• 한국수정란이식학회지
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• 제26권3호
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• pp.209-214
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• 2011

임신의 성립 및 유지에 중요한 자궁 내막과 호르몬의 변화는 생식기관에서 발현되는 K$_{2P}$ 통로의 발현을 변화시킬 수 있다. 본 연구는 한우의 임신 자궁 내막에서 K$_{2P}$ 통로의 발현 변화가 나타나는지 그리고 프로게스테론에 의해 그 발현량이 변화되는지를 확인하고자 수행하였다. 역전사중합효소 중합반응과 웨스턴블닷 분석을 통하여 임신한 한우의 자궁 내막에서 mRNA와 단백질의 발현 변화를 조사하였다. TREK-1을 제외한 K$_{2P}$ 통로의 mRNA 발현량이 임신 자궁 내막에서 변화되었다. mRNA가 크게 변화되는 TASK-3, TREK-2, TRAAK 및 TRESK의 단백발현 변화량을 임신 자궁 내막에서 확인하였는데, TREK-2와 TRESK만 mRNA 발현 변화 양상과 동일하게 임신 자궁 내막에서 각각 7.9배, 2배 증가하였다. 자궁 내막세포에 프로게스테론(10 ${\mu}g$/mL)을 처리하였을 때 TREK-2와 TRESK는 자궁 내막 조직에서 보여준 결과와 유사하게 단백 발현량이 각각 10배, 6배 증가하였다. 이상의 결과로부터 K$_{2P}$ 통로, 특히 TREK-2와 TRESK는 프로게스테론 변화에 의해 임신 자궁 내막에서 발현량이 증가할 것으로 생각된다. 그리고 증가된 TREK-2와 TRESK는 임신에 의해 유발되는 생리학적 변화를 조절하는데 기여할 것으로 생각된다.

• 한국수정란이식학회지
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• 제26권1호
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• pp.79-84
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• 2011

This study was performed to identify the differentially methylated region (DMR) and to examine the mRNA expression of the imprinted H19 gene in day 35 of SCNT pig fetuses. The fetus and placenta at day 35 of gestation fetuses after natural mating (Control) or of cloned pig by somatic cell nuclear transfer (SCNT) were isolated from a uterus. To investigate the mRNA expression and methylation patterns of H19 gene, tissues from fetal liver and placenta including endometrial and extraembryonic tissues were collected. The mRNA expression was evaluated by real-time PCR and methylation pattern was analyzed by bisulfite sequencing method. Bisulfite analyses demonstrated that the differentially methylated region (DMR) was located between -1694 bp to -1338 bp upstream from translation start site of the H19 gene. H19 DMR (-1694 bp to -1338 bp) exhibits a normal mono allelic methylation pattern, and heavily methylated in sperm, but not in oocyte. In contrast to these finding, the analysis of the endometrium and/or extraembryonic tissues from SCNT embryos revealed a complex methylation pattern. The DNA methylation status of DMR Region In porcine H19 gene upstream was hypo methylated in SCNT tissues but hypermethylated in control tissues. Furthermore, the mRNA expression of H19 gene in liver, endometrium, and extraembryonic tissues was significantly higher in SCNT than those of control (p<0.05). These results suggest that the aberrant mRNA expression and the abnormal methylation pattern of imprinted H19 gene might be closely related to the inadequate fetal development of a cloned fetus, contributing to the low efficiency of genomic reprogramming.

• Clinical and Experimental Reproductive Medicine
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• 제33권3호
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• pp.149-157
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• 2006

목 적: 본 연구는 인공 수정 시술에 있어서 정상 배란주기의 여성에서 clomiphene citrate와 GnRH antagonist를 병합하여 임선을 시도한 방법을 기존의 clomiphene citrate 단독으로 사용한 방법과 비교하여 그 임상적 유용성을 알아보고자 하였다. 연구방법: 116명의 환자들 중 43명에게는 자연 배란주기를 이용하여 인공 수정 시술을 시행하였고, 38명과 35명에게 각각 clomiphene citrate 단독 요법과 clomiphene citrate와 GnRH antagonist 병합 요법으로 인공 수정 시술을 시행하여 그 임상 결과를 비교 분석하였다. 결 과: 각각의 군에서 기저 LH, FSH, $E_2$와 배란 유도를 위해 hCG를 투여하는 날의 FSH, $E_2$는 차이를 보이지 않았으나 clomiphene citrate/GnRH antagonist 병합 요법군에서 hCG를 투여한 날의 LH와 자궁 내막 두께는 의미 있게 감소하였으며 유의한 임신율의 증가가 관찰되었다. 결 론: 자연 배란주기법군, clomiphene citrate 단독 요법 사용군에 비해 clomiphene citrate/GnRH antagonist 병합 요법 사용군에서 유의하게 임신율의 향상을 나타냈다.

• 한국가축번식학회지
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• 제18권2호
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• pp.141-150
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• 1994

전자현미경에 의해 자궁부착 전후의 돼지 수정란의 형태형성 및 분화에 따른 배발생 과정을 검토하였다. 돼지 초기배는 자궁이주후 균일하게 자궁에 배분되기전 약 2~3일간은 자궁각의 proximal portion에 존재하며, 임신 4일째에 할구와 할구의 경계를 상실하는 tight한 gap junction을 가진 상실배로 발달한다. 배반포를 형성하는 시기에 estradiol 17$\beta$는 compact한 상실배를 cavitated blastocyst로 발달을 촉진시키면서, steroid hormone이 이후의 배발생을 지배한다. Hatching의 시기는 교배후 6~7일경 zona pellucida을 둘러사고 있는 glycoprotein의 thinning과 lysis에 의해 이루워지는데, hatching 과정은 embryo의 세포수와 무관하였으며, 이때의 embryo의 직경은 0.5~1.0mm인 것을 본 실험에서 확인하였다. 12일경부터는 embryo는 prostaglandins, IGF-binding protein, retinol binding protein, plasminogen activator등의 단백질이 풍부해 이들 인자가 elongation 개시 후보로 고려될 수 있었다. 또한 이 시기의 embryo는 embryonic disc로 발달시 progesterone과 estrogen을 estradiol 17$\beta$로 전활할 수 있으며, 이러한 변화와 함께 spherical stage로부터 tubular 혹은 filamentous form으로 변형되었다. Estrogen이 임신을 통해 prostagladins의 분비를 uterine lumen에 지시하는지는 알 수 없으나 13일 경을 전후해 conceptus estrogen이 uterine arterial blood flow, uterine vasular permeability을 증가시키는 것으로 나타났으며, 자궁에서 protein과 calcium, PGF2$\alpha$, plasminogen inhibitor를 증가시키는 것으로 나타났다. 이 시기의 자궁 변화와 함께 embryo의 attachment는 trophoblast와 uterine membrane사이의 느슨한 결합에 의해 개시되었으며, 18일경 uterine과 trophoblastic microvili의 interdigitation에 의해 완성된다. 이 시기에 conceptus attachment를 위해 필요한 uterine microvili에서의 glycocalyx의 형성과 endometrial epithelium의 erosion을 야기하기 위해 plasminogen activator을 분비하였으며, 반면 자궁에서 plasminogen 역할을 하는 것은 estrogen이며, blastocyst cell 표면의 lectin binding이 attachment에 중요한 역할을 한다. 이상과 같은 일련의 과정을 거친 초기배는 성공적인 임신으로 유도된다고 본다. 따라서, 본 연구는 이상과 같이 착상을 전후한 시기의 배를 전자현미경에 의해 형태형성의 변화를 특히 착상을 전후해 배 취사율이 높은 시기를 대상으로 분석하였다. 이 분석 시기중 성공적인 착상성공율은 56%(71/126)였다.

• 대한한의학방제학회지
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• 제25권2호
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• pp.223-251
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• 2017

Objectives : A lot of experiment results of Yugmijihwangtang(YM) are reported in various kinds of journals. Many of them report on the new effects that are not recorded in the traditional medical texts. So it is necessary to take it into consideration that newly reported effects could be of help to clinical practice, because this process of comparison of Donguibogam and scientific experiment results will have basis to lead into the evidence based medicine. Methods : We compared the effects of in Donguibogam and the experiment results of YM. Results : The effects of YM in Donguibogam are to replenish essence and marrow, and to treat red wen, fatigue, treat hypouresis, urinary sediment, urinary urgency, hematuria, hydrocephalus, speech and movement retardation, yin-deficiency, diabetes mellitus, nonalcoholic fatty liver, melanoma, disability to see near and far sight, tinnitus, hearing loss, alopecia, angiogenesis, cough, cough at night, trachyphonia, and, infantile convulsion. The experiment results of YM since 2000 in both Korea and China are to inhibit atopic dermatitis, renal interstitial fibrosis, anti-oxidant, emphysema, stress, glomerulosclerosis, diabetic nephropathy, chronic glomerulonephritis, hemorrhage, plantar sweating, dermal aging, kidney aging, bone loss, breast cancer, pathological myocardial cell, primary liver cancer, thrombosis, osteoporosis, intrauterine growth retardation, chronic renal failure, IgA nepropathy, slow cerebral development, and hippocampal tissue lesions on the one hand, and to help bone formation, renin-angiotensin- aldosterone system, cerebral recovery, cognitive function and expression, osteoblast proliferation and differentiation, learning and memory, cold-tolerance and oxygen deficit-tolerance and anti-fatigue, endometrial formation, humoral and cell-mediated immunity, immune regulation effect, Hypothalamus-Pituitary-Ovary Axis, and spermatogenesis, on the other hand. Conclusion : When we compared the effects of YM with the experiment results of YM, there existed a considerable gap between them. So, from now on, it is expected that a great effort and consideration are needed to solve these gaps from an academic and clinical point of view.

• 한국가축번식학회지
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• 제17권2호
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• pp.141-148
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• 1993

This studies were carried out to investigate the effects of co-culture with cumulus cells, oviduct epithelial cells and uterine endometrium cells on the in-vitro fertilization and cleavage rate of bovine follicular oocytes. The ovaries were obtained from slaughtered Korean native cows. The follicular oocytes surrounded with cumulus cells were recovered by aspirating follicular fluids from the visible follicles of diameter 3~5 mm. The follicular oocytes were cultured in TCM-199 medium containing hormones and 10% FCS for 24~48 hrs in a incubator with 5% CO2 in air at 38.5$^{\circ}C$ and then matured oocytes were again cultured for 12~18 hrs with motile capacitated sperm by preincubation. The results obtained in these experiments were summarized as follows: 1. The in vitro maturatin and fertilization rate of bovine oocytes co-cultured with cumulus cells in TCM-199 medium were 64.0~74.1% and 40.0~58.6% respectively. And in-vitro fertilization rate of cumulus-enclosed oocytes(55.4%) were significantly(p<0.05) higher than cumulus-denuded oocytes(23.1%). 2. The in-vitro maturatin and fertilization rate of bovine oocytes co-cultured with 1$\times$104 cells/ml, 1$\times$106 cells/ml, 1$\times$108 cells/ml and 1$\times$1015 cells/ml oviduct epithelial cells in TCM-199 medium were 59.3% and 40.7%, 64.0% and 48.0%, 58.3% and 37.5%, 52.0% and 32.0%, respectively. 3. The in-vitro maturation and fertilization rate of bovine oocytes co-cultured with 1$\times$104 cells/ml, 1$\times$106 cells/ml, 1$\times$108 cells/ml and 1$\times$1015 cells/ml uterine endometrium cells in TCM-199 medium were 56.0% and 36.0%, 60.7% and 42.9%, 59.3% and 37.0%, 52.0% and 36.0%, respectively. 4. When the in-vitro fertilized oocytes were co-cultured with cumulus cells, oviduct epithelial cells and uterine endometrium cells, the development rate to be blastocyst was 12.2%, 15.6% and 11.7%, respectively and rates were higher than that of control, 2.1%(P<0.05).

• Clinical and Experimental Reproductive Medicine
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• 제16권1호
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• pp.69-79
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• 1989

Steroid hormone profiles during luteal phase of clomiphene citrate(CC)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin(hCG)-stimulated in vitro fertilization (IVF) cycles and of follicle-stimulating hormone(FSH)/hMG/hCG-stimulated IVF cycles were compared. In seventy three cycles stimulated with CC/hMG/hCG regimen, follicles were aspirated during exploratory laparotomy and yielded 7 pregnancies, and in 83 cycles stimulated with FSH/hMG/hCG regimen, follicles were aspirated by laparoscope and made 13 pregnancies. Serum estradiol($E_2$) and progesterone($P_4$) levels were determined on days 2, 5, 7, and 9 after follicle aspiration. The FSH/hMG/hCG regimen was more effective than the CC/hMG/hCG regimen in folliculogenesis, ie, ovarian stimulation, follicular phase $E_2$ peak levels, oocyte maturation, and the number of retrieved oocytes. There was no significant difference between luteal serum $P_4/E_2$ ratio of the two regimens, suggesting that secretory endometrial build-up ability for implantation may not differ each other. Several significant correlations were observed between follicular phase seum $E_2$ peak levels and luteal phase serum $E_2$ and $P_4$ levels in the FSH/hMG/hCG-stimulated cycles but any correlation was not significant in the CC/hMG/hCG-stimulated cycles, suggesting that somewhat more follicles may eventually fall in atresia even after attaining dominant stage in the CC/hMG/hCG-stimulated cycles than the FSH/hMG/hCG-stimulated cycles.

• Clinical and Experimental Reproductive Medicine
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• 제37권2호
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• pp.153-158
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• 2010

목 적: 과배란유도 후 자궁강내 인공수정 시술시 황체기 보강으로서 경구 미분화 프로게스테론 투여법과 경구 디드로게스테론 투여법의 임상적 효용성을 비교하고자 하였다. 연구방법: 2007년 1월부터 2009년 8월까지 시행된 과배란유도 후 자궁강내 인공수정 시술 183주기를 후향적으로 분석하였다. 과배란유도는 성선자극호르몬 단독 또는 클로미펜과 성선자극호르몬의 병합요법을 사용하였다. 136주기에서는 황체기 보강으로서 경구 미분화 프로게스테론을 하루 300 mg으로 투여하였고 47주기에서는 디드로게스테론을 일일 20 mg으로 투여하였다. 결 과: 여성의 연령, 불임 인자, 성숙난포수 ($\geq$16 mm), 총운동성정자수, triggering 날의 자궁내막 두께는 두 군간 유의한 차이가 없었다. 자궁내 태낭이 확인되는 임상적 임신율은 미분화 프로게스테론 투여군에서 21.3%, 디드로게 스테론 투여군에서 19.1%로 차이가 없었다 (p=0.92). 유산율은 미분화 프로게스테론 투여군에서 다소 높은 경향을 보였으나 통계학적인 차이는 없었다 (34.5% vs. 11.1%, p=0.36). 결 론: 황체기 보강으로서 경구 디드로게스테론 투여법은 경구 미분화 프로게스테론 투여에 비하여 비슷한 임신율과 유산율을 보였다. 그러나 상대적으로 디드로게스테론 투여군의 수가 적어 좀더 많은 환자를 대상으로 한 전향적 연구가 필요하다.

• Asian Pacific Journal of Cancer Prevention
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• 제15권16호
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• pp.6639-6641
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• 2014

Background: To evaluate the clinical significance of atypical squamous cells of undetermined significance (ASCUS) in PAP test in post-menopausal women and compare with reproductive age women. Materials and Methods: A total of 367 patients who referred to our gynecologic oncology clinic were included to the study between September 2012 and August 2013. Data for 164 post-menopausal (group 1) and 203 pre-menopausal (group 2) women with ASCUS cytology were evaluated retrospectively. Immediate colposcopy and endocervical curettage was performed for both groups and conization for all women with a result suggestive of CIN2-3. Histopathological results and demographic features of patients were compared between the two groups. Results: Mean age of the patients was $54.6{\pm}6.5$ years in group 1 and $38{\pm}6.6$ years in group 2. Some 14 (8.5%) of post-menopausal women and 36 (17.7%) of pre-menopausal women were current smokers (p=011). Totals of 38 (23.2%) post-menopausal and 64 (31.5%) pre-menopausal women were assessed for HPV-DNA. High risk HPV was detected in 7 (4.3%) and 21 (10.3%), respectively (p=0.029). Final histopathological results recorded were normal cervix, low grade cervical intra-epithelial neoplasia (CIN 1), and high grade cervical intra-epithelial neoplasia (CIN2-3). In group 1 results were 84.8%, 12.2% and 1.8%, respectively, and in group 2 were 71.9%, 23.2% and 4.9%. There were no cases of micro invasive or invasive cervical carcinoma in either group. Two cases were detected as endometrial carcinoma in the menopausal group (1.2%). Conclusions: In current study we found that preinvasive lesions were statistically significantly higher in pre-menopausal women than post-menopausal women with ASCUS. Cervicitis was more common in menopausal women. Therefore, we think that in case of ASCUS in a post-menopausal woman there is no need for radical management.

• Asian Pacific Journal of Cancer Prevention
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• 제16권18호
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• pp.8239-8245
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• 2016

The p53 gene is inactivated by the human papillomavirus (HPV) E6 protein in the majority of cervical cancers. Treatment of HeLa S3 cells with siRNA for HPV E6 permitted adenovirus-mediated transduction of a p53 gene linked to an upstream estrogen response element (ERE). Our previous study in non-siRNA treated HHUA cells, which are derived from an endometrial cancer and express estrogen receptor ${\beta}$, showed enhancing effects of an upstream ERE on adenovirus-mediated p53 gene transduction. In HeLa S3 cells treated with siRNA for HPV E6, adenovirus-mediated transduction was enhanced by an upstream ERE linked to a p53 gene carrying a proline variant at codon 72, but not for a p53 gene with arginine variant at codon 72. Expression levels of p53 mRNA and Coxsackie/adenovirus receptor (CAR) mRNA after adenovirus-mediated transfer of an ERE-linked p53 gene (proline variant at codon 72) were higher compared with those after non-ERE-linked p53 gene transfer in siRNA-treated HeLa S3 cells. Western blot analysis showed lower ${\beta}$-tubulin levels and comparatively higher p53/${\beta}$-tubulin or CAR/${\beta}$-tubulin ratios in siRNA-treated HeLa S3 cells after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with those in non-siRNA-treated cells. Apoptosis, as measured by annexin V binding, was higher after adenovirus-mediated ERE-linked p53 gene (proline variant at codon 72) transfer compared with that after non-ERE-linked p53 gene transfer in siRNA-treated cells.