• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.161-170
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• 1998

Protein expression patterns of matrix metalloproteinases (MMPs) were examined in mouse reproductive organs during estrous cycle. Estrous cycle was classified into diestrus, proestrus, estrus or metestus and MMP expression was analyzed by zymography using gelatin as a substrate. Uterine fluid (UF) obtained both at diestrus and proestrus exhibited 4 major MMPs including 106kDa, 64kDa, 62kDa and 59kDa gelatinases. However, in UF at estrus, the gelatinolytic activity of 64kDa MMP disappeared and that of 106kDa and 62kDa MMPs dramatically decreased. At metestrus, 64kDa MMP activity reappeared and 106kDa and 62kDa MMP exhibited increased activities such that the band intensity of 106kDa was comparable to that in UF at diestrus. Gelatinolytic activity of 59kDa MMP was not changed throughout the cycle. Both ovarian and oviductal tissue homogenate revealed 4 MMPs which corresponded to the 4 MMPs of UF. However, unlike UF MMPs, gelatinolytic activity of these MMPs did not show distinct changes throughout the cycle. Either an inhibitor of MMP, 1,10-phenanthroline, or a metal chelator, EDTA, abolished the appearance of the above MMP activities in gelatinated gel whereas a serine proteinase inhibitor, phcnylmethylsulfonyl fluoride, failed to inhibit the appearance of MMP activities, proving that gelatinolytic activity of the above reproductive tissues were due to the enzymatic activity of MMP. When gclatinolytic activity of mouse serum was examined, it revealed 5 MMPs (131kDa, 106kDa, 89kDa, 64kDa and 62kDa bands) and one gelatinase (84kDa) band. From these results, it is concluded that the protein expression of MMPs of mouse reproductive organs, particularly uterus, is temporally regulated during estrous cycle and uterine 106kDa, 64kDa and 62kDa MMPs are suggested to play an important role in cyclic tissue remodeling of mouse uterus.

• Clinical and Experimental Reproductive Medicine
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• 제12권1호
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• pp.41-46
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• 1985

Various immunoserologic and cellular immunity techniques have been used to explore the presence of antisperm antibodies in the serum and seminal plasma of male patients and in the blood and genital fluid of infertile women. Several recent comparative investigations using various assays to detect and quantitate levels of antibody to human spermatozoa have produced widely varying results. So the first WHO workshop on iso- and autonatibodies to human spermatozoa in 1974 tried to establish some unification in the techniques used. The purpose of this study is to compare the results of two methods-the Kibrick macro-agglutination test and the Isojima micro-immobilization test-using the same test materials based on recommandation from WHO workshop. The results are as follows: 1. Twenty normal controls showed negative reactions in all the 2 tests. Out of 25 patients, the positive sera were noted in 15 (60%) on the Kibrick test and 13 (51%) on the Isojima test. 2. Twelve (48%) out of 25 patients showed positive reactions in the two tests, and 16 (64%) out of 25 patients showed positive reaction in one or more tests. 3. The titers of the antisperm antibodies on the Kibrick test was higher than that on the Isojima test. Therefore, it seems to be possible to increase the chances of detection of the antisperm antibodies, if two tests are imployed.

• Clinical and Experimental Reproductive Medicine
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• 제16권2호
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• pp.179-185
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• 1989

불염여성에서 복강내액의 독성 유무를 알기 위해서 콤퓨터 정액 분석기를 이용하여 복강내액의 정자 활동성을 분석하였다. 연구대상은 자궁내막증 환자 14예와 자궁내막증이 없는 불염환자로부터의 복강내액 9예로 하였다. 연구방법은 건강인의 정자를 셰척 분리하여 대상환자의 복강내액과 Hams F10 배양액을 동량 흔합하여 (50%) 배양후 정자 활동성을 측정하였다. 정자 활동성의 감소는 배양전 0시간의 수치와 비교해서 1시간， 4시간 그리고 24시간후의 결과를 비교 분석하였다. 자궁내막증 제 1기 및 제 2기 환자와 자궁내막증이 없는 환자의 복수에서의 정자 활동성의 감소는 통계학적으로 의의있는 차이는 없었다. 그러나 자궁내막증이 심한 제 3기의 환자군에서는 의의있는 차이가 있었다(p<.05). 실험군간의 정자 활동성의 평균치 차이는 없었으나，각 군에서의 환자별 개인 성적은 정자 활동성이 현저히 감소한 독성있는 복강내액과 비독성의 복강내액을 쉽게 구별할 수 있었다. 결론적으로 정자 배양을 이용한 복강내액 독성 검사는 비교적 간단하고 경제적이므로 염상적으로 최근에 대두된 복강내 수정 치료법의 예비검사로서 유용할 것으로 사료된다.

• Clinical and Experimental Reproductive Medicine
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• 제18권2호
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• pp.123-131
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• 1991

In order to study the growth and maturation of ovarian follicle, the localization and activity of alkaline phosphatase(ALPase) and adenosine triphosphatase(ATPase) of the granulosa cells and theca layer were examined according to the follicle size, the follicle state and the ovarian cyclic phase in pig. Theca interna of the small follicles was more heavyly localized with reaction product by the activites of ALPase and ATPase than that of the large follicles. It is assumed that, as the follicles proceed to growth and maturation, antrum formation is the result of the follicular fluid accumulation by means of active transport by the activities of ALPase and ATPase in theca interna. The activities of ALPase and ATPase in atretic follicles were higher than those of normal follicles. These results imply that the mechanisms of follicle maturation and atresia are different according to the phase of ovarian cycle.

• Asian-Australasian Journal of Animal Sciences
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• 제19권3호
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• pp.335-340
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• 2006

A large number of toxicological substances and pharmacological and physical agents can cause reproductive intervention at the cellular and molecular level. The present study was designed to assess the effect of mercury ($HgCl_2$) at 50 to $550{\mu}M$ concentration ranges, in vitro, on the sperm membrane and DNA integrity, viability, and acrosomal status of normal bull spermatozoa. The samples were processed for sperm analyses using semen-diluting fluid (PBS, pH 7.2). We recorded a sharp increase in the lipid peroxidation/LPO rate; the highest was at $550{\mu}M$ mercury concentration, indicating a deleterious effect of mercury on the sperm membrane intactness. There was also a strong negative correlation between LPO rate and % viable spermatozoa (R = 0.987, p<0.001). Data obtained from a comet assay technique revealed that mercury is capable of inducing DNA breaks in the sperm nuclei. Interestingly, 92% of DNA breaks were double-stranded. The correlation between LPO rate and % DNA breaks was 0.984. Performing the gelatin test indicates that mercury is able to alter the integrity of acrosomal membranes showing an abnormal acrosome reaction. In this regard, a strong link was found between LPO rate and % halos (R = 0.990, p<0.001). Collectively, mercury proved to be a potent oxidant in the category of environmental factors affecting bull spermatozoa. Hence, considering the wide spread use of mercury and its compounds, these metals should be regarded with more concern.

• Clinical and Experimental Reproductive Medicine
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• 제20권1호
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• pp.19-29
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• 1993

Plasminogen activator (PA)-plasmin system in follicular fluid is involved in the process leading to follicular rupture at ovulation. It is well known that PA is closely associated with cellular differentiation and tissue remodeling on evidences from the study of normal and malignant tissues. This study was designed to ascertain a potential role of PA in the ovarian folliculogenesis. Immature Sprague-Dawley rats were injected with pregnant mare serum gonadotropin, followed by injection of serine protease inhibitor (SPI; mixture of 1 mol/L benzamidine and 1 mol/L amino-caproic acid) into the unilateral ovarian bursa. In the control study, mechanical effect of bursal injection and contralateral ovarian effect SPI were ruled out. Total antral follicular areas relative to total ovarian cross-sectional areas was siginificantly lower in SPI-injected ovary than in saline-injected ovary. SPI injection decreased the relative antral follicular area by 33 % respectively. Electron microscopic finding of granulosa cell in the atretic follicle showed the presence of pyknotic nucleus, blurring of neucleolemma, degeneration of mitochondria and dilation of endoplasmic reticulum. After induction of ovulation with hCG, the number of oocytes released was significantly decreased in SPI-injected oviduct than in saline-injected oviduct. From above results, author discussed that PA may play a role not only in ovulation but also in some processes of folliculogenesis.

• Clinical and Experimental Reproductive Medicine
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• 제15권2호
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• pp.157-164
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• 1988

We found reports of only about 20 cases of well documented ejaculatory duct obstruction presenting with infertility and treated by TUR. While infertility caused by ejaculatory duct obstruction is rare accurate diagnosis by vasoseminal vesiculography is important, since these obstructions may be treatable. In our 29 patients undergoing vasoseminal vesiculography for 1 year to demonstrate obstruction site in seminal tract the ejaculatory duct obstruction was found in 6 patients. The intial methods of treament were TUR of the ejaculatory duct in 2 patients, forceful lavage of ejaculatory duct through the vas deferens in 4 patients. One patient's wife who had undergone TUR of the ejaculatory duct delivered a normal female baby 15 months postoperatively. In one patient who had undergone forceful lavage of ejaculatory duct semen analysis returns to normal level except the motility. When initial methods of treatment fail to get the normal semen we have the plan to perform TUR of the ejaculatory duct or aspiration of seminal fluid in the seminal vesicle under the control of the ultrasonography for AIH according to the level of ejaculatory duct obstruction. Our report suggests that ejaculatory obstruction has been underdiagnosed and should receive more attention by urologist.

• 한국가축번식학회지
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• 제7권2호
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• pp.8-26
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• 1983

Various early pregnancy diagnostic methods have been developed in order to improve the reproductive efficiency in cow, mare, mule, sow, sheep, goat, dog, cat, rabbit, buffalo, camel, elephant, monkey, deer, lion, coipus and guinea pig. These methods include abdominal swelling, abdominal palpation, esturs cylce detection, Lupin test, gonadotropin assay, colostrum injection test, sperm motility assessment, cervical mucus viscosity test, Kaber chromagens method, estrogen test, A Scheim-Zond다 test, spectrophotometric detection of estrogen in urine and feces, boric acid crystraline formation test in urine, oxytocin injection test, diamino-oxidase test, PMSG HA test, behaviour test, Simolus iodine detection test, detection of tryptophane in urine, x-ray method, Cuboni and Lunaas method, vaginal biopsy method, Friedmann Schneider diagnostic method, electrode method, barium chloride detection method, ECG, Doptone method, ultrasound method, ultrasound scanning method, LDH method, rectal palpation method, CL palpation method, radioautography, serum creatine test, serum globulin test, chlormadine method, CAP method, Medata Do, pp.ers method, body fluid test, Plasma oCS detection method, ERIA, LHRH method, negative latex cogulation test and oestrone sulphate detection method. The most reliable methods with high a, pp.icability to farm animals such as sheep, mare, sow and cow are rectal palpation, ultrasound method and hormonal assay in blood and milk. However, they require complicated laboratory works for the early diagnosis of pregnancy and in most cases, the simple and economical methods which are described up to now need a long period of time after conception. Generally, it is possible to detect pregnancy after one estrus cycle, even though it varies depending on the species of animals. For improvement of the reproductive efficiency, it is required to develop a more accurate, economical, simple and early detectable method. It is anticipated that the result of a study on the detection method of EPF(early pregnancy factor) would be a, pp.icable to various animals within 6 hours after conception.

• Clinical and Experimental Reproductive Medicine
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• 제21권3호
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• pp.261-266
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• 1994

Semen preparation for Assisted Reproductive Technology(ART) involves the separation of the motile sperm from non-motile, dead sperm, cellular debris and seminal fluid. The aim of this study is to compare the motile sperm recovery rate and motility index of different sperm preparation method(swim-up method, 80% isotonic continuous percoll method, two-layer discontinuous percoll method, mini-percoll method). In Mini-Percoll method, pellets from patients were suspended in 0.3ml of medium and layerd on a discontinuous percoll gradient consisting of 0.3ml each of 50, 70, 95% isotonic percoll. All semen samples are divided into normal and subnormal sperm group(oligo-, astheno-, oligoasthenozoospermia). Especially, we evaluated the effect of mini-percoll method in subnormal sperm group. In normal sperm group, mini-percoll method and two layer discontinuous percoll method (40%/80%) allowed increasing of motile sperm recovery rate. But motility index was higher in swimup method than the other methods. In subnormal sperm group, mini-percoll method has advantages as compare with the other methods in motile sperm recovery rate and motility index. These results suggest that modified mini-percoll method could be certainly a valuable tool in some cases of severe male factor sperm.

• Clinical and Experimental Reproductive Medicine
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• 제33권1호
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• pp.53-60
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• 2006

목 적: 보조생식술에서의 정자처리법으로서 인간 난포액을 이용한 swim-down 방법의 유용성을 확인해 보고자 하였다. 연구방법: 분당서울대학교병원 산부인과에서 불임평가 목적으로 정액검사를 시행할 때 정자무력증 (asthenozoospermia, sperm motility < 50%)을 보이는 12명의 남성을 대상으로 하였다. 이들에서 검사 후 남은 정액을 100% 인간 난포액을 이용한 swim-down법과 SpermGrad를 이용한 밀도차 분리법을 적용하여 각각 처리하고 컴퓨터 정액분석기를 이용하여 정액검사를 시행하였다. 결 과: 두 군 모두에서 운동성과 빠른 운동성 정자의 비율, VCL (curvilinear velocity), ALH (amplitude of lateral head displacement)치 및 과활동성 정자의 비율이 통계적으로 유의하게 상승하였고 LIN (mean linearity)치는 유의하게 감소하였다. 100% 인간 난포액을 이용한 swim-down법에서 정자의 운동성이 SpermGrad를 이용한 밀도차분리법에 비하여 유의하게 높게 나타났으며 ($81.2{\pm}4.7$ vs. $67.6{\pm}2.3$, p=0.02), 다른 변수들은 두 군에서 차이를 보이지 않았다. 결 론: 100% 인간 난포액을 이용한 swim-down법은 정자무력증을 보이는 경우 유용한 정자 처리법으로 사료된다.