• Clinical and Experimental Reproductive Medicine
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• v.21 no.2
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• pp.137-147
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• 1994

Controlled ovarian hyperstimulation(COH) for in vitro fertilization and embryo transfer(IVFET) often results in the production of more embryos than can be efficaciously transferred at one time. However, embryo cryopreservation provides a mechanism by which additional embryos can be stored for later thawing and transfer. From November, 1990 to October, 1992, we completed 42 transfer cycles of cryopreserved pronucleus(PN) l-cell embryos using the fixed protocol of hormonal replacement therapy in a physiological manner regardless of individual ovarian function. Artificial endometrial stimulation was performed with only exogenous estradiol and progesterone(E-P) in 36 transfer cycles (Group I) and with gonadotropin-releasing hormone agonist(GnRHa) and exogenous estradiol and progesterone(GEEP) in 6 transfer cycles(Group II ). The results were as follows. 1. The Survival rate of total cryopreserved-thawed embryos was 64.9%(198/305): 64.9% (172/265) in Group I and 65.0% (26/40) in Group II. 2. Total 168 embryos were transferred with an average of 4.7 per ET in Group I and total 26 embryos were transferred with an average of 4.3 per ET in Group II. 3. The pregnancy rate(PR) per cryopreserved-thawed ET and the implantation rate was 33.3 %(14/42) and 6.7%(13/194), respectively. The PRs per cryopreserved-thawed ET were 30.6% (11/36) in Group I and 50.0% (3/6) in Group II without significant difference. 4. The take home baby rate was 11.1%(4/36) in Group I and 33.3% (2/6) in Group II.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.111-117
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• 2004

Objectives: To investigate the effect of LH receptor in folliculogenesis, we confirm the expression level of LH receptor (LH-R) mRNA in human granulosa cells (GCs) and its expression levels were analyzed by comparison to embryo developmental rate and pregnancy rate. Materials and Methods: GCs were obtained at the time of oocyte retrieval from the patients undergoing IVF-ET program. The patients were divided into two groups: Group I (n=20) is poor responder (retrieved oocyte(s)$\leq$3ea), Group II (n=80) is normal responder (retrieved oocytes>3ea). After the extraction of total RNA, semiquantitative RT-PCR was performed and the expression level of LH-R mRNA was normalized by $\beta$-actin. Statistical analysis was performed by using $X^2$ test, Student's t-test and Pearson correlation. Results: In Group II, the relative values of LH-R mRNA (0.680 vs. 0.463, p<0.005) and pregnancy rate (54.7% vs. 23.1%, p<0.05) were significantly higher than in Group I. Number of retrieved oocyte(s) was gradually increased when the expression of LH-R mRNA was increased (p<0.05). But the quality of retrieved oocyte and transferred embryo were not related with the expression of LH-R mRNA. When the pregnancy rate was compared with FSH only group and FSH combined with hMG group in the ovarian stimulation protocol, FSH combined with hMG group was significantly higher than FSH only group in Group I (37.5% vs. 0%), and the expression of LH-R mRNA was significantly higher in hMG combined group than FSH only group (p<0.05). Conclusion: Expression level of LH-R mRNA has important role in ovarian function related with the response to gonadotrophin in human folliculogenesis. Furthermore these data might provide the evidence that additional use of hMG is helpful to poor responders.

• Clinical and Experimental Reproductive Medicine
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• v.38 no.3
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• pp.164-167
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• 2011

Objective: There are limited data regarding the significance of elevated serum CA-125 level during IUI cycles, even though it is used widely during the initial evaluation of infertile patients. The aim of this study was to investigate the prognostic value of serum CA-125 levels during IUI cycles. Methods: Among the patients with controlled ovarian stimulation and IUI cycles at Seoul National University Hospital from Jan 2005 through Dec 2009, 92 cases with no identified endometriotic lesion, ovarian tumor, salpingeal lesion, or uterine myoma were selected. To compare the clinical characteristics between the pregnancy group and the non-pregnancy group, the Mann-Whitney U test and Fisher's exact test were used. Results: The overall pregnancy rate was 18.5% (17/92). The pregnancy group showed a higher number of follicles 16 mm in diameter ($p$=0.036), endometrial thickness ($p$ <0.001), ampules of gonadotropin ($p$=0.009), and higher body mass index ($p$=0.022) than the non-pregnancy group. No significant difference was observed in the serum CA-125 level or the proportion of patients with CA-125 exceeding 17 IU/mL between the two groups. Conclusion: The prognostic value of serum CA-125 level among infertile patients with IUI cycles is considered limited.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.433-436
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• 2004

Polycystic ovarian syndrome (PCOS) is a very common endocrine disorder in women of reproductive age. There are some evidences that nerve growth factor (NGF) is involved in the pathogenesis of PCOS. In this study, we investigated the effect of Korean red ginseng total saponin (GTS) on the ovarian morphology and NGF expressions in the ovaries, pituitary and hippocampus. The oil control animals were injected with 0.2 ml oil/rat. Animals in estradiol valerate (EV) control group were injected i.m. with 4 mg EV in 0.2 ml oil/rat. The GTS was administered (100 mg/kg) i.p. every other day for 60 days, beginning 1 day after the EV injection. PCO was induced by a single injection of EV (4 mg, i.m.). At day 60, the expressions of NGF were examined by immunohistochemistry. The main findings were as follows; PCO was fully developed with a single i.m. injection of EV, and PCO showed the increased expression of NGF, and GTS administration decreased NGF expressions in the ovaries without affecting pituitary and hippocampus significantly. The present results demonstrate that GTS attenuates PCOS by the stimulation of NGF expression.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.3
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• pp.169-173
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• 2016

Objective: To determine the preferred regimen for women with adenomyosis undergoing in vitro fertilization (IVF), we compared the IVF outcomes of fresh embryo transfer (ET) cycles with or without gonadotropin-releasing hormone (GnRH) agonist pretreatment and of frozenthawed embryo transfer (FET) cycles following GnRH agonist treatment. Methods: This retrospective study included 241 IVF cycles of women with adenomyosis from January 2006 to January 2012. Fresh ET cycles without (147 cycles, group A) or with (105 cycles, group B) GnRH agonist pretreatment, and FET cycles following GnRH agonist treatment (43 cycles, group C) were compared. Adenomyosis was identified by using transvaginal ultrasound at the initial workup and classified into focal and diffuse types. The IVF outcomes were also subanalyzed according to the adenomyotic region. Results: GnRH agonist pretreatment increased the stimulation duration ($11.5{\pm}2.1days$ vs. $9.9{\pm}2.0days$) and total dose of gonadotropin ($3,421{\pm}1,141IU$ vs. $2,588{\pm}1,192IU$), which resulted in a significantly higher number of retrieved oocytes ($10.0{\pm}8.2$ vs. $7.9{\pm}6.8$, p=0.013) in group B than in group A. Controlled ovarian stimulation for freezing resulted in a significantly higher number of retrieved oocytes ($14.3{\pm}9.2$ vs. $10.0{\pm}8.2$, p=0.022) with a lower dose of gonadotropin ($2,974{\pm}1,112IU$ vs. $3,421{\pm}1,141IU$, p=0.037) in group C than in group B. The clinical pregnancy rate in group C (39.5%) tended to be higher than those in groups B (30.5%) and A (25.2%) but without a significant difference. Conclusion: FET following GnRH agonist pretreatment tended to increase the pregnancy rate in patients with adenomyosis. Further largescale prospective studies are required to confirm this result.

• Animal Bioscience
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• v.34 no.8
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• pp.1290-1302
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• 2021

Objective: Follicle selection is an important process in chicken egg laying. Among several small yellow (SY) follicles, the one exhibiting the highest expression of follicle stimulation hormone receptor (FSHR) will be selected to become a hierarchal follicle. The role of lncRNA, miRNA and other non-coding RNA in chicken follicle selection is unclear. Methods: In this study, the whole transcriptome sequencing of SY follicles with different expression levels of FSHR in Jining Bairi hens was performed, and the expression of 30 randomly selected mRNAs, lncRNAs and miRNAs was validated by quantitative real-time polymerase chain reaction. Preliminary studies and bioinformatics analysis were performed on the selected mRNA, lncRNA, miRNA and their target genes. The effect of identified gene was examined in the granulosa cells of chicken follicles. Results: Integrated transcriptomic analysis on chicken SY follicles differing in FSHR expression revealed 467 differentially expressed mRNA genes, 134 differentially expressed lncRNA genes and 34 differentially expressed miRNA genes, and sosondowah ankyrin repeat domain family member A (SOWAHA) was the common target gene of three miRNAs and one lncRNA. SOWAHA was mainly expressed in small white (SW) and SY follicles and was affected by follicle stimulation hormone (FSH) treatment in the granulosa cells. Knockdown of SOWAHA inhibited the expression of Wnt family member 4 (Wnt4) and steroidogenic acute regulatory protein (StAR) in the granulosa cells of prehierarchal follicles, while stimulated Wnt4 in hierarchal follicles. Overexpression of SOWAHA increased the expression of Wnt4 in the granulosa cells of prehierarchal follicles, decreased that of StAR and cytochrome P450 family 11 subfamily A member 1 in the granulosa cells of hierarchal follicles and inhibited the proliferation of granulosa cells. Conclusion: Integrated analysis of chicken SY follicle transcriptomes identified SOWAHA as a network gene that is affected by FSH in granulosa cells of ovarian follicles. SOWAHA affected the expression of genes involved in chicken follicle selection and inhibited the proliferation of granulosa cells, suggesting an inhibitory role in chicken follicle selection.

• Journal of Embryo Transfer
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• v.24 no.3
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• pp.145-150
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• 2009

This study was conducted to examine the effects of human follicular fluid and gonadotropin (FSH+HCG+rhEGF) on in vitro maturation, fertilization and development of human immature oocytes. Cumulus-oocyte complexes (COCs) were collected following for in vitro fertilization and embryo transfer (IVF-ET) cycles of the patients. At the time of oocytes collection, oocytes were classified into MII, MI and GV in accordance with their appearance (MII: Fully mature oocyte at metaphase II of meiosis; MI: Nearly mature oocytes at metaphase I of meiosis; GV: Immature oocytes at prophase I of meiosis). After controlled ovarian stimulation using gonadotropin(FSH) and human chorionic gonadotropin (HCG) in 70 ICSI cycles, 158 MI to MII matured oocytes were intracytoplasmic sperm injection (ICSI) ${\sim}4$ h after in vitro culture and 553 MII oocytes were ICSI after denudation. The aspirated MI and GV oocytes were cultured in culture medium containing 10% (v/v) serum protein substitute (SPS), 10% (v/v) human follicular fluid (hFF) and 10% (v/v) serum protein substitute (SPS)+1 IU/ml FSH+10 IU/ml HCG+10 ng/ml recombinant human epidermal growth factor (rhEGF). The maturation rate of immature oocytes was similar among the three group. When maturation medium was supplemented with 10% SPS, 10% hFF or gonadotropins, the fertilization rate of in vitro matured oocytes was higher in 10% SPS (80.0%), but there was no statistical significance (78.2%; hFF, 76.9%; gonadotropin, p>0.05). The development rate of human embryos developed to $6{\sim}8$ cells were not significant difference in the medium containing SPS, hFF and gonadotropins (65.6%, 65.9% and 66.7%). The results of these study suggest that human follicular fluid and gonadotropins supplemented in the culture medium was not effected on the in vitro maturation, fertilization and development of human immature oocytes.

• The Korean Journal of Physiology
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• v.30 no.2
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• pp.197-208
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• 1996

Endosomes lower their internal pH by an ATP-driven proton pump, which is critical to dissociation of many receptor-ligand complexes, the first step in the intracellular sorting of internalized receptors and ligands. Endosomes are known to exhibit n great range of pH values that can vary between 5.0 and 7.0 within a single cell although the factors that regulate endosomal pH remain uncertain. To evaluate the morphological and topological differences of endosomes in the different stages, confocal microscopy was used. The early endosomes labeled with fluorescein isothiocyanate-dextran for 10 min at $37^{\circ}C$ were identifiable at the peripheral and tubule-vesicular endosome compartment. In contrast, the late endosomes formed by 10 min pulse and 20 min trace were located deeper in the cytoplasm and showed more vesicular features than early endosomes. For the purpose of determining whether ATP-dependent acidification was heterogeneous and whether the differences in acidification were attributed to differences in the activity of $Na^{+}-K^{+}$-ATPase and/or $Cl^{-}$ channel, endocytic compartments were fractionated into subpopulation using percoll gradient and measured ATP-dependent acidification. While all fractions exhibited ATP-dependent acidification activity, both the initial rate of acidification and extent of proton translocation were lower in early endosomes and gradually increased in late endosomes. Phosphorylation by PKA and ATP enhanced ATP-dependent acidification in both early and late endosomes, hut there was no difference in the degree of enhancement by phosphorylation between two subpopulations. When ATP-dependent acidification was determined in the presence or absence of vanadate ($Na_{3}VO_{4}$) or ouabain, only early endosomes exhibited the vanadate or ouabain dependent stimulation of acidification activity, suggesting the inhibition of $Na^{+}-K^{+}$-ATPase. Therefore, it seems probable that the inhibition of early endosome acidification by $Na^{+}-K^{+}$-ATPase observed in vitro at least in part plays a physiological role in controlling the acidification of early endosomes in vivo.

• Journal of Embryo Transfer
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• v.17 no.2
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• pp.123-128
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• 2002

The present study aimed at determining the effective dose of Folltropin, a follicle timulating hormone (FSH), on superovulation in indigenous cows of Bangladesh. Fifteen regularly cycling 5～7 years old dry cows, weighing 200～250 kg with 2.5～3.0 body condition scores (BCS) were divided into three groups (n＝5). Individual groups were superovulated with 100, 200 or 300 mg of Folltropin per animal. The superovulation treatment was initiated at Day 10 or Day 11 of the estrous cycle (Day 0＝day of estrus). Alfaprostol (6 mg) was injected to each cow 72 h after the initiation of superovulation treatment to induce eestrus. After confirming standing estrus, the cows were inseminated 2～3 times, 12 h apart, depending on the duration of estrus. At Day 6 or Day 7, individual horns of the uterus were flushed with 150～200 $m\ell$ of phosphate buffered saline supplemented with BSA (0.2％), penicillin (100 IU／$m\ell$) and streptomycin (100 $\mu\textrm{g}$／$m\ell$) using a two-way foley catheter. The embryos were concentrated, removing the excess medium through an embryo filter, and identified under a stereomicroscope. The identified embryos were collected, washed four times, evaluated and graded as excellent, good, fair or poor. The excellent, good and fair embryos were considered as transferable quality embryos. The mean (range). numbers of embryos collected vs. transferable quality embryos far 100, 200 and 300 mg of Folltropin were 4.5 (1～10) vs. 3.5 (1～8); 2.5 (1～4) vs. 1 (0～2) and 0.0 (0～0) vs. 0.0 (0～0), respectively, Folltropin at a dose of 100 or 200 mg produced suitable ovarian stimulation for superovulation in indigenous zebu cows of Bangladesh. A dose of 300 mg or more Folltropin consistently caused preovulatory corpora lutea formation in the ovaries and resulted in zero embryo recovery.

• Journal of Yeungnam Medical Science
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• v.37 no.1
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• pp.47-53
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• 2020

Background: This study was conducted to analyze clinical factors that can affect pregnancy rates in normal responders undergoing the freeze-all policy in in vitro fertilization. Methods: We evaluated 153 embryo transfer cycles in 89 infertile women with normal response to controlled ovarian stimulation (COS). After COS, all embryos were cultured to the blastocyst stage, and good quality blastocysts were vitrified for elective frozen-thawed embryo transfer (FET). Clinical variables associated with COS and the results of COS and culture, including the number of retrieved oocytes, fertilized oocytes, and frozen blastocysts were compared between the pregnant group and the non-pregnant group. Results: After a single cycle of COS for each patient, 52 patients became pregnant while 37 did not. Significant differences were observed in the number of matured oocytes, fertilized oocytes, frozen blastocysts, and transferred embryos. The number of frozen blastocysts in the pregnant group was almost twice that in the non-pregnant group (5.6±3.1 vs. 2.8±1.9, p<0.001). The area under the receiver operating characteristic curve for the 4 frozen blastocysts was 0.801 in the pregnant group. Conclusion: In the freeze-all policy, the number of matured oocytes, number of fertilized oocytes, and number of frozen blastocysts might be predictive factors for pregnancy.