• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.101-111
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• 2001

The present study was conducted to develop an in vitro culture system that would support bovine follicle growth from preantral to antral stage, oocyte maturation, fertilization, and embryonic development. Bovine preantral follicles (150$\pm$1.2 ${\mu}{\textrm}{m}$) surrounded by theca cell were isolated ezymatically and mechanically from ovarian cortical slides in Leibovitz L-l5 medium containing 1 mg/$m\ell$ collagenase and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium. The survival and growth rates of follicles cultured in the presence of FSH (10~150 ng/$m\ell$) were significantly higher than those of control group (P ＜ 0.001), but no significant differences were observed in survival and growth rates of follicles between the LH treatment groups (1~125 ng/$m\ell$) and the control. The survival (40%) and growth (244 $\pm$ 0.5 $\mu\textrm{g}$) of follicles cultured with FSH (90 ng/$m\ell$) and LH (25 ng/$m\ell$) were higher than those of control (25%, 160 $\pm$1.0 $\mu\textrm{g}$). Finally, 50% percent of healthy antral follicles were obtained, and almost 60% of them has complete meiotic division with 1st polar body (18.1%) and 10.0% have developed to the cleaved embryo and blastocyst stage. These results suggest that bovine preantral follicle with intact theca cell can grow to the antral stage using these culture conditions, and that oocytes from in vitro-matured bovine preantral follicle may acquire meiotic competence and can undergo fertilization and development.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.9
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• pp.1205-1209
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• 2000

The present study was undertaken to investigate the effect of stimulation of follicular development with eCG on the peripheral levels of inhibin and FSH in Murrah buffaloes. Estrus was synchronized in five normally cycling females by insertion of Crestar (Intervet, Boxmeer, Holland) implants for nine days. Estradiol valerate was administered i.m. on the day of implant insertion. On the 10th day of the induced estrous cycle a single dose of 3000 IU eCG (Folligon, Intervet, Boxmeer, Holland) was given, followed by treatment with 25 mg of $PGF_2$ alpha (Lutalyse, Upjohn, Belgium) 48 h later. Blood samples were obtained during the induced estrus, on cycle day 10 (luteal phase), at the superovulatory estrus (43 h after PGF) and during the periovulatory period (64 h after PGF). Ultrasonography was done daily to monitor follicular development. Plasma concentrations of inhibin and FSH were determined by specific radioimmunoassays. Differences between $mean{\pm}SEM$ values of different phases of the cycle were compared by ANOVA. The mean number of small (2-5 mm), medium (6-9 mm) and large (>10 mm) follicles observed two days after eCG treatment and on the day of superovulatory estrus was $2.8{\pm}0.31$, $5.2{\pm}0.30$ and $1.4{\pm}0.09$ and $1.9{\pm}0.21$, $2.8{\pm}0.40$ and $5.0{\pm}0.83$, respectively. The mean number of ovulations was $3.6{\pm}0.37$ and the mean number of unovulated follicles was $6.1{\pm}0.47$. Most of the follicles >10 mm in diameter had ovulated (72%). The mean ${\pm}SEM $ of plasma inhibin concentration $(2584.15{\pm}17.92pg/ml)$ during the superovulatory estrus was significantly higher $(p{\leq}0.05)$ than during the induced estrus $(749.87{\pm}17.29pg/ml)$, the luteal phase $(1099.54{\pm}24.98pg/ml)$ and periovulatory period $(1682.71{\pm}29.88pg/ml)$, respectively. $Mean{\pm}SEM$ plasma FSH concentration during the induced estrus $(10.35{\pm}0.41ng/ml)$ was not different from that during the superovulatory estrus $(8.52{\pm}0.39ng/ml)$, but was significantly higher $(p{\leq}0.05)$ than during the luteal phase $(2.81{\pm}0.42ng/ml)$ and periovulatory period $(5.7{\pm}0.28ng/ml)$. These data indicate that treatment with eCG in buffaloes for inducing superovulation results in a significant elevation in plasma inhibin levels and a decrease in plasma FSH levels during the superovulatory estrus. Thus, we suggest that the elevated plasma inhibin coming from fully developed follicles continued for a long time which results in inhibition of FSH leading to poor ovulation in the remaining follicles, which may be the cause of suboptimal superovulatory response.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.3
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• pp.340-350
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• 2007

The objective of this study was to determine whether peroxisome proliferator-activated receptor ${\gamma}$(PPAR${\gamma}$ is involved in the regulation of weaning to estrus of primiparous sows. Twelve sows composed of 6 groups of 2 full-sibs in a similar age (325.2 d), body weight (BW; 152.4 kg) and backfat thickness (BFT; 27.0 mm) at start of lactation, were allocated to accept 31 MJ (restricted group, R-group) or 53 MJ (control group, C-group) DE/d treatment, respectively. The experimental results indicated that the low energy intake resulted in excessive losses of BW and BFT during lactation in R-group sows, which may be related to decrease of serum 15-deoxy-${\Delta}^{12,14}$-prostaglandin $J_2$ (15d-$PGJ_2$), a ligand of PPAR${\gamma}$ The obvious peak and the frequency of LH, FSH and estradiol ($E_2$) were only observed in C-group sows. Except for $E_2$ at d 1 and 2, serum FSH, LH and $E_2$ concentrations in R-group were lower than those in C-group sows after weaning. However, the serum progesterone ($P_4$) level in R-group sows was always more than that in C-group. The expression abundances of PPAR${\gamma}$and GnRH receptor (GnRH-R) in pituitary, FSH receptor (FSH-R), LH receptor (LH-R), estrogen receptor (ES-R) and aromatase in ovary of anestrous sows were lower than those of estrous sows. Neither the BFT nor the BW was associated with the mRNA abundance of PPAR${\gamma}$in hypothalamus during lactation. Expressions of PPAR${\gamma}$in pituitary and ovary were affected evidently by the BFT changes and only by the loss of BW of sows during and after lactation. Furthermore, PPAR${\gamma}$mRNA level in ovary was significantly related to the expression abundances of GnRH-R, FSH-R, ES-R and aromatase, and GnRH-R was obviously associated with PPAR${\gamma}$expression in pituitary. However, PPAR${\gamma}$expression in hypothalamus likely has no effects on these genes expression and no obvious difference for all sows. Not serum $E_2$ or $P_4$ alone but the ratios of $E_2$ to $P_4$ and 15d-$PGJ_2$ to $P_4$, and serum FSH and LH were evidently related to PPAR${\gamma}$expression in pituitary and ovary. It is concluded that PPAR${\gamma}$is associated with body conditions, reproduction hormones and their receptor expression, which affected the functions of pituitary and ovary and ultimately the estrus after weaning of primiparous sows.

• Clinical and Experimental Pediatrics
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• v.52 no.12
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• pp.1377-1382
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• 2009

Purpose:The gonadotropin-releasing hormone (GnRH) test results of girls with precocious puberty were analyzed to determine whether this test can efficiently and clearly differentiate between central precocious puberty (CPP) and other disorders. Methods:Clinical and laboratory data of 54 girls with precocious pubertal signs were reviewed. Intravenous GnRH test was performed with blood samples obtained at 0, 30, 60, and 90 minutes. A peak luteinizing hormone (LH) level of ${\geq}5.0IU/L$ was indicative of CPP. Results:Of the 40 girls with CPP, 36 (90.0%), 3 (7.5%), and 1 (2.5%) showed peak LH levels at 30, 60, and 90 minutes, respectively. A percentage of girls whose peak LH ${\geq}5.0IU/L$ up to 30, 60, and 90 minutes was 92.5%, 100%, and 100%, respectively. The peak LH/follicle stimulating hormone (FSH) ratio of girls with CPP was 0.89${\pm}$0.49 and was <1 in 16 of the 40 girls (40.0%). Girls with peak LH/FSH ratio of >1.0 showed higher chronological age (CA) ($8.3{\pm}0.6$ vs. $7.7{\pm}1.0$ years, P=0.033), bone age (BA) ($10.9{\pm}0.8$ vs. $9.7{\pm}1.1$ years, P=0.001), and BA-CA difference ($2.6{\pm}0.7$ vs. $2.0{\pm}0.7$ years, P=0.009) than those of girls with peak LH/FSH ratio of ${\leq}1.0$. Higher percentage of girls with peak LH/FSH ratio of >1.0 showed advanced breast development (${\geq}TannerIII$) (93.7% vs. 41.7%, P=0.001). Conclusion:LH levels after 30 and 60 minutes of intravenous GnRH administration are the most useful for diagnosing CPP in girls.

• Journal of Embryo Transfer
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• v.13 no.2
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• pp.107-115
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• 1998

The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.0$\pm$11.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.7$\pm$1.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

• Clinical and Experimental Reproductive Medicine
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• v.41 no.2
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• pp.86-91
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• 2014

Objective: The aim of the present study was to investigate the relationship between ovarian follicle count and volume on ultrasonography and serum hormone levels including the levels of the anti-$M\ddot{u}llerian$ hormone (AMH) and gonadotropin in women with the polycystic ovary syndrome (PCOS). Methods: A total of 118 Korean women aged 18-35 years who were newly diagnosed with PCOS at a university hospital were included in this study. Serum LH, FSH, and AMH levels were measured in the early follicular phase, and the total antral follicle count (TFC) and the total ovarian volume (TOV) were assessed by ultrasonography. The correlations between serum hormonal parameters and ultrasonography characteristics in women with PCOS were evaluated using Pearson's correlation coefficients and a linear regression analysis. Results: Serum AMH levels were significantly correlated with serum LH levels and LH/FSH ratios, and TFC and TOV were significantly correlated with each other on ultrasonography. Serum AMH and LH levels and the LH/FSH ratio were significantly correlated with TFC. Statistically significant correlations between TOV and the LH level (r=0.208, p=0.024) and the LH/FSH ratio (r=0.237, p=0.010) were observed. However, the serum AMH level was not significantly correlated with the ovarian volume, and this result did not change after adjusting for age and body mass index. Conclusion: Serum AMH is not related to the ovarian volume in women with PCOS. My results suggest that serum LH level and the LH/FSH ratio may be more useful than the serum AMH level for representing the status of the ovarian volume in women with PCOS.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.7
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• pp.897-900
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• 2000

The experiment was conducted to study the effect of GnRH administration at induced estrus on pituitary and ovarian response in buffalo heifers. Eight Murrah river buffaloes of 12 to 13 months of age were treated with $PGF_{{2}{\alpha}}$ and eCG combination. GnRH (Fertagyl) 200 ug was injected (iv) at estrus in four heifers (treated group) while saline (2 ml, iv) was injected in remaining four heifers (control group). Blood was collected through jugular catheter to estimate plasma FSH, LH and estradiol level. The pretreatment plasma FSH, LH and estradiol values ranged from $8.46{\pm}1.97ng/ml$ to $12.31{\pm}1.30ng/ml$, $0.87{\pm}0.21ng/ml$ to $1.19{\pm}0.29ng/ml$ and $19.09{\pm}2.38pg/ml$ to $20.24{\pm}1.00pg/ml$ respectively. The plasma estradiol concentration elevated significantly (p<0.05) within 24 hr after eCG administration and reached its peak levels of $154.09{\pm}17.28pg/ml$ and $181.95{\pm}31.82pg/ml$ at estrus in respectively treatment and control groups. The plasma FSH and LH concentrations did not increase during follicular development after eCG administration while initial significant (p<0.05) increases in both plasma FSH and LH concentrations occured within 5 and 10 min, reaching peak levels of respectively $110.06{\pm}23.56ng/ml$ and $13.15{\pm}3.13ng/ml$ within 90 min after GnRH injection was detected. A sharp and significant decline in plasma estradiol concentration ($59.27{\pm}8.78pg/ml$) associated with synchronized ovulation within 24 hours after GnRH injection was recorded. The observation suggest that the hypophysis of prepubertal buffaloes treated with eCG have gonadotrophins awaiting the releasing factor to evoke release of gonadotrophin during the follicular phase to induce synchronized ovulation.

• The Journal of Korean Medicine
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• v.30 no.5
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• pp.137-145
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• 2009

Objectives: This study was performed to elicit the effectiveness of a herbal formula, Kamisoyo-san (Jiaweixiaoyaosan) on enhancing ovarian function in infertile woman with ovarian dysfunction. Methods: 28 patients who initially visited between November 2006 and February 2009 and were administered Kamisoyo-san until May 2009 were retrospectively evaluated for their ovarian function by means of basal FSH (b-FSH), menstrual cycle, body mass index (BMI), and body fat ratio. To identify the major factor of improving ovarian function, 28 patients were classified into two groups by criteria of patent factors, such as pre-administration b-FSH, patient age, and treatment duration, respectively. Two groups were compared in terms of pregnancy percentage, b-FSH, menstrual cycle, BMI, and body fat ratio. Results: Post-administration b-FSH significantly decreased in comparison with pre-administration (p=0.004). The higher group (b-FSH $\geq$ 25mIU/mL) in pre-administration b-FSH was more effective in decrease of post-administration b-FSH than the lower one (10mIU/mL < b-FSH < 25mIU/mL). Conclusion: Kamisoyo-san may have a therapeutic effect on the infertility of child bearing period woman with ovarian dysfunction.

• Reproductive and Developmental Biology
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• v.31 no.3
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• pp.199-205
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• 2007

The present study was carried out to assess the effect of superovulation response and efficiency of embryo production induced with injection of FSH dissolved in polyethylene glycol (PEG) in Hanwoo. Eighty-eight cows were divided into four groups. In group 1, cattle were intramuscularly treated with twice-daily administration of 50mg FSH for 4 days. Group 2 and 3 were subcutaneously single injection of 400mg and 200mg FSH dissolved in 30% PEG, respectively. Group 4 were subcutaneously single injection of 200 mg FSH dissolved in 30% PEG at 7 day after CIDR insertion. The number of corpus luteum (CL) in group 2 resulted in significantly (p<0.05) higher compared to group 1, 3 and 4 (18.5 vs. 11.2, 13.1 and 13.9, respectively). However, the number of total ova $(7.9{\sim}10.4)$, transferable embryos $(3.7{\sim}4.7)$, degenerate embryos $(1.9{\sim}3.5)$ and unfertilized ova $(1.8{\sim}2.7)$ did not differ among treatment groups. No difference was observed in pregnancy rate after transferring the recovered embryos among groups $(36.0{\sim}50.0%)$. In addition, blood progesterone concentrations at embryo recovery did not differ among all groups. In conclusion, although no differences were observed in the number of total ova, transferable embryos and pregnancy rate after transfer, a single injection of reduced dose of FSH (200mg FSH) at 7 day after CIDR insertion is more practical for superovulation treatments than frequent injection because of reduction of stress in Hanwoo and decreases of cost and laber.

• Development and Reproduction
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• v.13 no.4
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• pp.329-339
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• 2009

Cortisol is present in high concentration in the ovary and its receptor is expressed in the ovarian cells. Moreover, cortisol is known to have a role in steroid synthesis and cell metabolism in human granulosa and lutein cells. However, little is known of the role of cortisol presenting in high concentration in the follicles after LH surge on the granulosa-lutein cells. Therefore, the this study we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ in the granulosa-lutein cells that are obtained during oocyte-retrieval after treatment with 5, 50, and $500{\mu}g/m\ell$ cortisol and 1 IU/$m\ell$ FSH. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in 50 and $500{\mu}g/m\ell$ cortisol treated cells. We found, however, that FSH did not suppress the apoptosis of the cells induced by cortisol. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by cortisol treatment, whereas $E_2$ was not changed. We also demonstrated that FSH did not inhibit the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present study suggests that cortisol of high concentration could cause the apoptosis of human granulosa-lutein cells by suppressing the production of $P_4$. However, we need more studies to elucidate the mechanism by which cortisol induces apoptosis in human granulosa-lutein cells in view of the fact that our results are inconsistent with previous reported data.