Effects of oral contraceptives and metformin on the outcome of in vitro maturation in infertile women with polycystic ovary syndrome.

UNLABELLED: Abstract Objective: To evaluate the effects of oral contraceptives and metformin on the outcome of in vitro maturation (IVM) in infertile women with polycystic ovary syndrome (PCOS). METHODS: This is a retrospective study of 108 women with PCOS, subject to 152 cycles of IVM treatment. The study was held at the Reproductive Medicine Center of the First Affiliated Hospital of Wenzhou Medical College, People's Republic of China. Before entering IVM treatment, 54 patients who received oral contraceptive pill (marvelon, 0.15 mg desogestrel, and 0.03 mg ethinylestradiol), one tablet every day, and metformin 500 mg twice or three times per day were defined as the pretreated group, and another 64 patients who were not administered any drugs as the control group. The main outcome measures were the rates of oocyte maturation, fertilization, cleavage, miscarriage, clinical pregnancy, and live birth. RESULTS: There were no significant differences between the two groups in the rates of oocyte maturation, fertilization, cleavage, and clinical pregnancy (p > 0.05). A significantly lower miscarriage rate was obtained in the pretreated group than in the control group (16.13% vs 4.0%, p < 0.01). The live birth rate per embryo transfer seemed to be higher in the pretreated group than in the control group (37.70% vs 30.38%, p = 0.363), but was not statistically significant. CONCLUSIONS: Pretreatment with oral contraceptives and metformin improved the outcome of IVM related to the miscarriage rate and possibly also live birth rate.

[Outcome of pregnancy in women with polycystic ovary syndrome treated by in vitro maturation of immature oocytes].

OBJECTIVE: To investigate pregnancy outcome of infertility women with polycystic ovary syndrome (PCOS) treated by in vitro maturation (IVM) of immature oocytes. METHODS: From Nov. 2003 to Oct. 2007, medical documents of 118 women with PCOS underwent 140 IVM treatment cycles in Reproductive Medical Center of First Affiliated Hospital of Wenzhou Medical College were collected. Follow up of 62 pregnancies were performed by prenatal examination in hospital or telephone query to record perinatal monitoring and pregnancy outcome. RESULTS: There are 62 pregnant women including 5 biochemical pregnancies and 57 clinical pregnancies obtained in 140 transferred cycles, resulting in the pregnancy rate of 40.7% (57/140). The rates of singleton pregnancies, twin pregnancies and triplet pregnancies were 61.4% (35/57), 29.8% (17/57) and 5.3% (3/57), respectively. The rate of ectopic pregnancy was 3.5% (2/57). Seven (7/57, 12.3%) women underwent early abortion during 7 -14 weeks of pregnancy, and 1 case (1/57, 1.75%) with premature rupture of membranes occurred at 22 gestational week. One woman with twin pregnancy spontaneously reduced to singleton at 8 gestational weeks. Totally, the rate of pregnancy complications was 26.3% (15/57) including premature rupture of membranes (1 case), placenta previa (1 case), hypertensive disorder (1 case), preterm delivery (10 cases) and gestational diabetes mellitus (2 cases). Until now 47 women gave birth to 65 infants including 29 singleton infants and 18 twins. One female preterm neonate died after 6 days' delivery due to pneumonia, no malformation was observed on the other neonates. 21.3% (10/47) of deliveries were premature,76.6% (36/47) of deliveries were full-term, 2.1% (1/47) of deliveries were postterm. The mean birth weight was 2972 gram. The rate of infants with low weight was 26.2% (17/65). CONCLUSIONS: A relatively high clinical pregnancy rate has been achieved, the rates of early abortion, ectopic pregnancy, pregnancy complications, perinatal mortality, and neonatal malformation occurring after the treatment of IVM in women with PCOS are not mounting. However, the relative high rates of multiple pregnancies, low birth weight and preterm labor were increased.

[Correlation between serum inhibin B level after treatment with gonadotropin releasing hormone agonist and outcome of in vitro fertilization-embryo transfer].

OBJECTIVE: To evaluate the decreased level of serum inhibin B (INHB) treated by gonadotropin releasing hormone agonist (GNRH-a) in predicting ovarian response and pregnancy in in vitro fertilization-embryo transfer (IVF-ET). METHODS: The prospective study enrolled 124 women given by GnRH-a+ recombine follicle stimulating hormone (rFSH) + human chorionic gonadotrophin (hCG) long term stimulation protocol undergone their first cycle of IVF-ET treatment. The following predictive factors were collected and analyzed, such as age, basal level of follicle stimulating hormone (FSH), the ratio of FSH/ luteinizing hormone (LH), the concentration of INHB after down-regulation, total number of antral follicle count (AFC) and mean ovarian volume. Ovarian response was evaluated by the number of oocytes obtained. A multiple regression analysis and logistic regression model were used for all possible prognostic variables to evaluate the value of different hormones in predicting ovarian response and pregnancy after IVF-ET. Receiver operating characteristic (ROC) analysis was used to evaluate the level of INHB in predicting the number of oocytes obtained. The sensitivity and specificity were calculated at the discriminating cut-off point. RESULTS: The concentration of INHB after down-regulation showed a highly significant positive correlations with the number of oocytes obtained (r = 0.435, P < 0.01). The multiple regression analyses showed INHB was the most significant predictor of the number of retrieved oocytes, but INHB was not associated with IVF-ET outcome significantly (P > 0.05). ROC analyses showed INHB after down-regulation had the largest area under curve (AUC) 0.933 (95%CI: 0.878 - 0.988). When a threshold of 15 ng/L of INHB was established, 95.5% sensitivity and 50.0% specificity in ovarian response were observed. CONCLUSIONS: The level of INHB was the best factor in predicting ovarian response in IVF-ET. Decreased level of INHB was the early sign of ovarian reserve function failure, however, useless in predicting IVF-ET outcome.

In vitro maturation and fertilization of oocytes from unstimulated ovaries in infertile women with polycystic ovary syndrome.

OBJECTIVE: To evaluate the effects of in vitro maturation and fertilization of oocytes from unstimulated ovaries in infertile women with polycystic ovary syndrome (PCOS). DESIGN: Retrospective study. SETTING: Reproductive Medicine Center, First Affiliated Hospital of Wenzhou Medical College, People's Republic of China. PATIENT(S): One hundred eighteen women with PCOS undergoing 152 cycles of in vitro maturation treatment. INTERVENTION(S): Oocyte retrieval was carried out by ultrasound-guided puncture on days 9-14 of the cycle. The oocytes were cultured in vitro using maturation culture medium, which consisted of M-199 + 20% fetal bovine serum (FBS) + 75 mIU/mL recombinant FSH +/- 0.5 IU/mL hCG. After the oocytes had matured in vitro, fertilization and embryo transfer were performed. MAIN OUTCOME MEASURE(S): Rates of clinical pregnancy, multiple pregnancies, and live birth. RESULT(S): Relatively optimal laboratory results were obtained in this study. Embryo transfer was performed in 140 cycles, with a clinical pregnancy rate (PR) of 40.0% per transfer. Fifty-six babies have been born and there are 10 ongoing pregnancies. The overall multiple PR was 33.93%. CONCLUSION(S): Our results show that using in vitro matured oocytes from unstimulated ovaries could be offered as an alternative to conventional IVF in women with PCOS, and future work should address ways to decrease the incidence of multiple pregnancies.

[Clinical study on in vitro maturation of immature oocytes transferred from conventional in vitro fertilization.].

OBJECTIVE: To investigate clinical effect of in vitro maturation (IVM) of immature oocytes transferred from conventional in vitro fertilization embryo transfer (IVF-ET) cycles. METHODS: From January 2008 to June 2009, medical documents of 155 infertile patients underwent IVF-ET in the Reproductive Medical Center of First Affiliated Hospital of Wenzhou Medical College were analyzed retrospectively. If more than 20 oocytes were monitored after 5 - 7 days of ovulation induction or follicular developmental retardation were confirmed after 8 - 13 days of ovulation induction, according to patients' wish, IVM were transferred in 60 cycles (group A). In the mean time, IVF was continued in 95 cycles (group B). The mean dosage of gonadotropin, the cancellation rate of cycles, the mean numbers of oocytes retrieved and maturation, the rate of fertilization and excellent quality embryos, pregnancy outcome and the incidence rate of ovarian hyperstimulation syndrome (OHSS) were compared and analyzed. RESULTS: The rates of embryo transfer were 92% (55/60) in group A and 63% (60/95) in group B, which showed significant differences (P < 0.05). In group A, the mean dosage of the gonadotropin, the mean number of oocytes retrieved, the cleavage rate and OHSS rate were (1030 +/- 468) U, 10 +/- 6, 82.2% (231/281) and 0, and were (1544 +/- 338) U, 14 +/- 4, 94.0% (502/534) and 35% (21/60) in group B, respectively, all data above exhibited statistical difference between two groups (P < 0.05). However, the rates of fertilization and excellent quality embryos had no significant differences between two groups (P > 0.05). In group A, the rate of clinical pregnancy per transfer was 53% (29/55) and multiple pregnancy was 14% (4/29), and were 47% (28/60) and 32% (9/28) in group B, they all had no significant differences (P > 0.05). CONCLUSION: IVM of immature oocytes used in conventional IVF cycles not only obtained a high clinical pregnancy rate, but also reduced gonadotropin using dosage and avoided OHSS completely.

[Effect of desogestrel and ethinyl estradiol pretreatment in superovulation cycles with short protocol].

OBJECTIVE: To study the effect of desogestrel and ethinyl estradiol (DEE) pre-treatment combined with gonadotropin releasing hormone agonist (GnRH-a) stimulation in in vitro fertilization-embryo transplantation (IVF-ET). METHODS: A retrospective analysis was performed in 101 infertile women who received a short protocol of GnRH-a for IVF-ET treatment from June 2004 to June 2007 in the Reproductive Medicine Center of First Affiliated Hospital of Wenzhou Medical College. Patients had been pre-treated with oral contraceptive pill (OCP) for two months before GnRH-a combined with recombinant follicle stimulation hormone (r-FSH) treatment (study group, n = 42) or had not been pretreated with OCP (control group, n = 59). A statistical analysis of two groups was carried out for the assessment of ovulation stimulating effect of OCP and its influence on the IVF. RESULTS: Serum FSH was significantly decreased after OCP in the study group. Twelve pregnancies were obtained including 1 case of spontaneous abortion at 7 weeks in the study group, and 11 pregnancies were obtained including 2 cases of spontaneous abortion during 7 -9 weeks in control group. The clinical pregnancy rates in the study group (23%, 12/53) was higher than that in the control group (17%, 11/63), but the differences were not significant (P > 0.05). The miscarriage rate in the study group (8%, 1/12) was lower than that in the control group (18%, 2/11), however no significant differences were found between them (P > 0.05). The cycle cancellation rate in patients of the study group (5%, 3/56)was significantly lower than that in patients of the control group (17%, 13/76, P < 0.05). The differences between patients of the two groups with respect to age, basal level of FSH and luteinizing hormone (LH), antral follicle counts, the mean number of oocyte retrieval, the days of stimulation, total dose of r-FSH used, fertilization rate and embryo cleavage rate, however were insignificant. CONCLUSION: OCP pretreatment combined with short protocol of GnRH-a stimulation in IVF could significantly decrease the cycle cancellation rate, with a declining miscarriage rate and increasing pregnancy rate.

[HLA-A site genotyping on single blastomeres is studied by nest-PCR-SSP method].

OBJECTIVE: To assess the accuracy and reliability of the nest-PCR-sequence specific primer(SSP) method in HLA-A site genotyping of single blastomeres retrieved from human pre-implantation embryos. METHODS: By nest PCR on HLA-A exon 2, the success rate of first-round amplification was estimated for single blastomeres. Based on the first-round amplification, the HLA-A genotype of every single blastomeres was analyzed by commercially available PCR-SSP kits. RESULTS: The amplification of HLA-A exon 2 were performed to 120 blasotmeres retrieved from in vitro fertilization(IVF) surplus embryos donated by 10 couples. The average success rate of family 1-5 and 6-10 was 78.2%(43/55) and 93.8%(61/65), respectively. And 86.7%(104/120) in total. Eighty blastomeres were further tested by nest-PCR-SSP, among which 11 blastomeres failed to HLA-A exon 2 amplification and then failed to genotyping while the other 69 blastomeres succeed in HLA-A exon 2 amplification and succeed in genotyping. Except for 6 blastomeres that were uncertain for allele lost because of parents' homozygosity, the left 63 blastomeres had accurate HLA genotyping. Among these 63 blastomeres, 59 blastomeres had genotypes confirmed from their parents(93.6%), 3 blastomeres lost one of parents' alleles(4.8%), and only one blastomere had two more than parents' alleles(1.6%). CONCLUSION: The above research results indicated that based on the successful first round amplification of single blastomeres, nest-PCR-SSP strategy offers a convenient and reliable option for HLA genotyping on single blastomeres, which is a key process in pre-selecting HLA-identical sibling for allogeneic cord blood cell transplantation.

Bicyclo[2.2.2]octene-based "crab-like" molecules: synthesis, complexation, luminescence properties, and solid-state structures.

[structure, reaction: see text] The U-shaped, multifunctionalized tetraetheno-bridged dicyclopenta[b,i]anthracenediol 10 was synthesized to serve as a platform molecule. The molecule was prepared from the Diels-Alder adduct 5a of tricycloundecatriene 3 and bicyclo[2.2.2]octene-fused p-benzoquinone 4. Functionalization of 10 to construct crab-like molecules was achieved via the base-promoted bis-O-alkylation of two endo-oriented hydroxyl groups at termini in 10 with the following alkyl halides: allyl, propagyl, and benzyl bromides; 1-bromo- and 1-iodo-4-(bromomethyl)benzene; 9-(bromomethyl)anthracene; 1-(bromomethyl)pyrene; and isomeric bromomethylpyridines. Single-crystal X-ray structures were obtained for bis-phenyl (21) and bis-pyrenyl (25) crabs, and for the silver(I) complex (32 and 33) crabs. The silver(I) complex 32 from bis-o-pyridyl crab 30 is a [2+2] dimeric dimetallocyclophane, and 33 from bis-m-pyridyl crab 29 is a [1+1] metallo-bridged cyclophane. The self-assembled intramolecular pi-stacking of pyrenyl rings in 25 with an interplanar distance of 3.40 A and the consequent pi-pi interactions were revealed by the X-ray crystal structure and its luminescence property.

Synthesis and luminescence properties of U-shaped polycyclic molecules containing syn-facial functionalized quinoxaline rings.

[reaction: see text] Rigid U-shaped molecules containing syn-facially situated quinoxaline rings have been synthesized in three steps, by a combination of the Diels-Alder reaction, the ruthenium-catalyzed oxidation, and the Zn(OAc)(2)-catalyzed condensation of the resulting bis-alpha-diketones with benzene-1,2-diamines. The unsymmetric bis-quinoxalines, with electronically different substituents, and the quinoxaline ring-attached alpha-diketones were also prepared. Their luminescence properties were examined and described.

[Analysis of pregnancy outcomes after in vitro fertilization-embryo transfer and intracytoplasmic sperm injection].

OBJECTIVE: To compare pregnancy and perinatal outcomes between in vitro fertilization-embryo transfer (IVF-ET) and intracytoplasmic sperm injection (ICSI). METHODS: A retrospective study was carried out to measure pre-clinical and clinical abortion, ectopic pregnancies, multiple gestations, birth weight, gestational age, congenital malformation and perinatal mortality in patients receiving either IVF-ET (n = 143, group 1) or ICSI (n = 173, group 2) from January 1999 to June 2001. The outcomes of singleton and twin were compared separately. RESULTS: The maternal age, infertility duration, parity and the number of transferred embryo were comparable between the two groups. There were no significant differences in abortion rate (16.1% vs 13.3%), birth rate (65.7% vs 74.6%) between IVF-ET and ICSI groups (P > 0.05). In singleton, the rates of low birth weight, small for gestational age and pre-term birth were 1.8%, 7.3%, 5.5% respectively in IVF-ET group and 6.8%, 8.1%, 14.9% respectively in ICSI group. In twin, the rates of low birth weight, small for gestational age and pre-term birth were 34.2%, 30.3%, 42.1% respectively in IVF-ET group and 42.6%, 38.0%, 46.3% respectively in ICSI group. There were no significant differences between the two groups (P > 0.05). But the rates of low birth weight, small for gestational age and pre-term birth were higher in twin than in singleton (P < 0.01). The incidence of congenital malformation was 2.2% and 1.6% in IVF-ET and ICSI group respectively (P > 0.05). CONCLUSIONS: The pregnancy and perinatal outcomes are similar between IVF-ET and ICSI groups. Twin is the main cause of low birth weight, small for gestational age and pre-term birth.

[Effects of metformin on gonadotropin-induced ovulation in patients with polycystic ovary syndrome].

OBJECTIVE: To evaluate the effects of metformin on gonadotropin-induced ovulation in patients with polycystic ovary syndrome (PCOS). METHODS: Forty patients with PCOS (study group) and 20 women with normal weight and menstrual cycle (control group) were enrolled. Serum follicle stimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), fasting glucose (FG), fasting insulin (FINS) and fasting leptin were measured before and after treatment. In the study group, 20 cases (group A) were assigned to take 500 mg of metformin three times daily for 12 weeks, if pregnancy did not occur, high purified FSH (FSH-HP) was added for one cycle; another 20 cases (group B) were induced ovulation with FSH-HP alone for one cycle. RESULTS: There were significant high FINS and leptin levels in the study group as compared with the control group [(20 +/- 16) vs (12 +/- 6) nmol/L, P < 0.05; (14 +/- 16) vs (8 +/- 4) mg/L, P < 0.05]. The obese PCOS group had markedly higher serum FINS and leptin than the non-obese PCOS group [(24 +/- 18) vs (14 +/- 8) nmol/L, P < 0.05; (20 +/- 22) vs (8 +/- 4) mg/L, P < 0.05], but serum FINS and FG were not significantly different between the non-obese PCOS and the control group (P > 0.05). After administration of metformin for 12 weeks, serum LH, T, leptin and FINS decreased significantly (P < 0.05 - 0.01), serum FSH levels and body mass index showed a slight decrease, whereas no change was found in FG. In the study group, 3 cases conceived during metformin therapy, the remaining 37 were induced ovulation with FSH-HP or FSH-HP and metformin, 7 cases obtained pregnancy. The rates of ovulation and pregnancy in group A were higher than those in group B (88% vs 70%, 24% vs 15%), but no significant difference was found. CONCLUSIONS: Metformin therapy in PCOS can decrease the FINS and leptin levels, normalize the endocrine abnormalities, resumes ovulation and pregnancy in some patients, and may improve the ovarian response to gonadotropin.

[Endocrine and metabolic effects of metformin in combination with compound cyproterone acetate in women with polycystic ovarian syndrome].

OBJECTIVE: To study the endocrinologic and metabolic effects of metformin in combination with compound cyproterone acetate (CPA) on patients with polycystic ovarian syndrome (PCOS). METHODS: A prospective study involved total 45 PCOS patients as group A and 20 non-PCOS infertility patients as control (group B). Complete baseline work-up including body mass index (BMI), waist/hip ratio (WHR), ferriman-Gallwey score (FGS), gonadotrophin, testosterone (T), sex hormone-binding globulin (SHBG), dehydroepiandrosterone sulfate (Ds), insulin (FI) and glucose tolerance test, were performed in all patients. Patients in group A were treated with CPA alone (group A1), metformin alone (group A2) or combination of CPA with metformin (group A3), respectively by randomization. At the end of 12-week therapy, subjects were re-evaluated and above parameters were measured. RESULTS: Women in group A had significant increases in BMI, WHR, FGS, luteinizing hormone (LH), T, FI, insulin resistance, and significantly decrease in high-density lipoprotein (HDL)-C comparing with the control group (P < 0.01). No significant difference among A1, A2 and A3 was found at baseline. LH, T, free testosterone (FT) were significant decreased from (13.9 +/- 5.9) IU/L, (2.1 +/- 0.8) nmol/L and (2.8 +/- 2.3) nmol/L respectively to (5.8 +/- 2.2) IU/L, (1.2 +/- 0.4) nmol/L and (0.8 +/- 0.5) nmol/L respectively and SHBG was significant increased from (99 +/- 42) nmol/L to (187 +/- 64) nmol/L in group A3, when compared with LH, T and FT from (13.8 +/- 7.6) IU/L, (2.2 +/- 1.1) nmol/L and (2.5 +/- 1.9) nmol/L respectively to (11.8 +/- 6.5) IU/L, (1.8 +/- 0.8) nmol/L and (1.7 +/- 1.0) nmol/L respectively and SHBG from (99 +/- 40) nmol/L to (120 +/- 51) nmol/L in group A2 (P < 0.05 approximately 0.001). HDL-C were significantly increased from (1.5 +/- 0.3) mmol/L to (1.8 +/- 0.3) mmol/L in group A3 comparing with HDL-C from (1.5 +/- 0.4) mmol/L to (1.6 +/- 0.4) mmol/L in group A1 (P < 0.001). CONCLUSIONS: The PCOS patients treated with metformin in combination with compound cyproterone acetate may be more effective in inhibiting hyperandrogen and hypersecretion of LH than metformin alone and more obvious in improving lipid profiles than CPA alone.