• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.108-112
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• 2006

• Clinical and Experimental Reproductive Medicine
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• v.32 no.4
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• pp.293-300
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• 2005

연구목적: 단일 유전자 이상에 대한 착상전 유전진단을 성공적으로 시행하기 위해서는 효과적이고 신뢰도가 높은 PCR 방법의 확립이 중요하다. 본 연구에서는 alkaline lysis와 duplex nested PCR 방법을 단일 림프구와 할구의 유전자 분석에 적용하여 그 효용성을 확인하고자 하였다. 재료 및 방법: 단일 유전자의 이상이 확인된 Duchenne muscular dystrophy (DMD), ornithine transcarbamylase (OTC) 결핍증과 epidermolysis bullosa (EB) 가계의 대상자들에서 채취한 단일 림프구와 공여 받은 배아의 할구를 이용하여 각각 PCR, restriction fragment length polymorphism (RFLP)와 direct DNA sequencing 분석을 시행하였다. 이러한 분석에서 유전자 증폭률 (amplification rate)과 두개의 allele 중에서 하나의 allele이 증폭되지 않는 allele drop-out (ADO) 빈도에 대해 살펴보았다. 결 과: 단일 림프구와 할구를 이용한 PCR 방법의 유전자 증폭률은 DMD에서 91.1%와 81.8%, OTC 결핍증에서 96.0%와 78.1%, EB에서 91.3%와 90.0%를 각각 나타냈으며, ADO 빈도는 OTC 결핍증에서 13.3%, EB에서 16.8%로 관찰되었다. 결 론: 본 연구에서 적용한 alkaline lysis와 duplex nested PCR 방법은 단일 유전자에 대한 착상전 유전진단에 성공적으로 적용할 수 있는 방법으로 생각되며, ADO 빈도를 최소화할 수 있는 효율적인 방법의 개발에 대한 지속적인 연구가 필요하다.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.2
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• pp.127-133
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• 2003

Objective: The aim of this study was to investigate the morphological aspects of testicular tissue before and after freezing-thawing by light and transmission electron microscopy. Methods: Tissue biopsies were carried out on mouse testis for freezing. Samples in medium containing 20% glycerol were frozen by computer-controlled freezing program. The effect of freezing-thawing on the structural change of testicular tissues were examined by light and electron microscopy. Results: The freezing-thawing procedure had no significant effect on tubular diameter. However, it caused folding of the lamina propria, and notable damage to Sertoli cells, spermatogonia and spermatocytes. The cells were detached, desquamated from the basal lamina and had increased vacuolization. Round spermatids, elongated spermatids and spermatozoa were less affected, and most of them maintained their normal structure. Conclusions: The structure of spermatogonia, spermatocyte and basal compartments in seminiferous epithelium was significantly altered by freezing-thawing procedure of mouse testicular tissues. Thus, we need to develop a more reliable method for the cryopreservation of testicular tissues.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.3
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• pp.142-147
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• 2011

Objective: This study was performed to evaluate testicular sperm chromatin condensation using aniline blue-eosin (AB-E) staining and its effects on IVF-ET. Methods: Chromatin condensation was analyzed using AB-E staining in 27 cases of testicular sperm extraction. There were 19 cases of obstructive azoospermia (OA) and 8 cases of non-obstructive azoospermia (NOA) in IVF-ET. Mature sperm heads were stained red-pink whereas immature sperm heads were stained dark blue. The percentage of sperm chromatin condensation was calculated from the ratio of the number of red-pink sperm to the total number of sperm analyzed. Results: The overall percentages of chromatin condensation in OA and NOA were $31.1{\pm}11.2%$ and $26.3{\pm}14.4%$, respectively. The fertilization rate was significant higher in OA than NOA ($p$ <0.05); however, the rates of good embryos and clinical pregnancy did not show statistical differences. In OA and NOA, statistical differences were not observed in the rate of chromatin condensation, fertilization, good embryos, and clinical pregnancy between the pregnant group and non-pregnant group. Conclusion: Chromatin condensation is less stable than OA and showed a low fertilization rate in NOA. While there were no significant differences in chromatin condensation results between NOA and OA, we propose that a pattern of decreased chromatin condensation in NOA is one of the factors of low fertilization results requiring further study.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.2
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• pp.121-129
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• 2001

Objective: The aim of this study is to compare the efficiency of a method for the cryopreservation of mouse blastocyst.. Methods: Mouse embryos were obtained at 2-cell stage and cultured to blastocyst stage in T6 medium supplemented with 10% fetal bovine serum. Morphologically normal blastocysts were collected and randomly divided to one control and four experimental groups. In control group, blastocysts were cultured in vitro continuously for additional two days. In group 2, blastocysts were exposed to vitrification solution (ethylene glycol) only without cryopreservation (exposure only group). In group 3, 4 and 5, blastocysts were cryopreserved by slow-freezing procedure with glycerol (slow-fteezing group) or by vitrification procedure using EM grids (EM grids group) and cryoloop (cryoloop group), respectively. Frozen blastocysts were thawed and cultured for additional two days. Twenty four hours after thawing, some blastocysts were fixed and stained with Hoechst 33342 (bisbenzimide) and the number of nuclei in each blastocysts were counted to confirm the survival of bias to cysts in experimental groups. Results: Survival rate and hatching rate of the blastocysts in slow-freezing group (24 h: 72.4% and 66.0%, 48 h: 63.2% and 64.6%) and EM grids group (24 h: survival rate 77.3%, 48 h: 70.1% and 71.4%) were significantly lower ($X^2$-test p<0.05) than those of control group (24 h: 93.4% and 86.0%, 48 h: 88.5% and 90.7%). In contrast, the survival rate and hatching rate of the blastocysts in cryoloop group (24 h: 84.1% and 84.1%,48 h 79.3% and 87.7%) is well compared with those in the control group. The mean (${\pm}SD$) cell number of blastocyst in the exposure only ($89.2{\pm}11.5$), EM grids ($85.0{\pm}10.3$) and cryoloop ($89.0{\pm}11.0$) groups, except slow-freezing group ($79.0{\pm}10.0$), were not significantly different from that of control group ($93.1{\pm}13.9$) 24 h after thawing (Student's t-test). Conclusion: This study demonstrates that higher survival rate of vitrified-thawed mouse blastocyst can be obtained using cryoloop as the embryo container at freezing rather than slow-freezing or vitrification using EM grids. The results of this study suggest that vitrification using cryoloop (with ethylene glycol) may be a preferable procedure for mouse blastocyst cryopreservation and could be applied to the human blastocyst cryopreservation.

• Advances in Traditional Medicine
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• v.5 no.4
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• pp.347-351
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• 2005

In the present investigation twelve indigenous medicinal plants have been screened for their antiimplantation activity in albino rats. The plant material was subjected for soxhlation successively and separately from non-polar solvents to polar solvents i.e., petroleum ether benzene and ethanol. Out of these three extracts the petroleum ether extract of seeds of Citrus medica, aerial part of Oxalis corniculata and Tinospora cardifolia have showed maximum antiimplantation activity. Ethanol extract of leaves of Cardiospermum helicacabum, roots of Echinops echinatus, leaves of Melia azedarach, seeds of Momordica charantia and bark of Terminalia bellirica have shown maximum antiimplantation activity amongst the three extracts of each plant material screened. Though all the three extracts of seeds of Annona squamosa and leaves of Zizyphus jujube screened for antiimplantation activity, no extract has showed any loss in implantation. The details of the results obtained are discussed.

• Advances in Traditional Medicine
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• v.4 no.2
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• pp.70-76
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• 2004

Petroleum ether, benzene and alcohol extracts of the seeds of C. juncea were tested for antiimplantation and pregnancy interruption activities in female albino rats. Of these three extracts, the alcohol extract was found to be the most effective in causing antiimplantation and pregnancy interruption activities. These adverse effects on fertility are reversible upon withdrawal of the extract treatments. The alcohol extract was found to possess estrogenic activity. After subjecting to preliminary phytochemical screening, the alcohol extract showed positive tests for alkaloids, steroids, glycosides, saponins, flavonoides, fixed oils, phenols and tannins.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.207-213
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• 2010

Thousands of new chemicals have been introduced to environment during last decades. Many of them and common consumer products have been shown to be the endocrine disrupting chemicals. One such chemical group is the phthalates, used in soft poly vinyl chloride (PVC) material and in a huge number of consumer products. The prevalence of these modem chemicals have a remarkable increase. Approximately 3.5 million tons of the main phthalate, di-(2-ethylhexyl) phthalate (DEHP), are produced annually worldwide and indeed, DEHP is considered a ubiquitous environmental contaminant. It has been demonstrated that high doses of phthalate can adversely affect adult and developing animals. In this review, we critically discuss the conclusions of recently original research papers and provide an overview of studies on reproductive disrupting effects of phthalates. In addition, we review the reproductive toxicity data of phthalates in some in vitro research and in both male and female reproductive systems in experimental and domestic animals. Finally, we point out some critical issues that should be addressed in order to clarify the implication of phthalates for human reproduction.

• Development and Reproduction
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• v.27 no.3
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• pp.101-115
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• 2023

Environmental factors impact oyster growth, condition, and gonadal development, which is linked to gamete characteristics observed through histology. The reproductive cycle of bivalves is related to energy storage and utilization. Therefore, in this study, the year-round growth change and gonadal development of oysters were observed using histological analysis, and the biochemical composition changes were confirmed. The oysters used in this study are being nurtured in Gadeok-do, and 40 oysters were randomly sampled monthly from March 2021 to February 2022. Result of histological analysis of gonads, oysters were showed early development from December to February, late development from March and April, mature and ripe from May to July, spawned from August to October, and spent from November to December. Condition index values of oysters decreased in summer and autumn and increased again when entered the spent after spawning. The protein content of oysters was high in May, the maturity period, and the lipid content decreased during the spawning period. In addition, EPA and DHA, the major fatty acids of oysters, were low during the spawning period and high during the maturation period. As a result, this study suggested a close relationship between changes in oyster growth, biochemical composition, and the reproductive cycle.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.4
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• pp.293-298
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• 2003

목 적: 인간의 배아줄기세포는 전분화능과 영속성을 가지고 있어 발생 및 분화에 관련된 기초 연구 뿐 만 아니라 재생의학, 약물검색 등에서도 매우 유용한 재료로 이용될 수 있다.본 연구에서는 유전체의 변형이 배아줄기세포주의 확립 효율에 미치는 영향을 살펴보고자 비정상적인 포배기 배아에서 내세포괴를 분리하여 배양하였다. 연구 방법: 인간의 체외수정 및 배아이식술에서 공여 받은1개 또는3개의 전핵이 관찰되는 비정상 수정란 (n=20)과 착상전 유전진단에서 이수성이 확인된 배아 (n=27)를 대상으로 하였다. 일반적인 immunosurgery 방법으로 영양배엽세포들을 제거하고 내세포괴를 분리한 후 PMEF 혹은 STO feeder 세포위에서 배양하였다. 배아줄기세포의 배양시스템을 검증하기 위해서 이미 확립된 Miz-hES1 cell line을 동시에 같은 조건 하에서 계대배양하였다. 결 과: 비정상 수정란에서 발생된 포배기 배아에서 분리한 1개의 내세포괴가 배아줄기세포와 유사한 colony를 형성하였으나, 계대배양에는 실패하였다. 이수성 배아에서 발생된 포배기 배아의 내세포괴 배양에서는 두개의 colony가 계대배양 중에 영양배엽세포의 형태로 분화되어 미분화 상태를 유지하지 못하였다. 동일한 시기와 조건 하에서 계대배양된 Miz-hES1 cell line이 미분화상태로 유지됨을 karyotyping (46, XY)과 immunophenotyping (positive in SSEA-3 and -4)으로 확인하였다. 결 론: 본 연구의 결과에서 비정상 수정란과 이수성 배아에서 발생된 포배기 배아에서 유래한 내세포괴는 배아줄기세포주 확립 및 미분화 상태 유지 능력이 매우 저조한 것으로 여겨진다. 따라서, 인간의 배아줄기세포주를 확립하는데 있어 배아의 정상여부가 중요한 요소로 작용할 것으로 생각된다.