• Clinical and Experimental Reproductive Medicine
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• v.14 no.2
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• pp.109-118
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• 1987

Ninety-one patients with irreparable tubal disease underwent in Vitro Fertilization-Embryo Transfer (IVF-ET) in Seoul National University Hospital. Ovulation was stimulated in 104 cycles by human menopausal gonadotropin (HMG) or follicular stimulating hormone (FSH)/human chorionic gonadotropin (HCG). The patients were classified as high (>900 pg/ml), intermediate (400-900 pg/ml), or low (<400pg/ml) responder according to preovulatory $E_2$ response and four $E_2$ patterns were found. The overall pregnancy rate per cycle in this consecutive series was 11.5% (n=12). The number of preovulatory oocytes per cycle was higher significantly in intermediate and high responder group than in low responder group (P<0.01), While the number of immature oocytes per cycle significantly higher in low responder group than high and intermediate responder group (P<0.01). The pregnancy rate in each responder group was not signigicant. According to the $E_2$ pattern of response, there was no significant difference in number of the immature and preovulatory oocytes recovery per cycle. There was a apparently direct relationship between the preovulatory $E_2$ pattern and pre gnancy rate was noted.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.1
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• pp.83-88
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• 1994

This study was designed to investigate the relationship between insulin resistance and obesity in the pathogenesis of polycystic ovarian syndrome(PCO). Twenty-two women with PCO, of whom thirteen were non-obese with body mass index(BMI, kg/$m^2$) of <25 and nine were obese with BMI${\geq}$25 were studied. Eight non-obese control women and seven obese control women were studied. Serum concentrations of testosterone, lutenizing hormone(LH)/follicle-stimulating hormone(FSH) ratio, and insulin-like growth factor I (IGF-I) were found to be significantly higher(P<0.05) in PCO women compared with control women, which clearly is not related to obesity. Serum glucose, insulin, and C-peptide levels were measured during a 2-hour oral glucose tolerance test(OGTT). Non-obese and obese women with PCO both(P<0.05) compared with control women demonstrated significant hyperinsulinemia after OGTT. The degree of hyperinsulinemia was found to be significantly higher in the obese women with PCO compared with the non-obese women with PCO. We concluded that obesity may contribute to hyperinsulinemia, however may not playa central role in the pathogenesis of PCO.

• Journal of Embryo Transfer
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• v.14 no.2
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• pp.113-119
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• 1999

This study was performed to improve the development of the in vitro fertilized bovine embryos by the condition of in vitro maturation. COCs were matured in TCM 199 supplemented with 0.1% PVA, 10ng/ml EGF, Hormones (5$\mu\textrm{g}$/ml FSH, 10 IU hCG, 1 $\mu\textrm{g}$/ml estradiol 17-$\beta$) or granulsa cell+Hormones atmosphere 39$^{\circ}C$, 5% CO2, 95% air for 24hrs. Matured oocytes were fertilized with frozen-thawed semen capacitated with 5mM caffein in BO medium for 20 hrs. IVF embryos were cultured in TCM 199 containing with hormones(same as matured medium), 10% FBS and co-culture with bovine oviduct epitherial cells. Maturation rates of COCs were showed 73.8%, 78.5%, 83.2% and 87.6% respectively, and were significant differences between PVA, EGF, and Hormones, GC+Hormones(p<0.05). The cleavage rates of IVF embryos were revealed 72.5%, 78.4%, 82.3% and 84.2% and showed same tendency as maturation rates(p<0.05). The blastocysts matured by above maturation condition and cultured for 7~10 days after fertilization had 34.4, 43.6, 52.3 and 59.3 cells had no differences among the treatments. These results suggest that high molecules as a substitutes of serum and growth factor may induce nuclear resumption of COCs but we need more study to produce transferable IVF blastocysts by use of that agents.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.283-291
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• 1997

In vitro development of bovine embryos is affected by many factors such as energy substrates, amino acids, and some growth factors. It has been reported that mRNA of insulin, PDGF and their receptors are detected in cow embryos, and that some chelating agents such as EDTA and transferrin have beneficial role on mouse and bovine embryos. The author hypothesized that insulin, transferrin arid PDGF added to a culture medium increase in vitro development of bovine embryos by chelating toxic substance(s) or increasing cell growth and metabolism. Immature oocytes from slaughtered ovaries of Holstein cows and heifers were matured for 24 hours in a TCM199 containing 10% fetal calf serum, FSH, LH and estradiol with granulosa cells in vitro. Matured oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Embryos cleaved to 2- to 4-cell at 30 hours after IVF were selected and cultured in a 30-$\mu$l drop of a synthetic oviduct fluid medium (SOFM) containing 0.8% BSA, Minimum Essential Medium essential and non-essential amino acids, and insulin, transferrin or PDGF for 9 days. Supplementation of a SOFM with insulin, and /or transferrin did not increase develop-mental rate to expanding and hatching blastocyst of 2- to 4-cell bovine embryos compared with control. The highest developmental rate to hatching blastocyst was shown when PDGF was added at the concentration of 10 ng /ml among the supplementing doses tested in the present study (p<0.05). Addition of PDGF without insulin to a SOFM could not increase embrye development, but combined addition of PDGF with insulin significantly increased (p<0.05) embryo development to hatching blastocyst (50%) compared with control (38%). In conclusion, insulin and PDGF supplemented to a SOFM may act synergistically and have beneficial effect on in vitro development of 2- to 4-cell bovine embryos matured and fertilized in vitro.

• Korean Journal of Animal Reproduction
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• v.9 no.1
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• pp.9-30
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• 1985

The study was carried out to find out the changes of the sex hormone levels in the milk of Holstein cows during the reproductive stages such as the estrous cycle, pregnancy and periparturient period. The FSH, LH, estradiol-17$\beta$ and progesterone from the milk samples were assayed by radioimmunoassay methods. The results of this study were summarized as follows: 1. The levels of progesterone and estradiol-17$\beta$ were similar among inter-quarters, but they were higher in after milking than before milking times, with no statistical significance. 2. The milk progesterone levels during the estrous cycles reached a peak mean level of 3.55$\pm$0.26ng/$m\ell$ at 15 days after estrus and they did not show any differences among the length of estrous cycles. The estradiol-17$\beta$ levels during the estrous cycles showed a peak level of 36.40$\pm$2.38pg/$m\ell$ at estrus, and decreased(17.20$\pm$0.46 pg/$m\ell$ to 18.65$\pm$1.26pg/$m\ell$) at luteal phase. 3. The FSH levels during the estrous cycles ranged from 2.25$\pm$0.23mIU/$m\ell$ to 4.35$\pm$0.24mIU/$m\ell$ showing significant changes. The LH levels during the estrous cycles gradually increased and remained a peak level of 10.90$\pm$0.36mIU/$m\ell$ from 20 to 25 days after estrus. 4. The progesterone levels during the pregnancy were decreased from 30 to 60 days after artificial insemination, and therafter continuously increased until 240 days. The estradiol-17$\beta$ levels during the pregnancy were 24.56$\pm$1.19pg/$m\ell$ at day 30 after artificial inseminaton, and increased rapidly until 180 days. The levles were agagin decreased by 26.17$\pm$3.03pg/$m\ell$ until 210 days and markedly increased by 68.00$\pm$8.70pg/$m\ell$ until 240 days. 5. The prolactin levels during the pregnancy were 31.27$\pm$2.31ng/$m\ell$ and 42.60$\pm$2.37ng/$m\ell$ at day 150 and 240 after artificial insemination respectively. The LH levels during the pregnancy reached a peak of 27.47$\pm$7.90mIU/$m\ell$ at day 30 after artificial insemination, and thereafter gradually decreased. 6. The progesterone levels during the periparturient period reached a peak of 4.61$\pm$0.34ng/$m\ell$ at day 3 prepartum, and thereafter gradually decreased, and showed 2.05$\pm$0.60ng/$m\ell$ at day 7 postpartum. The estradiol-17$\beta$ levels during the periparturient period showed high level from 207.23$\pm$6.04pg/$m\ell$ at day 1 prepartum to 239.90$\pm$13.90pg/$m\ell$ at day 2 prepartum, and thereafter began to decline and reached 51.87$\pm$1.72pg/$m\ell$ at by 7 postpartum. 7. The prolactin levels during the periparturient period showed relatively higher level at the time of parturition. The LH levels during the periparturient period rnage from 6.32$\pm$0.32mIU/$m\ell$ to 13.90$\pm$1.37mIU/$m\ell$ showing significant changes. 8. The progesterone levels(4.6$\pm$0.8ng/$m\ell$) of the pregnant cows were significantly higher than those (1.84$\pm$1.4ng/$m\ell$) of nonpregnant cows. The cows of artificial insemination from 61 to 90 days after parturition showed higher progesterone levels. 9. During 20 to 25 days after artificial insemination, the accuracy of pregnancy diagnosis from milk progesterone levels were 94.4% for nonpregnant cows(<2.3ng/$m\ell$), and 75.0% for pregnant cows( 3.2ng/$m\ell$). The average overall accuracy of pregnancy prediction for nonpregnant and pregnant cows 83.3% 10. The results obtained this study suggest that the understanding of the endocrinological mechanisms by means of milk hormone analysis during the estrous cycle, pregnancy and parturition would give the basic information needed for increasing efficiency of reproduction. This study would not only provide an accurate method of the early pregnancy diagnosis by milk progesterone levels but also contribute to the research of providing the method of detecting of FSH levels in milk, which was difficult in blood serum.

• Archives of Plastic Surgery
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• v.32 no.6
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• pp.699-705
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• 2005

Transgender is the severe type of gender identity disorder. The prevalence rate of transgender is reported to occur to about 1 out of 50,000 men, and about 1 out of 10,000 women. As for Korea, it is estimated to have about 1400 transgender patients. Lately, not only the numbers of them are increasing but also they are influencing our society increasingly. As for female transgender patients, they take female hormone for a long term before and even after the operation to maintain their physical identity of female. We have analyzed insulin like growth factor-1(IGF-1), insulin like growth factor protein binding-3(IGFBP-3), female hormone, male hormone and thyroid hormone in female transgender patients who have undergone the gender reassignment operation. We examined the changes of hormone level due to having female hormone steadily, and also examined how the steady use of the hormone could affect body organs. As for IGF-1, it showed significantly low in the female transgender group compared to control ($319.30{\pm}37.4$ vs $539{\pm}55.0$, p<0.05). As for IGFBP-3, there was no significant difference ($2859{\pm}200.3$ vs $2607{\pm}262.5$, p>0.05). As for female hormone, there was no significant differences in FSH($13.42{\pm}13.8$ vs $8.95{\pm}3.5$, p>0.05), estradiol($104.41{\pm}97.1$ vs $121.68{\pm}60.2$, p>0.05), and LH($7.62{\pm}5.6$ vs $7.4{\pm}3.3$, p>0.05). Even in comparison of testosterone, there was no significant differences($0.23{\pm}0.09$ vs $0.33{\pm}1.33$, p>0.05). As for thyroid hormone, there was no significant differences in TSH and free T4($1.34{\pm}0.94$ vs $1.71{\pm}0.12$, $1.4{\pm}0.37$ vs $1.46{\pm}0.17$, p>0.05). Therefore, this study concludes that apart from the decreased level of IGF-1, the possible endocrine side-effect problem due to female hormone seems to be low because there was no differences of female, male, and thyroid hormone level compared with normal female. Further study will be required in metabolic change including bone metabolism occurred by decrease level of IGF-I.

• Journal of Embryo Transfer
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• v.23 no.1
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• pp.37-42
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• 2008

This study was performed to improve the pregnancy rates of recipients following transfer of bovine embryos produced in vivo. Superovulation response didn't showe significant differences between each season (4.18 in spring; 4.36 in summer; 5.50 in fall; 4.38 in winter). Pregnancy rate was significantly different (p<0.05) between fresh (43.4%) and frozen embryos (17.2%). In administration of hCG to recipients, the pregnancy rate of fresh embryos (45.7%) was slightly higher than that of control (35.3%), but the pregnancy rates of frozen embryos in control group (25.0%) was higher than that of hCG group (16.0%). When synchrony of recipient and embryo was -2, -1, 0 and 1, the pregnancy rates were 20.0, 45.0, 30.3 and 26.3%, respectively. The pregnancy rates of recipients synchronized by naturally or $PGF_{2{\alpha}}$, CIDR/$PGF_{2{\alpha}}$ and E/P/CIDR/$PGF_{2{\alpha}}$/E treatments were 35.3, 48.0, 29.0 and 40.0%, respectively. Gestation lengths and birth weights of female and male calf were 288 and 290.5 days, 28.3 and 30.0 kg, respectively. The results were showed that the superovulation response was not affected by seasons, and also pregnancy rate didn't increase by administration of hCG, synchrony of embryo and recipients, synchrony methods. Further study and concern should be focused on improving the embryo freezing and pregnancy rate for commercial embryo transfer.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.191-198
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• 2001

The purpose of this study was to investigate the effects of embryo sources such as in vivo vs. in vitro produced blastocyst, and culture systems on the membrane permeability and viability of bovine blastocyst following GMP vitrification. To produce in vivo embryos, six cows were superovulated by administration of follicle stimulation hormone (FSH) and prostaglandin $F_{2{\alpha}}$(PG $F_{2{\alpha}}$). in vitro embryos were produced by two different culture systems, oviduct co-culture (OCS) and defined culture system (HECM-6; DCS). Ovaries were picked up at a local slaughterhouse and transported to laboratory in 3$0^{\circ}C$ saline within 2 h. Ovaries were washed with same saline three times and then placed in saline on warm plate adjusted at 3$0^{\circ}C$ during aspiration. The blastocysts produced were assigned for membrane permeability and viability following GMP vitrification. The membrane permeability of blastocysts was checked in 0.5 M sucrose solution on warm plate at 35$^{\circ}C$ for 0, 2, 5 and 7 min, respectively. Then the diameters (width and length) of embryo cytoplasms were measured by a eyepiece meter, and they were converted to their volume by 4/3 $\pi$ $r^3$. The blastocysts were cryopreserved by GMP vitrification method, where they were sequentially placed into vitrification solution before being loaded into GMP vessels and immersed into L$N_2$ within 20 to 25 sec. Post-thaw blastocysts were serially washed in 0.25 and 0.15 M sucrose in HM and TCM-199 for 5 min each, and then cultured in TCM 199 supplemented with 10% FCS for 24 or 48 h. The volume change of in vivo blastocyst at 0, 2, 5 and 7 min (100, 37.1, 34.3 and 31.6%) was significantly more shrunk than those of in vitro blastocysts derived from OCS (100, 59.8, 48.9, 47.9%) and DCS (100, 57.2, 47.3 and 46.9%) (P＜0.05). The viability of post-thaw blastocyst derived from in vivo (93.6%) was also significantly different from those in OCS and DCS (81.9 and 83.6%; P＜0.05). In the present culture system, the morphology of embryos produced in vitro was similar to that of in vivo embryos, but the quality in membrane permeability and post-thaw viability showed a big difference from their sources as in vivo or in vitro derived from OCS and DCS. The results indicated that the quality of in vivo embryos in membrane permeability and post-thaw viability was better than those of in vitro embryos derived from OCS or DCS.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.211-220
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• 1989

The aim of this study was to compare the response and their outcome of superovulation induction protocol in IVF-ET program. One hundued seventy seven infertile women were stimulated by FSH/hMG(group I, N=128), clomiphene citrate/hMG(group II, N=51), and hMG(group III, N=18) for the purpose of ovulation induction. The results were as follows; 1. The mean ages of patients were $31.9{\pm}3.8$ in group I, $30.6{\pm}3.3$ in group II, and $29.3{\pm}2.5$ in group III. 2. The day of hCG administration was $11.1{\pm}1.8$ in group I, $12.1{\pm}2.0$ in group II, and $13.7{\pm}6.8$ in group III. The hCG administration day of group III was significantly delayed compared with that of group I (p<0.001). 3. The terminal E2 pattern in group II was different from those of group I and III, but there was no significant difference. 4. The mean numbers of mature eggs aspirated were $5.5{\pm}3.8$ in group I, $5.0{\pm}3.3$ in group II, and $5.6{\pm}5.4$ in group III. There was no significant differences in the mean numbers of mature eggs retrieved among the three groups. 5. The fertilization rate of eggs was significantly higher in group II (67.9%) than that of group I (52.2%)(p<0.001). 6. The cleavage rate of group I (67.0%) was significantly lower than those of group II (93.2%) and group III (95.8%) (p<0.0001). 7. The mean numbers of embryos transfered were $3.3{\pm}1.4$ in group I, $3.1{\pm}1.3$ in group II, and $3.2{\pm}1.6$ in group III and the ET rate was 69.0% in group I, 77.3% in group II, and 100% in group III. There was significant difference in ET rate between group I and group III (p<0.005). 8. The pregnancy rates per OPU cycle or ET cycle were not significantly different among the three groups, but delivered and ongoing pregnancy rates were significantly different between group I (36.8%) and group II (p<88.8%) (p<0.05).

• Journal of Embryo Transfer
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• v.10 no.2
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• pp.131-138
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• 1995

Essential and non-essential amino acids supplemented to culture medium stimulate mammalian embryo development in vitro. Amino acids such as glycine, taurine and alanine are concentrated in the lumen of oviduct and uterus and it can he thought that these amino acids may have physiological role on fertilization and embryo development. Our aim of this experiment was to investigate the effects of essential and non-essential amino acids, taurine or glycine supplemented to fertilization medium on the cleavage and subsequent in vitro development of bovine oocytes matured and fertilized in vitro. Immature oocytes were obtained from slaughtered Holstein cows and heifers and matured in TCM199 containing 10% fetal calf serum, 2.5 $\mu$g /mL of FSH and LH and 1 $\mu$g / mL of estradiol with granulosa cells in vitro. After maturation, oocytes were coincubated with sperm in fertilization medium supplemented with Minimum Essential Medium (MEM) essential and non-essential amino acids, taurine (3.75 mM) or glycine (10 mM) for 30 hours in vitro. Inseminated oocytes were cultured in synthetic oviduct fluid medium (SOEM) containing MEM essential, non-essential amino acids and 1 mM glutarnine up to 8 days after fertilization.Supplementation of fertilization medium with MEM essential and non-essential amino acids lowered significantly (p<0.05 and p<0.001) the cleavage rate after 30 hours of IVF (53.3%) and at Day 3 (62.7%: Day 0: the day of I VF) compared to control (64.3% and 77.3%, respectively). Subsequent developmental rates to morulae (Mo) and expanding blastocysts (ExBL) also significantly decreased (p<0.001 and p<0.05 for Mo and ExBL) when oocytes were coincubated with sperm in the medium containing MEM amino acids. Taurine added to fertilization medium have not increased the cleavage rate over the control, whereas glycine showed significantly lower (p<0.01) cleavage rate at Day 3 than that of taurine, but there was no significant difference in the developmental rates to Mo and ExBL of bovine embryos irrespective of the supplementation of taurine or glycine to fertilization medium. In conclusion, supplementation of fertilization medium with essential and non-essential amino acids, taurine or glycine has no beneficial effect on in vitro cleavage and development of bovine oocytes matured and fertilization in vitro.