal cells expressed only in the estrus phase. In the ovariectomized rats, a few of ER-$\alpha$, c-fos and c-jun immunoreactive cells were observed in the vaginal epithelium and no immunoreaction were found in the stromal cells. ER-$\alpha$ and c-fos immunoreaction fully expressed in the proestrus coincident with the cell proliferation, mutinous transformation and cornification of vaginal epithelium. These data indicate that vagina epithelium and stromal reals express multiple protein such as ER-$\alpha$, c-fos and c-iun by estrogen that may function in process of cells proliferation and differentiation of vagina epithelium.

• Journal of the Korean Society of Food Science and Nutrition
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• v.34 no.1
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• pp.27-35
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• 2005

This study was performed to determine the effect of three different concentrations of soy-isoflavones on excretions through urine and feces in either sham-operated or ovariectomized female rats. Seventy-two 16-week old Sprague-Dawley rats underwent sham operation or bilateral ovariectomy. They were provided diets containing different levels of soy isoflavones for 6 weeks: 50 ppm (low isoflavone intake; LI), 250 ppm (medium isoflavone intake; MI) and 500 ppm (high isoflavone intake; HI). The subsequent fecal and urinary excretions of daidzein and genistein were then measured. In the sham operated rats, body weight gains and food efficiency ratio of the MI and HI groups were significantly lower than control group, while food intake was not different. However, there was no significant difference in ovariectomized rats. The fecal excretion of daidzein was significantly higher in the HI group than the LI and MI groups, and that of genistein increased as dietary isoflavone intakes increased in both the sham operation and ovariectomy. The reverse tendency of fecal recovery was shown with dietary isoflavone dose only in genistein among ovariectomized rats. When dietary isoflavones were increased from 50 ppm to 500 ppm, the amounts of daidzein and genistein in the urine increased dose-dependently. The higher intakes of isoflavones leaded to lower recovery rates of daidzein and genistein in the urine in the sham-operated rats but not in the ovariectomized rats. The urinary recovery was significantly higher in the LI group than the MI and HI groups in the sham-operated rats. The excreted amounts and recovery of the two isoflavones in the urine were higher in the sham operated groups than in the ovariectomized groups, which implied an increased bioavailability of isoflavones by ovariectomy. Therefore, the results suggest that a more efficient use of soy isoflavones in ovariectomized conditions may occur, which indicates that a higher dose of soy isoflavones is necessary for the postmenopausal states.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.253-258
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• 1997

토끼에서는 수정란 이식과 같은 기본적인 번식공학적 방법의 효율성이 아직 생쥐와 같은 실험동물에 비해 떨어지고 있어 생물공학적인 기술을 응용하는데 큰 어려움이 있다. 특히 유전자 이식에 의한 형질전환 토끼의 생산과 같은 생물공학적인 기술을 실용화하는데 효율이 높은 수정란이식 기술의 개발이 필수적이라고 할 수 있다. 본 연구에서는 토끼에서 수정란 이식 기술의 첫 단계인 과배란 유도를 효율적으로 이용할 수 있는 방법을 정립하기 위해 반복적인 과배란 유도가 배란율 및 수정란의 질적인 면과 양적인 면에 미치는 영향을 조사하였다. 연구방법으로는 FSH와 HCG를 사용하여 과배란을 유도하였고 2.5 개월의 반복처리간격으로 3번의 반복적인 과배란 처리를 한 후 반복처리에 따른 배란율과 배란된 난자의 형태학적 상태, 배양에 의한 발생 능력 상태 등을 감소하였으며 배란수의 변이도 커지는 경향을 나타내었다(첫번째, 32.6(+-)2.5; 두번째 28.7(+-)3.7; 세번째 20.99(+-)3.8). 제 2극체의 돌출, 전핵의 형성, cummulus cell의 존재등에 의한 회수된 난자의 형태학적 관찰에 의한 방법으로 난자를 분류한 결과 과배란의 반복수가 증가함에 따라 다양한 모양의 난자가 회수되어 배란이 광범위한 시간대에 일어나고 있음을 나타내었다. 또한 과배란의 반복적인 유도에 의해 난소의 혈포수는 증가하였으나 채란된 난자의 채외배양에 의한 발육율에는 차이가 없었다. 그러므로 과배란의 반복적인 유도는 공란토의 난소반응에는 영향을 미쳤으나 난자의 질에는 영향을 미치지 않았음을 나타낸다.

• Journal of Embryo Transfer
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• v.7 no.2
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• pp.117-124
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• 1992

미성숙 래트의 외경정맥에 카테타를 장착하고, 다음날 (28일령) 대조군에는 4IU, 다배란 처치 군에는 20IU의 PMSG를 피하주사하였다. 각 실험동물은 혈중의 LH농도 변화를 측정하기 위하여 PMSG 투여 직전 (0시간), 투여후 12시간, 그 이후 6시간 간격으로 혈액을 채취하고 72시간에 희생시켰다. 그 결과 다배란 용량의 PMSG 투여는 먼저 배란반응 및 난소중량을 대조군에 비하여 각각 4,7배 및 2.1배나 현저하게 (P<0.05) 증가시켰다. 그리고 혈청 LH농도는 Radioimmunoassay(RIA)에 의하여 결정되었는데, 먼저 두 군 모두 두 개의 분명한 peak을 가진 경시적 변화관계를 보였다. 즉 이들 두 군네 있어서 LH농도 변화는 0-18시간대에 처음으로 완만한 증가와 54-60시간대에 두번째는 급격한 증가(surge)를 보였다. 그러나 두 군간에 LH농도의 크기는 현저하게 달라, 다배란처치군의 동물에 있어서는 두번째의 LH peak에 앞서 전반적인 LH농도가 대조군보다 현저하게 (P<0.001) 높았으며, 반대로 PMSG 투여후 60시간에 일어나는 peak에 있어서는 LH농도가 대조군보다 현저하게 (P<0.001) 54%나 낮았다. 덧붙여 두 peak간의 증가폭은 대조근에 비하여 다배란 처치군에서 훨씬 낮았다. 본 열구 결과는 PMSG 처치된 래트에 있어서 두 가지의 분명한LH peak의 존재를 정의하며, 다배란 처치에 따른 난소과잉 자극과 내인성 LH surge의 감소와의 연관성을 밝힌다.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.108-108
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• 2002

포유동물의 암컷 생식기관에 존재하는 다양한 종류의 matrix metallo-proteinase (MMP)는 난소와 자궁의 구성성분의 주기적인 변화를 조절하며 이중 난소의 MMP는 난포의 성장과 배란 그리고 퇴화 동안 조직재구성에 매우 중요한 역할을 한다고 알려져 있다. 본 실험에서는 근래에 새로 발견된 사람의 난포액에 존재하는 분자량 약 110kDa인 MMP-2 isoform GA110을 동정하고 자 하였다. 난포액으로부터 GA110 단백질을 분리하기 위하여 난포액에 5mM ethylenediaminetetraacetic acid(EDTA)를 처리한 후 DEAE Sepharose Fast Flow를 이용한 chromatography를 시행하였다. 그 결과, 난포액 단백질들은 0.2M NaCl 의 분획에서 GA110 활성을 나타내었고 anti-human MMP-2 antibody에 대한 면역반응도 뚜렷이 나타났다. DEAE Sepharose Fast Flow에서 얻은 분획 중 GA110의 활성과 면역반응을 모두 나타내는 분획만을 모아 Gelatin Sepharose 4B affinity chromatography로 다시 분리하였다 분리한 결과 resin에 흡착된 단백질 (eluate) 분획들에서 매우 뚜렷한 GA110 gelatinase 활성을 나타내었으며 면역반응 또한 관찰되었다. 이 분획들의 단백질을 농축한 후 zymography를 시행하여 나타난 GA110 단백질 band를 잘라 내었으며 이로부터 단백질을 electroelution하여 농축한 후 reducing agent인 2-mercaptoethanol를 처리하였다. 이를 전기영동 후 MMP-2 (propeptide region) antibody를 사용하여 immunoblotting 한 결과 70-72kDa의 단백질만이 면역반응을 나타내었다. 마지막으로 위와 같이 준비된 70-72kDa 단백질의 아미노산 서열을 Edman degradation 방법으로 분석하였다. 그 결과 이 단백질의 N 말단의 10개의 아미노산 배열 순서가 알려진 사람의 proMMP-2의 전체 배열순서 중 propetide domain의 N 말단에서부터 다섯 번째에서 시작하여 10개의 아미노산의 서열과 정확하게 일치하였다. 위 결과들로 미루어 사람의 난포액에 존재하는 MMP-2의 새로운 isoform인 GA110은 70-72kDa의 ProMMP-2가 disulfide bond를 통해 homodimer 구조를 이루고 있는 것으로 여겨진다.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.1
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• pp.69-76
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• 2008

Objective: The purposes of this study were to determine the distribution of follicle-stimulating hormone receptor (FSHR) genotypes in infertile Korean women and to evaluate the relationship between FSHR genotypes and clinical outcomes of IVF-ET cycles. Methods: Genomic DNA was extracted from peripheral blood in 1, 020 of infertile Korean women. Genotypes of FSHR at Thr307Ala (T/A) and Asn680Ser (N/S) were screened by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Clinical outcomes related to the genotypes of FSHR were evaluated in IVF-ET cycles (n=302) with controlled ovarian hyperstimulation (COH) of infertile women under 40 years old. Results: In a population of 1, 020 infertile Korean women, the frequency of TT/NN, TA/NS and AA/SS for the major variant Thr307Ala and Asn680Ser was 44.80%, 41.96% and 10.49%, respectively. There was no significant difference in characteristics of ovarian response and clinical pregnancy rate among the major genotypes of FSHR in IVF-ET cycles with COH. However, implantation rate of AA/SS patients was significantly higher than that of TT/NN patients (24.5% vs 15.7%, p<0.05). Conclusion: This study showed that FSHR genotype was not directly associated with ovarian response in IVF-ET cycles with COH. The relationship between clinical outcomes and FSHR genotypes of patients should be substantiated by further studies.

• Korean Journal of Veterinary Research
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• v.32 no.1
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• pp.15-24
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• 1992

The effects of CC the ovulatory response, oocyte normality, ovarian steroidogenesis and subsequent embryo developmental potential were examined in PMSG-treated rats. On Days of 25~27 of age, immature female Sprague Dawley rats were treated with three different doses(0.05, 0.1 or 1.0mg /day) of clomiphene citrate or vehicle. The females subsequently received 4IU PMSG on Day 28 and/or 10IU hCG on Day 30, and were killed on Day 31. Some females given 0.1mg CC or vehicle with 4IU PMSG were then mated and killed on Days 2, 3, 4 and 5 of pregnancy. Compared to vehicle(control) group, by increasing the doses of CC, there were a significant decrease in the ovulatory response as judged by both the proportion of rats ovulating and the mean number of oocytes per rat and a marked reduction of ovarian weight. The increasing doses of CC substantially promoted the degeneration(%) of oocytes ovulating in a dose-dependent manner. The CC-mediated inhibitions of the ovulatory response and ovarian weight were oompletely overcome by a subsequent treatment of hCG. Increasing doses of CC resulted in a siginificant elevation of serum estradiol with the decreased levels of progesterone and androgens. The additive treatment with hCG was effective to reduce the elevation of estradiol and to increase the reduction of progesterone produced by high dose(1.0mg) of CC. The preimplantation embryos recovered from 0.1mg CC-treated pregnant rats demonstrated a progressive early loss from Day 3 of pregnancy with a significant increase in the percentage of degeneration during all periods examined, compared to controls. The rate of progressive embryo cleavage in the CC-treated rats were slower than that in controls from Day 3 of pregnancy. Additionally, the percentage of the cleaved embryos recovered from the CC-treated rats remained significantly lower consistently from Day 2 of pregnancy, compared to control regimen. These results demonstrate a possible mechanism of CC-mediated inhibition of ovulatory response in the rats which may include the attenuation or blockade of the endogenous secretion of gonadotropins and also suggest that its detrimental effects observed on oocyte normality and embryonic development may be caused by abnormal follicular steroidogenesis( especially elevated estradiol) preceding fertilization.

• Korean Journal of Animal Reproduction
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• v.12 no.3
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• pp.167-173
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• 1988

The present experiment was carried out to investigate super-ovulating response, ovulation time and fertilizability in the angora rabbit that gonadotropin was treated. The results obtained were summarized as follows ; 1. Number of ovulating point and ovarian weight in PMSG 100I.U.(25${\pm}$2.45;1104.2${\pm}$110.6mg) and PMSG 200I.U.(30.6${\pm}$1.76, 1330.0${\pm}$153.9mg) treated group were significantly higher than natural mating(6.6${\pm}$1.49, 560.2${\pm}$60.6mg) and HCG treated group(9.8${\pm}$0.8;651.6${\pm}$55.1mg)(P<0.01). 2. Survival rate and recovery rate in natural mating, HCG and PMSG 100I.U. treated group were significantly higher than PMSG 200I.U. treated group.(P<0.01)(P<0.05). 3. Ovulation started at 10hrs and mostly finished at 16hrs after HCG injection. 4. The fertilizable life of egg ovulated was during 8hrs after ovulation.

• Journal of Veterinary Clinics
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• v.5 no.1
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• pp.31-35
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• 1988

The effects of inflammation on the changes in some blood metabolites and ovarian response to super-ovulation treatment in sheep were investigated. Inflammation was induced artificially with LPS(Lipopoly-saccharide E. coli； 0.2mg/kg). Changes in the levels of NEFA, GOT, ${\gamma}$-GTP and body temperature after LPS injection were observed. Superovulation was carried out about 2 months after LPS injection. The body temperature, the levels of NEFA and GOT increased, and the level of total cholesterol decreased transiently. However, the level of ${\gamma}$-GTP did not change significantly. Superovulation results were not different between LPS injection and control groups. It was indicated that superovulation results did not decrease in the sheep which showed regular estrous cycle even after the induction of inflammation.