Combination of Korean Red Ginseng Extract and Hydrogen-Rich Water Improves Spermatogenesis and Sperm Motility in Male Mice / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of hydrogen-rich Korean Red Ginseng (KRG) water (HRGW) mixture on the spermatogenesis and sperm motility of mice of different ages.@*METHODS@#Eighty young (3 month-old) and aged (12 month-old) male mice were randomly assigned to 4 groups (n =10 per group) including control group, hydrogen-rich water (HRW) group (10 mL/kg daily), KRG group (50 mg/kg daily) and HRGW group (10 mL/kg and 50 mg/kg daily) by an oral zoned needle for 4 weeks. Sperm count and motility were measured using sperm suspension released from cauda epididymis. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and reactive oxygen species (ROS) in serum have also been estimated. Tubular changes were examined through histological hematoxylin and eosin staining. Expression of antioxidation (PPx3, PPx4, GSTm5 and GPx4), spermatogenesis (inhibin-a, neptin-2 and CREM), antiaging (SIRT1 and SIRT2), and angiogenesis [visfatin and vascular endothelial growth factor (VEGF)] related genes were examined through real-time polymerase chain reaction.@*RESULTS@#HRW and KRG treatment stimulated spermatogenesis followed by increasing sperm production and sperm motility (P <0.05). These effects were strengthened synergistically by a HRGW mixture (P <0.05 or P <0.01). HRGW greatly increased the expressions of antioxidation, antiaging, spermatogenesis related genes and VEGF especially in aged mice (P <0.05). Serum testosterone and FSH levels also increased, while serum ROS level decreased (all P <0.05).@*CONCLUSION@#HRGW increases sperm production and motility by enhancing antioxidation and stimulating spermatogenesis and sex hormone production, particularly in aged mice.

Establishment of Effective Mouse Model of Premature Ovarian Failure Considering Treatment Duration of Anticancer Drugs and Natural Recovery Time

OBJECTIVES: This study was aimed to establish the most effective premature ovarian failure (POF) mouse model using Cyclophosphamide (CTX), busulfan (Bu), and cisplatin considering treatment duration of anticancer drugs and natural recovery time. METHODS: POF was induced by intraperitoneally injecting CTX (120 mg/kg)/Bu (12 mg/kg) for 1 to 4 weeks or cisplatin (2 mg/kg) for 3 to 14 days to C57BL/6 female mice aged 6 to 8 weeks. Controls were injected with equal volume of saline for the same periods. Body weight was measured every week, and ovarian and uterine weights were measured after the last injection of anticancer drug. To assess ovarian function, POF-induced mice were superovulated with pregnant mare serum gonadotropin and human chorionic gonadotropin, and then mated with male. After 18 hours, zygotes were retrieved and cultured for 4 days. Finally, the mice were left untreated for a period of times after the final injection of anticancer drug, and the time for natural recovery of ovarian function was evaluated. RESULTS: After 2 weeks of CTX/Bu injection, ovarian and uterine weights, and ovarian function were decreased sharply. Cisplatin treatment for 10 days resulted in a significant decrease in ovarian and uterine weight, and ovarian function. When POF was induced for at least 2 weeks for CTX/Bu and for at least 10 days for cisplatin, ovarian function did not recover naturally for 2 weeks and 1 week, respectively. CONCLUSIONS: These results suggest that CTX/Bu should be treated for at least 2 weeks and cisplatin for at least 10 days to establish the most effective primary ovarian insufficiency mouse model.

Effects of three-area laser-assisted zona thinning in 8-cell human embryos on pregnancy outcomes in vitro fertilization / 대한생식의학회지

OBJECTIVE: This study conducted a preliminary examination of the effects of three-area laser-assisted zona thinning (LAZT) during the cleavage stage of embryo development on the hatching process in human in vitro fertilization-embryo transfer (IVF-ET) with subjects of advanced female age or frozen-thawed (FT) embryos. METHODS: Eight-cell stage embryos were treated with LAZT in three areas of the zona pellucida at 120° intervals. The control group was embryos without LAZT. Of the 72 consecutive fresh cycles and the 28 FT embryo transfer cycles, the patients in 55 fresh cycles and 17 FT cycles declined LAZT, and those cycles were defined as the control group. RESULTS: In the fresh cycles, the pregnancy rates were similar in the LAZT and control groups. However, in the FT cycles, the pregnancy rate was significantly higher in the LAZT group than in the control group (45.5% in the LAZT group vs. 23.5% in the control group, p < 0.05). CONCLUSION: These results show that multi-area LAZT resulted in significantly improved pregnancy outcomes in human 8-cell embryos compared to controls.

Effect of Glucagon-like Peptide-1 on the Differentiation of Adipose-derived Stem Cells into Osteoblasts and Adipocytes

OBJECTIVES: Glucagon-like peptide-1 (GLP-1) is an intestinally secreted hormone and it plays an important role in the regulation of glucose homeostasis. However, the possible role of GLP-1 in the differentiation of adipose-derived stem cells (ADSCs) remains unknown. Therefore this study investigated the effect of GLP-1 on the differentiation of ADSCs into osteoblasts and adipocytes. METHODS: ADSCs were isolated from human adipose tissues of the abdomens, cultured and characterized by flow cytometry and multi-lineage potential assay. ADSCs were induced in osteogenic and adipogenic media treated with two different doses (10 and 100 nM) of GLP-1, and then the effect of GLP-1 on differentiation of ADSCs into osteoblast and adipocyte was examined. The signaling pathway involved in these processes was also examined. RESULTS: Isolated human ADSCs expressed mesenchymal stem cell (MSC) specific markers as well as GLP-1 receptor (GLP-1R) proteins. They also showed multiple-lineage potential of MSC. GLP-1 was upregulated the activity and mRNA expression of osteoblast-specific marker, alkaline phosphatase and the mineralization of calcium. In contrast, GLP-1 significantly suppressed the expression of adipocyte-specific markers, peroxisome proliferator-activated receptor gamma (PPAR-gamma), lipoprotein lipase (LPL) and adipocyte protein 2 (AP2). This decreased expression of adipocyte specific markers caused by GLP-1 was significantly reversed by the treatment of extracellular signal-regulated kinase (ERK) inhibitor, PD98059 (P < 0.05). CONCLUSION: This result demonstrates that GLP-1 stimulates osteoblast differentiation in ADSCs, whereas it inhibits adipocyte differentiation. The ERK signaling pathway seems to be involved in these differentiation processes mediated by GLP-1.

Effect of laser-assisted multi-point zona thinning on development and hatching of cleavage embryos in mice / 대한생식의학회지

OBJECTIVE: This study aimed to examine the effect of laser-assisted zona thinning (LAZT) at one or four-points on the blastocyst formation and hatching process in mice with respect to female age. METHODS: Eight-cell or morula embryos collected from superovulated C57BL female mice with different ages (6-11 and 28-31 weeks) were treated with LAZT at one-point (LAZT1) or four-points (LAZT4). The zona pellucida was thinned to more than 70% of its initial thickness by making two holes of 15-20 microm. RESULTS: In the young mice, LAZT resulted in a significant increase in early hatching and hatching rates compared to the control group (p<0.05). However, in the old mice, LAZT significantly increased blastocyst formation as well as early hatching and hatching compared to the controls (p<0.05). These effects were more remarkable in LAZT4 than in LAZT1 and in aged mice than in young ones. CONCLUSION: These results show that multi-point LAZT leads to a significant improvement of blastocyst formation and hatching in mice compared to controls.

Effect of Phorbol 12-Myristate 13-Acetate on the Differentiation of Adipose-Derived Stromal Cells from Different Subcutaneous Adipose Tissue Depots

Human adipose-tissue-derived stromal cells (hADSCs) are abundant in adipose tissue and can differentiate into multi-lineage cell types, including adipocytes, osteoblasts, and chondrocytes. In order to define the optimal harvest site of adipose tissue harvest site, we solated hADSCs from different subcutaneous sites (upper abdomen, lower abdomen, and thigh) and compared their proliferation and potential to differentiate into adipocytes and osteoblasts. In addition, this study examined the effect of phorbol 12-myristate 13-acetate (PMA), a protein kinase C (PKC) activator, on proliferation and differentiation of hADSCs to adipocytes or osteoblasts. hADSCs isolated from different subcutaneous depots have a similar growth rate. Fluorescence-activated cell sorting (FACS) analysis showed that the expression levels of CD73 and CD90 were similar between hADSCs from abdomen and thigh regions. However, the expression of CD105 was lower in hADSCs from the thigh than in those from the abdomen. Although the adipogenic differentiation potential of hADSCs from both tissue regions was similar, the osteogenic differentiation potential of hADSCs from the thigh was greater than that of hADSCs from the abdomen. Phorbol 12-myristate 13-acetate (PMA) treatment increased osteogenic differentiation and suppressed adipogenic differentiation of all hADSCs without affecting their growth rate and the treatment of Go6983, a general inhibitor of protein kinase C (PKC) blocked the PMA effect. These findings indicate that the thigh region might be a suitable source of hADSCs for bone regeneration and that the PKC signaling pathway may be involved in the adipogenic and osteogenic differentiation of hADSCs.

Expression of SDF-1alpha and leptin, and their effect on expression of angiogenic factors in mouse ovaries / 대한생식의학회지

OBJECTIVE: Ovarian angiogenesis plays an important role in folliculogenesis. However, little is known about the expression of angiogenic factors during follicular development according to female age. Stromal cell derived factor-1alpha (SDF-1alpha) plays a role in granulosa cell survival and embryo quality as an angiogenic chemokine. Leptin is also involved in folliculogenesis and angiogenesis. This study examined expression of SDF-1alpha and leptin, and their effects on the expression of angiogenic factors in the ovary during follicular development according to female age. METHODS: Ovaries were collected from C57BL mice of two age groups (6-9 weeks and 24-26 weeks) at 6, 12, 24, and 48 hours after 5 IU pregnant mare's serum gonadotropin (PMSG) injection. The expression of ovarian SDF-1alpha and leptin mRNA was evaluated by RT-PCR. In the organ culture experiment, the ovaries were cultured in transwell permeable supports with Waymouth's medium treated with various doses of SDF-1alpha (50-200 ng/mL) or leptin (0.01-1 microg/mL) for 7 days. Then, mRNA expression of vascular endothelial growth factor (VEGF), endothelial nitric oxide synthase (eNOS), and visfatin were examined in the cultured ovaries. RESULTS: Expression of SDF-1alpha and leptin in the ovary was significantly lower in the aged mouse group compared to the young mouse group (p<0.05). Expression of these two factors increased with follicular development after PMSG administration. SDF-1alpha treatment stimulated visfatin expression in a dose-dependent manner, while leptin treatment significantly increased eNOS expression. CONCLUSION: These results suggest that decrease of ovarian SDF-1alpha and leptin expression may be associated with aging-related reduction of ovarian function. SDF-1alpha and leptin may play a role in follicular development by regulating the expression of angiogenic factors in mouse ovaries.

Alendronate enhances osteoblastic differentiation with increased expression of Id-1 and Id-2 in pre-osteoblast cell line, MC3T3-E1 / 대한산부인과학회지

OBJECTIVE: Alendronate, a widely used bisphosphonates, acts to inhibit bone resorption by interfering with the activity of osteoclasts. Recently, it has been reported that alendronate also may increase bone proliferation and osteoblastic differentiation. However, little is known about mechanism of the action of alendronate on osteoblast differentiation, especially in transcription level. Inhibitors of DNA binding/ differentiation (Ids) are helix-loop-helix (HLH) transcription factors and play an important role in BMP-induced osteoblast lineage-specific differentiation. Therefore, this study was aimed to investigate the effect of alendronate on osteoblast differentiation and expression of Id-1 and Id-2. METHODS: MC3T3-E1, pre-osteoblast cell line, were treated with alendronate of various concentrations (10(-9) M-10(-4) M) and time periods (24, 48 and 72 hours). And then, the effect of alendronate on osteoblast differentiation was examined by alkaline phosphatase (ALP) activity and RT-PCR for osteoblast differentiation markers such as ALP, type 1 collagen (Col 1), and osteocalcin (OCN). The expressions of Id-1 and Id-2 were measured by RT-PCR. RESULTS: Alendronate treatment increased not only ALP activity, but also expressions of ALP, Col 1, and OCN. Also, alendronate treatment up-regulated the mRNA levels of Id-1 and Id-2 genes. This alendronate-induced osteoblastic differentiation is more effective in lower doses rather than high doses. CONCLUSION: This study shows that the expression of transcription factor Id-1 and Id-2 was increased in a dose-dependent manner during alendronate-induced osteoblast differentiation.

Effects of Leptin on Proliferation, Anabolic and Catabolic Metabolism in Chondrocytes / 대한정형외과연구학회지

PURPOSE: Leptin may play an important role in the pathophysiology of osteoarthritis. However, the effect of letpin on the anabolic and catabolic metabolisms in chondrocytes remains unclearly elucidated. Therefore, the purpose of this study was to investigate the effect of leptin on proliferation, anabolic and catabolic metabolism of chondrocyte using ATDC5 chondrogenic cell line. MATERIALS AND METHODS: The effects of leptin on chodnrocyte proliferation, anabolic and catabolic meatabolism were examined in ATDC5 cells treated with leptin at varying concentrations(10, 100, 300, 600 ng/ml) for 24, 48, and 72 hours. The cell proliferation was evaluated by MTT assay. The anabolic and catabolic activities were assayed by RT-PCR for transforming growth factor-beta(TNF-alpha), proteoglycan-4 (PRG4), type- I collagen (type- I Col) and tumor necrosis factor-beta(TNF-alpha), matrix metalloproteinase -2 (MMP-2), respectively. RESULTS: Leptin treatment did not influence cell proliferation of chondrocyte regardless of concentration. TGF-beta expression was increased until 48 hours of leptin treatment compared to controls. Especially, it was significantly increased in leptin of 10 ng/ml and 100 ng/ml (P<0.05). PRG4 expression was not different between letpin treatment and controls. Type-I Col expression was decreased in dose- and time-dependent manner. Leptin of 10ng/ml significantly inhibited MMP-2 and TNF-alpha expressions compared to controls (P<0.05). CONCLUSION: This study shows that leptin at low concentration increases TGF-beta expression, but inhibits the expression of TNF-alpha and MMP-2. Also this study shows that leptin do not affect the cell proliferation of chondrocytes. These results suggest that leptin at low or physiological level contributes to the prevention of cartilage damage by stimulating anabolic activity and inhibiting catabolic activity of chondrocyte rather than chondrocyte regeneration by increasing cell proliferation.

The effects of bisphosphonates on differentiation of human amniotic fluid-derived mesenchymal stem cells into osteoblast / 대한산부인과학회지

OBJECTIVE: It has been known that amniotic fluid (AF) is rich source of mesenchymal stem cells (MSCs). Bisphosphonates are widely used in clinical treatment of various metabolic bone diseases and their primary action is the inhibition of osteoclastic bone resorption. However, litter is known about whether bisphosphonates affect the differentiation into osteoblast, especially from AF-derived MSCs (AFMSCs). Therefore, the purpose of this study is to investigate whether these bisphosphonates influence in the process of AFMSCs differentiation into osteoblast. METHODS: AF samples were obtained by second trimester amniocentesis for fetal karyotyping from 6 pregnant women. Cells were treated with various concentration (0, 10(-10), 10(-8), 10(-6) M) of zoledronate and alendronate and analyzed over 21 days of culture. Differentiation into osteoblast was determined by cell staining and RT-PCR for alkaline phosphatase (ALP). RESULTS: It was observed that AFMSCs could differentiate into osteoblast. Alendronate had more potent effect than zoledronate in osteoblastic differentiation. ALP expression was increased with increasing concentration of zoledronate and it was highest in 10(-8) M alendronate. However, no effect of bisphosphonates was found in 14 days of culture. CONCLUSION: This study shows that AFMSCs can be differentiated into osteoblast. The induction of these differentiation following bisphosphonate treatment was appear to be drug type-, dose-, and culture time-dependent. However, further studies are needed to conclude a consistent outcome for the effects of bisphosphonate on differentiation potential of AFMSCs.

The Effect of Zoledronate on the Differentiation of Human Amniotic Fluid-Derived Mesencymal Stem Cells into Adipocytes / 대한수혈학회지

BACKGROUND: Amniotic fluid is a rich source of fetal mesenchymal stem cells (MSCs). However, little is known about whether bisphosphonates affect the differentiation into adipocytes. Therefore, this study was aimed to investigate whether zoledronate influences the differentiation of AFMSCs into adipocytes. METHODS: Amniotic fluid cells samples were obtained from 6 pregnant women by second trimester amniocentesis for performing fetal karyotyping. The cells were treated with various concentration (10(-10), 10(-8), 10(-6) M) of zoledronate and the cells were analyzed over 21 days of culture. Differentiation into adipocytes was determined by oil-red O staining and for fatty acid synthase (FAS), acetyl CoA carboxylase 1 (ACC1) and sterol regulatory elementary binding protein-1 (SREBP-1). RESULTS: Differentiation of AFMSCs into adipocytes was found by oil-red O staining. Zoledronate influenced the differentiation of AFMSCs into adipocytes in a dose- and time-dependent manner. At 7 days of culture, the expressions of FAS and SREBP-1 showed no significant differences compared to that of the control regardless of the dose of zoledronate. Very little ACC1 expression was found. However, the expressions of these three markers were remarkably increased at 14 days of culture. Of them, the ACC1 expression was significantly increased by 10(-8) M and 10(-6) M of zoledronate. At 21 days of culture, there were no effects of zoledronate on the expressions of FAS and SREBP-1. However, the ACC1 expression was decreased with an increasing dose of zoledronate (P<.05). CONCLUSION: This study shows that AFMSCs can be differentiated into adipocytes. The induction of this differentiation following zoledronate treatment appears to be dose dependent and time-of-culture dependent.

Relationship between Osteoarthritis and Leptin Concentrations in Synovial Fluid / 대한정형외과연구학회지

PURPOSE: Leptin may play an important role in the pathophysiology of osteoarthritis. This study investigated whether leptin concentration in synovial fluid is related to the radiographic severity of osteoarthritis. MATERIALS AND METHODS: Synovial fluids were obtained from 29 osteoarthritis patients who underwent knee surgery and 10 who had no abnormality on articular cartilage during arthoscopic examination. The progression of osteoarthritis was classified by Kellgren Lawrence grading scale. The concentrations of leptin was measured with commercial enzyme-linked immnosorbent assay kits. RESULTS: A significant increase in synovial fluid concentrations was observed in osteoarthritis patients (6.7+/-4.1 ng/ml) compared to the control (2.4+/-1.3 ng/ml). Leptin levels were increased with advancing osteoarthritis stage, resulting in the highest level in stage IV patients(10.7+/-4.9 ng/ml; range 4.7-15.8) compared to that of stage I patients (4.0+/-2.0 ng/ml; range 1.2-7.3). In osteoarthritis patients, age showed a significant correlation with leptin concentrations. CONCLUSION: This study shows that synovial fluid leptin concentrations were closely related to the radiographic severity of osteoarthritis, and suggests that the age of patient may influence synovial fluid leptin concentrations during osteoarthritis progression.

Effect of serum E2 levels on the pregnancy outcome of IVF-ET / 대한산부인과학회지

OBJECTIVE: To evaluate the effects of serum E2 levels on the day of hCG administration on the pregnancy outcome of IVF-ET after controlled ovarian hyperstimulation (COH). METHODS: Data of 455 cycles of fresh IVF-ET with COH were retrospectively investigated. Serum E2 levels on the day of hCG administration were categorized into 5 groups; A ( or =4,000 pg/mL). RESULTS: Mean E2 levels on the day of hCG administration were 3,745.3 pg/mL and mean number of retrieved oocytes were 10.1. Of 455 cycles, 148 (32.5%) cycles were clinically pregnant. Implantation rate was 12.2% and delivery rate was 18.7%. The number of obtained oocytes increased with increasing levels of serum E2. Pregnancy rate gradually increased as E2 levels increased up to the group D, but began to fall in the group E. In younger women ( or =38 yrs), pregnancy rate and delivery rate were significantly higher in the group C than other groups. CONCLUSIONS: This result shows that serum E2 levels have a concentration-dependent effect on the pregnancy outcome and there is an optimal range of E2 levels to achieve for a successful pregnancy. Excessive E2 levels seem more deleterious to the pregnancy outcome in older women aged > or =38 years.

Short Coasting of One or Two Days by Withholding Both Gonadotropins and GnRH Agonist Prevents Ovarian Hyperstimulation Syndrome without Compromising the Outcome / 대한불임학회지

OBJECTIVE: To evaluate the effect of short coasting, by withdrawing both gonadotropins and GnRH agonist (GnRHa), on the prevention in severe ovarian hyperstimulation syndrome (OHSS) without compromising pregnancy outcome. METHOD: Thirty-seven women who had been coasted during COH for IVF were coasted when > or =20 follicles > 15 mm with serum E2 level of 4,000 pg/ml were detected. Coasting was initiated for one or two days depending on the status of follicle on ultrasound and serum E2 level. Both gonadotropin and GnRHa were withheld for coasting. Retrospective study was carried and changes of serum E2 levels, number of oocytes retrieved, fertilization rate, pregnancy rate were compared and analyzed. RESULTS: The mean serum E2 level fell from 6,993 pg/ml on the onset of coasting to 3,396 pg/ml on the day of hCG administration. The mean number of oocytes retrieved and fertilization rate were 15.7 and 70.0%, respectively. Fifteen patients were pregnant (40.6%) and implantation rate was 15.2%. Twenty-six (70.3%) patients were coasted for one day and 11 (29.7%) were coasted for two days. The mean decrease rate of serum E2 level was 43% in one day coasting group and 15% (1st day) and 81% (2nd day) in two day coasting group. The pregnancy outcome was similar between the two groups. After coasting, no severe or moderate OHSS occurred in any patients and mild OHSS occurred in 3 (8.1%) patients. CONCLUSIONS: Coasting for one or two days can be used successfully in the prevention of OHSS without compromising IVF cycle outcome.

The Effect of Leptin on the Proliferation of the Endothelial Progenitor Cells from Peripheral Blood / 대한수혈학회지

BACKGROUND: Leptin has been found to be involved in the regulation of hematopoiesis processes and angiogenesis. Therefore, we investigated the effect of leptin in the proliferation and angiogenesis of peripheral blood (PB)-derived endothelial progenitor cells (EPCs). METHODS: Mononuclear cells were isolated from PB of healthy male volunteers and were cultured in endothelial cell growth medium-2 (EGM-2). After 6 days of culture, cells were treated with 50 ng/mL vascular endothelial growth factor (VEGF) and/or with various concentrations of leptin (10 ng/mL, 100 ng/mL, 1microgram/mL, and 10 microgram/mL) and were further cultured for one week. Proliferation of EPCs was examined by an assay measuring the uptake of 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbo cyanine-labeled acetylated LDL (Dil-ac-LDL) and tubule formation on a matrigel. The control group of cells was not treated with VEGF and/or leptin. RESULTS: The number of Dil-ac-LDL labeled-EPCs, tubule formation on matrigel and the number of cells present along tubules were significantly increased in the leptin-treated groups of cells as compared to the control group or VEGF treated group of cells (P<.05). The effect was synergistically increased in the group of cells co-treated with leptin and VEGF. The number of EPCs was increased in a leptin dose-dependent manner that was maximal at a concentration of 1microgram/mL leptin. CONCLUSION: This study shows that leptin increased in vitro proliferation and angiogenesis of EPCs derived from peripheral blood.

Two cases of spontaneous pregnancy in women with premature ovarian failure: Case report / 대한산부인과학회지

Premature ovarian failure (POF) is a syndrome defined as hypergonadotropic hypogonadism associated with amenorrhea, oligomenorrhea or other forms of menstrual irregularity for at least 3 consecutive months before the age of 40. The management of POF is approached by HRT, emotional support and infertility treatment. Women with premature ovarian failure who desire to become pregnant are best treated by assisted reproductive technology with donor oocyte. However, POF has the possibility of a 5-10% spontaneous pregnancy. The physician should recommend the patient to consult with their physician if they have any symptoms of pregnancy or no withdrawal bleeding after HRT. Therefore we report two cases of spontaneous pregnancies in women with premature ovarian failure.

Improvement of Pregnancy Rate by the Selection of Early Cleavage Embryos to 2-cell Stage in Human IVF / 대한불임학회지

OBJECTIVE: Evaluation of embryos using early cleavage to 2-cell stage has been proposed, but a critical time-point for selecting embryos is unclear. The aim of the present study is to provide a guideline including critical time-point in the selection of early cleaving embryo for the reduction of multiple pregnancies as well as the increase of pregnancy rate in human IVF. METHODS: This prospective study was performed in 116 cycles from 85 patients who underwent conventional IVF or ICSI at the infertility clinic of Good Moonhwa Hospital from January 2002 to December 2003. Early cleavage (EC) of embryos to 2-cell stage was assessed at 25 h and 27 h postinsemination/microinjection. Embryos that had early cleaved at each time point were designated as EC-1 and EC-2, respectively, while others were designated as non-early cleavage (NEC). RESULTS: At least one early cleavage embryo was observed in 54 (46.6%) for the EC-1 and 84 (72.4%) for the EC-2 of the 116 cycles assessed. Clinical pregnancy rates (PR) were significantly higher in the EC-1 group (66.7%) compared to the EC-2 group (53.6%) or the NEC group (31.2%) (p<0.05). Significant improvement of the pregnancy rate was found when at least two or more embryos were early cleaved at 25 h postinsemination or when the proportion of early cleavage embryo at 25 h postinsemination was higher than 20% (p<0.05). CONCLUSIONS: The critical time-point for the selection of early cleavage embryos with high implantation potential is more effective in 25 h postinsemination/microinjection compared to 27 h. The proportion as well as number of early cleavage embryos is also an important factor for the prediction of pregnancy outcome and the chance of multiple pregnancies. These results demonstrated that the evaluation of early cleavage embryos to 2-cell stage is an easy, simple, and objective method for the selection of good quality embryos suitable for embryo transfer.

A Case of Limb Reduction Defect after Chorionic Villus Sampling at 9 Gestational Weeks / 대한산부인과학회잡지

Since it has been proposed that chorionic villus sampling could be an etiology of limb reduction defect, the relation between limb defect and chorionic villus sampling remains controversial. We experienced a case of limb reduction defect after chorionic villus sampling at 9 gestational weeks and report the case with brief review of literature.

Effects of Metformin on Clomiphene-resistant Infertile Women with Polycystic Ovary Syndrome / 대한산부인과학회잡지

OBJECTIVE: The aim of this study was to evaluate the efficacy of metformin for ovualtion induction and pregnancy in clomiphene citrate (CC)-resistant women with polycystic ovary syndrome (PCOS). METHODS: From March 2001 to March 2002, 19 patients with PCOS who had at least two consecutive cycles of anovulation in response to CC treatment up to 250 mg/day at the Infertility Center of Moon Hwa Hospital were enrolled in this study. The participants were required to have tubal patency on hysterosalpingography and their husbands were required to have normal semen analysis. The mean age was 30.5 +/- 3.6 years, the body weight 62.7 +/- 10.1 kg, the duration of infertility 3.7 +/- 2.1 years and the BMI 24.7 +/- 3.6 kg/m2. For 19 patients, a total of 75 cycles were treated with 1) CC+gonadotropin (group 1; 24 cycles), 2) CC+metformin (group 2; 29 cycles), or 3) CC+gonadotropin+metformin (group 3; 22 cycles). As for gonadotropin, highly purified-follicle stimulating hormone (HP-FSH) or/and hMG were used from the 3rd day of CC treatment. In the first cycle, metformin (1,500 mg/day) was administered during 1-28 days of menstrual cycle. Metformin was discontinued when a pregnancy was confirmed. RESULTS: Among 19 patients, 17 patients were ovulated (89.5%) and 7 patients (36.8%) were pregnant. Of a total of 75 cycles, 51 cycles (68.0%) were ovulated successfully with one of three treatment methods. Metformin treatment had similar ovulation rate compared to gonadotropin treatment. There was no significant difference in ovulation rate among the three groups (70.8% vs 58.6% vs 63.7%). However, the pregnancy rate was significantly higher in group 3 (18.2%, 4 cycles) compared to group 1 (8.3%, 2 cycles) and group 2 (6.9%, 2 cycles). Of pregnant cycles, all 2 cycles from group 1 were spontaneously aborted. One cycle in group 2 and one cycle in group 3 were spontaneously aborted and all other pregnant cycles were normally delivered. CONCLUSION: With the combination therapy of metformin, the improvement in pregnancy rate among CC-resistant PCOS infertile women might be expected.

Learning Curve of Laparoscopic Myomectomy / 대한산부인과학회잡지

OBJECTIVE: To evaluate the effectiveness and feasibility of laparoscopic myomectomy compared to open myomectomy METHODS: A retrospective study of 85 cases of myomectomy was performed. Twenty six cases of open myomectomy (group I) and 59 cases of laparoscopic myomectomy (group II) were done by one main surgeon from 1996 to 2002 in the department of OBGYN at Moonhwa Hospital. Group II was divided into two subgroups, group IIA and group IIB. Group IIA included 17 cases of laparoscopic myomectomy done from 1996 to 1998 during learning period. Group IIB included 42 cases of laparoscopic myomectomy performed from 1999 to 2002 after learning period. RESULTS: There were no significant differences in age, parity, the number of myoma, and the size of myoma between groups I and II. The intensity of postoperative pain and febrile morbidity were significantly lower in group II than in group I (P<0.05). Mean operation time was significantly shorter in group I than in group II. However, after completing the learning curve, no significant difference was found in the operation time between group I and group IIB. Blood loss was significantly decreased in group II compared to group I (P<0.05). CONCLUSION: The learning curve for lasparoscopic myomectomy needed 17 cases and laparoscopic myomectomy could be an excellent minimally invasive method as an alternative of open myomectomy after learning curve.