Qingxue Zhang

Cryopreservation of human ovarian tissue by solid-surface vitrification

OBJECTIVEnTo cryopreserve human ovarian tissue using solid-surface vitrification (SSV) technique for the first time.nnnSTUDY DESIGNnHuman ovarian slices from each of 26 patients were randomly allocated to fresh, SSV, and slow-freezing groups, respectively. Histological observation and the TUNEL assay of the tissue were performed after cryopreservation. In vitro culture was done to study the initial recruitment of follicles and hormone production ability after SSV/slow-freezing.nnnRESULTSnThe majority of primordial follicles were maintained intact through either SSV or the slow-freezing method. No statistically significant destructive effect of SSV or slow-freezing for primordial follicles and stromal cells was found using the TUNEL assay. In the SSV and slow-freezing groups, estradiol and progesterone were secreted continuously during 10 days in culture, and the proportions of growing follicles increased significantly comparing to the uncultured fresh group. The follicular proportions and the concentrations of estradiol and progesterone exhibited no statistically significant differences between the SSV and slow-freezing groups.nnnCONCLUSIONnSSV is an effective, simple and inexpensive alternative for human ovarian tissue cryopreservation.

Increased GDF9 and BMP15 mRNA levels in cumulus granulosa cells correlate with oocyte maturation, fertilization, and embryo quality in humans.

BackgroundOocyte secreted factors (OSFs), including growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15), play an important role in the process of follicular development and oocyte maturation. Since OSFs are expressed in oocytes and cumulus granulosa cells, the aim of the present study was to explore whether the expression levels of GDF9 and BMP15 mRNAs in cumulus granulosa cells can be used as molecular markers for predicting oocyte developmental potential.MethodsCumulus cells of 2426 cumulus-oocyte complexes were collected from 196 female patients who underwent intracytoplasmic sperm injection (ICSI) and were used for mRNA detection on the egg retrieval day. Pearson correlation analysis was used to analyze the correlation between OSF expression and general physiological parameters. Partial correlation analysis was used to analyze the correlation between OSF expression and oocyte developmental potential. Covariance analysis was used to compare OSF expression among different groups. Receiver operating characteristic curves were used to examine the diagnostic value of GDF9 and BMP15 mRNA for predicting pregnancy.ResultsThe expression levels of GDF9 and BMP15 mRNAs were significantly associated with age, body mass index (BMI), oocyte maturation, normal fertilization, and cleavage rate (Pu2009<u20090.05). The expression levels of GDF9 and BMP15 mRNAs in the group with high-quality embryos were significantly higher than those in the group without high-quality embryos (Pu2009<u20090.05). The expression levels of GDF9 and BMP15 mRNAs in the pregnancy group were significantly higher than those in the nonpregnancy group (Pu2009<u20090.05). The cut-off value of GDF9 mRNA for predicting pregnancy was 4.82, with a sensitivity of 82% and a specificity of 64%. The cut-off value of BMP15 mRNA for predicting pregnancy was 2.60, with a sensitivity of 78% and a specificity of 52%.ConclusionsThe expression levels of GDF9 and BMP15 mRNAs were closely associated with oocyte maturation, fertilization, embryo quality, and pregnancy outcome; therefore, GDF9 and BMP15 mRNAs in cumulus granulosa cells may be considered as new molecular markers for predicting oocyte developmental potential.

Impact of overweight and underweight on IVF treatment in Chinese women.

Objective.u2003To investigate the effects of body mass index (BMI) on ovarian stimulation and the outcome of IVF or ICSI treatment in Asian population who have different characteristics of BMI from white people. Design.u2003In this retrospective study, the first fresh cycles of 1107 Chinese women were classified by BMI: underweight (BMIu2009<18.5u2009kg/m2), normal weight (BMI: 18.5–23.9u2009kg/m2) and overweight (BMIu2009≥u200924u2009kg/m2). Dose and days of ovarian simulation, cancelled cycles and number of oocytes collected, fertilisation and embryo utilisation rates and pregnancy outcome were compared among BMI groups. Results.u2003It showed that overweight women required more ampoules of gonadotrophin (Pu2009=u20090.002) and had lower peak oestradiol concentration (Pu2009=u20090.001), increased cycle cancellation due to insufficient follicle development (Pu2009=u20090.018) and a higher miscarriage rate (10.5 vs. 5.4%, Pu2009=u20090.018, ORu2009=u20092.006 (95% CI: 1.09–3.69, Pu2009=u20090.025)) compared with normal weight women. However, no differences were found in clinical pregnancy and live-birth rates. Underweight compared with normal weight women showed no differences in ovarian stimulation and IVF outcome although the clinical pregnancy rate was lower (31.1 vs. 37.3%). Conclusion.u2003At a lower cutoff point of BMI in Chinese women, overweight is associated with increased miscarriage risks and impaired response to ovarian stimulation after IVF and ICSI.

Ovarian volume and follicle number in the diagnosis of polycystic ovary syndrome in Chinese women

To assess the value of ovarian volume and follicle number in the diagnosis of polycystic ovary syndrome (PCOS) in a Chinese population.

Oral oestradiol supplementation as luteal support in IVF/ICSI cycles: a prospective, randomized controlled study.

OBJECTIVEnTo explore whether oral oestradiol (E2) supplementation (6 mg) in the luteal phase is beneficial to the outcome of patients undergoing gonadotrophin-releasing hormone agonist (GnRHa) long protocol in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) cycles.nnnSTUDY DESIGNnProspective, randomized, controlled study at the IVF Clinic, Sun Yat-sen Memorial Hospital. In total, 402 patients with an indication for IVF or ICSI were recruited. Patients were prospectively randomized to receive either progesterone injection plus oral E2 supplementation (Group A, n=202) or progesterone injection alone (Group B, n=200) as luteal support after oocyte retrieval. The main outcome measure was the clinical pregnancy rate.nnnRESULTSnNo significant difference in the clinical pregnancy rate or miscarriage rate was observed between Group A and Group B (50.9% vs 58.0%, 14.6% vs 11.2%; p>0.05). In different age subgroups (≤35 years and >35 years) all measurements were comparable in patients with or without E2 supplementation, as well as in subgroups with different E2 levels on the day of human chorionic gonadotrophin injection (E2≥3000 pg/ml and E2<3000 pg/ml).nnnCONCLUSIONnAdding E2 as luteal support did not increase the clinical pregnancy rate or reduce the miscarriage rate. Routine use of a combination of E2 and progesterone as luteal support in GnRHa long protocol IVF/ICSI cycles is not recommended.

Dichorionic quadramniotic quadruple gestation with monochorionic triamniotic triplets after two embryos transfer and selective reduction to twin pregnancy: case report.

OBJECTIVEnTo report a dichorionic quadramniotic quadruple gestation with monochorionic triamniotic triplets after intracytoplasmic sperm injection (ICSI) treatment and transfer of two embryos.nnnDESIGNnCase report.nnnSETTINGnAcademic reproductive medicine center.nnnPATIENT(S)nA 28-year-old nulligravida who underwent first ICSI cycle.nnnINTERVENTION(S)nAfter 4 years of primary infertility for andrologic subfertility, the patient underwent her first ICSI cycle, with transfer of two fresh embryos on day 3.nnnMAIN OUTCOME MEASURE(S)nTransvaginal sonogram performed at 5 weeks after embryo transfer showed a dichorionic quadramniotic quadruple gestation with monochorionic triamniotic triplets and a singleton.nnnRESULT(S)nAfter extensive counseling, the couple decided to proceed to a selective reduction of two of monochorionic triamniotic triplets and a twin pregnancy remained. Fetal reduction was successfully performed by intracardiac puncture and aspiration at 5 + 3/7 weeks after embryo transfer. Two healthy baby girls were born.nnnCONCLUSION(S)nSeveral factors have been suggested to explain the increase in monozygotic multiple pregnancies after assisted reproductive technique (ART): advanced maternal age, superovulation, and manipulation of the zona pellucida (ZP) including ICSI and assisted hatching, prolonged culture, and in vitro culture condition. All patients should be informed that monozygotic multiple pregnancies occur at a significantly higher rate after ART procedures and could be an important complication after IVF-ICSI treatment.

Clinical outcome of one-third-dose depot triptorelin is the same as half-dose depot triptorelin in the long protocol of controlled ovarian stimulation

OBJECTIVE: Appropriate dosage of the long-acting depot gonadotrophin releasing hormone (GnRH) agonist has not been determined in long protocol for IVF, and one-third-dose depot triptorelin was compared with half-dose in a luteal long protocol of in-vitro fertilization/ intra cytoplasmic sperm injection (IVF/ICSI) treatment in this study. MATERIALS AND METHODS: This is a prospective, randomized, open clinical trial. 100 patients were randomized into two groups. Group I received one-third-dose (1.25 mg) depot triptorelin. Group II received half-dose (1.87 mg). The clinical and experimental parameters were compared between the two groups. RESULTS: There was no premature luteinizing hormone (LH) surge in both groups. On Day 3–5 of menstrual cycle after down-regulation, fewer patients showed low-level LH (<1.0 IU/L) and estradiol (<30 pg/mL) in group I (P <0.05). There were fewer oocytes retrieved (P =0.086), fewer total embryos and available embryos for cryopreservation in Group I (P <0.05), while good-quality embryo rate was higher in group I (P <0.05). The length and dose of ovarian stimulation was lower in Group I, but not significantly. The clinical pregnancy (52% versus 40%), implantation (48% versus 37.5%), delivery (46% versus 32%), or live birth (42% versus 32%) rates and the abortion (8% versus 20%) rates showed no significant differences. CONCLUSION: Depot triptorelin 1.25 mg can be successfully used with reduced pituitary suppression and lower cost in a long protocol for in-vitro fertilization.

Triggering final oocyte maturation with reduced doses of hCG in IVF/ICSI: a prospective, randomized and controlled study

OBJECTIVEnTo compare the effectiveness of urinary human chorionic gonadotropin (u-hCG) at reduced doses of 4000 IU and 6000 IU in inducing final oocyte maturation during in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) cycles.nnnSTUDY DESIGNn164 patients with an indication for IVF or ICSI recruited in this randomized, single-blinded and controlled study in IVF clinic at the Sun Yat-sen Memorial Hospital. Patients were prospectively randomized to receive 4000 IU (Group A, n=83) and 6000 IU (Group B, n=81) of hCG for triggering final oocyte maturation. Number or percentage of mature oocytes retrieved per patient, fertilization rates, pregnancy rates were the main outcome measures.nnnRESULTSnNo evidence of statistically significant difference in the number or proportion of mature oocytes retrieved was observed in both groups. The lower fertilization rate and significantly lower clinical pregnancy rate were observed in Group A. The ovarian hyperstimulation syndrome (OHSS) rates in both groups were also similar. In the subgroup of BMI< 20 kg/m(2), fertilization rate were significantly higher in the administration group of hCG at the dose of 6000 IU when compared with the dose of 4000 IU (82.40% vs. 70.92%, P=0.017); in contrast, no significant difference in clinical pregnancy rates was observed in both groups. In the subgroup of BMI 20-25 kg/m(2), clinical pregnancy rates were significantly higher in patients treated with hCG at dose of 6000 IU than patients treated with hCG at dose of 4000 IU (65.3% vs. 35.0%, P=0.004); however, no significant difference in fertilization rates was observed.nnnCONCLUSIONnBoth doses of u-hCG revealed an equal effect on the induction of final oocyte maturation in the patients with moderate or high ovarian response; however, the reduced dose of hCG could result in an obvious impact on clinical pregnancy rates and did not exhibit an obvious effect on OHSS rates.

Vasoactive Intestinal Peptide Can Promote the Development of Neonatal Rat Primordial Follicles During In Vitro Culture

ABSTRACT Recruitment of primordial follicles is essential for female fertility. Some of the intraovarian growth factors involved in the initiation of primordial follicle growth have been identified, but the exact mechanisms regulating follicle activation are poorly understood. Strong evidence indicates that vasoactive intestinal peptide (VIP), a neuropeptide found in ovarian nerves, plays a role in the physiology of follicle development and function. The aim of the present study was to determine whether VIP might regulate the activation and growth of neonatal rat primordial follicles in an in vitro culture system. Ovaries from 4-day-old rats were cultured for 14 days in medium containing 10−7 M VIP. At the end of the culture, the developmental stages and viability of the follicles were evaluated using histological sections. Immunohistochemistry studies for proliferating cell nuclear antigen (PCNA) were performed to assess the mitotic activity of granulosa cells. In addition, the expression level of kit ligand (KL) mRNA was examined after culture. Histology showed that primordial follicles could survive and start to grow in vitro. The proportion of primordial follicles was decreased and the proportion of early primary follicles increased after in vitro culture with VIP. Immunolocalization of PCNA showed that follicle growth was initiated after VIP treatment. The expression level of KL mRNA was increased in the VIP treatment group. Thus, VIP can promote primordial follicle development, possibly mediated in part through upregulating the expression of KL.

Endometriosis fertility index score maybe more accurate for predicting the outcomes of in vitro fertilisation than r-AFS classification in women with endometriosis

BackgroundEndometriosis is a common disease. The most widely used staging system of endometriosis is the revised American Fertility Society classification (r-AFS classification) which has limited predictive ability for pregnancy after surgery. The endometriosis fertility index (EFI) is used to predict fecundity after endometriosis surgery. This diagnostic accuracy study was designed to compare the predictive value of the EFI with that of the r-AFS classification for IVF outcomes in patients with endometriosis.Methods199 women with endometriosis receiving IVF treatment after surgery were analysis. The EFI score and r-AFS classification in their ability to predict these IVF outcomes were compared in the same population. ROC curves were used to analyse the predictive values of the EFI and r-AFS indices for clinical pregnancy, and their accuracies were evaluated by sensitivity, specificity, and the Youden’s index.ResultsThe Area Under the Curve (AUC) of the EFI score (AUCu2009=u20090.641, Standard Error(SE)u2009=u20090.039, Pu2009=u20090.001, 95% CIu2009=u20090.564-0.717, cut-off scoreu2009=u20096) was significantly larger than that of the r-AFS classification (AUCu2009=u20090.445, SEu2009=u20090.041, Pu2009=u20090.184, and 95% CIu2009=u20090.364-0.526). The antral follicle count, oestradiol level on day of hCG, number of oocytes retrieved, number of oocytes fertilised, and number of cleaved embryos in the greater than or equal to 6 EFI score group was greater than that of the lower than or equal to 5 EFI score group, and the dose of gonadotropin of the greater than or equal to 6 EFI score group were less than that in the lower than or equal to 5 EFI score group. Implantation rate, clinical pregnancy rate, and cumulative pregnancy rate in the greater than or equal to 6 EFI score group were higher than in the lower than or equal to 5 EFI score group.ConclusionsIt suggests that the EFI has more predictive power for IVF outcomes in endometriosis patients than the r-AFS classification. The clinical pregnancy rate was higher in patients with EFI greater than or equal to 6 score than with EFI lower than or equal to 5 score.