• Clinical and Experimental Reproductive Medicine
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• v.31 no.3
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• pp.155-168
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• 2004

Objective: Melatonin, which is secreted by pineal gland play an important role in the regulation of ovarian function via seasonal rhythm and sleep in most mammals. It also has a role in the protection of cells by removing toxic oxygen free radicals brought about by metabolism. In the present study, effects of melatonin on the mouse oocyte maturation were examined using two different culture conditions provided with 5% or 21% oxygen concentration. Material and Method: Immature mouse oocytes were obtained from the ovarian follicles of $3{\sim}4$ weeks old ICR strain mice intraperitoneally injected with 5 I.U. PMSG 44 hour before. Under stereomicroscope, morphologically healthy oocytes with distinct germinal vesicle (GV) were liberated from the graafian follicles and collected using mouth-controlled micropipette. They were then cultured for 17 hour at $37^{circ}C$, 5% $CO_2$ and 21% $O_2$ (95% air) or 5% $CO_2$, 5% $O_2$ and 90% $N_2$. New modified Hank's balanced salt solution (New MHBS) was used as a culture medium throughout the experiments. Effects of melatonin were examined at a concentration of $0.0001{\mu}M$, $0.01{\mu}M$ or $1.0{\mu}M$. For the prevention of spontaneous maturation of immature oocytes during culture, dibutyryl cyclic AMP (dbcAMP) and/or hypoxanthine were included in the medium. Results: Under 21% oxygen condition, oocytes cultured in the presence of $0.01{\mu}M$ melatonin showed a significantly higher maturation rates, in terms of germinal vesicle breakdown (95.0% vs 89.0%) and polar body formation (88.1% vs 75.4%), compared to those cultured with $0.0001{\mu}M$ or $1.0{\mu}M$ melatonin. However, no difference was observed in oocytes cultured under 5% oxygen whether they were treated with melatonin or not. In the presence of $0.01{\mu}M$ melatonin, oocytes either cultured under 21% or 5% oxygen exhibited no difference in the polar body formation (85.6% vs 86.7%). However, in the absence of melatonin, oocytes cultured under 21% oxygen exhibited lower polar body formation (74.7%). When oocytes were cultured in the presence of dbcAMP alone or with varying concentrations of melatonin, those treated with both compounds always showed better maturation, i.e., germinal vesicle breakdown and polar body formation, compared to those cultured with dbcAMP alone. At the same concentration of melatonin, however, oocytes exposed to 21% oxygen showed poor maturation than those to 5% oxygen. Similar results were obtained from the experiments using hypoxanthine instead of dbcAMP. Conclusion: Based upon these results, it is suggested that melatonin could enhance the meiotic maturation of mouse oocytes under 21% oxygen concentration, and release oocytes from the meiotic arrest by dbcAMP or hypoxanthine regardless of the concentration of oxygen, probably via the removal of oxygen free radicals.

• The Journal of Korean Obstetrics and Gynecology
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• v.25 no.2
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• pp.43-65
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• 2012

Objectives: This study was designed to investigate the effects of Hyeolbuchukeotang( HCT) on the Estradiol Valerate(EV) induced Polycystic Ovaries and the Implantation of rats. Methods: PCO was induced by single intramuscular(IM) injection with EV in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for 60 days. Control group(n=8) were injected with EV and orally administrated distilled water for 60 days. HCT treated group(n=8) were injected with EV and orally administrated HCT for 60 days. At the end day of experiment, we measured weights of body, ovaries, adrenal glands, uterus and contents of serum LH, FSH, ADD. The histomorphometrical changes of ovaries were also evaluated. And we observed the NGF and CRF expression by immunohistochemistry. Also we observed the mating and count the number of implantation. Results: - The weights(mg) of ovaries in HCT treated group($58.9{\pm}6.5$) were significantly increased (p<0.01) compared with control group ($42.3{\pm}8.5$). - The number of mature follicles in HCT treated group($9.8{\pm}2.6$) was significantly increased (p<0.01) compared with control group($6.1{\pm}2.1$). - The number of cystic follicles in HCT treated group($1.6{\pm}1.2$) was significantly decreased (p<0.01) compared with control group($3.8{\pm}1.5$). - The value of serum FSH(mIU/ml) in HCT treated group($3.870{\pm}2.151$) was significantly decreased(p<0.05) compared with control group($10.476{\pm}6.294$). - The expression of NGF-immunoreactive cells in the ovarian cells in HCT treated group was lesser observed than control group. - The number of implantation in HCT treated group($7.4{\pm}1.8$) was significantly increased (p<0.05) compared with control group($3.1{\pm}4.4$). Conclusions: Hyeolbuchukeo-tang(HCT) is effect on polycystic ovaries and the implantation of rats by EV-induced.

• Journal of Embryo Transfer
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• v.23 no.3
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• pp.189-195
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• 2008

The objective of this study is to investigate the changes in concentrations of leptin and insulin in serum of Korean cattle (Hanwoo) with reproductive disorders and to examine the relationship among leptin, insulin, and body condition score (BCS). The concentration of leptin in serum of pregnant Hanwoo showed insignificant difference from that in serum of Hanwoo with reproductive disorder, such as repeat breeding, follicular cyst, corpus luteum cyst, ovarian atrophy, and feeble estrus (p>0.05). However, the concentrations of leptin and insulin in serum were changed with different BCS value. In emaciated Hanwoo (BCS $2.0\sim2.9$), they were significantly decreased compared to BCS $3.0\sim3.4$ (p<0.05). The leptin showed different genotypes with different BCS value. In BCS $2.0\sim2.9$, C/T genotype was expressed (83.3%) more than C/C (16.7%) or T/T (0%) genotype, whereas C/C genotype was expressed (62.5%) more than C/T (25.0%) or T/T (12.5%) genotype in BCS $3.5\sim4.0$. The insulin concentration in follicular fluid obtained from ovary with follicular cyst which has follicles having diameter of $25\sim40 mm$ was significantly higher (p<0.05) than those in normal follicle fluid which has follicles having diameter of $3\sim10 mm$. These results showed that concentration of leptin and insulin in serum were related to BCS value and follicular size and suggest that the changes in concentration of leptin and/or insulin in serum could be a potent biomarker for diagnosis of bovine reproductive disorder.

• Journal of Embryo Transfer
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• v.29 no.1
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• pp.43-50
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• 2014

To describe the macroscopic anatomy and ovarian-physiological difference of the genital organs of the female Korean water deer, organs from captured animals in a wild area of Korea were dissected. The ovary of estrus group was about $1.10{\pm}0.02mm$ long and weighed about $0.50{\pm}0.02g$. And pregnant group was about $1.3{\pm}0.10mm$ long and weighed about $0.40{\pm}0.05g$. And the crowns of corpora lutea were found in the estrus group, but we couldn't find crowns at the pregnant group. Especially, the estrus ovaries tended (p=0.04) to be heavier than the ovaries during pregnancy. The MMP-9 activity was higher at the Graafian follicles of pregnant group than that in estrus group. However, with regard to follicles of estrus group, MMP-2 level was higher than that in pregnant group. Furthermore, apoptosis detection marker (Casp-3) was highly expressed in Graafian follicle of the pregnant group and the corpora lutea of estrus group. Thus, the differential expression of MMPs observed in this study suggests that the reflected the mechanisms underlying of monovulatory in estrus and/or pregnancy. Our results may be very useful as it provides with information that may be considered for the development of reproductive biotechnologies in endangered animals.

• The Journal of Korean Obstetrics and Gynecology
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• v.23 no.4
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• pp.35-46
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• 2010

Purpose: This study was designed to investigate the effects of Cyperi Rhizoma (CR) on changes in weights morphological and histopathological observations in Polycystic Ovary(PCO) rats. Methods: PCO was induced by single intramuscular injection with EV(2mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for five weeks. PCO control group(n=8) were injected with EV and orally administrated distilled water for five weeks. CR treated group(n=8) were injected with EV and orally administrated CR for five weeks. Then we measured weight of body, ovaries. The histopathology changes of ovaries, and the expression of nerve growth factor(NGF) were also evaluated. Results: Single injection of EV induced suppression of weight gain, formation of cysts, increase of NGF expression. Oral administration of CR prevent suppression of weight gain shown in control group. In addition CR group showed upward tendency of ovarian size. Formation of cystic follicles induced by EV injection is suppressed by CR treatment. In additon, CR treatment lowered expression levels of nerve growth factor(NGF), which were elevated by induction of PCOS. Conclusion: These results suggest that CR can be used for patients with PCOS to prevent formation of cystic follicles and malfunction of ovary. And also suggest that the related mechanisms are involved in suppression of NGF expression.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.77-82
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• 2000

This study was conducted to establish the methods of ultrasound-guided transvaginal retrieval of oocytes (ovum pick-up) in Korean native, Hanwoo Heifers. To obtain the basic data about Hanwoo, the size of ovaries in luteal and follicular phases were measured and the number of follicles in ovaries during the estrus cycle was counted by using ultrasound. And to determine the effective anesthetic to Hanwoo, various mixture of anesthetic compounds, Rompun(equation omitted), lidocaine, Monzal(equation omitted), and Domosedan(equation omitted), were treated. The size of Hanwoo ovaries were not significant differently between luteal and follicular phases. The number of medium and small follicles were peak on day 3 and 12 of the estrous cycle, and this result suggested that Hanwoo has 2 follicular growth waves per estrus cycle. The most effective anesthetic method was intramuscular injection of a.3m! Rompun(equation omitted), epidural injection of 5$m\ell$ lidocaine and sprayed cervix by 2$m\ell$ lidocaine.

• Journal of Animal Science and Technology
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• v.64 no.3
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• pp.432-442
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• 2022

The purpose of this study was to analyse the effects of light colour on rabbit reproductive performance and the expression of key follicular development genes. Rabbits (n = 1,068, 5 months old, 3.6-4.4 kg live body weight) were divided randomly into four groups, housed individually in wire mesh cages and exposed to red, green, blue, and white light-emitting diode (LED) light (control). The lighting schedule was 16 L : 8 D-15 d / 150 lx / 6:00 am-22:00 pm (3 d preartificial insemination to 12 d postartificial insemination). Red light and white light affected the conception rate and kindling rate and increased the total litter size at birth (p < 0.05). The effects of red light on litter size at weaning, litter weight at weaning, and individual weight at weaning increased compared with the green and blue groups. The effects of red light on live litter size at birth were increased compared with those in the blue group (p < 0.05). Compared to white light, green and blue light reduced the number of secondary follicles (p < 0.05). Compared to red light, green and blue light reduced the number of tertiary follicles (p < 0.05). Compared with white light, red LED light resulted in greater ovarian follicle stimulating hormone receptor and luteinizing hormone receptor mRNA expression (p < 0.05). Compared with green and blue LED light, red LED light resulted in greater B-cell lymphom-2 mRNA expression (p < 0.05). Compared with green LED light, red LED light inhibited FOXO1 mRNA expression in rabbit ovaries (p < 0.05). Red light can affect the reproductive performance of female rabbits and the expression of key genes for follicular development.

• Clinical and Experimental Reproductive Medicine
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• v.28 no.3
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• pp.225-233
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• 2001

Objective : To evaluate the efficacy of low-dose aspirin on IVF outcome and endometrium in patients undergoing IVF-ET. Materials and Methods : From February, 2001 to Jun, 2001, 60 infertile patients were randomly divided into study group (28 cycles) and control group (32 cycles). The study group received a daily oral dose of 25 mg of aspirin for at least 2 weeks from first visiting day. Controlled ovarian hyperstimulation was initiated in all patients with the GnRH agonist starting in the midluteal phase of the previous cycle. Results: There were no significant differences in age of the patients, basal serum E2, LH, FSH level and endometrial thickness among two groups. There were no statistically significant differences between the study group and the control group respectively in dosage ($26.5{\pm}4.8$ vs $26.2{\pm}5.3$ amples) and duration ($10.4{\pm}4.2$ vs $9.8{\pm}5.3$ days) of gonadotropin administration, serum E2 level on the hCG administration day ($1823{\pm}342$ vs $1854{\pm}543$), LH ($14.5{\pm}2.7$ vs $14.8{\pm}3.1$), FSH ($16.7{\pm}3.4$ vs $18.3{\pm}4.7$), the number of follicles > 15 mm ($13.2{\pm}6.3$ vs $12.8{\pm}5.9$), the number of oocytes retrieved ($9.2{\pm}2.4$ vs $8.4{\pm}1.7$), the number of embryos transferred ($4.7{\pm}2.0$ vs $4.7{\pm}2.0$), fertilization rate (68.4% vs 64.5%), implantation rate (21.3% vs 17.6%), and clinical pregnancy rate (28.4% vs 26.2%). The endometrial thickness and the percentage of endometrial trilaminar pattern on hCG day were significantly higher in study group than control group ($12.9{\pm}3.7mm$ vs $10.4{\pm}2.8mm$, 78.3% vs 64.5%). Conclusion: Many reports suggest that low-dose aspirin improve ovarian response, implantation rate, fertilization rate, implantation rate, and pregnancy rate by increasing the blood flow, but we couldn't prove the significant effect of low-dose aspirin on the IVF outcome except on endometrium. This may be affected by dose of aspirin, duration, and number of patients studied. This trial is small, so our results highlight the need for a large randomized controlled trial to identify the effect of low-dose as pirin on IVF-ET outcome.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.273-281
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• 1996

Previously, we have proposed a two-cell type model for follicular steroidogenesis inamphibians with Rana nigromacu lota. Present experiments were carried out to ascertain whether the model Is applicable to R. dybowskii. The role of theca layer were also reassessed by using granulosa cell-free pure theca layer (P-THEP). Theca/epithelium (THEP) layers, P-THEP layers, and granulosa cell enclosed-oocytes () were obtained from ovarian follicles of R. dybowskii by microdissection. Intact follicles (IFs) and different types of tissues were cultured for 6 hour in amphibian Ringer's m the presence or absence of FPII (0.05 gland/mi) or various steroid precursor (100 ng/ml). The amounts of product steroids converted by the components were measured by RIA. Exogenously added pregnenolone (P5) resulted in a marked increase in progesterone (P$_4$) by GCEOs (2143 pg/follicle) and IFs (2346 pg/follicle) but a smaller increase in P4 by THEP layer (495 pg/follicle). Addition of P$_4$ increased 17 a-hydroxyprogesterone (17 $\alpha$-OHP$_4$) levels by GCEOs (1118 pg/follicle) and IFs (1333 pg/follicle) but less by THEP layer (290 pg/follicle). However, much less amounts of P$_4$ or 17 $\alpha$-OHP$_4$ were producad by P-THEP layers than THEP in the presence of P5. Exogenous 1 7$\alpha$-OIIP$_4$ increased androstenedione (AD) levels by GCEOs (1415 pg/follicle) and IFs (561 pg/follicle) but not by THEP layers. In contrast, addition of AD resulted m a marked increase in testosterone (T) levels by TIIEP (2594 pg/follicle) and IFs (2223 pg/follide) but much less by GCEOs (339 pg/follicle). Exogenous T increased estradiol (E$_2$) levels by GCEOs (551pg/follicle) and IFs (887 pg/follicle), but not by THEP layer (<10 pg/follicle). Without addition of FPH or steroid precursors, very low or nondetectable levels of steroids were produced (< 20 pg/follicle) by all the types of follicular components examined. The data presented here indicate that the two-cell type model based on the study with R. nigromacu Iota is applicable to R. dybowskii and also suggest that the minor pathway, which convert P5 to 17$\alpha$-OHP$_4$, is not present in theca layer.

• Clinical and Experimental Reproductive Medicine
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• v.14 no.2
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• pp.119-126
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• 1987

Exponential rise rate(ERR) of serum estradiol concentrations during active follicular phase was calculated for 49 ovulation induction cycles by human menopausal gonadotropin to know the ovulation induction outcome according to ERR classified into 3 groups with low, moderate and high ERR values(Group I${\leqq}$0.3, 0.3${\leqq}$0.6，Group III>0.6). The results were summarised as follows : 1. No significant difference in the dosage of human menopausal gonadotropin was identified in each group. 2. The mean serum estradiol concentration at the day of human chorionic gonadotropin injection in Group II and Group ill was significantly higher than that in Group I . 3. The mean diameter of leading follicles at the day of human chorionic gonadotropin injection showed no significant difference in each group. 4. No significant difference in the ovulation rate was observed in relation to ERR. How ever, 20% and 20.8% of pregnancy rate in Group I and Group II were achieved while no pregnancy was occurred in Group III. 5. The ovarian hyperstimulation frequency was significantly higher in Group ill that in Group I and Group II. In conclusion, the study suggests that exponential rise rate of serum estradiol is a useful tool in HMG ovulation induction by preventing ovarian hyperstimulation without reducing pregnancy success rate.