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Objective: To detect ZP (zona pellucida) gene (ZP1-ZP4) mutations in patients with oocyte anomalies.Design: Case-control genetic study.Setting: University-based reproductive medicine center.Patient(s): A total of 92 infertile patients with repeated cycles of oocyte maturation arrest (group I, n = 49) or oocyte morphologic defect (group II, n = 43) as well as 373 healthy controls.Intervention(s): Genomic DNA extracted from peripheral blood and coding regions of ZP genes amplified by polymerase chain reaction and sequenced by a DNA analyzer.Main Outcome Measure(s): Variant prediction of ZP genes with software.Result(s): In group I with oocyte maturation arrest, no novel variants were found. In group II with oocyte morphologic defects, four novel variants, two in the ZP1 gene [c.247T>C (p.W83R) and c.1413G>A (p.W471X)] and two in the ZP2 gene [c.1599G>T (p.R533S) and c.1696T>C (p.C566R)] were detected in 4 of 43 patients (approximately 9%) but were absent from the controls. Protein alignments showed that the four variants were highly conserved among different species, and all four variants were predicted to be deleterious by gene software predictions.Conclusion(s): ZP gene variants may account for patients with oocyte morphologic abnormalities but not for those with oocyte maturation arrest. [ABSTRACT FROM AUTHOR]

The ricefield eel is becoming an important aquaculture species in China. However, the regulation of oocyte growth and maturation in ricefield eels is poorly understood. In this study, the development of ricefield eel ovarian follicles was classified into nine stages, namely the primary growth/previtellogenic (PG/PV), early vitellogenic (EV), mid-vitellogenic (MV), late vitellogenic (LV), full-grown (GV0), germinal vesicle migration 1 (GV1), germinal vesicle migration 2 (GV2), germinal vesicle migration 3 (GV3), and germinal vesicle breakdown (GVBD) stages, which are based on follicle morphologies and sizes, vitellogenic states, and the positions of germinal vesicles. Comparative transcriptomic analysis was performed on isolated ovarian follicles from the MV to the GVBD stages. A total of 438,410 high-quality reads were obtained after filtering from six cDNA libraries of ovarian follicles at the MV, LV, GV0, GV1, GV2, and GVBD stages, 92.09% of which were perfectly mapped to the ricefield eel genome. Real-time PCR analysis showed that the expression profiles of 20 representative genes related to oocyte growth and maturation largely paralleled with those inferred from transcriptome data, which indicated the reliability of RNA-Seq data for gene expression analysis in the present study. The estradiol biosynthesis pathway was highly activated in ovarian follicles at the LV stage, which may be related to the stimulation of vitellogenin synthesis and/or meiotic arrest. The high expression of the Vtg receptor gene vldlr in ovarian follicles at MV and LV is consistent with active vitellogenesis at these stages. Thirteen cathepsin-encoding genes were highly expressed in ovarian follicles with different expression profiles during development from MV to GVBD, which implied their differential roles in vitellogenin hydrolysis during oocyte growth and maturation. Many cell cycle-related genes showed the lowest expression at GV0 and then increased expression at GV1 and GV2, which indicated that the resumption of meiosis may be initiated at GV0. The highest expression of the adenyl cyclase genes adcy3a , adcy5 , adcy6 , adcy6-like , and adcy7 at MV and/or LV, while that of phosphodiesterase genes pde1c , pde4b , pde4b-like , pde4c , pde7a, pde8b , pde10 , and of the cyclic AMP-dependent transcription factor atf-3 gene observed at GV0 indicated that cAMP signals may play important roles in oocyte meiotic arrest and resumption in ricefield eels. Parallel expression profiles of urok , mmp2 , mmp9 , mmp11 , mmp14a , mmp14b , mmp17a , mmp19 , and mmp25 suggest their potential roles in tissue remodeling during oocyte maturation and ovulation in ricefield eels. Collectively, the data of the present study will aid in further understanding the molecular mechanisms underlying oocyte growth and maturation in ricefield eels as well as in other teleosts. • The oocyte maturation stage was subdivided into GV1, GV2, GV3, and GVBD. • Transcriptomes of ovarian follicles were obtained to clarify molecular mechanisms. • Multiple cathepsins may be involved in vitellogenin hydrolysis in oocytes. • PDEs including Pde4b-like and Pde10 are possibly involved in meiotic resumption. [ABSTRACT FROM AUTHOR]

Study Question: Is a dual ovulation trigger with a combination of GnRH agonist (GnRHa) and hCG superior to single hCG and/or single GnRHa trigger in improving treatment outcomes in advanced-age women (aged ≥ 35 years) undergoing IVF/ICSI treatment?, Summary Answer: Co-administration of GnRHa and hCG as a dual trigger increases the number of good-quality embryos but it is not associated with a higher number of oocytes retrieved, compared with single hCG or GnRHa trigger., What Is Known Already: Many studies have demonstrated that a dual trigger has positive impact on oocyte maturation, retrieval rate and pregnancy rate without increasing the risk of ovarian hyperstimulation syndrome (OHSS) in some groups of IVF patients, when compared with single hCG trigger. Few studies have however been conducted to compare a dual trigger with a single GnRHa trigger, and insufficient evidence exists to support which trigger can achieve the best outcomes in IVF patients aged ≥35 years., Study Design, Size, Duration: This was an open-label randomized controlled trial of 510 participants conducted at single reproductive medical center from January 2019 to December 2021. After a sample size calculation performed by retrospectively analyzing our previous clinical data, we planned to recruit 170 patients in each group and 510 patients in total for the study., Participants/materials, Setting, Methods: Women aged ≥35 years undergoing IVF/ICSI treatment, receiving a non-pituitary down-regulation protocol, and with low risk of OHSS, were enrolled in this trial. On the trigger day, patients were randomized into three groups: hCG alone (who received 6000 IU of hCG), GnRHa alone (who received 0.2 mg of triptorelin) and dual trigger (who received 0.2 mg of triptorelin plus 2000 IU of hCG) groups. The primary outcome parameter was the number of retrieved oocytes. The secondary outcome parameters included, among others, the number and rates of mature oocytes, two pronuclei (2PN) embryos and good-quality embryos, as the rates of OHSS, clinical pregnancy, miscarriage and live birth., Main Results and the Role of Chance: There were no significant differences in the baseline demographic characteristics among the three groups. The dual trigger was associated with a higher retrieval rate (87.9% vs 84.1% in the hCG group, P = 0.031; 87.9% vs 83.6% in the GnRHa group, P = 0.014). However, the number of retrieved oocytes in the dual trigger group was comparable with those in the hCG group (4.08 ± 2.79 vs 3.60 ± 2.71, P = 0.080) and the GnRHa group (4.08 ± 2.79 vs 3.81 ± 3.38, P = 0.101); comparable data between the groups were also found when analyzing the number of 2PN embryos and the 2PN rate. In the dual trigger group, the numbers of good-quality embryos and viable embryos were both significantly higher than in the hCG group (1.74 ± 1.90 vs 1.19 ± 1.45, P = 0.016 and 2.19 ± 2.11 vs 1.56 ± 1.66, P = 0.008, respectively) and the GnRHa group (1.74 ± 1.90 vs 1.20 ± 1.67, P = 0.003 and 2.19 ± 2.11 vs 1.45 ± 1.75, P = 0.001, respectively). Pregnancy outcomes after fresh embryo transfer (ET) were comparable between the groups. The live birth rate and ongoing pregnancy rate after frozen ET in the dual trigger group were significantly higher than those in the GnRHa group (32.6% vs 14.1%, P = 0.007 and 34.8% vs 17.6%, P = 0.013, respectively), but not superior to those in the hCG group (32.6% vs 27.9%, P = 0.537 and 34.8% vs 27.9%, P = 0.358, respectively)., Limitations, Reasons for Caution: Women of advanced age are quite a heterogeneous population and overlap with poor ovarian responders or patients with diminished ovarian reserve. We therefore could not entirely exclude selection biases or confounding factors. This study was also not a double-blinded trial; the patients in the GnRHa and dual trigger groups could have been affected by the placebo effect., Wider Implications of the Findings: The results of this study suggest that in advanced-age women with low risk of OHSS, a dual trigger or even a single hCG trigger may be a better choice than a single GnRHa trigger., Study Funding/competing Interest(s): This study was supported by the Shanghai Municipal Health Commission of Science and Research Fund (20184Y0289). The authors declare no conflict of interest., Trial Registration Number: This trial was registered in the Chinese Clinical Trial Registry (ChiCTR-1800016285)., Trial Registration Date: 24 May 2018., Date of First Patient’s Enrolment: 2 January 2019., (© The Author(s) 2022. Published by Oxford University Press on behalf of European Society of Human Reproduction and Embryology. All rights reserved. For permissions, please email: journals.permissions@oup.com.)

The Chinese mitten crab (Eriocheir sinensis) is an economically important aquaculture species in China, with distinct differences in ovarian maturation status between crabs fed with natural diets and artificial diets during the listing period, thus, leading to selling price differentiation. Our previous study showed that dietary supplementation with 100 mg/kg icariin can effectively promote ovarian development of E. sinensis. However, the internal molecular mechanism has not yet been elucidated because of a lack of comprehensive genome sequence information. We compared the ovary transcriptomes of E. sinensis fed with two diets containing 0 and 100 mg/kg ICA using the BGISEQ-500 platform. This yielded 12.54 Gb clean bases and 54,794 unigenes, 13,832 of which were found to be differentially expressed after icariin exposure. Twenty pathways closely related to gonadal development were selected through KEGG analysis. Seven differentially expressed genes relevant to vitellogenesis and oocyte maturation (serine/threonine-protein kinase mos-like, Eg2, cytoplasmic polyadenylation element-binding protein, cyclin B, vitellogenin 1, cathepsin D, and juvenile hormone esterase-like carboxylesterase 1) were validated by qRT-PCR, and four proteins (MEK1/2, ERK1/2, Cyclin B and Cdc2) associated with the progesterone mediated oocyte maturation pathway (i.e., MAPK/MPF pathway) were analyzed by western-blot. The results showed that icariin could promote the synthesis, processing and deposition of vitellogenin in oocytes, and that it also has the potential to promote oocyte maturation (resumption of Meiosis I) by altering the expression of the relevant genes and proteins. Unlabelled Image • Icariin promotes the synthesis, processing and deposition of vitellin in oocyte. • Icariin can positively affect the progesterone mediated oocyte maturation pathway. • Icariin promotes the dephosphorylation of Cdc2 and activates Cyclin B expression. • Icariin activates the MAPK pathway by raising the phosphorylation of MEK and ERK. [ABSTRACT FROM AUTHOR]

Background: Influence of pre-retrieval human chorionic gonadotropin (HCG) priming on outcomes of in vitro maturation (IVM) remains controversial. This study aimed to evaluate the effect of HCG priming before oocyte retrieval on clinical outcomes of IVM cycles in patients with polycystic ovarian syndrome (PCOS). Methods: This was a retrospective cohort study analyzing data from the first IVM cycles of unstimulated PCOS patients in a reproductive center of university affiliated hospital from January 2006 to December 2017. Patients received HCG injection before oocyte retrieval were assigned to HCG priming group and those without HCG administration were categorized as none HCG priming (Non-HCG) group. Main outcomes included oocyte maturation rate, number of embryos available, clinical pregnancy rate, and live birth rate. Candidate factors of clinical pregnancy rate was explored by univariate analysis and multivariate logistic regression analysis. Results: There were 324 patients meeting the inclusion and exclusion criteria. Among them, 129 women received HCG priming and 195 other did not. Women in HCG group had significantly lower basal FSH level (5.17 ± 1.63 vs. 5.80 ± 2.38) than Non-HCG group. Both FSH levels were <10 IU/L and the absolute difference was 0.63 IU/L. Other basic characteristics were similar between groups with or without HCG priming. Oocyte maturation rate was trend to be higher in HCG group (52.68 vs. 48.56%) but no statistical significance was found ( P = 0.097). No significant difference in clinical pregnancy rate was found between HCG and Non-HCG groups (31.37 vs. 35.67%). Miscarriage rates (31.25 vs. 34.43%) and live birth rates were also similar between groups. HCG priming was not correlated with clinical pregnancy rate in both univariate analysis ( P = 0.468) and multivariate logistic regression analysis ( P = 0.538; OR = 1.212; 95%CI: 0.657-2.237). Conclusion: HCG priming before oocyte retrieval may not improve clinical outcomes of IVM in patients with PCOS., (Copyright © 2020 Lin, Zheng, Ma, Li, Zhang, Yang, Xu and Zhu.)