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.

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.

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.

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.

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.

Successful correction of posthysterectomy vaginal vault prolapse by repair of fascial defects and by using uterosacral ligament suspension / 대한산부인과학회잡지

OBJECTIVE: To report the method and efficacy of the site specific-defects repair in the endopelvic fascia and uterosacral ligament suspension for the correction of posthysterectomy vaginal vault prolapse. METHODS: This study was performed in 3 patients with posthyterectomy vaginal vault prolapse who underwent the surgery for the reconstruction of endopelvic fascia and the correction of vault prolapsed vagina by using uterosacral ligament suspension at the department of Obstetrics and Gynecology in Moon-Hwa Hospital. Among them, 1 patient was treated by laparotomy, and 2 patients were treated by laparoscope. RESULTS: The age of patients ranged from 49 to 67 years. The mean operation time was 111 minutes (105-120 minutes). The status of postoperative vagina was normal in all 3 patients, who were discharged 9 days after operation without symptoms. No complication or recurrence were found in them despite long-term follow-up for 24 months, 12 months, 7 months, respectively after operation. One patient has been treated with HRT therapy. CONCLUSION: The reconstruction of the defect of endopelvic fascia and uterosacral ligament fixation may be very safe, effective, and functional in the correction of vaginal vault prolapse. This study reports the treatment of vaginal vault prolapse using uterosacral ligament within Korea for the first time.

Detection of HPV in tissue of cervical lesion: Comparative study between in situ hybridization and PCR in situ hybridization / 대한산부인과학회잡지

OBJECTIVE: The aim of this study was to identify human papilloma virus(HPV) infection in cervical lesions by using polymerase chain reaction in situ hybridization(PCR-ISH) and to compare the results of PCR-ISH with conventional in situ hybridization(ISH) METHODS: Forty-seven randomly chosen neutral-buffered formalhyde-fixed cervical biopsies in which cervical intraepithelial neoplasia and invasive cervical cancer had been diagnosed were tested for HPV DNA by PCR-ISH and ISH. The method of PCR-ISH includes deparaffinization of specimens, PCR amplification of DNA, in situ hybridization and detection of amplified products. RESULTS: The positivity rate of ISH was 36% for all biopsies and 26%, 40% for CIN and invasive cancer respectively. By using PCR-ISH the positivity was 80%, 84% respectively. All PCR-ISH positive cases were ISH positive and no ISH positive was PCR-ISH negative. CONCLUSION: In our study HPV DNA, undetectable by standard in situ hybridization, was detectable by PCR in situ hybridization. Our study shows that PCR in situ hybridization is highly sensitive method for detection of HPV in cervical neoplastic specimens with morphological evidence.

c-erbB-2 Oncoprotein Assay in Ovarian Carcinoma and Its Clinical Correlation with Prognostic Factors / 대한부인종양콜포스코피학회잡지

c-erbB-2 oncogene is a gene that encodes a growth factor receptor-like molecule with tyrosine kinase activity and has a structure similar to that of the epidermal growth factor receptor. Overexpression of the c-erbB-2 oncoprotein is detected in human adenocarcinoma of the breast, cervix, and salivary gland, in all of which the association between overexpression of the c-erbB-2 and poor prognosis of the disease has been reported. The role of c-erbB-2 oncoprotein in the tumorigenesis of the human ovary has been poorly understood and remains controversial. In order to explore the relationship between c-erbB-2 oncoprotein status and ovarian carcinoma, tissue from 32 patients, each of whom had invasive ovarian carcinoma prior to treatment, 10 patients with benign cyst and 10 control case who underwent hysterectomy due to gynecological disease at Yonsei University Colloge of Medicine were analyzed. We measured c-erbB-2 oncoprotein with an enzyme-linked immunosorbent assay(ELISA) which was a sandwich type. Patients with invasive ovarian cancer were found to have significantly higher median c-erbB-2 oncoprotein expression than patients with either benign ovarian cyst(P<0.05) or control group(P<0.05), respectively. However there was no significant difference in c-erbB-2 oncoprotein status between benign cyst and control group. verexpression of c-erbB-2 oncoprotein was found in 7 of 32(21.9%) epithelial ovarian cancer. In invasive epithelial cancer, no significant difference in c-erbB-2 oncoprotein levels was noted when stratified according to age, menopausal status, histological cell type, tumor size or surgical stage. Our results were consistent with the concept that c-erbB-2 oncoprotein may play an important role in malignant and tumorigenesis in epithelial ovarian cancer.