Declining fertility rates, mainly due to reduced sperm quality, challenge the health of mankind. Thus, there is an urgent need for advanced male infertility diagnostics. Despite the known importance of sperm intracellular pH (pHi) in sperm activity during fertilization, the absence of tools to detect subtle changes in spermatozoa limits the knowledge and clinical application of sperm pHi. Here, we developed RCPH, a novel dual-color fluorescent probe, demonstrating high precision (ΔpHi < 0.1) in monitoring spermatozoa pHi with spatial-temporal response. In a cohort of 79 healthy individuals and 35 asthenospermia patients, RCPH detection accurately reflects clinical parameters. With an unprecedented resolution, RCPH revealed that the sperm pHi in asthenospermia patients was 0.1 units lower compared to healthy controls. RCPH also proved effective in drug screenings that enhance pHi, thereby improving sperm motility. RCPH not only advances our knowledge of sperm pHi but also offers diagnostic potential to predict fertility outcomes and enhance in vitro fertilization (IVF) success for reproductive medicine and etiology-based interventions.

BackgroundInfertility affects approximately 15% of couples worldwide, with male factors accounting for nearly 50% of cases. Intracytoplasmic sperm injection (ICSI) has become the standard treatment for male factor infertility, but outcomes vary significantly among couples. While conventional genetic testing using blood samples is common in reproductive medicine, the genetic composition of sperm may differ significantly from somatic cells due to mosaicism and de novo mutations during spermatogenesis.MethodsWe collected semen samples from 11 couples with varying ICSI outcomes: successful clinical pregnancy (n = 6), implantation failure (n = 3), and early pregnancy loss (n = 2). Sperm DNA was extracted using magnetic-activated cell separation and whole-exome sequencing was performed. The sequencing data were aligned to the GRCh37/hg19 reference genome and analyzed for potentially pathogenic mutations. Semen analysis and karyotype were also evaluated.ResultsSemen analysis showed no significant differences between groups except for sperm morphology. Whole-exome sequencing identified distinct mutation patterns between groups. Mutations in USP9X, SPAG6 and ADGRG2 were observed in the clinical pregnancy group. Implantation failure and pregnancy loss were associated with mutations in genes involved in embryo adhesion, immune regulation, and genomic stability, including MAGEC1, MUC4 and SERPINA2.ConclusionThis pilot study suggests that direct sperm exome sequencing may reveal genetic variants associated with different ICSI outcomes. While our findings require validation in larger cohorts, they generate hypotheses about sperm-specific factors that might influence post-fertilization developmental events and pregnancy outcomes.

On the day of fresh oocyte retrieval in in vitro fertilization (IVF) cycles, a novel portable artificial intelligence optical microscope (AIOM) was employed to assist in the assessment of semen parameters. This study analyzed the correlation between sperm kinetic and morphological parameters with short-term IVF outcomes. Additionally, it explored whether these parameters could serve as predictive indicators for rescue intracytoplasmic sperm injection (R-ICSI) in IVF patients. A retrospective analysis was conducted on patients undergoing short-term IVF at the West China Second Hospital of Sichuan University between May 2021 and May 2024. Based on fertilization outcomes, the short-term IVF patients were categorized into a successful fertilization group (group A, n = 281) and a group requiring R-ICSI after failed fertilization (group B, n = 49). AIOM was utilized to analyze semen parameters including pH, sperm concentration, sperm motility parameters, sperm movement trajectory parameters, and sperm morphological parameters. The study further investigated the correlation between these short-term IVF fertilization-related laboratory indicators and IVF outcomes. No statistically significant difference was observed in semen pH between the two groups. However, there were significant differences in sperm concentration and the majority of motility parameters. Specifically, compared to group A, patients in group B exhibited lower sperm concentration (p = 0.01), motility (p = 0.01), local motility (p = 0.01), progressive motility (PR) (p = 0.00), total motility (p = 0.01), and amplitude of lateral head displacement (ALH) (p < 0.01), along with higher immotility (p = 0.00). No statistically significant differences were found between the two groups in other sperm motility, velocity, or trajectory parameters. Additionally, sperm morphological parameters were also associated with short-term IVF fertilization outcomes. Compared to group A, group B had higher sperm head length mean (p < 0.01), head perimeter mean (p < 0.01), and head area mean (p = 0.01), as well as lower tail length mean (p = 0.01). Multivariate regression analysis of fertilization outcomes indicated that higher immotility (p = 0.01) and head length mean (p < 0.01), along with lower tail length mean (p = 0.04), were independent risk factors affecting successful short-term IVF fertilization. Notably, head length mean showed a significant negative correlation with polyspermy rate (p < 0.01), whereas tail length mean was significantly positively correlated with polyspermy rate (p < 0.01). Optimization of semen parameters with AIOM at the time of fertilization is significantly associated with short-term IVF fertilization outcomes. Abnormal semen parameters at fertilization—specifically, higher immotility and head length mean, along with lower tail length mean—can be considered risk factors for fertilization failure and may serve as predictive indicators for potential R-ICSI.

This study aimed to explore the roles of spermatozoal sialidase NEU1 and NEU3 in affecting the fertilization rate of in vitro fertilization (IVF), and whether sperm sialidase NEU1 and NEU3 can be used as predictors of IVF outcome. This was a prospective cohort study that collected semen samples from 194 IVF couples between January 2024 and April 2024. Patients were grouped based on oocyte maturity and routine semen analysis. Detection of sperm NEU1 and NEU3 was carried out in semen from the same ejaculate for IVF by using flow cytometry, followed by correlation analysis with fertilization rates. NEU1 and NEU3 are independent indicators separate from regular semen parameters. With a metaphase II (MII) rate ≥80

Purpose: To clarify the prognostic value of lymph node regression (LNR) status including the lymph node regression grade (LNRG) and N downstaging in patients with esophageal cancer receiving neoadjuvant therapy based on available evidence. Methods: Several databases were searched up to 25 March 2024. The main outcomes included overall survival (OS), disease-free survival (DFS) and cancer-specific survival (CSS). Hazard ratios (HRs) and 95% confidence intervals (CIs) were combined. Subgroup analyses based on the neoadjuvant therapy and pathological type were also conducted. Results: In total, 14 retrospective studies with 3,212 participants were included. Nine and five studies explored the relationship between LNRG and N downstaging and survival, respectively. Pooled results indicated that complete LNR predicted significantly improved OS (HR = 0.47, 95% CI: 0.41-0.55, P < 0.001) and DFS (HR = 0.42, 95% CI: 0.32-0.55, P < 0.001) and subgroup analysis based on neoadjuvant therapy and pathological type manifested similar results. Besides, N downstaging was also significantly related to improved OS (HR = 0.40, 95% CI: 0.21-0.77, P = 0.006) and CSS (HR = 0.27, 95% CI: 0.12-0.60, P < 0.001). Conclusion: LNR could serve as a novel and reliable prognostic factor in patients with esophageal cancer receiving neoadjuvant therapy and complete LNR and N downstaging predict better survival.

ABSTRACT Objective Currently, the most commonly used methods for linkage analysis of pre‐implantation genetic testing for monogenic disorders (PGT‐M) are next generation sequencing (NGS) and SNP array. We aim to investigate whether the application efficacy of Asian screening array (ASA) in PGT‐M preclinical workup for the Chinese population is superior to NGS based single nucleotide polymorphism (SNP) panels. Methods We conducted a retrospective analysis by reviewing 294 couples from a single center over the past 4 years and compared the detection results between NGS‐based SNP panels and ASA. Using the numbers of informative SNPs upstream and downstream flanking of variants, we assessed the detection efficiency of both methods in monogenic diseases, chromosomal microdeletion syndrome and males with de novo variants, among other scenarios. Results Results indicate that ASA offers a greater number of informative SNPs compared with NGS‐based SNP panels. Additionally, data analysis for ASA is generally more straightforward and may require less computational resources. While ASA can address most PGT‐M challenges, we have also identified certain genes in previous tests that are not suitable for PGT‐M using ASA. Conclusion The application of ASA in PGT‐M preclinical workup for Chinese populations has good practical value as it can perform linkage analysis for most genetic variants. However, for certain variants, NGS or other testing methods, such as mutated allele revealed by sequencing with aneuploidy and linkage analysis (MARSALA), may still be necessary for completion.

BACKGROUND:Many studies have reported that electronic health (e-health) care helps health professionals manage patients undergoing assisted reproductive technology (ART) and improves their reproductive outcomes and psychological distress. However, little is known about the effectiveness of e-health care on the health outcomes of patients undergoing ART. OBJECTIVES:This study aimed to evaluate the effectiveness of e-health care on patient-centered health outcomes, such as live birth rate, pregnancy rate, time to pregnancy, etc. as well as psychological distress (i.e., infertility distress and anxiety) among individuals undergoing ART. DESIGN:A systematic review with random-effects or fixed-effects meta-analysis was conducted to compare e-health interventions with usual care in patients undergoing ART. METHOD:Electronic database, including Medline, EMBASE, Web of Science, CINHAL, and CENTRAL, were systematically searched from the inception to December 01, 2023. The authors independently reviewed the articles based on inclusion and exclusion criteria, extracted data, and assessed risk of bias used the Cochrane Risk of Bias tool version 2.0. Heterogeneity was evaluated with I2 and Chi-square. We pooled data from each study using fixed-effects meta-analysis if heterogeneity was low. Random-effects meta-analysis was used to pool data with high heterogeneity. Subgroup analysis (i.e., data collection time point) and sensitivity analysis was performed. RESULT:Data were synthesized from 21 articles covering a total of 6,749 participants (female:6,227; male:522). Pooled analysis showed that e-health care may not increase live birth rate (RR 1.44 95 %CI0.78-2.67, P = 0.25). The clinical pregnancy rate was increased to 1.57 times in the e-health care group compared with the control group (Z = 2.19, P = 0.03) and the e-health care group had an increase in time to pregnancy by 17.40 days than that of the control group (Z = 2.13, P = 0.03). Lower score of Fertility Problem Inventory-social subscale was found in the e-health care group. Subgroup analysis showed that the risk ratio of clinical pregnancy was 3.07 (95 %CI 1.60-5.89) in < 3 months group, 1.21 (95 %CI 0.93-1.59) in ≥3 months group. The fertility-related knowledge level in e-health group was higher than that of the control group (Z = 2.01, P = 0.04). CONCLUSION:Low certainty evidence suggests that e-health care increases the clinical pregnancy rate after the intervention. Additionally, e-health care benefits in improving perceived infertility-related stress specific to social and the level of infertility-related knowledge. Future studies are needed to establish core outcome measures for e-health intervention targeting infertile individuals.

SETD3 is a member of SET domain-containing proteins. It has been discovered as the first metazoan protein (actin) histidine methyltransferase. In addition to this well-characterized molecular function of SETD3, it has been clearly shown to be involved in multiple biological processes, such as cell differentiation, tumorigenesis and viral infection. Here, we summarize the current knowledge on the roles of SETD3 beyond its histidine methyltransferase activity, and outline its cellular and molecular modes of action, as well as the upstream regulation on SETD3, therefore providing insights for the molecular basis of how SETD3 fine regulates multiple physiological and pathological processes.

The purpose of this study was to introduce the surgical process of Sommerlad-Furlow modified (S-F) palatoplasty and compare its surgical and functional outcomes with conventional Sommerlad (S) palatoplasty.Patients with non-syndromic cleft palate who had undergone either S-F palatoplasty or S palato-plasty were retrospectively reviewed. Data on the outcomes of velopharyngeal function and postsurgical palatal fistula incidence were collected for all patients. Data for preselected factors, including gender, age at palatoplasty, and cleft type, were also collected. Chi-square tests were conducted.1254 patients were included. The postsurgical velopharyngeal competence (VPC) rate after S-F palatoplasty was significantly higher than after S palatoplasty (total, 70.5% vs 57.9%, p < 0.0001; age < 1, 87.0% vs 69.2%, p < 0.0001; 1 < age < 2, 78.3% vs 69.3%, p = 0.0479). With regard to different types of cleft palate, the postsurgical VPC rates after S-F palatoplasty were all significantly higher than for S palatoplasty in all patients younger than 2 years of age (complete cleft palate, 78.7% vs 62.4%, p = 0.0016; hard and soft palate cleft, 84.4% vs 74.8%, p = 0.0172; submucosal cleft and soft palate cleft, 96.6% vs 68.4%, p = 0.0114). The postoperative fistula rate after S-F palatoplasty was 4.3%.This modified palatoplasty technique provided adequate cleft palate closure, with satisfactory speech outcomes and low fistula rates, while older age at palatoplasty may affect the postsurgical outcomes. Within the limitations of the study it seems that the Sommerlad-Furlow modified technique is an option for cleft palate repair.& COPY; 2023 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

AIMS:Polycystic ovary syndrome (PCOS) is a common endocrine disorder in the women of childbearing age. It is characterized by hyperandrogenism and abnormal follicular growth and ovulation. The polyol pathway is a glucose metabolism bypass pathway initiated by aldose reductase (ADR). Androgen induces the expression of ADR in the male reproductive tract, which has a general physiological significance for male reproductive function. Here we investigate whether hyperandrogenemia in PCOS leads to increased flux of the polyol pathway in ovarian tissue, which in turn affects follicular maturation and ovulation through oxidative stress.MAIN METHODS:We used clinical epidemiological methods to collect serum and granulosa cells from clinical subjects for a clinical case-control study. At the same time, cell biology and molecular biology techniques were used to conduct animal and cell experiments to further explore the mechanism of hyperandrogen-induced ovarian polyol pathway hyperactivity and damage to ovarian function.KEY FINDINGS:Here, we find that hyperandrogenism of PCOS can induce the expression of ovarian aldose reductase, which leads to the increase of the polyol pathway flux, and affects ovarian function through excessive oxidative stress.SIGNIFICANCE:Our research has enriched the pathological mechanism of PCOS and may provide a new clue for the clinical treatment of PCOS.

RNA modifications implicate pathological and prognosis significance in cancer development and progression, of which, m6A and m5C are representative regulators. These RNA modifications could produce effects on the function of other RNA by regulating gene expression. Thus, in this study, we aimed to explore the correlation between m6A/m5C regulators and early-stage lung adenocarcinoma (LUAD). Only the early-stage LUAD samples were included in this investigation, and the RNA-seq dataset of The Cancer Genome Atlas (TCGA) cohort was utilized to evaluate the expression of 37 m6A/m5C regulated genes. Based on the expression level of these 37 genes, early-stage LUAD patients were divided into 2 clusters, which were performed by consensus clustering, and the m6A/m5C subtypes had significantly different prognostic outcomes (p < 0.001). Cluster1, which has a better prognosis, was characterized by the C3 (inflammatory) immune subtype, low immune infiltration, chemokine expression, major histocompatibility complex (MHC) expression, and immune checkpoint molecule expression. Furthermore, compared with cluster1, cluster2 showed a T cell exhaustion state, characterized by a high expression of immune checkpoint genes, and immune cells, such as T cells, CD8+ T cells, cytotoxic lymphocytes, NK cells, and so on. In addition, patients in cluster2 were with high tumor mutational burden (TMB) and numerous significant mutated oncogene and tumor suppressor genes, such as WNT10B, ERBB4, SMARCA4, TP53, and CDKN2A (p < 0.001). A total of 19 genes were mostly related to the prognosis of LUAD and were upregulated in cluster2 (p < 0.05), showing a positive correlation with the mRNA expression of 37 m6A/m5C regulated genes. The predictive risk model was constructed using Cox and LASSO (least absolute shrinkage and selection operator) regression analysis. Finally, a seven-gene m6A/m5C risk model, comprising of METTL3, NPLOC4, RBM15, YTHDF1, IGF2BP1, NSUN3, and NSUN7, was constructed to stratify the prognosis of early-stage LUAD (p = 0.0049, AUC = 0.791). The high-risk score was associated with a poorer prognosis. This model was also validated using two additional GEO datasets: GSE72094 (p = 0.011, AUC = 0.736) and GSE50081 (p = 0.012, AUC = 0.628). In summary, it was established that the m6A/m5C-regulated genes performed a crosstalk function in the mRNA expression of early-stage LUAD. By interacting with other mRNA genes, m6A/m5C modification disturbs DNA replication and the tumor immune microenvironment (TIME). The seven-gene risk model may be a critical tool for the prognostic assessment of early-stage LUAD.

Abstract Objective In observational studies, testosterone has been reported to be associated with some types of cancers. However, the direction and magnitude of the causal association between testosterone and different types of cancer remain unclear. This Mendelian randomization study assessed the causal associations of total testosterone (TT) and bioavailable testosterone (BT) with cancer risk in men. Methods We performed two-sample Mendelian randomization using publicly available GWAS summary statistics to investigate the genetically causal association between testosterone and the risk of 22 kinds of cancers in men. Causal estimates were calculated by the inverse variance weighted method. We also performed additional sensitivity tests to evaluate the validity of the casualty. Results Genetically predicted BT level were significantly associated with an increased risk of prostate cancer [odds ratio (OR) = 1.17 95% confidence interval (CI): 1.09–1.26, P = 2.51E−05] in the MR analysis with the IVW method. TT was found to be the suggestive protective factor against stomach cancer (OR = 0.66, 95% CI: 0.48–0.93, P = 0.0116) as well as pancreatic cancer (OR = 0.59, 95% CI: 0.36–0.96, P = 0.0346). A suggestive association was found between TT and the occurrence of small intestine cancer (OR = 1.0004, 95% CI: 1.0001–1.0007, P = 0.0116). However, testosterone had no significant association with other cancers. Conclusion This study investigated the role of testosterone in the development of prostate cancer, stomach cancer, pancreatic cancer, and small intestine cancer but found no strong association with the other cancers in men.

Introduction Morphological evaluation is used to select embryos for in vitro fertilisation. However, it does not fully reflect the implantation potential. Preimplantation genetic testing for aneuploidies (PGT-A) can detect embryonic aneuploidy, but biopsy procedure is invasive. Currently, a non-invasive PGT (ni-PGT) approach using spent medium is being evaluated. However, the clinical benefit of ni-PGT has not been clearly demonstrated. A multicentre randomised trial is needed to verify whether ni-PGT can be an new effective tool for evaluating embryos. Methods and analysis Overall, 1148 couples aged 35~42 (women) receiving in vitro fertilization–intracytoplasmic sperm injection are planned to be enrolled. Couples will be digitally randomised to (1) ni-PGT and (2) conventional morphology groups at a 1:1 treatment ratio. The primary outcome will be the ongoing pregnancy rate related to the first transfer cycle within 6 months after oocyte retrieval. Ethics and dissemination The study protocol is approved by the Ethics Committee of Peking University Third Hospital and the participating hospitals. The results will be disseminated through international conferences and scientific journals. Trial registration number NCT04339166 .

The association between polycystic ovary syndrome (PCOS) and endometrial cancer remains unclear. We aimed to investigate the causal association between genetically predicted PCOS and endometrial cancer risk in two ethnic groups through a two-sample Mendelian randomization (MR) approach. Our study includes 13 single nucleotide polymorphisms (SNPs) as instrumental variables (IVs) for PCOS in Europeans, and another 13 SNPs are used as IVs for PCOS in Asians. Outcome data were obtained from the largest published meta-GWAS of European ancestry to date, as well as from the BioBank Japan Project of Asian ancestry. Our study demonstrates that genetically predicted PCOS is not causally associated with the risk of overall endometrial cancer in either Europeans or Asians (odds ratio (OR) = 0.93, 95% confidence interval (CI) = 0.85–1.01, p = 0.09 and OR = 0.98, 95% CI 0.84–1.13, p = 0.75, respectively). Subgroup analyses according to histotype further illustrate that PCOS might not be associated with the risk of either endometrioid endometrial cancer or non-endometrioid endometrial cancer in European ancestry. No pleiotropy is found in our study, and a sensitivity analysis shows similar results. Our results indicate that genetically predicted PCOS might not be associated with the risk of endometrial cancer.

Polycystic ovary syndrome (PCOS) is one of the most common endocrine and metabolic diseases among women of reproductive age. Inflammation may be involved in the pathogenesis of PCOS, but its exact relationship with PCOS remains unclear. Herein, we investigate the causal association between systemic inflammatory regulators and PCOS risk through a two-sample Mendelian randomization (MR) approach based on the latest and largest genome-wide association study (GWAS) of 41 systemic inflammatory regulators in 8293 Finnish participants and a GWAS meta-analysis consisting of 10,074 PCOS cases and 103,164 controls of European ancestry. Our results suggest that higher levels of IL-17 and SDF1a, as well as lower levels of SCGFb and IL-4, are associated with an increased risk of PCOS (OR = 1.794, 95% CI = 1.150 - 2.801, P = 0.010; OR = 1.563, 95% CI = 1.055 - 2.315, P = 0.026; OR = 0.838, 95% CI = 0.712 - 0.986, P = 0.034; and OR = 0.637, 95% CI = 0.413 - 0.983, P = 0.042, respectively). In addition, genetically predicted PCOS is related to increased levels of IL-2 and VEGF (OR = 1.257, 95% CI = 1.022 - 1.546, P = 0.030 and OR = 1.112, 95% CI = 1.006 - 1.229, P = 0.038, respectively). Our results indicate the essential role of cytokines in the pathogenesis of PCOS. Further studies are warranted to assess the possibility of these biomarkers as targets for PCOS prevention and treatment.

BACKGROUND We developed a hand-made silver container as a closed vitrification system (CVS) which avoid bacterial or viral contamination. OBJECTIVE The aim of this study was to evaluate the preservation outcomes by comparing silver CVS and slow freezing (SF) method. MATERIALS AND METHODS Donated human ovarian tissues were collected from nine patients. The fragments from each patient were randomly and evenly assigned to three groups: fresh control, silver CVS and SF group. The histology of the thawed ovarian tissue was investigated and the levels of secretion of estradiol (E2) and progesterone (P4) in the culture media were used to evaluate the development and function of thawed ovarian tissue. RESULTS The results showed that the proportion of morphologically normal primordial follicles was higher in silver CVS group than in SF (P < 0.005). E2 and P4 concentrations were significantly higher in the silver CVS group than in the SF group at any time point after day 6 (E2: P < 0.05; P4: P < 0.05). CONCLUSION This study showed that human ovarian tissue cryopreserved with silver CVS had better morphology and higher E2 and P4 levels during in vitro ovarian tissue growth compared with SF. It implies that silver CVS has a better potential for ovarian tissue development in future clinical use.

BACKGROUND:The clinical significance of signet ring cells (SRCs) in surgical esophageal and esophagogastric junction adenocarcinoma (EEGJA) remains unclear now.AIM:To explore the association between the presence of SRCs and the clinicopathological and prognostic characteristics in surgical EEGJA patients by combining and analyzing relevant studies.METHODS:The PubMed, Web of Science, and EMBASE electronic databases were searched for the relevant literature up to March 28, 2021. The relative risk (RR) with 95% confidence interval (CI) was calculated to assess the relationship between SRCs and clinicopathological parameters of surgical EEGJA patients, and the hazard ratio (HR) with 95%CI was calculated to explore the impact of SRC on the prognosis. All statistical analyses were conducted with STATA 12.0 software.RESULTS:A total of ten articles were included, involving 30322 EEGJA patients. The pooled results indicated that the presence of SRCs was significantly associated with tumor location (RR: 0.76, 95%CI: 0.61-0.96, P = 0.022; I 2 = 49.4%, P = 0.160) and tumor-node-metastasis stage (RR: 1.30, 95%CI: 1.02-1.65, P = 0.031; I 2 = 73.1%, P = 0.002). Meanwhile, the presence of SRCs in surgical EEGJA patients predicted a poor overall survival (HR: 1.36, 95%CI: 1.12-1.65, P = 0.002; I 2 = 85.7%, P < 0.001) and disease-specific survival (HR: 1.86, 95%CI: 1.55-2.25, P < 0.001; I 2 = 63.1%, P = 0.043).CONCLUSION:The presence of SRCs is related with advanced tumor stage and poor prognosis and could serve as a reliable and effective parameter for the prediction of postoperative survival and formulation of therapy strategy in EEGJA patients. However, more high-quality studies are still needed to verify the above findings.

With the application of hematopoietic stem cell transplantation, Subacute or acute increase in the incidence of hepatic veno-occlusive disease (HVOD) becomes more common and it can lead to fatal complications. This article characterizes a mouse model of HVOD induced by monocrotaline. After gavage with monocrotaline was performed on BALB/c mice, On the 3rd, 4th, 6th, 8th and 10th days, mice were anesthetized, blood was collected and the liver was removed. Liver slices were processed by HE stain, Masson's trichrome stain or immunohistochemical stain. From days 3 through 4, histopathology and cytokine changes were determined as severe, early HVOD. From days 6 through 8, the changes were considered to represent late HVOD. On the 10th day, the above changes showed that late HVOD gradually improved.