A modified flexible GnRH antagonist protocol using antagonist early cessation and a gonadotropin step-down approach improves live birth rates in fresh cycles: a randomized controlled trial.

STUDY QUESTION: Can pregnancy outcomes following fresh elective single embryo transfer (eSET) in gonadotropin-releasing hormone (GnRH) antagonist protocols increase using a gonadotropin (Gn) step-down approach with cessation of GnRH antagonist on the day of hCG administration (hCG day) in patients with normal ovarian response? SUMMARY ANSWER: The modified GnRH antagonist protocol using the Gn step-down approach and cessation of GnRH antagonist on the hCG day is effective in improving live birth rates (LBRs) per fresh eSET cycle. WHAT IS KNOWN ALREADY: Currently, there is no consensus on optimal GnRH antagonist regimens. Studies have shown that fresh GnRH antagonist cycles result in poorer pregnancy outcomes than the long GnRH agonist (GnRHa) protocol. Endometrial receptivity is a key factor that contributes to this phenomenon. STUDY DESIGN, SIZE, DURATION: An open label randomized controlled trial (RCT) was performed between November 2021 and August 2022. There were 546 patients allocated to either the modified GnRH antagonist or the conventional antagonist protocol at a 1:1 ratio. PARTICIPANTS/MATERIALS, SETTING, METHODS: Both IVF and ICSI cycles were included, and the sperm samples used were either fresh or frozen from the partner, or from frozen donor ejaculates. The primary outcome was the LBRs per fresh SET cycle. Secondary outcomes included rates of implantation, clinical and ongoing pregnancy, miscarriage, and ovarian hyperstimulation syndrome (OHSS), as well as clinical outcomes of ovarian stimulation. MAIN RESULTS AND THE ROLE OF CHANCE: Baseline demographic features were not significantly different between the two ovarian stimulation groups. However, in the intention-to-treat (ITT) population, the LBRs in the modified antagonist group were significantly higher than in the conventional group (38.1% [104/273] vs. 27.5% [75/273], relative risk 1.39 [95% CI, 1.09-1.77], P = 0.008). Using a per-protocol (PP) analysis which included all the patients who received an embryo transfer, the LBRs in the modified antagonist group were also significantly higher than in the conventional group (48.6% [103/212] vs. 36.8% [74/201], relative risk 1.32 [95% CI, 1.05-1.66], P = 0.016). The modified antagonist group achieved significantly higher implantation rates, and clinical and ongoing pregnancy rates than the conventional group in both the ITT and PP analyses (P < 0.05). The two groups did not show significant differences between the number of oocytes retrieved or mature oocytes, two-pronuclear zygote (2PN) rates, the number of embryos obtained, blastocyst progression and good-quality embryo rates, early miscarriage rates, or OHSS incidence rates (P > 0.05). LIMITATIONS, REASONS FOR CAUTION: A limitation of our study was that the subjects were not blinded to the treatment allocation in the RCT trial. Only women under 40 years of age who had a good prognosis were included in the analysis. Therefore, use of the modified antagonist protocol in older patients with a low ovarian reserve remains to be investigated. In addition, the sample size for Day 5 elective SET was small, so larger trials will be required to strengthen these findings. WIDER IMPLICATIONS OF THE FINDINGS: The modified GnRH antagonist protocol using the Gn step-down approach and cessation of GnRH antagonist on hCG day improved the LBRs per fresh eSET cycle in normal responders. STUDY FUNDING/COMPETING INTEREST(S): This project was funded by grant 2022YFC2702503 from the National Key Research & Development Program of China and grant 2021140 from the Beijing Health Promotion Association. The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER: The RCT was registered in the Chinese Clinical Trial Registry; Study Number: ChiCTR2100053453. TRIAL REGISTRATION DATE: 21 November 2021. DATE OF FIRST PATIENT'S ENROLLMENT: 23 November 2021.

Detection of chromosome aberrations in 17 054 individuals with fertility problems and their subsequent assisted reproductive technology treatments in Central China.

STUDY QUESTION: How do the types and frequency of chromosome aberrations in couples in central China affect fertility and ART treatment? SUMMARY ANSWER: Men with chromosome aberrations or polymorphisms have an increased risk of semen quality impairment and infertility, and couples affected by reciprocal translocations had a lower pregnancy rate compared with other chromosome aberrations. WHAT IS KNOWN ALREADY: Karyotyping is crucial for patients affected by infertility as chromosome aberrations play an important role in the etiology of male infertility. However, the influence of chromosome aberrations and polymorphisms on sperm motility and morphology remains controversial. Data on ART treatment outcomes in infertile couples affected by chromosome aberrations are insufficient. STUDY DESIGN, SIZE, DURATION: We conducted a retrospective study involving 17 054 patients affected by infertility who underwent karyotyping in our center between January 2020 and May 2022. PARTICIPANTS/MATERIALS, SETTING, METHODS: Karyotyping was performed on 17 054 patients with reproductive failure. All patients were from the central regions of China. The following data were collected from a medical records system using patient identification numbers: couples' ages, history of pregnancy and childbirth, type of infertility, years of infertility, cause of infertility, chromosome karyotypes, semen analysis results, assisted reproductive techniques performed, and treatment outcomes of ART. MAIN RESULTS AND THE ROLE OF CHANCE: The incidence of chromosome aberrations was 2.04%; 2.49% in men and 1.57% in women. By analyzing the relationships between chromosome aberrations/polymorphisms and abnormal semen parameters, we found that there were significantly higher rates of asthenozoospermia, oligospermia, and teratozoospermia among men with Robertsonian translocations and sex chromosomal structural aberrations compared with those with normal karyotypes. Higher rates of asthenozoospermia and teratozoospermia were also observed among men with autosomal reciprocal translocations. The incidence of chromosome aberrations in azoospermic men (13.75%), and in men with cryptozoospermia or severe oligospermia (6.97%) was significantly higher than that in men with mild oligospermia or normospermia (0.88-2.12%). In addition, we found that the progressive movement of sperm is impaired in men with Chromosome 21 polymorphisms compared with men with normal karyotypes (39.46% ± 20.51% vs 48.61% ± 18.76%, P = 0.026). The percentage of morphologically normal forms was lower in the chromosomal polymorphism group than in the normal karyotype group (5.01% ± 2.41% vs 5.59% ± 2.14%, P = 0.001), especially in men with polymorphisms on Chromosome 9 (enlarged Chromosome 9 heterochromatin [9qh+]: 4.48% ± 2.22% vs 5.59% ± 2.14%, P = 0.006; pericentric inversion of Chromosome 9 [inv(9)]: 5.09% ± 3.11% vs 5.59% ± 2.14%, P = 0.008). ART treatment was successful in 36.00% of couples affected by chromosome aberrations. However, couples affected by reciprocal translocations achieved a lower pregnancy rate (24.07%), which may be due to the lower euploidy rates (27.31%) when compared with that in other chromosome aberrations. LIMITATIONS, REASONS FOR CAUTION: First, although the initial cohort was large, chromosome aberrations were identified in a small number of patients. Second, the observational nature of the study design is limiting. Third, the couples affected by infertility in this study were all outpatients that did not undergo identical comprehensive examinations except for karyotyping, leading to the incomplete collection of medical records. Also, the population included in this study mainly focused on couples affected by infertility, which may not be included in the European Association of Urology (EAU) recommendation on male infertility. WIDER IMPLICATIONS OF THE FINDINGS: Men with chromosome aberrations or polymorphisms have an increased risk of semen quality impairment and infertility. Constitutional chromosome analysis is recommended for men affected by infertility and severe oligospermia or azoospermia to facilitate early and appropriate guidance for the most suitable treatment. Carriers of chromosome aberrations can achieve acceptable pregnancy outcomes through IVF. However, couples affected by reciprocal translocations have lower pregnancy rates, and more treatment cycles are needed before a successful pregnancy. A possible explanation may be the fewer euploid embryos obtained. STUDY FUNDING/COMPETING INTEREST(S): This work was supported by Grant 2021YFC2700603 from the National Key Research & Development Program of China. The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.

Clinical and genetic analysis of cytochrome P450 oxidoreductase (POR) deficiency in a female and the analysis of a novel POR intron mutation causing alternative mRNA splicing : Overall analysis of a female with POR deficiency.

OBJECTIVE: To characterize the clinical features of a female with P450 oxidoreductase (POR) deficiency and to investigate the underlying mechanisms of POR inactivation. METHODS: The proband was a 35-year-old woman with primary infertility and menstrual irregularity. The reproductive endocrine profile was evaluated. DNA sequencing was conducted for the identification of POR gene mutation. RT-PCR was performed to confirm the impact of the mutation on POR mRNA. A molecular model was built for the structural analysis of mutant POR protein. RESULTS: The evaluation of reproductive endocrine profile revealed elevation of serum follicle-stimulating hormone (11.48 mIU/ml), progesterone (11.00 ng/ml), 17α-hydroxyprogesterone (24.24 nmol/l), dehydroepiandrosterone (6300 nmol/l), and androstenedione (3.89 nmol/l) and decreased estradiol (36.02 pg/ml). Sequencing of the POR gene showed the female was a compound heterozygote of the paternal P399_E401 deletion and a novel maternal IVS14-1G>C mutation. Functional analysis revealed IVS14-1G>C mutation caused alternative splicing of POR mRNA, with the loss of 12 nucleotides in exon 15 (c.1898_1909delGTCTACGTCCAG). Also, the resulting mutant POR protein had a V603_Q606 deletion, which inactivated the nucleotide-binding domain of NADPH in POR protein (K602_Q606). CONCLUSION: The mutation IVS14-1G>C of the POR gene could cause alternative splicing of POR mRNA and dysfunction of the resulting POR protein. Under proper IVF strategy with glucocorticoid therapy and endometrial preparation, females with mild POR deficiency still have the opportunity to have a live birth.

The depot GnRH agonist protocol improves the live birth rate per fresh embryo transfer cycle, but not the cumulative live birth rate in normal responders: a randomized controlled trial and molecular mechanism study.

STUDY QUESTION: Do cumulative live birth rates (CLBRs) after one complete ART cycle differ between the three commonly used controlled ovarian stimulation (COS) protocols (GnRH antagonist, depot GnRHa (GnRH agonist) and long GnRHa) in normal responders undergoing IVF/ICSI? SUMMARY ANSWER: There were similar CLBRs between the GnRH antagonist, depot GnRHa and long GnRHa protocols. WHAT IS KNOWN ALREADY: There is no consensus on which COS protocol is the most optimal in women with normal ovarian response. The CLBR provides the final success rate after one complete ART cycle, including the fresh and all subsequent frozen-thawed embryo transfer (ET) cycles. We suggest that the CLBR measure would allow for better comparisons between the different treatment protocols. STUDY DESIGN, SIZE, DURATION: A prospective controlled, randomized, open label trial was performed between May 2016 and May 2017. A total of 819 patients were allocated to the GnRH antagonist, depot GnRHa or long GnRHa protocol in a 1:1:1 ratio. The minimum follow-up time from the first IVF cycle was 2 years. To further investigate the potential effect of COS with the GnRH antagonist, depot GnRHa or long GnRHa protocol on endometrial receptivity, the expression of homeobox A10 (HOXA10), myeloid ecotropic viral integration site 1 (MEIS1) and leukemia inhibitory factor (LIF) endometrial receptivity markers was evaluated in endometrial tissue from patients treated with the different COS protocols. PARTICIPANTS/MATERIALS, SETTING, METHODS: Infertile women with normal ovarian response (n = 819) undergoing IVF/ICSI treatment were randomized to the GnRH antagonist, depot GnRHa or long GnRHa protocol. Both IVF and ICSI cycles were included, and the sperm samples used were either fresh or frozen partner ejaculates or frozen donor ejaculates. The primary outcome was the live birth rate (LBR) per fresh ET cycle, and the CLBR after one complete ART cycle, until the birth of a first child (after 28 weeks) or until all frozen embryos were used, whichever occurred first. Pipelle endometrial biopsies from 34 female patients were obtained on Days 7-8 after oocyte retrieval or spontaneous ovulation in natural cycles, respectively, and HOXA10, MEIS1 and LIF mRNA and protein expression levels in the human endometrium was determined by quantitative real-time PCR and western blot, respectively. MAIN RESULTS AND THE ROLE OF CHANCE: There were no significant differences in CLBRs between the GnRH antagonist, depot GnRHa or long GnRHa protocol (71.4 versus 75.5 versus 72.2%, respectively). However, there was a significantly higher LBR per fresh ET cycle in the depot GnRHa protocol than in the long GnRHa and GnRH antagonist protocols (62.6 versus 52.1% versus 45.6%, P < 0.05). Furthermore, HOXA10, MEIS1 and LIF mRNA and protein expression in endometrium all showed significantly higher in the depot GnRHa protocol than in the long GnRHa and GnRH antagonist protocols (P < 0.05). LIMITATIONS, REASONS FOR CAUTION: A limitation of our study was that both our clinicians and patients were not blinded to the randomization for the randomized controlled trial (RCT). An inclusion criterion for the current retrospective cohort study was based on the 'actual ovarian response' during COS treatment, while the included population for the RCT was 'expected normal responders' based on maternal age and ovarian reserve test. In addition, the analysis was restricted to patients under 40 years of age undergoing their first IVF cycle. Furthermore, the endometrial tissue was collected from patients who cancelled the fresh ET, which may include some patients at risk for ovarian hyperstimulation syndrome, however only patients with 4-19 oocytes retrieved were included in the molecular study. WIDER IMPLICATIONS OF THE FINDINGS: The depot GnRH agonist protocol improves the live birth rate per fresh ET cycle, but not the cumulative live birth rate in normal responders. A possible explanation for the improved LBR after fresh ET in the depot GnRHa protocol could be molecular signalling at the level of endometrial receptivity. STUDY FUNDING/COMPETING INTEREST(S): This project was funded by Grant 81571439 from the National Natural Sciences Foundation of China and Grant 2016YFC1000206-5 from the National Key Research & Development Program of China. The authors declare no conflict of interest. TRIAL REGISTRATION NUMBER: The RCT trial was registered at the Chinese Clinical Trial Registry, Study Number: ChiCTR-INR-16008220. TRIAL REGISTRATION DATE: 5 April 2016. DATE OF FIRST PATIENT'S ENROLLMENT: 12 May 2016.

Association between TSH Level and Pregnancy Outcomes in Euthyroid Women Undergoing IVF/ICSI: A Retrospective Study and Meta-analysis.

The aim of this study was to determine the association between thyroid-stimulating hormone (TSH) level and pregnancy outcomes in euthyroid women undergoing in vitro fertilization (IVF)/intra-cytoplasmic sperm injection (ICSI). A total of 1185 women were enrolled in the retrospective study, and 12 studies with a total of 6624 women were included in the meta-analysis (including the data of the present retrospective study). Participants in the retrospective study were divided into two groups in terms of their serum TSH levels: TSH ≤2.5 mIU/L group (n=830) and TSH >2.5 mIU/L group (n=355). They were monitored for the status of clinical pregnancy or miscarriage. In the TSH ≤2.5 mIU/L group, 441 (53.1%) women achieved clinical pregnancy, while 48 (5.8%) had early pregnancy loss and 12 (1.4%) had ectopic pregnancy. In the TSH >2.5 mIU/L group, 175 (49.3%) women achieved clinical pregnancy, while 21 (5.9%) had early pregnancy loss and 3 (0.8%) had ectopic pregnancy. No significant differences were observed between the two groups in pregnancy outcomes (P=0.126, P=0.512, P=0.297). The meta-analysis also revealed no significant difference in the clinical pregnancy rate and the miscarriage rate between women with serum TSH ≤2.5 mIU/L and those with serum TSH >2.5 mIU/L. In conclusion, high TSH levels (TSH level >2.5 mIU/L) did not affect clinical pregnancy rate or increase miscarriage rate in euthyroid women undergoing IVF/ICSI.

Developmental Potential and Clinical Value of Embryos with Abnormal Cleavage Rate.

To investigate the developmental potential and clinical value of embryos with abnormal cleavage rate, a retrospective analysis was performed on 66 635 2-prokaryotic (2PN) and 1-pronuclear (1PN) embryos. The embryos were given conventionally in vitro fertilization (IVF) treatment and continuously cultured on the day 3 (D3) at the Reproductive Medicine Center, Tongji Medical College, Huazhong University of Science and Technology from January 2016 to December 2017. The embryos were separated into the day-2 (D2) undivided group with 106 cases, the arrested development group with 3482 cases, the blastomere reduction group with 541 cases, and the control group with 62 506 cases, respectively. The blastocyst utilization rates of these three abnormal groups were 2.83%, 10.86% and 6.84%, respectively, which were significantly different from that in control group (39.46%). Furthermore, 2 cases of anabiosis and 1 case of live birth were found in D2 undivided group. In arrested development group, there were 55 cases of anabiosis, 11 cases of clinical pregnancy in single-embryo transplantation (including 6 cases of live birth), and 25 cases of clinical pregnancy in combination with one normal embryo transplantation (including 23 cases of live births and 15 cases of dizygotic twins under B-ultrasound). There were 13 case of anabiosis in blastomere reduction group: there was 1 case of single embryo transplantation and clinical pregnancy was obtained; there were also 6 cases of clinical pregnancy in combination with one single normal embryo transplantation (including 5 cases of live births and 2 cases of dizygotic twins under B-ultrasound). In conclusion, embryos with abnormal cleavage rate still have the potential to continue to develop, and have certain blastocyst utilization rate and live birth.

Assisted conception does not increase the risk for mother-to-child transmission of hepatitis B virus, compared with natural conception: a prospective cohort study.

OBJECTIVE: To determine whether assisted conception increases the risk for mother-to-child transmission of hepatitis B virus (HBV) infection compared with natural conception. DESIGN: Prospective cohort study. SETTING: Research laboratory. PATIENT(S): A total of 305 children, 176 born with assisted conception and 129 born with natural conception, were born to a total of 251 hepatitis B surface antigen- (HBsAg-) positive women. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): The seropositive rates of HBsAg in children at birth and HBV infection rates at 9-15 months of age. RESULT(S): Overall, 7.5% (23/305) of children were HBsAg-positive at birth. The rate of HBsAg-positive children at birth did not significantly differ between children in the assisted conception group compared with those in the natural conception group (6.3% [11/176] vs. 9.3% [12/129]). Multivariate logistic regression analysis showed that conception method is not related to the rate of HBsAg-positive children at birth. All children who were positive for HBsAg at birth and were followed up for 9-15 months became negative for HBsAg after hepatitis B immunization. CONCLUSION(S): Assisted conception does not increase the risk for mother-to-child transmission of HBV compared with natural conception.

Cumulative live birth rates in more than 3,000 patients with poor ovarian response: a 15-year survey of final in vitro fertilization outcome.

OBJECTIVE: To estimate the cumulative live birth rates (CLBRs) in women with poor ovarian response (POR) diagnosed according to the Bologna criteria. DESIGN: A 15-year population-based observational cohort study. SETTING: Teaching hospital. PATIENT(S): Between 2002 and 2016 a total of 3,391 women with POR were followed from their first fresh, nondonor IVF cycle until they had a live birth or discontinued treatment. All IVF and intracytoplasmic sperm injection (ICSI) cycles and cryocycles were included. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Live birth rates per initiated cycle, the conservative and optimistic CLBR for multiple IVF cycles. RESULT(S): The CLBRs after six IVF cycles were 14.9% for the conservative and 35.3% for the optimistic estimate. The CLBR decreased from 22% for women ≤30 years to 18.3% for women aged 31-34 years, 17.2% for 35-37 years, 13.5% for 38-40 years, 10.5% for 41-43 years, and 4.4% among women >43 years in the conservative analysis. There was a significant decreased CLBR starting at age 38 years compared with women <35 years. After adjusting for age, antral follicle count, basal FSH level, and IVF cycle number, natural cycles were associated with the lowest CLBR among all the protocols, and this difference was significant compared with the other protocols. CONCLUSION(S): For women with POR, the CLBR declined with increasing age. Women with advanced age (≥38 years) achieved a significantly lower CLBR than young poor responders (<35 years). Very low CLBR was associated with women aged >43 years old. Natural cycle IVF is of no benefit for these patients.

Combined analysis of endometrial thickness and pattern in predicting clinical outcomes of frozen embryo transfer cycles with morphological good-quality blastocyst: A retrospective cohort study.

To evaluate the combined effect of endometrial thickness and pattern on clinical outcomes in females following in vitro fertilization/intracytoplasmic sperm injection and frozen-thawed embryo transfer (IVF/ICSI-FET).FET cycles using at least 1 morphological good-quality blastocyst conducted between 2012 and 2013 at a university-based reproductive center were reviewed retrospectively. Endometrial ultrasonographic characteristics were recorded on the day of progesterone supplementation in FET cycles. In the combined analysis, endometrial thickness groups (group 1: equal or < 8 mm; group 2: >8 mm) were subdivided into 2 endometrial patterns (pattern A: triple-line; pattern B: no-triple line). Clinical pregnancy rate, spontaneous abortion rate, and live birth rate in different groups were analyzed.A total of 1512 cycles were reviewed. The results showed that significant differences in endometrial thickness and pattern were observed between the pregnant group (n = 1009) and no pregnant group (n = 503) (P < .05), while no significant differences were found between the live birth group (n = 844) and no live birth group (n = 668). Combined analysis revealed those with endometrial thickness > 8 mm and triple-line endometrial pattern had significant higher clinical pregnancy rates, while spontaneous abortion rates and live birth rates showed no significant differences among these subgroups.This study suggested neither individual nor combined analysis of endometrial thickness and pattern had predicting effects on live birth following IVF treatments, and embryo quality might be the one that really has effects.

Endometrial thickness as a predictor of the reproductive outcomes in fresh and frozen embryo transfer cycles: A retrospective cohort study of 1512 IVF cycles with morphologically good-quality blastocyst.

To evaluate the relationship between endometrial thickness during fresh in vitro fertilization (IVF) cycles and the clinical outcomes of subsequent frozen embryo transfer (FET) cycles.FET cycles using at least one morphological good-quality blastocyst conducted between 2012 and 2013 at a university-based reproductive center were reviewed retrospectively. Endometrial ultrasonographic characteristics were recorded both on the oocyte retrieval day and on the day of progesterone supplementation in FET cycles. Clinical pregnancy rate, spontaneous abortion rate, and live birth rate were analyzed.One thousand five hundred twelve FET cycles was included. The results showed that significant difference in endometrial thickness on day of oocyte retrieval (P = .03) was observed between the live birth group (n = 844) and no live birth group (n = 668), while no significant difference in FET endometrial thickness was found (P = .261) between the live birth group and no live birth group. For endometrial thickness on oocyte retrieval day, clinical pregnancy rate ranged from 50.0% among patients with an endometrial thickness of ≤6 mm to 84.2% among patients with an endometrial thickness of >16 mm, with live birth rate from 33.3% to 63.2%. Multiple logistic regression analysis of factors related to live birth indicated endometrial thickness on oocyte retrieval day was associated with improved live birth rate (OR was 1.069, 95% CI: 1.011-1.130, P = .019), while FET endometrial thickness did not contribute significantly to pregnancy outcomes following FET cycles. The ROC curves revealed the cut-off points of endometrial thickness on oocyte retrieval day was 8.75 mm for live birth.Endometrial thickness during fresh IVF cycles was a better predictor of endometrial receptivity in subsequent FET cycles than FET cycle endometrial thickness. For those females with thin endometrium in fresh cycles, additional estradiol stimulation might be helpful for adequate endometrial development.

Oxidative stress in granulosa cells contributes to poor oocyte quality and IVF-ET outcomes in women with polycystic ovary syndrome.

The increased levels of intracellular reactive oxygen species (ROS) in granulosa cells (GCs) may affect the pregnancy results in women with polycystic ovary syndrome (PCOS). In this study, we compared the in vitro fertilization and embryo transfer (IVF-ET) results of 22 patients with PCOS and 25 patients with tubal factor infertility and detected the ROS levels in the GCs of these two groups. Results showed that the PCOS group had significantly larger follicles on the administration day for human chorionic gonadotropin than the tubal factor group (P < 0.05); however, the number of retrieved oocytes was not significantly different between the two groups (P > 0.05). PCOS group had slightly lower fertilization, cleavage, grade I/II embryo, clinical pregnancy, and implantation rates and higher miscarriage rate than the tubal factor group (P > 0.05). We further found a significantly higher ROS level of GCs in the PCOS group than in the tubal factor group (P < 0.05). The increased ROS levels in GCs caused GC apoptosis, whereas NADPH oxidase 2 (NOX2) specific inhibitors (diphenyleneiodonium and apocynin) significantly reduced the ROS production in the PCOS group. In conclusion, the increased ROS expression levels in PCOS GCs greatly induced cell apoptosis, which further affected the oocyte quality and reduced the positive IVF-ET pregnancy results of women with PCOS. NADPH oxidase pathway may be involved in the mechanism of ROS production in GCs of women with PCOS.

Hepatitis B surface antigen in oocytes and embryos may not result in vertical transmission to offspring of hepatitis B virus carriers.

OBJECTIVE: To investigate the vertical transmission of hepatitis B virus (HBV) via embryos to children and whether HBV in embryos has an effect on the development of the fetus and pregnancy outcomes of in vitro fertilization and embryo transfer (IVF-ET). DESIGN: Long-term follow-up study. SETTING: Research laboratory. PATIENT(S): Thirty-one couples with a hepatitis B surface antigen (HBsAg)-negative woman and HBsAg-positive man, and 41 couples with a HBsAg-positive woman and HBsAg-negative man, whose unfertilized oocytes and nonviable embryos were tested for HBV DNA, RNA, or HBsAg. INTERVENTION(S): HBV DNA, RNA, or HBsAg analyses in unfertilized oocytes and nonviable embryos. MAIN OUTCOME MEASURE(S): HBV serologic markers analyses. RESULT(S): We obtained follow-up data for 71 couples. A total of 24 babies were born, and no newborns exhibited defects at birth. Twelve babies were born to couples with HBV-positive oocytes and/or embryos. The pregnancy outcomes were not associated with the presence of HBV in oocytes and embryos. Three patterns of HBV serologic markers were screened. Twenty babies were anti-HBs-positive. Three babies were negative for HBsAg, antibody to hepatitis B surface antigen (anti-HBs), antibody to hepatitis B core antigen (anti-HBc), HBeAg, and antibody to hepatitis B e antigen (anti-HBe). One baby was seropositive for anti-HBs, anti-HBc, and anti-HBe at 6 months of age but seroconverted from anti-HBe-positive to anti-HBe-negative at 9 months of age. CONCLUSION(S): The presence of HBsAg in oocytes and embryos may not result in the vertical transmission of HBV in the offspring of HBV carriers.

The criteria for optimal down-regulation with gonadotropin-releasing hormone-agonist: a retrospective cohort study.

OBJECTIVE: The objective of this study is to investigate whether the degree of down-regulation using GnRH-agonists is associated with pregnancy outcomes. STUDY DESIGN: This retrospective analysis was performed on 2708 cycles from 2514 patients undergoing down-regulation with the luteal phase long protocol. The serum oestradiol (E2D) and luteinising hormone (LHD) levels, the diameter of the largest follicle (DLFD) and the endometrial-thickness (ENTD) after down-regulation were used to evaluate the degree of down-regulation. One-way analysis of variance with the Bonferroni adjustment, the chi-square test and multivariate logistic regression analyses were used for the statistical analysis. RESULTS: The cumulative clinical pregnancy rates (CCPR) and the cumulative live birth rates (CLBR) were higher in the cycles with E2D < 30 pg/ml (63.7%, OR = 1.405 (1.055-1.870) and 56.8%, OR = 1.372 (1.039-1.813)) and 30-55pg/ml (66.8%, OR = 1.439 (1.104-1.874) and 59.8%, OR = 1.397 (1.080-1.806)) than in those with E2D > 55 pg/ml (62.8% and 54.7%). There was a trend towards lower CCPRs and CLBRs in the cycles with DLFD > 10 mm or ENTD ≥ 6 mm; however, this difference was not significant. CONCLUSION: The degree of down-regulation is associated with ovarian response, pregnancy, and live birth. We propose the following criteria for optimal down-regulation: E2D 30-55 pg/ml, ENTD < 6 mm, and no apparent ovarian activity.

Neonatal outcomes after early rescue intracytoplasmic sperm injection: an analysis of a 5-year period.

OBJECTIVE: To evaluate the safety and efficacy of early rescue intracytoplasmic sperm injection (ICSI). DESIGN: Retrospective cohort study. SETTING: Teaching hospital. PATIENT(S): There were 13,232 ovarian stimulation cycles (IVF, n = 9,631; ICSI, n = 2,871; early rescue ICSI, n = 730) that resulted in the delivery of 5,001 babies (IVF, n = 3,670; ICSI, n = 1,095; early rescue ICSI, n = 246) from August 2008 to August 2013. INTERVENTION(S): Early rescue ICSI. MAIN OUTCOME MEASURE(S): Clinical pregnancy rates, neonatal outcomes, and congenital birth defects were analyzed. RESULT(S): The early rescue ICSI cycles did not seem to have a negative effect on the clinical pregnancy rate (43.42%) when IVF cycles (45.33%) were compared with ICSI cycles (44.39%). In the early rescue ICSI group, a total of 254 clinical pregnancies were achieved: 197 (33.67%) live births, 38 (6.49%) miscarriages, 2 (0.79%) induced abortions, 3 (1.18%) fetal deaths, and 4 pregnancies (1.57%) without completion at follow-up. Overall, the multiple gestations, the delivery method, mean gestational age, preterm deliveries, mean birth weight, and rate of congenital birth defects of the early rescue ICSI group were similar to those in the conventional IVF and ICSI groups. CONCLUSION(S): Early rescue ICSI had similar clinical pregnancy rates when compared with conventional IVF and ICSI, in addition to the delivery of healthy children. The clinical evidence from the early rescue ICSI group did not show an elevated rate of malformations. Early rescue ICSI seems to be a safe alternative method for individuals with total fertilization failure or near total fertilization failure when compared with conventional IVF treatment.

Is frozen embryo transfer cycle associated with a significantly lower incidence of ectopic pregnancy? An analysis of more than 30,000 cycles.

OBJECTIVE: To analyze the incidence of ectopic pregnancy (EP) in fresh compared with frozen-thawed cycles. DESIGN: Retrospective cohort study. SETTING: Teaching hospital. PATIENT(S): Thirty-one thousand nine hundred twenty-five women undergoing in vitro fertilization-embryo transfer (IVF-ET) from January 2006 to December 2013. INTERVENTION(S): Fresh IVF-ET compared with frozen-thawed ET (FET). MAIN OUTCOME MEASURE(S): Incidence of EP with fresh IVF-ET compared with frozen-thawed ET cycles, clinical pregnancy rate, and rate of EP per clinical pregnancy. RESULT(S): For the fresh IVF cycles, 19,173 patients underwent oocyte retrieval; 15,042 had an ET, 6,431 of these patients (42.7%) had a clinical pregnancy, and among these 297 (1.97%) appeared to have an EP. The group of patients undergoing frozen-thawed ET (12,752 patients) included 12,255; there were 5,564 pregnancies (45.4%) and 124 ectopic implants (1.01%). The incidence of an EP per clinical pregnancy was 4.62% for the fresh transfer group compared with 2.22% for the frozen-thawed cycle group; this difference was statistically significant. In addition, the fresh ET cycles had the highest risk of EP, followed by day-3 embryo FET cycles; blastocyst FET cycles had the lowest risk of EP, and the differences were all statistically significant. CONCLUSION(S): Frozen-thawed ET cycles were associated with a statistically significantly lower risk of EP when compared with fresh cycles. These findings are consistent with ovarian stimulation being associated with an increased risk of EP.

Regulation of myeloid ecotropic viral integration site 1 and its expression in normal and abnormal endometrium.

OBJECTIVE: To identify the expression profile and sex steroid regulation pattern of myeloid ecotropic viral integration site 1 (MEIS1) in endometrium. DESIGN: Molecular studies in human and animal tissue. SETTING: Reproductive medicine center of a university hospital. PATIENT(S) AND ANIMAL(S): Women with normal menstrual cycles for male infertility and female infertility with endometriosis. Sexually mature female mice (Kunming White strain). INTERVENTION(S): Primary cultured endometrial stromal cells, Ishikawa cells, and oophorectomized mice were treated with sex steroid. MAIN OUTCOME MEASURE(S): MEIS1 expression in the human endometrium during the menstrual cycle, mouse uterus during the peri-implantation period of pregnancy, and eutopic endometrium from patients with endometriosis was analyzed by immunohistochemistry staining and western blot. In addition, MEIS1 expression in response to sex steroid was examined both in vitro and in vivo by immunohistochemistry staining and western blot. RESULT(S): MEIS1 expression was markedly increased in endometrium during the implantation period, and in decidualizing stromal cells in human endometrium and murine uterus. Steroid hormones increased MEIS1 expression in primary cultured endometrial stromal cells, Ishikawa cells, and endometrium of oophorectomized mice. The effects of estrogen and progesterone were more marked in oophorectomized mice and were additive. MEIS1 expression was significantly lower in eutopic endometrium compared with normal endometrium in the midsecretory stage. CONCLUSION(S): MEIS1 is likely a key mediator between sex steroid and genes for uterine receptivity. Diminished endometrium MEIS1 expression may contribute to implantation failure in endometriosis.

Effect of dehydroepiandrosterone administration in patients with poor ovarian response according to the Bologna criteria.

BACKGROUND: Dehydroepiandrosterone (DHEA) is now widely used as an adjuvant to IVF treatment protocols in poor responders. However, clinical evidence for DHEA on improvement of ovarian response and IVF outcome is still limited, the validity of the results of the earlier studies, especially the varied inclusion criteria, is a subject of debate. Recently, the ESHRE Working Group developed a new definition, the Bologna criteria. The aim of the current study was to investigate the potential effect of DHEA treatment on in vitro fertilization (IVF) outcome of poor ovarian responders that fulfill the Bologna criteria. METHODS: This study investigated 386 poor ovarian responders that fulfill the Bologna criteria. Patients underwent IVF-ET treatment with the GnRH antagonist protocol. The study group contained 189 patients, who received 75 mg of DHEA daily (25 mg three times daily) before the IVF cycle. The control group was composed of 197 patients who received infertility treatment, but did not receive DHEA. The IVF outcome parameters in each group were compared. RESULTS: The study and control groups did not show statistically significant differences in terms of patient demographics characteristics, mean numbers of oocytes retrieved, mature oocytes, fertilization rate, cleavage rate, or embryo availability. While the DHEA group demonstrated significantly higher implantation rates (18.7% vs. 10.1%; P<0.01) and ongoing PRs (26.7% vs. 15.8%; P<0.05) as compared with the control. CONCLUSIONS: DHEA pre-treatment does not significantly increase oocyte yield. However, the ongoing PRs in this subgroup of women are significantly higher after DHEA administration, suggesting that DHEA may increase IVF results by improving oocyte and embryo quality.

Progesterone elevation on the day of HCG administration may affect rescue ICSI.

To investigate the relationship between serum progesterone concentration on the day of human chorionic gonadotrophin (HCG) administration and rescue intracytoplasmic sperm injection (ICSI), a total of 9858 patients who underwent IVF or rescue ICSI were retrospectively analysed. The results showed a significant difference in serum progesterone concentration on the day of HCG administration between the IVF group and rescue ICSI group (P < 0.01). Multivariate logistic regression showed that progesterone concentration was positively and significantly associated with rescue ICSI (OR 1.297, 95% CI 1.153-1.460, P < 0.001). Moreover, an increased rescue ICSI rate was associated with progressively higher progesterone concentrations in all cycles. In addition, patients with progesterone >1.5 ng/ml demonstrated a significantly higher rescue ICSI rate compared with patients with progesterone concentration ≤1.5 ng/ml (P < 0.05). In conclusion, elevated progesterone on the day of HCG administration had an adverse effect on oocyte fertilization; thus, greater attention should be paid to these patients in an attempt to avoid fertilization failure, especially when progesterone is >1.50 ng/ml. For the issue of oocytes fertilization, most literatures have found the presence of a negative association between P elevation and fertilization. They suggested that P elevation may only influence the endometrium, leading to impaired endometrial receptivity and had no adverse effect on the fertilization of oocytes. On the contrary, we enrolled 9,858 fresh cycles and found elevated P had an adverse effect on the oocytes fertilization, especially if the P concentration >1.50 ng/mL. It is the first report about the relationship between the rescue ICSI and serum P levels.

Regulation of endometrial receptivity by the highly expressed HOXA9, HOXA11 and HOXD10 HOX-class homeobox genes.

STUDY QUESTION: Are other HOX genes, in addition to HOXA10, involved in endometrial receptivity? SUMMARY ANSWER: The highly expressed HOXA9, HOXA11 and HOXD10 genes also appear to be involved in endometrial receptivity. WHAT IS KNOWN ALREADY: Within the HOX family of homeobox transcription factor genes are the leading candidates for the regulation of embryonic implantation. A crucial role of HOXA10 in endometrial receptivity has been well established. STUDY DESIGN, SIZE, DURATION: To identify HOX candidate genes, we performed data mining on all 39 human HOX genes in the 'Human body index' gene expression database of normal human tissue. The temporal and spatial expression pattern of four highly expressed HOX genes in the human endometrium was determined. To further investigate the function of these Hox genes, we used a robust in vivo mouse model in which we blocked maternal Hox gene expression. PARTICIPANTS/MATERIALS, SETTING AND METHODS: Analysis of a gene expression profile set in the public domain consisting of 504 samples representing 95 different normal human tissues, showed that in addition to HOXA10, also HOXA9, HOXA11, HOXB6 and HOXD10 mRNA showed increased expression in the human endometrium (16 samples). The temporal and spatial expression pattern of these four HOX genes throughout the menstrual cycle was determined in the endometrium from 27 female patients eligible for IVF-embryo transfer with a normal cycle by quantitative real-time PCR (qRT-PCR), western blot and immunohistochemistry. The role of maternal Hoxa9, Hoxa11 and Hoxd10 was assessed in a mouse implantation model by expression knockdown using RNA interference. Forty mice were transfected with Hoxa9-, Hoxa11- or Hoxd10-specific small hairpin RNA (shRNA) constructs or a vector control by injection into the uterine horn at Day 2 after vaginal plug detection (Day 1) (160 mice in total). The effects were examined by qRT-PCR and western blot at Day 4 and litter sizes counted at Day 9 of pregnancy. MAIN RESULTS AND THE ROLE OF CHANCE: HOXA10, HOXA9, HOXA11 and HOXD10 all showed increased expression during the mid-secretory phase of the menstrual cycle (P < 0.01). Knockdown of Hoxa9, Hoxa11 and Hoxd10 in the murine uterus resulted in significantly reduced average implantation rates (P < 0.01) and, with regard to four Hox target genes, also correlated with a significantly increased empty spiracles homolog 2 (Emx2) and insulin-like growth factor binding protein-1 (Igfbp1), and decreased integrin ß3 (Itgb3) and leukemia inhibitory factor (Lif), expression (P < 0.01). LIMITATIONS, REASONS FOR CAUTION: Menstrual cycle stage was not confirmed by serum hormone analysis. We verified the absence of significant differences in stage-specific expression of the reference genes used in our study (ACTB/Actb and GAPDH/Gapdh) and therefore possible limitations of this approach were minimized. In addition, the translatability of our data from a mouse model to patients needs to be investigated further. WIDER IMPLICATIONS OF THE FINDINGS: We provide evidence that three other HOX genes in addition to HOXA10 are involved in endometrial receptivity, and that part of their function is asserted through several known HOX target genes, suggesting the presence of a central HOX signal transduction pathway.

Effect of the site of assisted hatching on vitrified-warmed blastocyst transfer cycles: a prospective randomized study.

PURPOSE: To assess the effect of assisted hatching (AH) site on the clinical outcomes in vitrified-warmed blastocyst transfer cycles. METHODS: A total of 160 women who underwent vitrified-warmed blastocyst transfer cycles were randomized to either the ICM group (AH performing at the site near the inner cell mess, ICM), or the TE group (AH performing at the site opposite to the ICM). AH with laser zona drilling was performed 20 or 30 min after thawing once the ICM can be detected. Clinical pregnancy rate, implantation rate, live birth rate and the occurrence rate of monozygotic twins (MZT) pregnancy after transfer of these two groups were compared. RESULTS: No significant difference was found in the clinical pregnancy rate (63.8% vs. 67.5%), implantation rate (51.7% vs. 53.6%) and live birth rate (57.5% vs. 62.5%) between the ICM group and the TE group. The occurrence rate of MZT was comparable between the two groups (3.9% vs. 5.6%). CONCLUSIONS: The site of assisted hatching has no influence on the implantation, pregnancy and live birth rate in human vitrified-warmed blastocyst transfer cycles.