Rhesus (Rh) family B glycoprotein (RHBG) is a conserved ammonium transporter of the Rhesus family with established roles in systemic acid-base regulation. Although Rh family members are expressed in the male reproductive tract, the physiological role of RHBG in spermatogenesis and sperm function remains unclear. To investigate its role in male reproduction, Rhbg-/- mice were generated using clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9 (CRISPR/Cas9)-mediated gene editing, and male fertility and reproductive phenotypes were assessed by breeding assays, histological analyses of the testes and epididymides, sperm morphology evaluation, computer-assisted sperm analysis during capacitation, and transmission electron microscopy of flagellar ultrastructure. Rhbg-/- male mice showed largely preserved fertility and produced offspring numbers comparable to those of wild-type controls, although a modest reduction in litter size was observed. Consistently, testicular architecture, spermatogenesis, and epididymal morphology were normal, and transmission electron microscopy confirmed intact flagellar ultrastructure. Computer-assisted sperm analysis further showed normal progressive motility under both basal and capacitating conditions. However, under hyperactivation-associated conditions, Rhbg-/- sperm exhibited a mild reduction in total motility, accompanied by subtle decreases in straight-line velocity and straightness. In addition, a slight increase in tail-coiling abnormalities was observed, whereas overall sperm morphology remained largely normal. Together, these findings indicate that Rhbg is not essential for spermatogenesis or male fertility in mice, but its absence causes minor, context-dependent alterations in sperm trajectory efficiency during capacitation without affecting core motility or ultrastructural integrity. Thus, RHBG may contribute to the fine regulation of sperm motility behavior rather than acting as a major determinant of reproductive capacity.
This prospective cohort study investigated the association between dyslipidemia and prostate volume progression in middle-aged Chinese men. A total of 5607 men aged >40 years at The Second Xiangya Hospital of Central South University (Changsha, China), between January 2020 and January 2023, who did not have baseline benign prostatic hyperplasia, were followed for three years. Participants were categorized by their baseline lipid status according to Chinese guidelines. Prostate volume was measured annually, using transrectal ultrasound, with enlargement defined as a volume of ≥31 ml. The results showed that 48.0% of men with dyslipidemia developed prostatic enlargement, compared to 36.5% with normal lipid levels. Elevated ratios of low-density lipoprotein cholesterol to high-density lipoprotein cholesterol and total cholesterol to high-density lipoprotein cholesterol were significantly associated with an increased risk in a dose-response manner. This association was independent of age, body mass index, and fasting glucose levels. High-density lipoprotein cholesterol demonstrated a protective effect. These findings suggest that dyslipidemia, particularly as reflected in specific lipid ratios, is an independent and modifiable risk factor for accelerated prostatic enlargement. This indicates that lipid assessment could be valuable for early risk stratification and prevention.
Androgens and the androgen receptor (AR) play a central role in the development and progression of prostate cancer. While biochanin A (BCA) has been reported to suppress androgen synthesis and overcome resistance to abiraterone and enzalutamide, its direct effects on AR, particularly mutant AR, remain unclear. In this study, we systematically investigated the binding affinity and functional impact of BCA on a panel of clinically relevant AR mutations to identify patient subgroups that may benefit from BCA treatment. Our results demonstrated that BCA exhibits high-binding affinity toward the ART877A and ARW741C mutants. Our findings reveal a previously unrecognized mechanism by which BCA directly targets specific AR mutants, in addition to inhibiting androgen biosynthesis. This study provides mechanistic insights into the mutation-selective activity of BCA and supports its potential clinical application in molecularly stratified prostate cancer.
Paternal environmental factors can influence offspring health intergenerationally through alterations in sperm epigenetics. Exercise, as a key modifiable lifestyle factor, may similarly affect epigenetic inheritance. Emerging evidence from animal and human studies indicates that exercise modulates sperm DNA methylation and noncoding RNA expression. In animal models, paternal exercise has also been linked to changes in offspring phenotypes. However, inconsistencies exist across studies because of variations in exercise modalities and the specific epigenetic markers analyzed. More importantly, the mechanisms underlying the relationship between sperm epigenetic alterations and offspring phenotypes remain unclear. This review integrates evidence from January 2010 to December 2025 to summarize exercise-induced changes in sperm epigenetics and their associations with offspring metabolic and behavioral outcomes. It critically discusses major inconsistencies in the current literature and highlights underexplored areas, including exercise-induced changes in histone modifications and chromatin architecture in sperm, as well as a striking lack of human data on offspring health. We propose that future rigorous exercise standardization, multidimensional epigenetic profiling, well-designed human studies, and functional validation are essential to establish causality and translate findings into effective interventional strategies.
Varicocele is the most common cause of reversible male infertility. Although abnormalities in basic semen parameters are frequently observed, they offer limited value for assessing fertility potential. Elevated sperm DNA fragmentation (SDF) has been consistently reported in varicocele patients, but conventional SDF methods are hindered by technical complexity and poor standardization, limiting clinical usability. This study evaluated an automated platform integrating basic semen analysis, global SDF assessment (SCD-X12), and double-strand break (DSB) detection (SDFR-X12) for varicocele fertility assessment. Seventy varicocele patients and thirty normozoospermic controls were enrolled at the Department of Urology, University of Foggia (Foggia, Italy), from July 2023 to January 2025. All semen samples were analyzed using the LensHooke® X12 system, with additional aliquots assessed by terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) test. Inter-test agreement between SCD-X12 and TUNEL was good in healthy men (accuracy: 77.0%) but poor in varicocele patients (accuracy: 61.0%), indicating that global SDF assays perform reliably under normal conditions but lose interpretability when DNA damage becomes heterogeneous, as seen in varicocele patients. Although DSBs theoretically contribute to global DNA fragmentation, agreement between DSB levels (SDFR-X12) and global SDF was modest in healthy subjects (prevalence-adjusted bias-adjusted kappa [PABAK]: 0.4) and negligible in varicocele patients (PABAK: 0.06), indicating that DSB-related damage is distinct and varicocele-specific. In conclusion, global SDF assays (TUNEL and SCD-X12) are informative in non-varicocele men. In contrast, the SDFR-X12 module identifies elevated DSBs as a varicocele-specific signature, offering clearer characterization of varicocele-associated DNA damage. By integrating semen parameters, global SDF, and DSB assessment, the automated X12 system enhances scenario-specific testing and supports improved management of varicocele-related male infertility.
Sperm maturation in the epididymis is a complex and critical process that enables spermatozoa to acquire their motility and fertilizing ability. The epididymis is essential for sperm maturation and storage and provides a unique microenvironment to support these physiological processes. In recent years, epididymis-derived extracellular vesicles have attracted considerable attention for their role in controlling sperm quality. To date, the specific molecular mechanisms by which epididymal extracellular vesicles mediate sperm maturation remain inadequately elucidated. Several studies have hinted that extracellular vesicles mediate intercellular communication and cellular development, mainly through biomolecules such as transport proteins, lipids, and non-coding RNAs. This review constructs the framework of the molecular composition of epididymal vesicles and explores their potential functional implication in regulating sperm maturation. We also focus on the possible mechanisms through which these vesicles contribute to fertilization and early embryonic development and evaluate recent advances and application prospects of epididymal vesicles as biomarkers for male infertility.
This multicenter retrospective study assessed real-world treatment patterns and outcomes in de novo metastatic hormone-sensitive prostate cancer (mHSPC) across five urological centers in Hong Kong (China; from 2016 to 2023). Of 3308 patients, 814 (24.6%) presented with de novo mHSPC. High-volume and high-risk disease were present in 59.1% and 53.8% of patients, respectively. Despite guidelines favoring intensified therapy, androgen deprivation therapy (ADT) monotherapy remained the most common treatment (52.0%), while only 28.7% received upfront chemotherapy (20.5%) or androgen receptor pathway inhibitors (ARPI; 8.2%). During a median 40-month follow-up period, 48.4% of patients died and 60.1% progressed to castration-resistant prostate cancer (CRPC) within a median of 19 months. Five-year overall survival rates were highest for ARPI (64.9%), followed by chemotherapy (53.5%) and monotherapy (38.7%), with corresponding 5-year CRPC-free survival rates of 73.5%, 7.7%, and 15.0%, respectively. Upfront chemotherapy and ARPI demonstrated significant reductions in cancer-specific mortality compared to ADT monotherapy (chemotherapy hazard ratio [HR]: 0.58, 95% confidence interval [CI]: 0.41-0.82, P = 0.002; ARPI HR: 0.32, 95% CI: 0.17-0.62, P < 0.001). High-volume disease (HR: 2.14, 95% CI: 1.52-3.01, P < 0.001) and ISUP Grade 4 or 5 (HR: 5.70, 95% CI: 1.82-17.87, P = 0.003) conferred increased risk. Upfront chemotherapy and ARPI were independently associated with improved overall survival (chemotherapy HR: 0.56, 95% CI: 0.41-0.78, P < 0.001; ARPI HR: 0.41, 95% CI: 0.24-0.69, P < 0.001). High-volume mHSPC (HR: 1.65, 95% CI: 1.24-2.21, P < 0.001) and ISUP Grade 4 or 5 (HR: 2.38, 95% CI: 1.22-4.65, P = 0.011) were significant adverse prognostic factors. Advanced age at diagnosis was associated with increased all-cause mortality (HR: 1.02, 95% CI: 1.00-1.03, P = 0.037). These findings highlight an urgent need to improve adoption of intensified treatment strategies for optimal patient outcomes.
Erectile dysfunction (ED) is a prevalent complication of both type 1 and type 2 diabetes mellitus (DM), but the shared molecular mechanisms underlying this complication remain unclear. This study employed an integrative multiomics approach to identify conserved pathways in diabetic ED. A type 2 diabetes mellitus-related erectile dysfunction (T2DM-ED) rat model was established and validated functionally. Transcriptomic analysis of cavernous tissue identified differentially expressed genes (DEGs), which were cross-referenced with a public type 1 diabetes mellitus-related erectile dysfunction (T1DM-ED) dataset, revealing 141 shared DEGs enriched in extracellular matrix (ECM)-receptor interaction, focal adhesion, phosphatidylinositol 3-kinase (PI3K)-protein kinase B (Akt), and advanced glycation end products (AGE)-receptor for advanced glycation end products (RAGE) signaling. Machine learning prioritized five consistently downregulated hub genes (periostin [POSTN], elastin [ELN], collagen type VI alpha 3 chain [COL6A3], collagen type V alpha 2 chain [COL5A2], and secreted protein acidic and rich in cysteine [SPARC]), as validated by quantitative polymerase chain reaction (qPCR) and Western blot. Their encoded proteins (periostin, elastin, collagen VI, collagen V, and SPARC) were significantly suppressed. Further analysis revealed downregulation of the expression of the focal adhesion kinase (FAK)-PI3K-Akt survival pathway and brain-derived neurotrophic factor (BDNF) and increased apoptosis (cleaved caspase-3) and CD68+ macrophage infiltration. Single-cell RNA sequencing (scRNA-seq) mapping localized these hub genes to stromal and neural cells. These findings reveal a pathological microenvironment of ECM imbalance, impaired survival signaling, and inflammation, which are common to both diabetic ED subtypes. We conclude that macrophage-driven inflammation and ECM disruption converge to inhibit FAK-PI3K-Akt signaling, accelerating structural and functional decline. This inflammatory-ECM signaling axis represents a promising therapeutic target for diabetic ED.
Androgens act through the androgen receptor (AR), which regulates nearly a thousand genes. The human AR gene contains polymorphic repeats, including (CAG)n and (GGN)n, which affect AR transactivation. This study investigated their independent and combined effects on reproductive and general health. The study included 866 patients with male factor infertility (mean age: 32.8 years, and standard deviation: 6.8 years). Standard protocols were followed for semen analysis, phenotyping, and laboratory data collection. Repeat numbers of (CAG)n and (GGN)n polymorphisms were detected simultaneously using an established genotyping assay. Significantly lower sperm counts were observed in carriers of the AR gene with ≥24 compared to ≤22 GGN repeats (median: 13.5 × 106 vs 18.2 × 106 per ejaculate, P < 0.01). The meta-analysis with the Baltic young men cohort confirmed this association (n = 1843; linear regression: β = -0.38 × 106 [95% confidence interval, 95% CI: -0.75 × 106 to -0.01 × 106] per ejaculate, P = 0.044). The effect was further enhanced by long AR (CAG)n tract (≥25 repeats). The lowest sperm counts (median: 13.6 × 106 per ejaculate) and concentrations (3.5 × 106 ml-1) were detected in carriers of the AR haplotype combining ≥24 GGN and ≥25 CAG repeats (6.8% of patients). For AR (CAG)n repeats, a positive association was observed only with body mass index (BMI; P = 0.02). Neither AR repeat stretch affected semen volume, serum follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, glucose, lipids, uric acid, or C-reactive protein (CRP) levels. In conclusion, an increased AR (GGN)n repeat number exerts a pronounced negative modulatory effect on sperm parameters. To date, only a limited number of common genetic variants have been reported to be associated with quantitative sperm parameters.
Male preconception lifestyle factors, such as smoking, alcohol consumption, and body mass index, may affect semen quality, but their dose-dependent and nonlinear effects remain unclear. We retrospectively analyzed 3336 men attending the Department of Andrology, West China Second University Hospital, Sichuan University (Chengdu, China) between January 2019 and June 2023 and collected their demographic information, lifestyle behaviors, and semen parameters. Associations were examined using multivariable regression models with natural cubic splines, with adjustment for age, abstinence period, and season. Smoking showed a clear dose-dependent adverse effect on semen quality as shown by a decline in sperm concentrations, total sperm count, and sperm motility in men who smoked more than 20 cigarettes each day or for longer than 10 years. Former drinkers showed limited improvement in sperm motility. The body mass index showed complex nonlinear associations with semen volume and sperm morphology, and some parameters peaked in mildly overweight men, while obesity remained associated with impaired semen quality. These findings highlight the substantial role of lifestyle factors in male reproductive health during the preconception period and emphasize the importance of smoking cessation and weight management. Even modest changes in body weight may meaningfully improve semen parameters, supporting targeted lifestyle guidance in preconception care.
Epidemiological studies have reported a progressive decline in parameters of male reproductive health. Increasing evidence has linked this decline to environmental contaminants. Per- and polyfluoroalkyl substances (PFAS), a major class of synthetic organic chemicals containing fully or partially fluorinated alkyl chains, are one of important environmental contaminants. In China, hexafluoropropylene oxide dimer acid (GenX) and chlorinated polyfluoroalkyl ether sulfonic acids (F-53B) are predominant PFAS. Reproductive toxicity of PFAS has been well-supported by evidence. However, evidence on male reproductive toxicity induced by GenX and F-53B remains limited. The present study aimed to explore the potential molecular targets and mechanisms of male reproductive toxicity induced by GenX and F-53B. Utilizing network toxicology, the intersection of targets between GenX and F-53B and male reproductive toxicity was identified. Their Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment pathways were further analyzed. Next, four core genes, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), tumor necrosis factor (TNF), MYC proto-oncogene (MYC), and estrogen receptor 1 (ESR1), were selected, and their relationships with the primary enrichment results were examined. Molecular docking and binding energy analysis were used to support strong interactions between the compounds and the core genes. The involvement of the core genes in toxic effects caused by GenX and F-53B was further supported by in vivo evidence. Overall, an integrated framework combining network toxicology, molecular docking, and in vivo validation was used to demonstrate male reproductive toxicity associated with GenX and F-53B. Mechanistic hypotheses were generated that may inform future investigations and clinical risk mitigation strategies.
Somatic cell reprogramming technology can reverse fate determinations of the differentiated cells by regulating their epigenetic and gene expression programs. This reprogramming enables the acquisition of pluripotency to differentiate into mature and functional cells, which provides sufficient cells for regenerative and reproductive medicine. Significant progress has recently been made by peers and us in reprogramming somatic cells, e.g., Sertoli cells, fibroblasts, and peripheral blood mononuclear cells into functional stem cells. In this review, we systematically summarize the development and optimization of multiple core methods for cell reprogramming, including somatic cell nuclear transfer (SCNT), transcription factor-induced reprogramming, chemical reprogramming, and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (CRISPR/Cas)-based reprogramming. We also address the epigenetic remodeling mechanisms that are involved in somatic cell reprogramming, including the dynamic processes of chromatin accessibility regulation, DNA demethylation, histone modifications, and the regulation of non-coding RNAs (ncRNAs). Moreover, we discuss current challenges and perspectives in this field. Significantly, stem cells derived from somatic cells have great applications in regenerative medicine for treating various kinds of diseases, such as infertility, diabetes, neurodegenerative diseases, and chimeric antigen receptor T-cell (CAR-T) immunotherapy with attention to cell maturity, heterogeneity, and safety. Our in-depth understanding of approaches, mechanisms, and applications of somatic cell reprogramming into stem cells is essential for cell therapy and tissue engineering.
Non-obstructive azoospermia (NOA) and obstructive azoospermia (OA) are the main classifications of severe male infertility, but the molecular mechanism of NOA remains poorly understood. This study aimed to identify potential biomarkers and pathological mechanisms by comparing the proteomic differences in testicular tissues of NOA and OA patients. Through proteomic analysis based on liquid chromatography-tandem mass spectrometry (LC-MS/MS) of testicular samples from 5 NOA patients and 5 OA patients, we identified 5264 proteins, among which 717 differentially expressed proteins (DEPs) were found between the two groups (242 upregulated and 475 downregulated in NOA). Bioinformatics analysis indicated that these DEPs were significantly associated with reproductive development, gametogenesis, and cell structural stability. On the basis of this, six candidate proteins, including dysferlin (DYSF), myoferlin (MYOF), mitsugumin 53 (MG53), cluster of differentiation 63 (CD63), caveolin-3 (CAV3), and calpain-3 (CAPN3), were selected from the DEPs and verified in an expanded sample set (37 NOA cases and 28 OA cases) through quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, confirming their dysregulation in NOA. These findings provide new proteomic insights into NOA, highlighting the disruption of membrane repair and structural pathways, and offer potential biomarkers for understanding its pathogenesis.
Primary ciliary dyskinesia (PCD) is a severe sperm defect, leading to male infertility. PCD affects both respiratory function and sperm motility, as motile cilia and sperm flagella rely on axonemal architecture. Multiple morphological abnormalities of the sperm flagella (MMAF) is a distinct form of asthenoteratozoospermia, characterized by a heterogeneous spectrum of flagellar defects. In recent years, coiled-coil domain-containing ( CCDC ) genes have been shown to play crucial roles in both MMAF and PCD. In this study, a homozygous mutation in CCDC39 , c.1528-2A>G, was identified in a patient of a consanguineous Chinese family presenting a typical PCD phenotype. Quantitative real-time polymerase chain reaction (qPCR) and immunofluorescence demonstrated a significant reduction in CCDC39 mRNA levels and loss of the expression of CCDC39 and other axoneme dynein proteins, respectively. Diff-Quik staining and semen analysis from the patient revealed severely reduced sperm motility, in addition to a pronounced MMAF phenotype. Severe axonemal disorganization and ultrastructural defects were consistent with the PCD phenotype in the patient, further suggesting that CCDC39 deficiency is linked to both infertility and systemic ciliary dysfunction. After intracytoplasmic sperm injection (ICSI) treatment, the CCDC39-deficient patient achieved a successful pregnancy. Overall, our findings clearly indicate that the c.1528-2A>G mutation in CCDC39 is associated with the pathogenesis of both MMAF and PCD, thereby advancing genetic diagnosis, treatment, and prognosis related to in vitro fertilization (IVF) outcomes associated with the MMAF phenotype in PCD patients.
Infertility affects approximately 15% of couples of reproductive age worldwide, with male factors contributing to nearly 40%-50% of cases. Because semen quality is a fundamental indicator of male fertility, understanding its determinants is essential for clinical assessment. In this retrospective cohort study involving 131 183 patients, we compared semen parameters between infertile men and healthy controls and explored the relationships among age, infectious microorganisms, and semen quality. Our analysis demonstrated that infertile men had significantly poorer semen quality than controls did. Age also played a critical role, with the lowest risk of male infertility observed in the 25-29 years age group; across all age groups, risk followed a U-shaped pattern. Furthermore, the presence of antisperm antibodies, Neisseria gonorrhea, Chlamydia trachomatis, Ureaplasma urealyticum , and/or Mycoplasma genitalium was associated with reduced semen quality and increased infertility risk. In contrast, although herpes simplex virus II was detected in some semen samples, its overall prevalence was low, and no significant impact on semen parameters was detected. These findings emphasize the complex interplay among age, infection, and immune factors in shaping male reproductive potential. They also highlight the need for targeted prevention and treatment strategies to mitigate the burden of male infertility and to improve future therapeutic outcomes.
Biallelic variants in fibrous sheath-interacting protein 2 ( FSIP2 ) gene are a known cause of multiple morphological abnormalities of the sperm flagella (MMAF). This study aimed to identify novel FSIP2 variants and evaluate their impact on sperm ultrastructure and intracytoplasmic sperm injection (ICSI) outcomes. Whole-exome sequencing (WES) was employed to screen a cohort of 92 MMAF patients, with candidate variants validated via Sanger sequencing and third-generation sequencing. We identified one homozygous variant in a proband from a consanguineous family and two pairs of compound heterozygous variants in two unrelated, non-consanguineous families. Routine semen analysis demonstrated markedly reduced motility across all probands. Detailed morphological and ultrastructural assessments using Papanicolaou staining, scanning electron microscopy (SEM), and transmission electron microscopy (TEM) demonstrated that approximately 80.0% of spermatozoa exhibited pathological elongation of the mitochondrial sheath in the midpiece. Furthermore, 50.0%-70.0% of spermatozoa displayed fibrous sheath dysplasia or loss in the principal piece. Immunofluorescence assays and Western blotting confirmed that FSIP2 protein localization was disrupted, and the expression of key axonemal assembly factors was dysregulated. Notably, successful pregnancies were achieved via ICSI in the partners of two probands. This study expands the mutational spectrum of FSIP2 in both consanguineous and non-consanguineous populations. Ultrastructural abnormalities, such as mitochondrial sheath elongation and fibrous sheath disassembly, highlight FSIP2 's critical role in flagellar assembly. Clinical results further support ICSI as an effective therapeutic intervention for affected individuals.
Sperm quality influences fertility and offspring health through both genomic inheritance and epigenetic inheritance. Thus, for use in clinical-assisted reproductive technology (ART), spermatozoa must have optimal genomic and epigenetic structures. In patients with retrograde ejaculation, spermatozoa are usually recovered from urine and then cryopreserved for ART. However, the effects of urine exposure and subsequent freeze-thaw cycles on sperm quality remain unclear. This is particularly true for epigenetic changes and their underlying mechanisms. In this study, we examined how different durations of urine exposure (10 min and 40 min) followed by freeze-thaw cycles affected sperm motility, DNA integrity, and methylation levels of imprinting genes (H19-imprinted maternally expressed transcript [ H19 ], mesoderm-specific transcript [ MEST ], and the transposable element Alu [ Alu ]). As the duration of urine exposure increased, sperm motility (median [interquartile range]) decreased from 48.0% (39.0%-52.5%) to 1.0% (1.0%-5.0%), the DNA fragmentation index (DFI; median [interquartile range]) increased from 12.0% (9.3%-19.9%) to 23.5% (13.9%-33.9%), the MEST methylation level (mean ± standard deviation [s.d.]) increased from 3.8% ± 1.5% to 11.5 ± 1.2%, and the H19 methylation level (mean ± s.d.) decreased from 86.9% ± 0.9% to 82.1% ± 0.5%. The freeze-thaw process further reduced sperm motility, while the DFI and methylation levels of MEST and H19 did not significantly change. The Alu methylation level remained stable. These findings demonstrate that urine exposure affects sperm motility, DNA integrity, and methylation levels of some imprinting genes. These effects intensify over time. In contrast, the freeze-thaw process impacts only sperm motility. In clinical practice, minimizing exposure to urine might improve sperm quality.
Multiple morphological abnormalities of the sperm flagella (MMAF) compromise intracytoplasmic sperm injection (ICSI) outcomes. Testicular sperm aspiration (TESA) may be beneficial, but its impact on early embryonic development and pregnancy in MMAF remains uncertain. This study evaluated the effect of TESA on fertilization, cleavage, on-time Day-3 embryo development, and clinical pregnancy and explored modification by female age and MMAF genotype. In this retrospective cohort, 196 MMAF patients undergoing ICSI (January 2019-December 2024) without female-factor infertility were analyzed: TESA group ( n = 65) versus non-TESA group ( n = 131). Only fresh embryo transfer cycles were included. Outcomes were fertilization, cleavage, on-time Day-3 embryo development, and pregnancy. Multivariable regression was complemented by inverse probability of treatment weighting, propensity score matching, and stratified analyses by female age and genotype. Robustness was examined using bootstrap resampling and outlier-sensitivity analyses. Notably, TESA was associated with higher fertilization rates (adjusted coefficient: +5.63%, P = 0.022; inverse probability of treatment weighting average treatment effect [IPTW-ATE]: +5.9%, P = 0.022; Cohen's d: 0.371). Cleavage gains were smaller (IPTW-ATE: +3.8%, P = 0.013; Cohen's d: 0.246) and method-sensitive. No significant differences were found for on-time Day-3 development or pregnancy; power was limited for these endpoints. TESA mostly improved fertilization in women aged 30-35 years (IPTW-ATE: +9.5%, P = 0.017) and cleavage in genotype-negative patients (IPTW-ATE: +10.1%, P = 0.012). Propensity models showed good balance (area under the receiver operating characteristic curve: 0.638, and standardized differences <0.1); power was adequate for fertilization (0.825). Overall, TESA is associated with higher fertilization rates in MMAF-ICSI, with modest and method-sensitive evidence for cleavage, supporting selective use. Prospective, multi-center validation is needed. Limited downstream effects suggest a role primarily at early stages.
The aim of this study was to explore the predictors of International Society of Urological Pathology (ISUP) Grade Group (GG) upgrading in nonclinically significant prostate cancer (non-csPCa) biopsy specimens guided by transrectal ultrasound (TRUS) following radical prostatectomy (RP). A retrospective analysis was conducted on clinical data from 231 patients who underwent TRUS-guided biopsy and RP between January 2018 and December 2023. Univariate and multivariate logistic regression analyses were employed to identify independent predictors of postoperative ISUP GG upgrading. The diagnostic efficiency of these predictors was evaluated using receiver operating characteristic (ROC) curves. Additionally, ROC analysis was used to compare the predictive performance of different positive target (PT) imaging methods across prostate volume (PV) subgroups. The study included 111 (48.1%) patients in the GG concordance group and 120 (51.9%) patients in the GG upgrading group. Univariate analysis revealed that PV, prostate-specific antigen density, Prostate Imaging Reporting and Data System version 2.1 (PI-RADS v2.1), magnetic resonance imaging (MRI) PT, contrast-enhanced ultrasound (CEUS) PT, and clinical T stage significantly influenced GG upgrading (all P < 0.05). Multivariate logistic regression revealed PV (OR = 0.974; 95% CI: 0.955-0.994; P = 0.010), MRI PT (OR = 3.902; 95% CI: 1.507-10.102; P = 0.005), and CEUS PT (OR = 2.280; 95% CI: 1.073-4.845; P = 0.032) as independent predictors. The prediction model demonstrated the highest diagnostic performance (area under the receiver operating characteristic curve [AUC] = 0.738, P < 0.001). MRI PT showed superior predictive efficacy compared with CEUS PT in patients with a PV <30.13 ml. In conclusion, PV, CEUS PT, and MRI PT were identified as independent predictors of postoperative ISUP GG upgrading. The predictive model exhibited the best overall performance, and compared with CEUS PT, MRI PT demonstrated greater predictive accuracy for smaller prostates.
