Introduction:Although advanced paternal age (APA) is increasingly scrutinized in reproductive medicine, its independent impact on embryo development and clinical outcomes remains contentious, particularly when controlling for maternal age and embryo ploidy. Methods:This retrospective cohort study analyzed 357 preimplantation genetic testing for aneuploidy (PGT-A) cycles from couples with non-advanced maternal age (≤35 years). Cycles were stratified by paternal age into three groups: <35, 35-39, and ≥40 years. We compared sperm DNA fragmentation index (DFI), embryo development metrics, and clinical outcomes across these groups. Results:Men aged ≥40 years exhibited significantly higher levels of sperm DFI compared to both younger groups (both P < 0.05). While no significant differences were observed in normal fertilization, high-quality embryo rates, or euploid blastocyst rates across paternal age groups, blastocyst development was notably impaired in the APA group. Specifically, the ≥40-year group demonstrated significantly reduced blastocyst formation rates (57.3% vs. 68.6% and 67.2%) and high-quality blastocyst formation rates (33.2% vs. 41.3% and 40.2%) compared to the <35 and 35-39 groups, respectively. Crucially, multivariate regression analysis identified DFI as an independent factor, with higher DFI significantly associated with a reduced likelihood of forming high-quality blastocysts (OR = 0.987, P = 0.046) and achieving a clinical pregnancy (OR = 0.961, P = 0.036). The sensitivity analysis demonstrates that even when examining a population of very young women (≤32 years) where the influence of maternal age on oocyte quality is expected to be minimal and uniform, the negative association between sperm DFI and embryo development potential persists (aOR = 0.980, P = 0.009). Conclusion:Our findings indicate that APA itself does not directly affect blastocyst ploidy status but is associated with significantly elevated sperm DNA fragmentation. Despite the lack of direct evidence, the detrimental effects of APA on high-quality blastocyst formation and clinical pregnancy rates are probably associated with this increase in DFI. This study underscores the critical role of sperm DNA integrity in reproductive success and suggests that DFI assessment should be considered in the clinical evaluation of older men undergoing infertility treatments.

IntroductionDiminished ovarian reserve (DOR) is known to reduce the likelihood of achieving pregnancy or live births in women undergoing in vitro fertilization and embryo transfer (IVF-ET). Acupuncture maybe effective for DOR, but current evidence remains limited and inconclusive.Methods and analysisThis study is a multicenter, randomized, placebo-controlled trial conducted on a large scale. A total of 300 women with DOR preparing for IVF-ET will be randomized 1:1 to acupuncture or placebo acupuncture. Interventions will be administered from the second menstrual cycle preceding the IVF cycle until the day of oocyte retrieval. The primary outcome is the clinical pregnancy rate (CPR) following the first embryo transfer. Secondary outcomes include various IVF-ET indicators, such as the number of follicles ≥ 14 mm, estradiol (E2) levels, and endometrial thickness on the day of human chorionic gonadotropin (hCG) administration; the number of retrieved oocytes, metaphase II (MII) oocytes, and two pronuclei (2PN) fertilizations; as well as the rates of 2PN fertilization, available embryos, high-quality embryos, implantation, cycle cancellation, biochemical pregnancy, pregnancy loss, sustained pregnancy, and live birth. Additionally, ovarian reserve indicators—including antral follicle count (AFC), basal serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and E2, and anti-Müllerian hormone (AMH)—along with scores from the Self-Rating Anxiety Scale (SAS) will also be evaluated.Ethics and disseminationThe trial has been approved by the ethics committees of all participating centers. The results will be disseminated in academic journals.Clinical Trial Registrationhttp://itmctr.ccebtcm.org.cn/, identifier ID: ITMCTR2024000021.

Ovarian aging, characterized by declines in both oocyte quantity and quality, represents a major challenge to female reproductive health. However, the age-associated transcriptional alterations underlying human oocyte aging remain incompletely understood. Differential expression analysis between young and aged oocytes identified 63 age-associated differentially expressed genes, including increased expression of ribosome-associated genes, oxidative-phosphorylation-related genes such as COX5B and UQCRC2, the ubiquitin-associated gene UBA52, and mtDNA-encoded OXPHOS transcripts including MT-ND4 and MT-ND4L, together with reduced expression of the antioxidant-response gene GSTA1 and the mitochondrial respiratory-chain gene NDUFA9. Functional enrichment analyses revealed biological processes related mainly to translation, protein targeting, ribosomal functions, and oxidative phosphorylation. Further subcluster analysis identified an aged-enriched oocyte population with broader mitochondrial-related transcriptional alterations and enrichment of DNA-damage-response pathways. Subsequent PPI analysis of the intersecting genes identified two exploratory functional modules related to mitochondrial respiratory-chain processes and ribosome/ubiquitin-associated proteostasis. These findings highlight age-associated transcriptional signatures in human oocytes involving mitochondrial metabolism and proteostasis-associated processes, and reveal molecular heterogeneity among aged oocyte populations. Together, these results provide insights into the molecular changes associated with human oocyte aging and establish a transcriptomic framework for future studies of reproductive aging.

Objective Malignant neoplasms represent a major public health threat to women of child-bearing age(WCBA)worldwide,with incidence rates significantly higher than those in men of comparable age.However,systematic research remains lacking regarding the long-term evolution patterns of WCBA-specific cancer disease burden and their socioeconomic determinants across extended timeframes.This study aims to comprehensively evaluate the disease burden and spatiotemporal trends of 30 cancer types among global WCBA from 1990 to 2021,providing scientific evidence for developing targeted precision prevention and control strategies for female malignancies.Methods A cross-sectional design was adopted in this study,and the data from the Global Burden of Disease Study(GBD)2021 on the incidence,prevalence,and disability-adjusted life-years(DALYs)of 30 cancers among WCBA from 1990 to 2021 were extracted.Stratification was performed by age groups(15 to 29 and 30 to 49 years old)and socio-demographic index(SDI).Incidence rates,DALYs rates,and their estimated annual percentage change(EAPC)with 95%CI were calculated.Spearman correlation analysis and restricted cubic spline(RCS)models were applied to fit the non-linear relationship between disease burden and SDI(significance level P<0.05).Results In 2021,the 3 leading cancers by incidence among global WCBA were breast cancer(28.81/100 000,95%UI:26.84 to 30.94),cervical cancer(15.77/100 000,95%UI:14.40 to 17.23),and ovarian cancer(4.40/100 000,95%UI:3.86 to 4.88).From 1990 to 2021,the incidence of colorectal cancer demonstrated a significant upward trend(EAPC=0.48,95%CI:0.42 to 0.55,P<0.05),whereas the burden of stomach cancer(EAPC=-2.01,95%CI:-2.07 to-1.95,P<0.05)and leukemia(EAPC=-0.90,95%CI:-0.97 to-0.83,P<0.05)declined significantly.RCS fitting revealed significant non-linear correlations between disease burden of tracheal,bronchial,and lung cancer and SDI(P for non-linearity<0.05),with the incidence peaking at an SDI of approximately 0.8.Age-stratified analysis indicated that the 15 to 29 years age group was predominantly affected by leukemia and central nervous system tumors,while the burden in the 30 to 49 years age group concentrated on breast cancer,cervical cancer,and colorectal cancer.Conclusion Between 1990 and 2021,the overall cancer burden among global WCBA remained substantial,with significant evolutionary shifts in the disease spectrum.Cancer incidences demonstrate close associations with socio-economic development levels,characterized by high incidence in high-SDI regions coupled with low DALYs.This study provides macro-level data support for developing intervention strategies that integrate cancer prevention and control with reproductive health management.Countermeasures Health policymakers are recommended to implement stratified precision prevention and control policies based on age and regional SDI levels:for the 15-to 29-years-old population,priority should be given to promoting HPV vaccination and implementing fertility preservation measures during antineoplastic therapy;for those aged 30 to 49 years,coverage of combined breast and cervical cancer screening should be expanded,and early intervention for metabolic-related tumors such as colorectal cancer should be incorporated into workplace and community health management systems.

[This corrects the article DOI: 10.3389/fendo.2024.1518288.].

Ovarian dysfunction is closely associated with reproductive aging and systemic metabolic disturbances; however, the underlying microbial and metabolic mechanisms remain unclear. In this study, we analyzed fecal microbiome and serum metabolome profiles in young (7-week-old) and aged (12-month-old) female C57BL/6J mice using shotgun metagenomic sequencing and untargeted ultra-high-performance liquid chromatography-tandem mass spectrometry. Microbial and metabolic data were processed using QIIME2, HUMAnN, and MetaboAnalyst 5.0. Differential taxa and metabolites were identified using DESeq2 and linear discriminant analysis effect size (LEfSe), and their associations were evaluated using Spearman's correlation analysis. Our results showed that aged mice exhibited significant alterations in gut microbiota composition, including a decreased abundance of Firmicutes and an increased abundance of Bacteroidetes, along with enrichment of the genera Alistipes and Akkermansia. Serum metabolomic profiling identified 246 differential metabolites, primarily involved in amino acid and energy metabolism pathways. Integrated analysis revealed that tryptophan metabolism represents a key pathway linking microbial dysbiosis with systemic metabolic alterations. Notably, enriched microbial taxa, including Akkermansia muciniphila and species within the genus Alistipes, were strongly correlated with tryptophan-related metabolites. These findings indicate that ovarian failure is associated with coordinated alterations in the gut microbiome and serum metabolome, converging on tryptophan metabolism. This study provides new insights into host-microbiome-metabolite interactions in ovarian failure and highlights potential microbial and metabolic targets for therapeutic intervention.

Objective To investigate the bidirectional causal relationship between different subtypes of endometriosis and female infertility using Mendelian randomization(MR).Methods A two-sample MR analysis was performed on the data sourced from publicly available genome-wide association study(GWAS)of endometriosis and infertility.For each subtype of endometriosis,instrumental variables(IVs)satisfying MR assumptions were selected,with inverse variance weighting(IVW)as primary method.Sensitivity analyses,including Cochran's Q test,MR-Egger regression,and leave-one-out analysis,were conducted to verify the robustness of the results.Statistical significance was set at P<0.01(Bonferroni correction),and P<0.05 was considered as suggestive significance.Results Among different locations of endometriosis,ovary(IVW:OR=1.26,95％CI:1.17～1.35,P<0.001),pelvic peritoneum(IVW:OR=1.26,95％CI:1.20～1.33,P<0.001),rectovaginal septum(IVW:OR=1.11,95％CI:1.06～1.16,P<0.001),adenomyosis(IVW:OR=1.16,95％CI:1.07～1.25,P<0.001),and other types of endometriosis(IVW:OR=1.10,95％CI:1.03～1.17,P=0.004)were significantly associated with infertility.But no such association was observed in fallopian tube and intestinal endometriosis with infertility(P>0.05).Reverse causality analysis indicated that infertility might lead to fallopian tube endometriosis(IVW:OR=1.46,95％CI:1.11～1.93,P=0.007),and was associated with ovarian(IVW:OR=1.56,95％CI:1.27～1.93,P<0.001),pelvic peritoneal(IVW:OR=1.34,95％CI:1.11～1.62,P=0.003),rectovaginal(IVW:OR=1.46,95％CI:1.11～1.93,P=0.007),and other types of endometriosis(IVW:OR=1.61,95％CI:1.21～2.13,P=0.001).Suggestive significance was observed statistically in infertility promoting adenomyosis(IVW:OR=1.16,95％CI:1.07～1.25,P<0.05).It had not been proven yet that infertility could lead to intestinal endometriosis(P>0.05).Sensitivity analyses confirmed the robustness of the results.Conclusion Bidirectional causal relationships exist between different subtypes of endometriosis and infertility.Our findings based on MR analysis validate the clinical observation that endometriosis can lead to infertility,and we exploratively propose an important conclusion that infertility itself may also induce endometriosis.However,due to limited data,the causal relationships between intestinal/tubal endometriosis and infertility require further investigation.

Fertility in women decreases with age, but the molecular basis for age-related, unexplained infertility remains elusive. Here, we reveal distinct transcriptome changes in oocytes and surrounding cumulus cells from women in their mid-thirties, as evidenced by notably increased transcription of ribosome genes. Additionally, meiosis genes and actin and cohesin components are downregulated in oocytes with age. Lysosomes and proteostasis are also disrupted in cumulus cells. Moreover, DNA hypomethylation and altered heterochromatin deposition at specific genomic loci are linked to increased transcription of ribosome genes. Rapamycin effectively reduces translation and promotes protein homeostasis in cumulus cells. Remarkably, short-term rapamycin allows patients who fail repeated in vitro fertilization cycles with embryo developmental arrest to achieve high-quality blastocysts that yield successful pregnancy and live birth. These data suggest a causal role for elevated transcription of ribosome genes in aging oocytes and cumulus cells and identify rapamycin as a promising treatment for age-related infertility. This study is registered at Chinese Clinical Trial Registry (ChiCTR2300069828).

IntroductionPremature ovarian insufficiency (POI) is a condition characterized by ovarian dysfunction occurring before the age of 40, and its etiology is multifactorial, including genetic, immunological, infectious, environmental, and iatrogenic factors, with over half of the cases remaining unexplained. Whether the microbial communities and metabolites in follicular fluid, which is the direct microenvironment for oocyte survival, are related to POI has not been reported.MethodsIn this study, Follicular fluid samples of 26 patients with POI and 27 controls with a normal ovarian reserve were collected and analyzed using 16S rDNA sequencing and untargeted metabolomics. Conjoint analysis was performed to identify key microbial communities and metabolites that might be involved in POI.ResultsPatients with POI exhibited significant alterations in microbial richness and diversity and metabolic profile in their follicular fluid. The downregulation of ABC transporters and upregulation of the citrate cycle (TCA cycle) might be critical for the development and progression of POI. G-Rhodopseudomonas and g-Caulobacter were identified as key microbial genera, while L-aspartic acid, citrate, isoleucine, and cytidine were identified as key metabolites.DiscussionThese findings offer novel insights into the pathogenesis of POI and might pave the way for improved clinical outcomes for individuals with POI.

There is a considerable proportion of false positive results when the result of preimplantation genetic testing for aneuploidy is chromosomal mosaicism. The aim of this study was to assess the concordance among clinical trophectoderm biopsy, second trophectoderm biopsy, and inner cell mass biopsy in mosaic human embryos with segmental deletion and evaluate a modified biopsy protocol. This retrospective study included 100 pretested blastocysts, which were classified as segmental deletion mosaics with one to two chromosomes were affected, donated by 84 couples in a single, high-volume fertility center in Beijing China. Re-biopsy was taken from the inner cell mass and two symmetrical trophectoderm sites near the inner cell mass in each embryo. Samples from the inner cell mass and one of the two trophectoderm were prepared for modified protocol, which isolated portions into single cells, the cells with normal morphology were collected and cell debris were removed. Re-biopsy samples were analyzed by next-generation sequencing. Main outcome measures were concordance between clinical biopsy and blastocyst, and segmental mosaicism in modified protocol. Only six (6.5

The outrageous development of modernization has greatly improved the life expectancy of human beings, but aging severely affects the quality of life, which is an urgent clinical challenge that needs to be addressed. As a common medicinal herb/health food for both medicine and foodstuff, jujube has attracted extensive attention due to its effects on improving sleep quality and prolonging life. Unfortunately, the chemical and biological characteristics of jujube have not been fully characterized, it is necessary to systematically characterize the component profile and metabolic background of jujube, and the preliminary regulatory mechanism of jujube to exert anti-aging effect was further explored. Herein, we proposed a component characterization combined with metabolite profiling based on mass spectrometry (VIP >= 1, p < 0.05, FC >= 1.5 or FC <= 1/1.5) for identification, quality assessment and functional analysis of Jujube. The results suggested that in 12 batches of jujube samples from 3 regions of China, 43 differential components were identified to mediate the dynamic changes of 1571 endogenous metabolites, and then regulate 4 main pathways which are ascorbate and aldarate metabolism, sphingolipid metabolism, pyruvate metabolism and energy metabolism, to play a high edible and medicinal value of jujube. Subsequently, we employed molecular docking technology in conjunction with microscale thermophoresis affinity detection (60 % LED excitation power and 40 % MST power in NT. Control) to measure the binding affinities of differential components to specific targets. As anticipated, unique constituents in jujube, including glabrolide, betulinic acid, magnoflorine, and cytochalasin B, exhibited significant binding affinities to target proteins associated with the regulation of immune responses in the body. This study further revealed the potential regulatory mechanism of long-term consumption of jujube to "prolong life and improve health", and provided a theoretical basis for the quality control, classification processing and functional research of jujube.

The complement system, comprising over 30 proteins, is integral to the immune system, and the coagulation system is critical for vascular homeostasis. The activation of the complement and coagulation systems involves an organized proteolytic cascade, and the overactivation of these systems is a central pathogenic mechanism in several diseases. This review describes the role of complement and coagulation system activation in critical illness, particularly sepsis. The complexities of sepsis reveal significant knowledge gaps that can be compared to a profound abyss, highlighting the urgent need for further investigation and exploration. It is well recognized that the inflammatory network, coagulation, and complement systems are integral mechanisms through which multiple factors contribute to increased susceptibility to infection and may result in a disordered immune response during septic events in patients. Given the overlapping pathogenic mechanisms in sepsis, immunomodulatory therapies currently under development may be particularly beneficial for patients with sepsis who have concurrent infections. Herein, we present recent findings regarding the molecular relationships between the coagulation and complement pathways in the advancement of sepsis, and propose potential intervention targets related to the crosstalk between coagulation and complement, aiming to provide more valuable treatment of sepsis.

Objective: Infertility has emerged as a significant global public health concern, with a multitude of complex underlying causes. Epidemiological evidence indicates that immunological factors are significant contributors to the aetiology of infertility. However, previous studies on the relationship between immune inflammation and infertility have yielded inconclusive results. Methods: Mendelian randomisation (MR) is an emerging statistical method that employs exposure-related genetic variation as an instrumental variable (IV) to infer causal relationships between immune cells and infertility by modelling the principle of random assignment in Mendelian genetics. In this study, MR was employed to assess the causal relationship between 731 immune cell signatures and infertility. The data utilized in this study were obtained from publicly available genome-wide association studies (GWAS) and validated IVs, which were employed to fulfil the essential assumptions of MR analysis. Results: The Mendelian randomisation analysis revealed a total of 27 statistically significant immune cell phenotypes out of 731. The risk factor with the largest odds ratio (OR) was CD28(-) CD25(++) CD8(+) %T cell [OR, 1.21; 95% confidence interval (CI), 1.04-1.42], while the protective factor with the largest OR was activated and resting Treg AC (OR, 0.89; 95% CI, 0.82-0.97). Conclusion: The present study has demonstrated a correlation between certain characteristics of immune cells and female infertility. These results provide clues for further research into the immune mechanisms of infertility and may inform the development of novel therapeutic strategies.

Abstract Objective Premature ovarian insufficiency (POI) is a condition characterized by ovarian dysfunction occurring before the age of 40, with a prevalence ranging from 1–4%. The increasing incidence of this disease has a significant impact on women's reproductive health, as well as their metabolic, skeletal, neurological, and cardiovascular systems. The causes of premature ovarian insufficiency are multifactorial, including genetic, immunological, infectious, environmental, and iatrogenic factors, with over half of the cases remaining unexplained. Whether the microbial communities and metabolites in follicular fluid, which is the direct microenvironment for oocyte survival, are related to POI has not been reported. Methods This study included 26 patients with premature ovarian insufficiency and 27 controls with a normal ovarian reserve who underwent in vitro fertilization/embryo transfer. Follicular fluid samples were collected and analyzed using 16S rDNA sequencing and nontargeted metabolomics. Correlations between microbial communities and the metabolic profile of follicular fluid were analyzed to identify key microbial communities and metabolites that might be involved in the development and progression of POI. Results Patients with POI exhibited significant alterations in microbial richness and diversity and metabolic profiles in their follicular fluid. The downregulation of ABC transporters and upregulation of the citrate cycle (TCA cycle) might be critical for the development and progression of POI. G-Rhodopseudomonas and g-Caulobacter were identified as key microbial genera, while L-aspartic acid, citrate, isoleucine, and cytidine were identified as key metabolites. Conclusion This study revealed a distinctive microbial community metabolic profile in the follicular fluid of patients with POI. These findings offer novel insights into the pathogenesis of POI and might pave the way for improved clinical outcomes for individuals suffering from this condition.

ObjectiveThis study aims to evaluate the utility of polygenic risk scores (PRS) in women with early menopause (EM) and to investigate the clinical characteristics and risk factors associated with EM based on genetic risk.Study designGenotyping data and clinical data from women with EM and women with normal age of menopause retrieved from UK Biobank were used for early menopause risk prediction model establishment. Subsequently, 99 women diagnosed with EM and 1027 control women underwent PGT-M were recruited for model validation from across eight hospitals in China. According to PRS percentiles, these participants were further classified into high risk and intermediate risk groups. Characteristics among women at different risk levels were compared, and risk factors with early menopause were also statistical analyzed.Main outcome measuresThe proportion of women at high risk in EM and control groups; Characteristics with significant difference among women at different risk levels; risk factors associated with EM.ResultsThe proportion of high-risk women in the EM group was significantly higher than that in control women underwent PGT-M (Group PGT-M) (OR = 3.78), and that in women with normal age menopause from UK Biobank (Group UKB) (OR = 5.11). Notably, the women with high risk of EM exhibited distinct characteristics compared to women with the intermediate-risk of EM, and identified several risk factors associated with EM.ConclusionsWe established a PRS model to serves as a valuable instrument for EM risk prediction. The exploratory analysis revealed that women with high risk of EM exhibited a higher height, suggesting EM related genetic loci may also influence growth and development level. Several risk factors were found to be potentially associated with EM, such as excessive familial contentment, COVID-19 vaccination, staying up late, and the husband’s engagement in smoking and alcohol abuse.

OBJECTIVE:Observational studies suggest a link between blood lipid levels and endometriosis risk, but clinical evidence is limited and causality has not been established. This study aims to clarify this relationship using NHANES data (1999-2006) and bidirectional Mendelian Randomization (MR) analysis. METHODS:We analyzed NHANES data to explore the relationship between blood lipids and endometriosis risk using multivariable logistic regression, nonlinear testing, and trend analysis. For causal inference, MR was performed using GWAS data from the UK Biobank (lipid levels) and the Finnish Endometriosis Database (endometriosis). RESULTS:In NHANES (n=2,201), elevated triglycerides (TG) were significantly associated with increased endometriosis risk (OR, 2.10; 95% CI, 1.00-3.49; p < 0.05). MR analysis confirmed this association (OR, 1.19; 95% CI, 1.07-1.32; p < 0.006) and suggested HDL may protect against endometriosis (OR, 0.79; 95% CI, 0.63-0.99; p < 0.05). Additionally, bidirectional MR indicated that endometriosis may also contribute to TG dysregulation (OR, 1.02; 95% CI, 1.01-1.03; p < 0.006). CONCLUSION:Our findings suggest that elevated TG levels may increase endometriosis risk, while HDL may provide a protective effect. These results offer new insights and may inform future management strategies for endometriosis. However, large-scale longitudinal studies are needed to further validate these associations.

The endometrium undergoes dynamic changes throughout the menstrual cycle and pregnancy, which is unique to primates. Endometrium remodeling is essential for the implantation and nutritional support of the conceptus. Despite this, the role of uterine glands in driving endometrial tissue remodeling is still poorly understood. To address this, a 3-dimensional culture system was used to generate endometrial epithelial organoids from human endometrium biopsies. These organoids are genetically stable, long-term expandability. They reproduce some functions of uterine glands in vivo. The epithelial organoids exhibit characteristics of stem cells, with the proportion of stem cells increasing with culture time and passage number. Long-term maintenance of organoids strongly expressed stemness related genes accompanied by a decrease expression in mature epithelial gene, which suggests the organoids had switched from a mature stage to a progenitor stage. Thus we proposed the possible markers for epithelial progenitors. Meanwhile, long-term cultured organoids exhibit an increase in the proportion of luminal epithelial stem cells, accompanied by a decrease of glandular epithelial stem cells. Organoids also show hormone responsiveness, reflecting the various stages of the menstrual cycle and early pregnancy.

STUDY QUESTION:Is the chromosome copy number of the trophectoderm (TE) of a human reconstituted embryos after spindle transfer (ST) representative of the inner cell mass (ICM)?SUMMARY ANSWER:Single-cell multi-omics sequencing revealed that ST blastocysts have a higher proportion of cell lineages exhibiting intermediate mosaicism than conventional ICSI blastocysts, and that the TE of ST blastocysts does not represent the chromosome copy number of ICM.WHAT IS KNOWN ALREADY:Preimplantation genetic testing for aneuploidy (PGT-A) assumes that TE biopsies are representative of the ICM, but the TE and ICM originate from different cell lineages, and concordance between TE and ICM is not well-studied, especially in ST embryos.STUDY DESIGN, SIZE, DURATION:We recruited 30 infertile women who received treatment at our clinic and obtained 45 usable blastocysts (22 from conventional ICSI and 23 reconstituted embryos after ST). We performed single-cell multi-omics sequencing on all blastocysts to predict and verify copy number variations (CNVs) in each cell. We determined the chromosome copy number of each embryo by analysing the proportion of abnormal cells in each blastocyst. We used the Bland-Altman concordance and the Kappa test to evaluate the concordance between TE and ICM in the both groups.PARTICIPANTS/MATERIALS, SETTING, METHODS:The study was conducted at a public tertiary hospital in China, where all the embryo operations, including oocytes retrieval, ST, and ICSI, were performed in the embryo laboratory. We utilized single-cell multi-omics sequencing technology at the Biomedical Pioneering Innovation Center, School of Life Sciences, Peking University, to analyse the blastocysts. Transcriptome sequencing was used to predict the CNV of each cell through bioinformatics analysis, and the results were validated using the DNA methylation library of each cell to confirm chromosomal normalcy. We conducted statistical analysis and graphical plotting using R 4.2.1, SPSS 27, and GraphPad Prism 9.3.MAIN RESULTS AND THE ROLE OF CHANCE:Mean age of the volunteers, the blastocyst morphology, and the developmental ratewere similar in ST and ICSI groups. The blastocysts in the ST group had some additional chromosomal types that were prone to variations beyond those enriched in the blastocysts of the ICSI group. Finally, both Bland-Altman concordance test and kappa concordancetest showed good chromosomal concordance between TE and ICM in the ICSI blastocysts (kappa = 0.659, P < 0.05), but not in ST blastocysts (P = 1.000), suggesting that the TE in reconstituted embryos is not representative of ICM. Gene functional annotation (GO and KEGG analyses) suggests that there may be new or additional pathways for CNV generation in ST embryos compared to ICSI embryos.LIMITATIONS, REASONS FOR CAUTION:This study was mainly limited by the small sample size and the limitations of single-cell multi-omics sequencing technology. To select eligible single cells, some cells of the embryos were eliminated or not labelled, resulting in a loss of information about them. The findings of this study are innovative and exploratory. A larger sample size of human embryos (especially ST embryos) and more accurate molecular genetics techniques for detecting CNV in single cells are needed to validate our results.WIDER IMPLICATIONS OF THE FINDINGS:Our study justifies the routine clinical use of PGT-A in ICSI blastocysts, as we found that the TE is a good substitute for ICM in predicting chromosomal abnormalities. While PGT-A is not entirely accurate, our data demonstrate good clinical feasibility. This trial was able to provide correct genetic counselling to patients regarding the reliability of PGT-A. Regarding ST blastocysts, the increased mosaicism rate and the inability of the TE to represent the chromosomal copy number of the ICM are both biological characteristics that differentiate them from ICSI blastocysts. Currently, ST is not used clinically on a large scale to produce blastocysts. However, if ST becomes more widely used in the future, our study will be the first to demonstrate that the use of PGT-A in ST blastocysts may not be as accurate as PGT-A for ICSI blastocysts.STUDY FUNDING/COMPETING INTEREST(S):This study was supported by grants from the National Key R&D Program of China (2018YFA0107601) and the National Key R&D Program of China (2018YFC1003003). The authors declare no conflict of interest.TRIAL REGISTRATION NUMBER:N/A.

IntroductionEmbryo implantation provides an efficient way for patients with repeated implantation failure (RIF) to achieve pregnancy. The aim of this study is to compare the implantation outcomes of RIF patients in artificial cycle to those in natural cycle, both were treated with RNA sequencing endometrial receptivity test (rsERT) based personalized embryo implantation.MethodsThe endometrial receptivity (ER) analysis was performed using rsERT followed by personalized embryo transfer at optimal window of implantation (WOI). The implantation rate (IR), clinical pregnancy rate (CPR) and live birth rate (LBR) were calculated. The expression levels of biomarkers involved in pregnancy process in the patients detected as in receptivity status were also analyzed.ResultsThe rsERT shown that 44.8% (natural cycle) and 47.8% (artificial cycle) patients were in non-receptive status, which indicated a WOI displacement. After personalized embryo transfer, the IR of patients in artificial cycle was higher than those in natural cycle (52.2% vs 27.6%). The expressions of FKBP52, MUC1 and LPAR3 were significantly lower in artificial cycle than in natural cycle.ConclusionUsing artificial cycle for personalized embryo transfer based on rsERT may yield better pregnancy outcomes for RIF patients. A gene expression analysis of FKBP52, MUC1 and LPAR3 provided a potential way to increase implantation outcomes for RIF patients.

Auditory frequency coding is place-specific, which depends on the mechanical coupling of the basilar membrane-outer hair cell (OHC)-tectorial membrane network. Prestin-based OHC electromotility improves cochlear frequency selectivity and sensitivity. Cochlear amplification determines the frequency coding wherein discrete sound frequencies find a 'best' place along the cochlear length. Loss of OHC is the leading cause of age-related hearing loss (ARHL) and is the most common cause of sensorineural hearing loss and compromised speech perception. Lipid interaction with Prestin impacts OHC function. It has been established that high-fat diet (HFD) is associated with ARHL. To determine whether genetic background and metabolism preserve cochlear frequency place coding, we examined the effect of HFD in C57BL/6J (B6) and CBA/CaJ (CBA) on ARHL.We found a significant rescuing effect on ARHL in aged B6 HFD cohort. Prestin levels and cell sizes were better maintained in the experimental B6-HFD group. We also found that distortion product otoacoustic emission (DPOAE) group delay measurement was preserved, which suggested stable frequency place coding. In contrast, the response to HFD in the CBA cohort was modest with no appreciable benefit to hearing threshold. Notably, group delay was shortened with age along with the control. In addition, the frequency dependent OHC nonlinear capacitance gradient was most pronounced at young age but decreased with age. Cochlear RNA-seq analysis revealed differential TRPV1 expression and lipid homeostasis. Activation of TRPV1 and downregulation of arachidonic acid led to downregulation of inflammatory response in B6 HFD, which protects the cochlea from ARHL. The genetic background and metabolic state-derived changes in OHC morphology and function collectively contribute to a redefined cochlear frequency place coding and improved age-related pitch perception.