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To evaluate the safety and effectiveness of recombinant human follicle-stimulating hormone (rhFSH [Follitrope™]) in infertile women undergoing in vitro fertilization (IVF). To identify predictors of ovarian response that induce optimal clinical outcomes. This multicenter prospective study enrolled infertile women who were scheduled to undergo IVF after ovarian stimulation with rhFSH (Follitrope™) following the gonadotropin-releasing hormone (GnRH) agonist or GnRH antagonist protocol. Predictive factors for ovarian response were identified in the GnRH antagonist group based on the number of oocytes retrieved. A total of 516 infertile women were enrolled, among whom 136 (except one who withdrew before administration) received rhFSH using the GnRH agonist protocol and 379 using the antagonist protocol. The mean number of oocytes retrieved was 13.4 in the GnRH agonist group and 13.6 in the GnRH antagonist group. The clinical pregnancy rates were 32.3% (30/93) and 39.9% (115/288) in the GnRH agonist and antagonist groups, respectively. The incidence of ovarian hyperstimulation syndrome was 1.8% and 3.4% in the GnRH agonist and antagonist groups, respectively. No other significant safety risks associated with rhFSH administration were identified. Body mass index, basal serum FSH and anti-Müllerian hormone levels, and antral follicle count were identified as predictors of ovarian response by multiple regression with backward elimination, and the final regression model accounted for 26.5% of the response variability. In real-world practice, rhFSH (Follitrope™) is safe and effective in inducing ovarian stimulation in infertile women. Patient characteristics identified as predictors can be considered to be highly related to optimal clinical outcomes.

Objective To investigate the correlation between bladder wall thickness (BWT) measured by ultrasonography and lower urinary tract dysfunction (LUTD) in patients with lower urinary tract symptoms (LUTS). Methods Forty-eight women with LUTS who underwent urodynamic study and BWT by ultrasonography as outpatients were studied. We assessed LUTS during a medical examination by interview. The thinnest part of the bladder wall was measured by a transabdominal ultrasonography. We excluded patients who had visited another hospital previously because we did not know what treatment they had received, including medications, behavioral therapy, or other treatments. We constructed receiver operating characteristic (ROC) curves for diagnosis of LUTD and also determined reliable BWT criteria by calculating the area under the curve. Statistical analyses were performed using the Kolmogorov-Smirnov method and Student's t-test. Results The mean age, body mass index, and duration of symptoms were 59.9±9.7 years, 26.06±3.4 kg/m2, and 53.4±38.2 months, respectively. Urodynamic study parameters (Valsalva leak point pressure, maximal urethral closure pressure, functional length, and postvoid residual volume) were lower in patients with BWT

PURPOSE Progesterone resistance is thought to be a major factor that contributes to progression of endometriosis. However, it is not clear what causes progesterone resistance in endometriosis. This study aimed to assess whether cytokines or peritoneal fluid can affect progesterone receptor (PR) expression in endometrial cells and to verify whether PR expression is reduced in endometriosis. MATERIALS AND METHODS The PR-B/A ratio was measured via real-time polymerase chain reaction after in vitro culture, in which endometrial cells were treated with either tumor necrosis factor-alpha (TNF-α), interleukin-1 beta, or peritoneal fluid obtained from women with advanced-stage endometriosis. Immunohistochemistry was performed to compare PR-B expression between eutopic and ectopic endometrial tissues from women with and without advanced-stage endometriosis. RESULTS The PR-B/A ratio was significantly decreased by treatment with either TNF-α (p=0.011) or peritoneal fluid from women with advanced-stage endometriosis (p=0.027). Immunoreactivity of PR-B expression was significantly lower during the secretory phase than during the proliferative phase in endometrial tissues from control subjects (p

Secretory leukocyte peptidase inhibitor (SLPI) plays a role in proliferation and differentiation via the autocrine and paracrine systems. SLPI's expression is well-documented in the reproductive tract, but it remains unclear whether it is active during early embryonic development. In this study, the expression and role of Slpi in the early embryo were evaluated. In vitro embryo cultures in chemically defined simple medium resulted in a reduction in developmental speed from the 8-cell stage, as well as implantation rate compared with in vivo embryos. SLPI protein was localized to the membrane or submembrane cytoplasm in an embryonic stage-dependent manner. In vitro cultured embryos exhibited lower levels of Slpi mRNA expression than in vivo embryos. Slpi knockdown by antisense oligonucleotides attenuated the developmental speed and implantation rate compared with Slpi sense oligonucleotide-transfected embryos and in vitro controls. The critical period for the attenuation of developmental speed occurred after the 8-cell stage. SLPI treatment accelerated development, increased implantation rate, and ameliorated the suppressive effects of Slpi knockdown. Slpi knockdown did not induce changes in the total cell number or inner cell number in blastocysts. Meanwhile, SLPI upregulated the expression of the developmental factors matrix metalloproteinase-14, neutrophil elastase, and tissue inhibitor of metalloproteinase-1. Together, these results suggest that SLPI is an effective regulator of developmental speed and implantation competence in an autocrine and paracrine manner, respectively, and plays a role in controlling the expression of embryonic development factors, such as MMP family members.

This report describes the use of a simple transvaginal surgical method to connect the uterus with the lower vagina in patients with cervicovaginal atresia. We report two girls presenting with primary amenorrhea and cyclic abdominal pain. The girls had similar magnetic resonance imaging findings that revealed markedly enlarged uteri containing blood and no structures resembling a cervix or upper vagina. We performed transvaginal uterovaginal anastomosis with no perioperative or postoperative complications. After surgery, the patients had regular menstrual cycles and one started sexual activities with no complaints. The remarkable finding was the natural increase in the vaginal depth after surgery. This simplified transvaginal uterovaginal anastomosis technique, with its promising anatomical results, might be a treatment for cervicovaginal atresia. © 2016 Japan Society of Obstetrics and Gynecology.

The transobturator tape (TOT) method is the recent minimally invasive midurethral sling surgery. The TOT method was invented to reduce complication rate of surgical technique for female stress urinary incontinence. Pelvic bleeding following TOT procedure, although extremely rare, could be occurred. We presented three cases which treat pelvic arterial bleeding after midurethral sling (TOT and tension-free vaginal tape Secur) surgery via pelvic artery embolization. Therefore we report our cases with brief review of the literature.

Study question Does interleukin-32 (IL-32) play a role in the pathogenesis of endometriosis? Summary answer IL-32 might be involved in the pathogenesis of endometriosis through increased viability, proliferation and invasion of endometrial cells. What is known already Endometriosis is characterized as a chronic inflammatory disease and several proinflammatory cytokines are suggested to be involved in its pathogenesis and pathophysiology. IL-32, recognized as a new proinflammatory cytokine and a strong inducer of other proinflammatory cytokines, has been shown to serve as a key modulator in several chronic inflammatory diseases. Study design, size, duration This study included comparison of IL-32 levels in the peritoneal fluids between women with and without endometriosis, in-vitro experiments using Ishikawa cells and endometrial stromal cells (ESCs), and experiments on IL-32 transgenic mice and wild-type mice with induced endometriosis. Participants/materials, setting, methods IL-32 levels in the peritoneal fluids were measured using enzyme-linked immunosorbent assays. Cell viability, expression of proliferating cell nuclear antigen (PCNA), and cellular invasiveness were analyzed following in-vitro treatment of Ishikawa cells and ESCs with recombinant IL-32 alpha (α) and gamma (γ). Ectopic endometriotic lesions were compared between IL-32 transgenic mice and wild-type mice after autologous endometrial transplantation with immunohistochemistry for Ki-67 antigen and PCNA. Main results and the role of chance The peritoneal fluid concentration of IL-32 was significantly higher in patients with advanced stage endometriosis compared with the controls. In-vitro treatment with IL-32 α and γ caused significant increases in cellular viability, PCNA expression, and invasiveness in Ishikawa cells and ESCs. The IL-32 transgenic mice had a significantly larger size of the ectopic endometrial lesions with higher expression of Ki-67 antigen and PCNA compared with wild-type mice. Large scale data N/A. Limitations, reasons for caution It is still unclear whether IL-32 is a main regulator, or one of several downstream proinflammatory cytokines, causing establishment and/or progression of endometriosis. Wider implications of the findings Further investigation on IL-32 signaling pathways may contribute to development a more effective treatment of endometriosis. Study funding/competing interest(s) This research was supported by a grant of the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (Grant number: HI16C1682). None of the authors has anything to disclose.

This study was performed to investigate the effect of of transdermal testosterone gel (TTG) on controlled ovarian stimulation (COS) and IVF outcomes and ovarian morphology according to pretreatment duration in poor responders. A total of 120 women were recruited for this pilot study. They were randomized into control, 2 weeks, 3 weeks or 4 weeks TTG treatment groups. For three TTG treatment groups, 12.5 mg TTG was applied daily for 2 weeks, 3 weeks or 4 weeks in preceding period of study stimulation cycle. After 3 weeks of TTG pretreatment, significant increase of antral follicle count (AFC) and significant decreases of mean follicular diameter (MFD) and resistance index (RI) value of ovarian stromal artery were observed (p=0.026, p

Type 1 citrullinemia (CTLN1) is an autosomal recessive inherited metabolic disorder caused by anargininosuccinicnate synthetase deficiency. The patient was a 38-year-old Korean woman who is a carrier for CTLN1 and her first baby was diagnosed with CTLN1. Preimplantation genetic diagnosis (PGD) for CTLN1 in day 3 embryos using polymerase chain reaction was performed for live birth of healthy baby who is no affected with CTLN1. One unaffected blastocyst was transferred. This resulted in a clinical pregnancy and the live birth of healthy male twin. They were confirmed to be unaffected with CTNL1 by post natal diagnosis. This is the first case report of the use of PGD for CTNL1.

Objective To identify the most important factor in predicting ovarian reserve after laparoscopic ovarian cystectomy and to evaluate whether there is any difference in the postoperative decline of ovarian reserve between women with endometrioma and those with other benign ovarian cysts. Design Prospective cohort study. Setting University hospital. Patient(s) A total of 100 women who had undergone laparoscopic ovarian cystectomy for endometrioma (n = 68) or other benign ovarian cysts (n = 32). Intervention(s) Serum antimullerian hormone (AMH) levels measured by enzyme immunoassay preoperatively and at 3 months after surgery. Main Outcome Measure(s) Rate of AMH decline after surgery and follicle numbers retained in cystectomy specimens. Result(s) Serum AMH levels were obviously decreased at 3 months after the surgery (4.97 ± 2.83 vs. 3.33 ± 2.08 ng/mL, mean ± standard deviation). Adjusting for several parameters, we could see that bilaterality of the ovarian cyst was the only significant factor in predicting the rate of postoperative decline of AMH levels. The rate of AMH decline did not differ between the endometrioma group and the other benign ovarian cyst group. Conclusion(s) Bilaterality of the ovarian cyst is the only significant factor in predicting the rate of decline of AMH level after laparoscopic ovarian cystectomy. The rate of decline of AMH levels after surgery was similar between the endometrioma group and the other benign ovarian cyst group.

This study was performed to investigate the effect of peroxisome proliferators activated receptor-γ (PPAR-γ) ligand, pioglitazone, on production of regulated upon activation normal T-cell expressed and secreted (RANTES) and in vitro fertilization (IVF) outcome in infertile patients with endometriosis. Sixty-four infertile patients with stage III or IV endometriosis undergoing IVF were randomly allocated to the study or the control group. The long protocol of GnRH agonist (GnRH-a) was used for controlled ovarian stimulation (COS) in all patients. Patients in the study group were treated with pioglitazone at a dose of 15 mg/day orally from the starting day of GnRH-a treatment to the day of hCG injection. Blood samples were drawn for serologic assay of RANTES on the first day of GnRH-a treatment and the day of hCG injection. There were no differences between the study and control groups in patient characteristics. There were also no differences between the two groups in COS duration, and the numbers of retrieved oocytes, fertilized oocytes and embryos transferred. The clinical pregnancy rate per cycle was higher in the study group, but this difference was not statistically significant. However, embryo implantation rate was significantly higher in the study group of 12.5% compared with 8.6% in the control group (P

This study was performed to investigate the expression of cathepsin B mRNA and protein in eutopic and ectopic endometrial tissues of patients with endometriosis and in normal endometrial tissues and to clarify the association between the cathepsin B expression and endometriosis. A total of 40 women with histologically confirmed endometriosis were recruited for study group. For controls, 20 women undergoing operative treatment for uterine myoma, cervical intraepithelial neoplasia (CIN) or benign gynecologic conditions other than endometriosis were recruited. Eutopic endometrial tissues of both groups and ectopic endometrial tissue of study group were collected during the operations. We employed real time reverse transcriptase - polymerase chain reaction (RT-PCR) to quantify mRNA levels of cathepsin B in these tissues. Then, we performed western blot analysis to measure the protein levels of cathepsin B. The expressions of cathepsin B mRNA and protein were significantly higher in both eutopic and ectopic endometrial tissues of women with endometriosis than in endometrial tissues of controls. These data suggest that the higher expression of cathepsin B in the endometrial tissues might be associated with the development of endometriosis. In addition, eutopic endometrium itself with higher expression cathepsin B may play a pivotal role in the histogenesis of endometriosis.

Objective To evaluate the efficacy and safety of dienogest treatment in patients who had received dienogest for 12 months or more to treat endometriosis. Methods We analyzed the clinical data of 188 women with endometriosis who had been treated with 2 mg of dienogest once a day for 12 months or more at a single institute. We evaluated changes in endometriosis-associated pain and endometrioma size, recurrence rate, and adverse events following dienogest administration. Bone mineral density (BMD) was measured in patients who were prescribed dienogest for more than 18 months. Results Pain was significantly reduced at 12 months after dienogest medication. In those treated with dienogest due to recurrent endometrioma, the size of the endometrioma was significantly decreased at the 12-month and 18-month follow-ups. We found only one case of sonographic recurrence during dienogest administration among those who were treated postoperatively to prevent recurrence (1 of 114, 0.9%). The most common adverse drug reaction was uterine bleeding (3.2%), and other adverse events were generally tolerable and associated with low discontinuation rates (5.2%). Among the 50 patients in whom BMD was measured, 10 patients (20%) had a Z-score below the expected range for age. Conclusion The administration of dienogest for a year or more seems to be highly effective in preventing recurrence after surgery, reducing endometriosis-associated pain, and decreasing the size of recurrent endometrioma, with a favorable safety and tolerability profile. However, BMD should be checked in patients on long-term medication due to possible bone loss in some women.

The aim of the present study was to investigate the influence of vascular endothelial growth factor (VEGF) on the expression of insulin-like growth factor (IGF)-II, insulin-like growth factor binding protein (IGFBP)-2 and in cultured human luteinized granulosa cells (LGCs). Human LGCs were obtained from the follicular fluid by transvaginal oocyte aspiration from 30 infertile patients undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). The cells were cultured for 72 h with VEGF at concentrations of 0.1, 1.0, and 10.0 ng/ml. The cells not treated with VEGF served as controls. Reverse transcription-polymerase chain reaction (RT-PCR) was used to examine the expression of IGF-II, IGFBP-2, and 5 mRNA. The expression of IGF-II mRNA in the 10.0 ng/ml of VEGF group was significantly higher than that in the control group. Treatment with 10.0 ng/ml of VEGF significantly increased the expression of IGFBP-5 mRNA than all other groups. There were no statistically significant differences in the expression of IGFBP-2 mRNA among all the groups. VEGF may play a regulator role in human ovarian physiology by modulating the expression of IGF-II and IGFBP-5 in LGCs.

Aim: This study was performed to assess the expression levels of vascular endothelial growth factor receptor (VEGFR)-1, -2 and -3 mRNA in eutopic endometrial tissue and their effects on in vitro fertilization–embryo transfer (IVF-ET) outcomes in women with endometriosis. Materials and Methods: Twenty women with endometriosis III or IV (study group) and 20 with tubal infertility only (control group) were evaluated. Controlled ovarian stimulation in all patients consisted of the long protocol using gonadotrophin-releasing hormone agonist. Prior to the administration of 0.1 mg triptorelin daily in the mid-luteal phase of the preceding menstrual cycle, a biopsy specimen was taken from the endometrial cavity, and the expression levels of VEGFR-1, -2 and -3 mRNA were analyzed with real-time reverse transcription-polymerase chain reaction. Results: There were no significant differences in the numbers of retrieved oocytes and mature oocytes, fertilization rate and clinical pregnancy rate between the study and control groups. However, the implantation rate was significantly lower in the study group (P

Objective To investigate whether levothyroxine (LT4) treatment has beneficial effects on IVF results and pregnancy outcome in infertile patients with subclinical hypothyroidism undergoing IVF/intracytoplasmic sperm injection (ICSI). Design Prospective, randomized trial. Setting University-affiliated infertility clinic. Patient(s) A total of 64 infertile patients with subclinical hypothyroidism, defined as an elevated serum TSH level associated with a normal free T4 level and without frank symptoms of hypothyroidism. Intervention(s) Patients were randomized into an LT4 treatment group or control group. For the LT4 treatment group, 50 μg LT4 was administered from the first day of controlled ovarian stimulation for IVF/ICSI. Main Outcome Measure(s) Results of IVF and pregnancy outcome. Result(s) There were no differences in patient characteristics between the two groups. Total dose and days of recombinant human FSH used for controlled ovarian stimulation were also similar. The number of grade I or II embryos was significantly higher in the LT4 treatment group than in the control group. There was no significant difference in the clinical pregnancy rate per cycle between the two groups. However, the miscarriage rate was significantly lower in the LT4 treatment group than in the control group. Embryo implantation rate and live birth rate were significantly higher in the LT4 treatment group. In the control group, both thyroid peroxidase antibody and thyroglobulin antibody levels were significantly higher in the miscarried subgroup than in the delivered subgroup. Conclusion(s) LT4 treatment can improve embryo quality and pregnancy outcome in subclinical hypothyroid women undergoing IVF/ICSI.

Objective: To investigate the effectiveness of treatment with transdermal testosterone gel (TTG) before controlled ovarian stimulation (COS) using GnRH antagonist multiple-dose protocol (MDP) in low responders undergoing IVF/intracytoplasmic sperm injection (ICSI). Design: Prospective randomized controlled trial. Setting: University-affiliated infertility clinic. Patient(s): A total of 110 low responders, who were defined as patients who failed to produce %3 follicles with a mean diameter of R16 mm with the result that %3 oocytes were retrieved despite the use of a high gonadotropin dose (>2,500 IU) in a previous failed IVF/ICSI cycle. Intervention(s): Patients were randomized into TTG pretreatment group or control group. For TTG pretreatment group, 12.5 mg TTG were applied daily for 21 days in the cycle preceding COS for IVF. Main Outcome Measure(s): COS results and IVF outcome. Result(s): There were no differences in patients’ characteristics between the two groups. Total dose and days of rhFSH used were significantly fewer in the TTG pretreatment group than in the control group. The numbers of oocytes retrieved, mature oocytes, fertilized oocytes, and good-quality embryos were significantly higher in the TTG pretreatment group. Embryo implantation rate and clinical pregnancy rate per cycle initiated also were significantly higher in the women pretreated with TTG. No patient reported adverse effects attributed to TTG use. Conclusion(s): TTG pretreatment might be beneficial in improving both response to COS and IVF outcome in low responders undergoing IVF/ICSI. (Fertil Steril� 2011;95:679‐83. � 2011 by American Society for Reproductive Medicine.)

The aim of this study was to estimate whether coronary artery calcification is associated with serum estradiol (E2) level in postmenopausal women.We performed a multidetector CT scan of the heart and measured the coronary artery calcium score (CACS) in 436 postmenopausal women who did not take postmenopausal hormone therapy. Women were divided into two groups according to the CACS (≥100 or100). Serum E2 level, lipid profile, bone mineral densities of the lumbar vertebrae and femoral neck, current statin treatment status, and other coronary risk factors were analyzed in these two groups.The proportion of women with a higher serum E2 level (or =20 pg/mL) was significantly higher in the lower-CACS (100) group compared with the higher-CACS ((or =100) group (34.0% vs 12.5%; P0.05). The distribution of CACS was significantly different between women with higher and lower (20 pg/mL) serum E2 levels (P0.05), and the CACS was significantly lower in the group with a higher serum E2 level (P = 0.002). After adjusting various risk factors by weighted logistic regression model using inverse probability of treatment weighting, women with a higher serum E2 level had a reduced chance of having a higher CACS (crude odds ratio, 0.28; 95% CI, 0.08-0.95; P = 0.04; adjusted odds ratio, 0.25; 95% CI, 0.07-0.86; P = 0.03).Postmenopausal women with a higher serum E2 level had a reduced CACS independent of age and other coronary risk factors. These retrospective analyses might suggest that a higher level of E2 possibly lowers the calcified-plaque burden of coronary arteries in postmenopausal women.

PROBLEM The intracellular antioxidant system, based on glutathione (GSH), plays a key role in endometrial detoxification reactions and has been proposed to be involved in the pathogenesis endometriosis. This study was designed to evaluate whether estradiol (E(2)) and proinflammatory cytokines have any effects on expression of glutathione in endometrial stromal cells (ESCs). METHOD OF STUDY Glutathione levels were measured utilizing high-performance liquid chromatography following in vitro culture and treatment of ESCs with estradiol, tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). RESULTS The GSH level in E(2) (10(-8) m) treatment group was significantly higher than in the control group at 48 h (P < 0.05). In vitro treatment of ESCs with TNF-alpha 10 ng/mL as well as E(2) (10(-8) m) plus TNF-alpha 10 ng/mL for 48 hr also led to a significant increase in GSH level (P < 0.05; P < 0.05, respectively). Both IL-1beta 10 ng/mL and E(2) (10(-8) m) plus IL-1beta 10 ng/mL for 48 hr increased GSH level significantly (P < 0.05; P < 0.05, respectively) as well. CONCLUSIONS These findings might suggest that increased production of estradiol and proinflammatory cytokines in the peritoneal cavity possibly leads to the establishment of endometriosis through increased level of GSH.

BACKGROUND: P21-activated kinase I (PakI) integrates various signaling pathways that are vital to cell survival and function. This study was performed to evaluate whether sex steroids may regulate the expression of Pak I in endometrial cells as well as whether its expression is increased in the eutopic endometrium of women with endometriosis. METHODS: Following in vitro estradiol (E 2 ) and/or medroxyprogesterone acetate (MPA) treatment of Ishikawa cells and endometrial stromal cells (ESCs), PakI protein was analyzed utilizing western blot analysis and immunocytochemistry. Immunohistochemistry was performed to evaluate PakI immunoreactivity semiquantitatively in women with endometriosis and in controls. To assess the role of PakI on endometrial cell viability, crystal violet assay was performed following transfection of Ishikawa cells with PakI small interfering RNA (siRNA). RESULTS: In vitro treatment with E 2 plus MPA or MPA alone led to a significant decrease of PakI protein in Ishikawa cells and ESCs (both P < 0.05 versus control). Immunohistochemistry also revealed that PakI protein is significantly decreased during the secretory phase in both epithelial and stromal cells in the control subjects (P < 0.001 and P < 0.01, respectively). The immunoreactivity of PakI in glandular cells was significantly increased in the eutopic endometrium of women with endometriosis compared with the controls during the secretory phase (P < 0.01). Crystal violet assay has shown that transfection of Ishikawa cells with PakI siRNA led to a significant decrease of cellular viability (P < 0.05). CONCLUSIONS: These findings suggest that PakI is down-regulated by progesterone during the secretory phase in normal endometrium and increased PakI activity during the secretory phase might lead to establishment of endometriosis.

Angiogenin, a potent inducer of angiogenesis, is expressed in human endometrium. This study was performed to compare the expression of angiogenin mRNA level in the eutopic endometrium from women with and without endometriosis. Thirty-two women with advanced stage endometriosis and 29 control women were recruited. Following isolation of total RNA from endometrial tissue and reverse transcription, cDNA samples were amplified by real time polymerase chain reaction to quantify the expression of angiogenin genes. In selected patients, immunohistochemical staining was utilized to localize the area of angiogenin expression. Angiogenin mRNA level was significantly lower in the endometriosis group than in the control group during the secretory phase, especially the mid-secretory phase, and the decline was observed mainly in the women who presented with infertility. Within the endometriosis group, angiogenin mRNA levels did not differ between the proliferative and secretory phases, but, in the control group, the level in the secretory phase was higher than that during the proliferative phase. Immunohistochemistry showed that the glandular epithelial cell layer was decorated positively in both groups. These findings suggest that the relative deficiency of angiogenin expression in the secretory endometrium could impair implantation in women with advanced stage endometriosis.

Diabetes mellitus (DM) disturbs normal functions from the level of cells to the level of organs. In this study, the authors explore the detrimental effects of type 1 diabetes on meiotic regulation depending on the duration of DM. In non-diabetes-prone BioBreeding (BBdr) control rats, most of the large follicles had germinal vesicle (GV)-intact oocytes. Conversely, a decrease of intact GV that was dependent on the duration of diabetic symptoms was observed; only 54% of the large follicles of diabetes-prone BB (BBdp) rats had GV-intact oocytes at 6 weeks after diabetes induction. Furthermore, some of the secondary follicles in BBdp rats also had germinal vesicle breakdown (GVB) oocytes. The nuclear status of the euglycemia BBdp rat was similar to those of the BBdr rat. In BBdp rats, the rate of meiotic progression to the metaphase II stage was significantly lower; however, the rate of segregated oocytes was significantly increased compared with controls during induction of in vitro maturation. The rate of segregated oocytes was not affected by the presence of the cumulus after chronic symptoms. These results indicate that chronic DM has a detrimental effect on meiotic regulation during folliculogenesis and results in a reduced number of competent oocytes. In addition, these data suggest that the follicle cells can resume supporting the meiotic regulation under euglycemia through insulin administration, independent of the duration of DM.

Aim: The objective of the present study was to compare the levels of expression of epidermal growth factor (EGF), fibroblast growth factor-2 (FGF-2), and platelet-derived growth factor-A (PDGF-A) mRNA in the eutopic endometrium of women with and without endometriosis. Methods: Thirty-five women with advanced stage endometriosis and 31 control women were recruited. Following isolation of total RNA from endometrial tissue and reverse transcription, cDNA samples were amplified to quantify the level of expression of EGF, FGF-2, PDGF-A. Results: While the levels of mRNA of EGF and FGF-2 were not different between the two groups, the PDGF-A mRNA level was significantly lower in the endometriosis group than in the control group during the secretory phase (6.12 ± 0.51 vs 12.50 ± 0.99, mean ± SEM, P

Objective The objective was to investigate the influence of tumor necrosis factor (TNF)-α on estradiol, progesterone, insulin-like growth factor (IGF)-II, and insulin-like growth factor binding protein (IGFBP)-1, 2, and 3 in cultured human luteinized granulosa cells. Study design Human luteinized granulosa cells were obtained from follicular fluid by transvaginal oocyte aspiration from infertile patients undergoing controlled ovarian hyperstimulation (COH) for in vitro fertilization (IVF). The cells were cultured for 72 h with TNF-α at concentrations of 1.0, 10.0, and 100.0 ng/ml. The cells not treated with TNF-α served as controls. Radioimmunoassay (RIA) and reverse transcription-polymerase chain reaction (RT-PCR) were used to examine the influence of TNF-α on estradiol, progesterone, IGF-II, and IGFBP-1, 2, and 3. Results were analyzed using the Kolmogorov–Smirnov test and analysis of variance (ANOVA). Statistical significance was defined as p Results The concentrations of progesterone seemed to decrease as the concentrations of TNF-α increased and the concentration of progesterone in the 100.0 ng/ml TNF-α group was significantly lower than that in the control and other TNF-α groups. The expressions of IGF-II mRNA in the 10.0 and 100.0 ng/ml TNF-α groups were significantly lower than that in the control group. The expressions of IGFBP-2 mRNA seemed to be decreased in the 10.0 and 100.0 ng/ml TNF-α groups compared with that in the control group, but there were no statistical significances. Conclusion TNF-α may play a role as a regulator of human ovarian physiology by modulating the IGF systems in luteinized granulosa cells.

In this retrospective cohort study, the effect of endometrial polypectomy carried out on the day of oocyte retrieval and on the first day of ovarian stimulation in patients with a large (≥10 mm) endometrial polyp undergoing IVF and intractyoplasmic sperm injection (ICSI) was investigated and compared. A total of 74 eligible IVF-ICSI cycles in 74 women who underwent endometrial polypectomy on either the day of oocyte pick-up (late polypectomy group, 39 cycles) or the first day of ovarian stimulation (early polypectomy group, 35 cycles) between January 2007 and July 2012 were included in this study. Patient characteristics between early and late polypectomy groups were similar. Total dose and days of recombinant human FSH administered, numbers of retrieved oocytes, mature oocytes, fertilized oocytes, grade 1 or 2 embryos and embryos transferred between the two groups were also similar, as was clinical pregnancy rate per cycle, embryo implantation rate and spontaneous abortion rate between the two groups. Therefore, endometrial polypectomy on the day of oocyte retrieval could be a more patient-friendly option for patients with a large endometrial polyp undergoing IVF-ICSI.

Although phthalates were shown to have several negative effects on reproductive function in animals, its role in the pathogenesis of endometriosis remains to be elucidated.We aimed to investigate the in vitro and in vivo effects of di-(2-ethylhexyl)-phthalate (DEHP) and to compare the urinary levels of several phthalate metabolites between women with and without endometriosis.For experimental studies, we used endometrial cell culture and nonobese diabetic/severe combined immunodeficiency (NOD/SCID) mouse models. We also performed a prospective case-control study for human sample analyses.The study was conducted at an academic center.The activities of matrix metalloproteinase (MMP)-2 and 9, cellular invasiveness, phosphorylation of extracellular signal-regulated kinase (Erk), and expression of p21-activated kinase 4 were analyzed in endometrial cells treated with DEHP. The implant size was compared between NOD/SCID mice fed with and without DEHP. Urinary concentrations of several phthalate metabolites were compared between women with and without endometriosis.In vitro treatment of endometrial cells with DEHP led to significant increases of MMP-2 and 9 activities, cellular invasiveness, Erk phosphorylation, and p21-activated kinase 4 expression. The size of the endometrial implant was significantly larger in the NOD/SCID mice fed with DEHP compared with those fed with vehicle. The urinary concentration of mono (2-ethyl-5-hydroxyhexyl) phthalate, mono (2-ethyl-5-oxohexyl) phthalate, and mono (2-ethyl-5-carboxyphentyl) phthalate were significantly higher in women with endometriosis compared with controls.These findings strongly suggest that exposure to phthalate may lead to establishment of endometriosis by enhancing invasive and proliferative activities of endometrial cells.

Twin pregnancy consisting of complete hydatidiform mole (H-mole) and a coexisting fetus occurs with an estimated incidence of 1 per 22,000–100,000 pregnancies. The incidence of this unusual twin pregnancy with complete H-mole and a coexisting fetus after in vitro fertilization and embryo transfer (IVF–ET) is not thought to be greater than that of general population. We present an unusual twin pregnancy with complete H-mole and a coexisting fetus that occurred following IVF–ET, which was terminated at 21 weeks of gestation and developed into nonmetastatic gestational trophoblastic tumor.

We performed the present prospective case-control study to evaluate whether the plasma concentrations of phthalate esters are elevated in women with advanced-stage endometriosis in a Korean population. Measuring plasma levels of monoethylhexyl phthalate and di-(2-ethylhexyl) phthalate in 97 women with advanced-stage endometriosis and 169 control women by liquid chromatography-tandem mass spectrometry, we found that the concentrations of monoethylhexyl phthalate, as well as di-(2-ethylhexyl) phthalate, are significantly higher in those with advanced-stage endometriosis, which supports the hypothesis that exposure to phthalate might play a role in the establishment of endometriosis.

Objective To investigate the expression of p21-activated kinase 4 (Pak4) in both adenomyotic foci and the eutopic endometrium of women with adenomyosis, and whether the activities of matrix metalloproteinases (MMPs)-2 and -9 are regulated by Pak4 in endometrial cells. Design Experimental study using human samples and cell lines. Setting University hospital. Patient(s) Thirty-nine patients with histologic evidence of adenomyosis, and 34 patients with carcinoma in situ of the uterine cervix without adenomyosis or endometriosis. Intervention(s) Immunohistochemistry, zymography after transfection with Pak4 small interfering RNA (siRNA), and western blot analyses after nuclear factor kappa-B (NF-кB) inhibitor treatment. Main Outcome Measure(s) The Pak4 immunoreactivity of women with vs. without adenomyosis was compared semiquantitatively. The activities of MMP-2 and -9 were analyzed in eutopic endometrial stromal cells and Ishikawa cells after transfection with Pak4 siRNA. The Pak4 expression was evaluated in endometrial cells after treatment with NF-кB inhibitor. Result(s) Pak4 immunoreactivity was increased in adenomyotic foci and in the eutopic endometrium of women with adenomyosis. Transfection of endometrial cells with Pak4 siRNA led to significant decreases of MMP-2 and -9 activities. In vitro treatment of endometrial cells with tumor necrosis factor-alpha caused a significant increase of NF-кB activation and Pak4 expression, which was obviously decreased by the NF-кB inhibitor pyrrolidinedithiocarbamate. Conclusion(s) Our results suggest that Pak4 is regulated by NF-кB and that increased Pak4 expression can lead to development of adenomyosis by enhancing the invasiveness of endometrial cells through regulation of MMP-2 and -9 activities.

To evaluate the effects of combined administration of gonadotropin-releasing hormone (GnRH) agonist with human chorionic gonadotropin (hCG)for final oocyte maturation in GnRH antagonist cycles for in vitro fertilization (IVF).A total of 120 infertile women undergoing GnRH antagonist multiple-dose protocol for controlled ovarian stimulation were recruited and randomized into 2 groups: a study group (n=60) and a control group (n=60). For the study group, both GnRH agonist and recombinant hCG (rhCG) were injected concomitantly for final oocyte maturation when 1 or more follicles reached a mean diameter of 18 mm. For the control group, rhCG alone was administered for final oocyte maturation.There were no significant differences in patient characteristics. The 2 groups were also similar with respect to the number of oocytes retrieved, fertilized oocytes and good-quality embryos. Embryo implantation rate (24.7% vs. 14.9%), clinical pregnancy rate per cycle (53.3% vs. 33.3%), and live birth rate (50.0% vs. 30.0%) were significantly higher in the study group than in the control group (p = 0.006, p = 0.027, and p = 0.025, respectively).Combined administration of GnRH agonist with rhCG may be beneficial in improving endometrial receptivity and pregnancy rate in GnRH antagonist cycles for IVF.

It has been known that most cases of ovarian hyperstimulation syndrome (OHSS) are associated with the use of exogenous gonadotropins to induce multiple ovulation. However, OHSS is infrequently associated with a spontaneous ovulatory cycle, usually in the case of multiple gestations, hypothyroidism, or polycystic ovarian syndrome. We report a case of severe OHSS in a spontaneously pregnant woman with no underlying disease.

Objectives: The purpose of the present study is to evaluate the relationship between endometrial concentrations of estrogen receptor (ER) and progesterone receptor (PR), and sonographic endometrial findings in the preovulatory phase of menstrual cycle. Study Design: In 45 cycles of 45 infertile women with tubal factor only, transvaginal sonographic assessments and biopsy for immunohistochemical staining of the endometrium were made in the preovulatory phase of unstimulated, normal menstrual cycle. Immunohistochemical localization of ER and PR was scored according to intensity of staining and proportion of cells specifically stained in glandular epithelium and stroma, and the results were analysed according to the sonographic endometrial thickness ( 10 mm) and patterns. Endometrial patterns were classified as A, centrally hyperechogenic triple-line pattern or non-A, not triple-line. Results: There were no significant differences in the endometrial thickness, serum estradiol level and serum progesterone level between A and non-A groups. The receptor scores of epithelial and stromal ER and epithelial PR were comparable in A and non-A groups. However, the receptor score of stromal PR was significantly higher in A group, with 4.8 ± 1.4 compared with 2.7 + 1.7 in non-A group (p < 0.001). There were no differences in the receptor scores of epithelial ER, epithelial PR, stromal ER and stromal PR among the 3 groups according to the endometrial thickness. Conclusions: This study suggests that high PR expression in endometrial stroma could be related to the sonographic triple-line or multilayered pattern of endometrium in the preovulatory period.

Objective: To investigate the effect of pyridostigmine, an acetylcholinesterase inhibitor, as cotreatment for controlled ovarian hyperstimulation (COH) in low responders. Design: Randomized, double-blind, placebo-controlled study. Setting: A reproductive medicine unit in a university hospital. Patient(s): Seventy infertile women with a history of low ovarian response to COH using a GnRH agonist as part of a long stimulation protocol in previous IVF-ET cycles. Intervention(s): Sixty milligrams of pyridostigmine or placebo was administered orally twice daily from the first day of COH until the day of hCG injection in patients undergoing IVF-ET cycles. Main Outcome Measure(s): In vitro fertilization results, pregnancy outcome, and serum and intrafollicular concentrations of GH and insulin-like growth factor-1. Result(s): Pyridostigmine cotreatment was associated with significant decreases in the amount of gonadotropins and the duration of stimulation required. The clinical pregnancy rate was higher in the pyridostigmine group, but this difference was not statistically significant (25.7% vs. 11.4%). The serum GH level on the day of hCG injection was significantly higher in the pyridostigmine group than in the placebo group. Follicular fluid concentrations of GH and insulin-like growth factor-1 were significantly higher in the pyridostigmine group. Conclusion(s): This study suggests that pyridostigmine cotreatment for COH could affect the serum and intrafollicular GH and insulin-like growth factor-1 concentrations and, hence, improve the ovarian response to COH and the results of IVF in low responders undergoing IVF-ET.

Objective: To investigate the influence of epidermal growth factor (EGF) on preimplantation development, implantation, and expression of epidermal growth factor receptor (EGFR) itself in mouse embryos. Materials and Method: Eight-cell stage mouse embryos were cultured for 48 hours with EGF at concentrations of 0.1, 1.0, 10 and 100 ng/m/. Embryos not treated with EGF were served as control. The percentages of embryos which developed to the expanded, hatched blastocyst stage and in vitro implantation at 48 hours were determined. Reverse transcription-polymerase chain reaction (RT-PCR) has been used to examine the expression of EGFR in developed hatched blastocysts. Following reverse transcription, strategically designed nested primers, optimized for specificity, were used for amplification from the cDNA equivalent of a single embryo. The products were then verified by restriction enzyme digestion and sequence analysis. Results were analyzed with X2 test and Student's t-test as apropriate, and statistical significance was defined as p < 0.05. Results: The percentages of fully expanded blastocysts at 48 hours in all the EGF treated group were not significantly different from the control. The percentages of hatched blastocysts were significantly higher in the EGF treatment group at 0.1 ng/m/ (90.5 ± 9.8%) compared to the control (82.1 ± 7.2%), 1.0 ng/m/ (82.2 ± 12.7%), and 100 ng/m/ (81.9 ± 11.8%) (p < 0.05, p < 0.05, p < 0.05, respectively). The percentages of hatched blastocysts were significantly higher in the EGF treatment group at 10 ng/m/ (89.4 ± 7.5%) compared to the control, and 100 ng/m/ (p < 0.05, p < 0.05, respectively). The percentages of attached blastocysts in vitro were significantly higher following incubation with EGF at concentrations of 0.1 ng/m/ (37.0 ± 17.0%), 1.0 ng/m/ (32.0 ± 14.3%), 10 ng/m/ (21.3 ± 7.2%) compared to the control (9.5 ± 7.7%) (p < 0.05, p < 0.05, p < 0.05, respectively). The attachment rates in 0.1 ng/m/ and 1.0 ng/m/ EGF treatment groups were also significantly higher than those in other EGF treatment groups. Embryo development and attachment were not significantly inhibited or enhanced in cultures supplemented with 100 ng/m/ EGF compared to the control. The mRNA concentration of EGFR in embryos treated with 0.1 ng/m/ of EGF was significantly higher than those of the control and other EGF treatment groups. Conclusion: EGF may have a stimulatory role in later stage embryonic development, implantation and expression of EGFR in hatched blastocyst itself at the specific concentration.

It is well known that excessive thyroid hormone in the body is associated with bone loss. However, the mechanism by which thyroid hormone affects bone turnover remains unclear. It has been shown that it stimulates osteoclastic bone resorption indirectly via unknown mediators secreted by osteoblasts. To determine if interleukin-6 (IL-6) or interleukin-11 (IL-11) could be the mediator(s) of thyroid hormone-induced bone loss, we studied the effects of 3,5,3'-tri-iodothyronine (T3) on basal and interleukin-1 (IL-1)-stimulated IL-6/IL-11 production in primary cultured human bone marrow stromal cells. T3 at 10(-12)-10(-8) M concentration significantly increased basal IL-6 production in a dose-dependent manner. It also had an additive effect on IL-1-stimulated IL-6 production, but failed to elicit a detectable effect on basal or IL-1-stimulated IL-11 production. Treatment with 17beta-estradiol (10(-8) M) did not affect the action of T3 on IL-6/IL-11 production. These results suggest that thyroid hormone may stimulate bone resorption by increasing basal and IL-1-induced IL-6 production from osteoblast-lineage cells, and these effects are independent of estrogen status.