Abstract Objective: To explore the primary factors influencing monozygotic twinning (MZT) in patients undergoing assisted reproductive technology (ART) at a single center to clarify the mechanisms of MZT formation. Methods: This retrospective cohort study analyzed 2952 elective single embryo transfer (eSET) cycles conducted between December 2014 and December 2023 at the First Affiliated Hospital of Xiamen University, which resulted in 1363 clinical pregnancies. The examined variables included parental age, fertilization method, cycle type (fresh vs . frozen-thawed), and embryo stage (cleavage vs . blastocyst). Clinical pregnancy was diagnosed through human chorionic gonadotropin (hCG) levels 12–14 days post-transfer and confirmed via transvaginal ultrasound at 6–7 weeks of gestation. MZT was defined by the presence of additional gestational sacs (dichorionic) or two embryos within a single sac (monochorionic), while a monozygotic singleton (MZS) was defined as one embryo per sac. Results: The overall MZT incidence was 0.85% (25/2952) per cycle and 1.83% (25/1363) per pregnancy (1 morula, 4 cleavage-stage, and 20 blastocyst transfers). After excluding the morula case, 24 MZT pregnancies were matched 1:4 with singletons. After matching, embryo stage and cycle type (16 fresh and 8 frozen-thawed MZT cases) demonstrated no significant effect on MZT rates. When the analysis was restricted to blastocyst transfer (20 MZTs and 80 singletons), neither parental age nor cycle type showed a significant correlation with MZT occurrence, whereas the fertilization method was significantly associated with MZT frequency. Regarding embryo morphology, all 20 MZT blastocysts featured high-quality (grade A or B) inner cell masses and trophectoderms, with no grade C embryos observed. Multivariate logistic regression confirmed that the use of intracytoplasmic sperm injection (ICSI) significantly increased the risk of MZT, whereas parental age and cycle type did not. Conclusion: The fertilization method affects MZT incidence, with the ICSI procedure and sperm quality serving as key correlates. Furthermore, high-quality blastocysts may possess a higher potential for MZT, providing new insights for the development of individualized embryo transfer strategies.

Polycystic ovary syndrome (PCOS) is a common endocrine and metabolic disorder, accounting for approximately 80%–95% of anovulatory infertility cases. Patients with PCOS face various challenges during in vitro fertilization and embryo transfer (IVF-ET), including ovarian hyperstimulation, low fertilization rates, and an increased risk of miscarriage. The clinical manifestations of PCOS are diverse, with common symptoms including menstrual irregularities, hyperandrogenism, and polycystic ovarian morphology. Notably, approximately 50%–75% of patients with PCOS are affected by obesity, which not only exacerbates the clinical manifestations of the disease but also potentially affects reproductive outcomes. Body mass index (BMI) significantly influences IVF-ET outcomes in patients with PCOS by affecting ovarian function, oocyte quality, and endometrial receptivity. This review aims to explore the impact and underlying mechanisms of BMI on IVF treatment outcomes in patients with PCOS and emphasize the importance of weight management, with the goal of providing evidence-based support for optimizing assisted reproductive strategies, thereby improving pregnancy rates and reducing the risk of complications.

[This corrects the article DOI: 10.1097/RD9.0000000000000140.].

Objective: To study the effect of embryo cryopreservation duration on pregnancy and neonatal outcomes in women transferred with high-quality blastocyst during frozen embryo transfer (FET) cycles. Design: Multicenter Retrospective cohort study. Setting: Three tertiary-care academic medical centers. Patients: This retrospective study included a total of 24,101 women who underwent single high-quality blastocyst transfer during their first FET cycles at three tertiary academic medical centers between January 2016 and June 2023.Intervention(s) Women were categorized into two groups according to the duration of embryo cryopreservation: the short Cryo group consisted of 23,933 women with a storage time of 0-5 years, while the long Cryo group included 168 women with a storage time > 5 years. Women in the long Cryo group were matched to those in the short Cryo group using propensity score matching with a 1:4 ratio. Main outcome measure(s): The pregnancy outcomes and the neonatal outcomes. Result(s): After adjusting for potential confounding factors, no significant differences were observed between the two groups in pregnancy outcomes, including biochemical pregnancy (adjust odds ratio [aOR] 1.04, 95% confidence interval [CI], 0.70-1.56; P = 0.831), clinical pregnancy (aOR 1.10, 95% CI, 0.75-1.60; P = 0.638), ectopic pregnancy (aOR 2.15, 95% CI, 0.37-12.56; P = 0.394), miscarriage (aOR 0.95, 95% CI, 0.54-1.69; P = 0.871), and live birth (aOR 1.09, 95% CI, 0.76-1.55; P = 0.646). In addition, no significant differences were observed in neonatal outcomes, including very preterm birth, preterm birth, very low birth weight, low birth weight, high birth weight, birth weight, and gestational age. Conclusion(s): Our analysis found no evidence of significant associations between prolonged cryopreservation of high-quality blastocysts and adverse pregnancy or neonatal outcomes.

At present, the known genetic causes of abnormal oocyte development can only account for a minority of female infertility. In our previous study, the mutation c.1101 C > G, p.Tyr367* in discs large-associated protein 5 (DLGAP5) was identified as a novel genetic cause of human oocyte maturation abnormality and female infertility. The present study aimed to validate the function of DLGAP5 in oocyte maturation and further explore the underlying mechanism by which DLGAP5 regulates oocyte meiosis. Cell experiments elucidated that DLGAP5 participates in cell division, and its depletion induced G2/M arrest. The depletion of DLGAP5 in human oocytes by microinjection of siRNAs resulted in abnormal spindle morphology and oocyte maturation defects, exhibiting reduced germinal vesicle breakdown and polar body 1 extrusion rate. In addition, a similar phenotype of abnormal oocyte development was observed in Dlgap5-deficient mouse oocytes, which could be rescued by DLGAP5 cRNA microinjection. Furthermore, single-cell RNA Sequencing showed Dlgap5 knockout altered expression of genes involving in meiosis process in oocytes and deactivated PI3K-AKT signaling pathway. And PI3K-AKT activators facilitated the oocyte maturation resumption in Dlgap5-deficient mice. DLGAP5 has been demonstrated to regulate the process of oocyte maturation via the activation of PI3K-AKT pathway. It reinforces the significant role of DLGAP5 function in oocyte maturation regulation and reveals the underlying mechanism. It provides crucial insights into clinical consultation, genetic diagnosis, and treatment strategies among infertile patients.

Objective: Advanced maternal age is associated with reduced ovarian reserve and lower oocyte quality. Whether body mass index (BMI) further influences assisted reproductive technology (ART) outcomes in this population remains unclear. Methods: This retrospective study analyzed 222 patients aged >= 37 years who underwent in vitro fertilization or intracytoplasmic sperm injection with embryo transfer (IVF/ICSI-ET) at the First Affiliated Hospital of Xiamen University between January 2019 and December 2023. Patients were classified by BMI as underweight (<18.5 kg/m(2); n = 13), normal (18.5-24.9 kg/m(2); n = 154), or overweight (>24.9 kg/m(2); n = 55). Controlled ovarian stimulation protocols were individualized according to ovarian reserve. Laboratory and clinical outcomes were compared across groups, and multivariable logistic regression was used to identify factors associated with clinical pregnancy and live birth. Results: BMI, total gonadotropin dose, and use of IVF vs. ICSI differed among groups (all P <0.05). However, oocyte retrieval rate, metaphase II (MII) oocyte rate, cleavage rate, available embryo rate, high-quality embryo rate, biochemical pregnancy rate, clinical pregnancy rate, live birth rate, and miscarriage rate showed no significant differences (all P >0.05). In multivariable analysis, age, infertility duration, and progesterone level on the day of human chorionic gonadotropin (hCG) trigger were independent predictors of clinical pregnancy. Moreover, age and progesterone level on the hCG day were independently associated predictors of live birth (all P <0.05). Conclusions: Among women aged >= 37 years, higher BMI did not significantly affect key laboratory or pregnancy outcomes after IVF/ICSI-ET. Clinical results were mainly influenced by age, infertility duration, and progesterone level on the hCG trigger day.

Nicotinamide phosphoribosyltransferase (NAMPT) is an enzyme that involves into NMN-NAD + synthesis which involves into cellular metabolism related with aging, immune function, and neurodegeneration. However, its roles in early embryo development are still unclear. In present study we disturbed the NAMPT activity and employed immunofluorescence staining and live cell imaging to explore its roles during early embryo development. We showed that NAMPT mRNA level was stable during mouse early embryo development, and NAMPT accumulated in the nucleus of blastomeres in mouse embryos. The loss of NAMPT activity disturbed the early cleavage from zygote to 2-cell, 4-cell to morula formation in the dose-dependent manner. We found that NAMPT inhibition disrupted mitochondria function in 2-cell embryos, showing decreased mitochondria number and aberrant accumulation in the blastomeres, which further disturb mitochondrial membrane potential level and elevated ROS level in embryos, indicating the occurrence of oxidative stress. Moreover, NAMPT inhibition also increased the apoptotic index, showing with increased Annexin-V signals and apoptotic gene expression. Taken together, our study provided the evidence that NAMPT was essential for the mitochondria function to control oxidative stress and apoptosis during mouse early embryo development.

Objective: This study aimed to investigate the impact of Jinfeng Pills on the receptivity of a thin endometrium in rats and elucidate its mechanism of action. Methods: A thin endometrial model was established in female Sprague-Dawley rats. The rats were randomly assigned to the control, model, estradiol valerate, and Jinfeng Pill groups. Histological evaluation using hematoxylin and eosin staining was performed to assess morphological changes in the endometrium under light microscopy. Enzyme-linked immunosorbent assay was used to measure vascular endothelial growth factor (VEGF), estrogen, and progesterone levels in rat serum. Immunohistochemical analysis was used to examine morphological alterations in the endometrium. Immunofluorescence and quantitative polymerase chain reaction were employed to analyze the expression of VEGF, platelet endothelial cell adhesion molecule (CD31), beta-catenin, leukemia inhibitory factor (LIF), and homeobox gene A10 (HOXA10) proteins and mRNA in endometrial tissue. Results: Compared to the model group, the Jinfeng Pill group exhibited a thicker endometrium characterized by pronounced undulating architecture, increased glandular and vascular density, and greater endometrial volume. There was a statistically significant increase in serum VEGF levels in the Jinfeng Pill group (P <0.05). Additionally, protein levels and mRNA expression of VEGF, CD31, HOXA10, beta-catenin, and LIF were significantly high in the endometrium of the Jinfeng Pill group (P <0.05). Conclusion: Jinfeng Pills enhance the receptivity of a thin endometrium in rats by upregulating protein levels and mRNA expression of VEGF, CD31, HOXA10, beta-catenin, and LIF in the endometrium.

To explore the impact of high body mass index (BMI) on the embryo quality and clinical outcomes of polycystic ovary syndrome (PCOS) patients, and the possible genes involved. Patients who underwent in-vitro fertilization (IVF) treatment and embryo transfer in our center from November 2014 to September 2023, were divided into low BMI PCOS (LBP) group, high BMI PCOS (HBP) group, and high BMI control (HBC) group. Transcriptome sequencing was performed in eight PCOS patients’ granulosa cells (GCs). A total of 812 IVF/intracytoplasmic sperm injection (ICSI) cycles in the embryo part; and 489 fresh, 634 frozen-warmed embryo transfer (FET) cycles from the clinical part were included. The ICSI normal fertilization rate of HBP group was decreased compared to LBP and HBC groups (p = 0.013 0.008). The IVF blastocyst development rate in HBP group was lower than LBP group (p = 0.01). The preterm birth rate in HBP group was higher than in LBP (30.66

There is infertility treatment named natural cycle in vitro fertilization (IVF) combined with in vitro maturation (IVM) of immature oocytes (natural IVF/M). The objective of this study is to investigate the clinical outcomes of hCG triggering at the different sizes of dominant follicle during the treatment. A total of 705 patients with 1,011 natural IVF/M treatment cycles were included in this study. Among these 64 patients had repeated cycles, while 641 patients had only a single cycle. The cycles were divided into 4 groups according to the size of dominant follicle on the day of hCG injection (Group1:≤11.9 mm; Group2:12.0–13.9 mm༛Group3:14.0–15.9 mm༛Group4:≥16.0 mm).Through multivariate logistic regression analysis, candidate factors of number of mature oocytes, available and good quality embryos as well as clinical pregnancy rates were explored. Oocytes maturation, fertilization, cleavage, available and good quality embryo rates (p > 0.05) were no difference among 4 groups. The rates for implantation, clinical pregnancy, miscarriage, premature delivery, and live birth were similar among the groups. The presence and different sizes of the dominant follicle in the ovaries during natural cycle IVF/M does not affect clinical outcomes.

Obesity is associated with impaired spermatogenesis and decreased sperm quality, in part through reducing Sertoli cells (SCs) lactate and nicotinamide adenine dinucleotide (NAD+) production. It is not known whether nicotinamide mononucleotide (NMN) treatment improves spermatogenesis. In the present study, NMN improved lipid metabolism and enhanced spermatogenesis in obese mice, alleviated SCs dysfunction in vivo and in vitro. Label-free quantitative acetylomics analysis of mouse testes suggested that protein acetylation influenced both the structural and functional properties of metabolic proteins. The beneficial effects of NMN were due in part to changes in the acetylation of glycolysis-related proteins. Furthermore, multi-omics and correlation analyses demonstrated that interactions among the gut microbiota, metabolites, spermatogenesis, and LDHC acetylation mediated the beneficial effects of NMN. Importantly, we found that NMN treatment reduced acetylation of the lysine residues 5, 17, and 90 of LDHC, which plays a critical role in SC lactate production in obesity. Collectively, our findings show that NMN supplementation improves sperm quality in obese mice by decreasing LDHC acetylation, thereby increasing Sertoli cell lactate and NAD+ production.

Asthenozoospermia and teratozoospermia are common causes of male infertility. Despite their prevalence, the underlying metabolic mechanisms remain poorly understood. In this study, we conducted targeted metabolomic profiling of sperm samples from 131 Chinese reproductive-age men (48 normozoospermic controls, 40 asthenozoospermic patients, and 43 teratozoospermic patients) to identify distinct metabolic signatures associated with these conditions. We identified 47 significantly altered metabolites in asthenozoospermia compared to normozoospermia (18 downregulated and 29 upregulated). In teratozoospermia, 25 metabolites showed significant changes compared to normozoospermia (10 downregulated, 15 upregulated). These differential metabolites, along with others identified through machine learning as relevant, are implicated in key pathways including energy metabolism, lipid metabolism, and amino acid metabolism. Alterations, such as those in corticosterone and hypoxanthine, suggest potential impacts on redox balance and stress responses. Using machine learning approaches, we developed diagnostic models for asthenozoospermia and teratozoospermia diagnosis, with the Glmnet model showing strong performance (AUC = 0.99 for asthenozoospermia, AUC = 0.9997 for teratozoospermia), although these results require external validation. Two metabolites emerged as candidate diagnostic biomarkers: corticosterone reveals common metabolic disturbances in both pathological conditions, and arachidate specifically indicates impaired lipid metabolism in asthenozoospermia. These findings improve our understanding of the metabolic basis of sperm dysfunction, and highlight the potential of sperm metabolomics as a powerful tool for diagnosing and managing male infertility.

Unexplained recurrent spontaneous abortion (URSA) is a prevalent reproductive issue but its etiology remains obscure. Male exposure to environmental chemicals is suggested to elevate URSA risk in female partners. Herein, a case-control design set out to investigate associations between metal levels in human seminal plasma with URSA risk, plus to determine evidence of mediating effects by oxidative stress. Levels of 15 metal elements and oxidative stress marker malondialdehyde (MDA) in seminal plasma were measured in 125 male spouses of URSA cases compared to 108 male partners of women with successful pregnancy outcomes. The associations of single or mixed metals on URSA risk were analyzed using logistic regression and Bayesian kernel machine regression (BKMR), respectively. BKMR analyses reveal a joint effect of metal co-exposures on URSA risk. Through multiple statistical approaches, titanium (Ti), cadmium (Cd) or magnesium (Mg) were major contributors to metal mixtures elevating URSA risk. MDA was significantly and positively associated with URSA risk. Mediation analysis shows that the associations of Ti, Cd or Mg with URSA risk appear to be mediated by MDA at rates of 23.30%, 16.26% or 34.48%, respectively. In vitro experiments confirmed the seminal plasma relevant dose Ti, Cd or Mg exposure induced male mouse spermatocyte-derived GC-2 cells oxidative stress. Metal mixtures in seminal plasma are associated with increased URSA risk in female spouses, with Ti, Cd or Mg being significant contributors, potentially via oxidative stress, providing further insights into URSA etiology.

The aim of the current study was to investigate the relationship between environmental factors and metabolic gene genotypes related to semen quality. A total of 341 men were recruited and classified into normal or abnormal semen quality groups according to the World Health Organization’s 2010 criteria. Alcohol and tobacco use among men was self-reported. Pb (lead), As (arsenic), Ti (titanium), and Zn (zinc) metal elements in seminal plasma were measured using inductively coupled plasma mass spectrometry (ICP-MS). The ALDH2 rs671 and GSTP1 rs1695 polymorphism were detected using high-resolution melting (HRM) PCR. Individual environmental factors, including smoking, drinking, and exposure to Pb, As, Ti, or Zn, did not significantly associate with the risk of semen abnormalities. The ALDH2 GA/AA mutation genotype increased the risk of semen abnormalities in smoking males (AOR = 1.27; 95

To investigate the effects of chronic HBV infection on the outcome of in vitro fertilisation-embryo transfer and clinical characteristics of newborns, as well as the factors influencing different outcomes of in vitro fertilisation-embryo transfer (IVF-ET). In this study, a total of 3900 couples undergoing IVF-ET were collected and divided into four groups according to the different HBsAg carrier status of each couple, comparing the general demographic data and clinical characteristics between the four groups, analysing the differences in IVF-ET outcomes between the groups, and using multifactorial analysis of factors influencing their IVF-ET outcomes. The results showed that no significant differences (p > 0.05) were found in IVF-ET outcomes among the four groups, but multifactorial logistic regression showed that male and female age, low literacy level of men, total number of eggs acquired, LH value and P value on HCG day may affect the success rate of different IVF-ET outcomes (embryo outcome, pregnancy outcome and perinatal outcome) to different degrees. We also analysed the clinical data of 952 newborns and there were no statistically significant differences (p > 0.05) in variables including sex distribution, length, weight, health status and Apgar score. Therefore, our study suggests that neither uniparental infection nor biparental HBV infection may affect the outcome of IVF-ET or the clinical characteristics of the newborns, but the outcome of IVF-ET is affected by different factors.

Background Endometriosis (EM) is a major cause of infertility, but the pathogenesis and mechanisms are not yet fully elucidated. MiR-19b-3p is involved in many diseases, but its functional role in EM-associated infertility remains unexplored. This study aimed to examine miR-19b-3p abundance and IGF1 concentration in cumulus cells (CCs) and follicular fluid of EM-associated infertility patients, and to investigate the potential role of miR-19b-3p in KGN cells by identifying its target and elucidating the underlying mechanisms.Results The results from the case-control study indicated that, compared to the control group consisting of patients with tubal infertility, patients with EM-associated infertility exhibited a lower percentage of mature oocytes. MiR-19b-3p level was elevated in CCs from EM-associated infertility patients. IGF1 was identified as a direct target of miR-19b-3p and was negatively regulated by miR-19b-3p in KGN cells. Overexpression of miR-19b-3p significantly inhibited cell viability and proliferation, promoted apoptosis, and arrested the cell cycle at G0/G1 phase in KGN cells. The effects of miR-19b-3p were reversed by co-transfection of IGF1, and the biological effects of miR-19b-3p in KGN cells were mediated by IGF1. Additionally, miR-19b-3p targeted IGF1 to down-regulate AKT phosphorylation and participate in the apoptotic pathway in KGN cells.Conclusions This study demonstrates that miR-19b-3p level is elevated in CCs and IGF1 concentration is decreased in follicular fluid in patients with EM-associated infertility. MiR-19b-3p regulates the biological effects of KGN cells by targeting IGF1.

Objective: This study aimed to assess the effects of the Jinfeng pill on ovarian volume and endocrine function in rats with poor ovarian response and to elucidate the gene expression changes that contribute to the Jinfeng pill’s role in improving ovarian function. Methods: Thirty 8-week-old specific pathogen-free (SPF) female rats were randomly assigned to five groups: control, model, estradiol, Jinfeng pill, and Jinfeng pill + estradiol, with six rats in each group. All groups, except the control were administered Tripterygium glycoside tablets. After establishing the poor ovarian response model, the three intervention groups (estradiol, Jinfeng pill, and Jinfeng pill + estradiol) received their respective drug interventions. Hematoxylin-eosin staining was employed to assess follicle distribution, and immunohistochemistry was used to evaluate the expression of vascular endothelial growth factor and endostatin in rat ovaries. Hormone levels were measured using ELISA. Results: The uterine and total ovarian masses in the three intervention groups were higher than those in the model group; however, the differences were not statistically significant. The ovaries of the Jinfeng pill and Jinfeng pill + estradiol groups contained more follicles at various developmental stages. The three intervention groups showed significantly increased vascular endothelial growth factor expression and decreased endostatin expression compared with the model group, with the Jinfeng pill + estradiol group exhibiting the most pronounced differences. Anti-Müllerian hormone levels were slightly elevated in the Jinfeng pill group compared to the model group. However, only the Jinfeng pill + estradiol group showed a statistically significant decrease in follicle-stimulating hormone levels compared to the model group (P = 0.01). Conclusions: Jinfeng pill modulates ovarian protein expression, increases ovarian volume, and improves blood circulation. Additionally, it may elevate hormone levels and enhance ovarian response function in rats with poor ovarian function.

Abstract Background Primary Sjögren syndrome (pSS) is often related to adverse neonatal outcomes. But it’s currently controversial whether pSS has an adverse effect on female fertility and clinical pregnancy condition. More importantly, it’s unclear regarding the role of pSS in oocyte and embryonic development. There is a lack of comprehensive understanding and evaluation of fertility in pSS patients. Objective This study aimed to investigate oocyte and embryonic development, ovarian reserve, and clinical pregnancy outcomes in Primary Sjögren syndrome (pSS) patients during in vitro fertilization (IVF) treatment from multi-IVF centers. Methods We performed a muti-central retrospective cohort study overall evaluating the baseline characteristics, ovarian reserve, IVF laboratory outcomes, and clinical pregnancy outcomes between the pSS patients and control patients who were matched by Propensity Score Matching. Results Following PSM matching, baseline characteristics generally coincided between the two groups. Ovarian reserve including anti-müllerian hormone (AMH) and antral follicle counting (AFC) were significantly lower in the pSS group vs comparison (0.8 vs. 2.9 ng/mL, P < 0.001; 6.0 vs. 10.0, P < 0.001, respectively). The pSS group performed significant reductions in numbers of large follicles, oocytes retrieved and MII oocytes. Additionally, pSS patients exhibited obviously deteriorate rates of oocyte maturation, 2PN cleavage, D3 good-quality embryo, and blastocyst formation compared to comparison. As for clinical pregnancy, notable decrease was found in implantation rate (37.9% vs. 54.9%, P = 0.022). The cumulative live birth rate (CLBR) following every embryo-transfer procedure was distinctly lower in the pSS group, and the conservative and optimal CLBRs following every complete cycle procedure were also significantly reduced in the pSS group. Lastly, the gestational weeks of the newborns in pSS group were distinctly early vs comparison. Conclusion Patients with pSS exhibit worse conditions in terms of female fertility and clinical pregnancy, notably accompanied with deteriorate oocyte and embryo development. Individualized fertility evaluation and early fertility guidance are essential for these special patients.