Background Monosodium glutamate (MSG) is a widely used food additive, yet concerns persist regarding its potential reproductive toxicity. Previous studies have associated MSG exposure with oxidative stress, inflammation and apoptosis, which may contribute to testicular damage and impaired spermatogenesis. However, its dose‐dependent effects on histological, biochemical and inflammatory parameters of the testes remain underexplored. Objective This study investigates the dose‐dependent effects of chronic MSG exposure on testicular structure, histomorphometric alterations, inflammatory markers (tumour necrosis factor‐alpha [TNF‐α]) and apoptotic activity (Caspase‐3) in male Wistar rats. Methods Thirty male Wistar rats were assigned to five groups ( n = 6). Group A (control) received distilled water, while Groups B, C, D and E received MSG at 30 mg/kg, 100 mg/kg, 300 mg/kg and 1000 mg/kg, respectively, for 60 days. Histological assessment, testicular protein concentration, TNF‐α and Caspase‐3 expression were analysed. Results Histological analysis revealed progressive seminiferous tubule disorganisation, germinal epithelium atrophy and expanded interstitial spaces in higher MSG doses. Combined H&E and reticulin staining enabled detection of cellular changes and reticular fibre alterations not fully apparent with routine histology alone. Luminal and tubular diameters, cross‐sectional area and germinal epithelium height were significantly reduced ( p < 0.05). TNF‐α levels increased dose‐dependently, indicating inflammation, while Caspase‐3 expression was significantly elevated ( p < 0.05), suggesting increased apoptosis. Conclusion Chronic MSG exposure is associated with dose‐dependent testicular damage, characterised by histological degeneration, inflammation and apoptosis. These changes suggest potential impairment of spermatogenic function. Public awareness and further research on protective strategies against MSG‐induced testicular toxicity are recommended.
Background The decline in glandular function among older males may contribute to the high prevalence of metabolic syndrome (MetS); however, research on the relationship between sex hormone levels and the progression of MetS in older men is scarce. This study aimed to elucidate the significance of serum total testosterone (T), estradiol (E2), and the T /E2 ratio in older men with MetS, with a specific focus on age‐specific variations. While low T is a known risk factor, the role of E2 and the T /E2 balance across different age cohorts remains poorly defined. Men aged ≥ 46 years who attended Harrison International Peace Hospital between June 2022 and December 2024 were enrolled in the study. A total of 405 patients were screened for a confirmed diagnosis of MetS (178 cases) and no MetS‐related component abnormalities (227 cases) and stratified by age into three groups: low age (46–64 years), middle age (65–74 years), and high age (≥ 75 years). Propensity score matching (PSM) was used to balance age, body mass index (BMI), cigarette smoking, alcohol consumption, and exercise habits, resulting in 125 pairs of participants successfully matched in a 1:1 ratio between the two groups in each age stratum (tolerance: 0.03). Comparisons were made between the T, E2, and T/E2 ratios across groups. Correlations between these three indicators and individual components of MetS were analyzed. The predictive roles of these three indicators combined with other high‐risk factors in the occurrence of MetS among middle‐aged and older men across different age groups were also explored. Results After PSM, the MetS group demonstrated significantly lower serum total testosterone (T) and T/E2 ratio compared to the non‐MetS group across all age strata (all p < 0.05), while estradiol (E2) levels showed no significant difference. A significant age‐dependent decline in T and T/E2 was observed within the MetS group. Correlation analyses revealed that T and T/E2 ratio were negatively associated with key MetS components (e.g., triglycerides [TG], fasting blood glucose), with the strength of these correlations attenuating in the oldest cohort (≥ 75 years). Multivariable models identified the T/E2 ratio as an independent risk factor, and receiver operating characteristic (ROC) analysis established age‐specific thresholds (e.g., T/E2 >12.33 for 46–64 years) with high predictive value (AUCs up to 0.866). Conclusion The T/E2 ratio provides significant predictive information beyond serum T alone for MetS in older men, particularly within the 46–74 year age window. These findings advocate for the incorporation of age‐specific assessment of the T/E2 ratio into clinical strategies for risk stratification and early intervention of MetS.
At present, coffee and tea have become the most consumed beverages worldwide. A defining characteristic of these drinks is their significant caffeine content, which is a methylxanthine alkaloid recognized for its central nervous system stimulant properties. Therefore, the impact of caffeine on human health remains an evolving area of research. One important route in this dynamic research is the impact of caffeine on male fertility. Even though, according to the PubMed database, over 500 articles have been published linking caffeine to sperm, the biochemical mechanisms through which caffeine affects semen quality are still complex and did not receive sufficient analyses. In this review, we aimed to map the current mechanisms based on the available published literature. In addition, we sought to summarize the evidence regarding the safety of caffeine consumption on human semen quality. To achieve this, we searched all published articles using the keywords “caffeine” combined with “sperm” and “semen.” A focused search was also performed using these terms along with “human” to filter for studies conducted specifically on human subjects. In conclusion, caffeine has been reported to exert positive effects on semen quality. Known mechanisms by which caffeine affects semen quality are via inhibiting phosphodiesterase (PDE) activity and increasing levels of cyclic adenosine monophosphate (cAMP), enhancing seminal creatine kinase activity, modulating sperm capacitation, inducing sperm hyperactivation independent of protein kinase A (PKA) pathway, reducing oxidative stress, and influencing steroidogenic enzyme activities. However, caffeine may induce potential toxicity to sperm when used at high doses.
Introduction Erectile dysfunction (ED) has a high incidence and complex etiology. Traditional assessments, based on intercourse scenarios, often neglect erectile function during masturbation, possibly leading to misdiagnosis. This study aimed to comprehensively assess ED based on erection during masturbation. Method A single‐center cross‐sectional study was conducted at the First Affiliated Hospital of Anhui Medical University, with the study period spanning from June 2024 to June 2025. The International Index of Erectile Function‐5 (IIEF‐5) scale (which was administered separately for intercourse and masturbation to capture scenario‐specific function) and nocturnal penile tumescence and rigidity (NPTR) monitoring were adopted to classify the ED subtype (Group A: NPTR (abnormal) + ED during masturbation; Group B: NPTR (abnormal) + no ED during masturbation; Group C: NPTR (normal) + ED during masturbation; Group D: NPTR (normal) + no ED during masturbation). A structured questionnaire survey was implemented to gather information on potential risk factors of the study participants, followed by statistical analysis of the association. Results Among the 525 participants we studied, 371 suffered from ED. These patients were generally older, had a higher body mass index (BMI), a higher smoking rate, and lower physical activity levels. Subtypes differed in IIEF‐5 scores, comorbidities, and related factors: Group A represents the most severe ED, and its IIEF‐5 score is also the lowest (8.16 ± 3.87). Group A was associated with organic diseases; Group B with sexual desire disorder; Groups C and D with psychological disorders. Conclusion This is the first study to use masturbation erection to classify ED types, each with unique characteristics and causes, providing a basis for personalized diagnosis and treatment.
The quality of the spermatozoa likely influences embryo development and subsequent pregnancy outcomes. Advanced sperm selection techniques choose viable, mature, and structurally intact sperm with high DNA integrity for fertilization. This review study aimed to determine whether there is evidence on the overall benefit of these techniques on the morphokinetic parameters of embryos and clinical outcomes in ICSI. The protocol was registered with PROSPERO, and comprehensive searches were carried out in the Cochrane, PubMed, Google Scholar, and Scopus databases. After removal of duplicates, 35 articles underwent title and abstract screening, of which 16 were excluded. Full‐text assessment was performed on the remaining studies, resulting in the inclusion of seven randomized controlled trials for qualitative synthesis. Most of the included studies were assessed as having a low risk of bias. Studies have reported higher fertilization, improved embryo morphology scores, and increased clinical pregnancy rates using advanced selection techniques compared to density gradient centrifugation or swim‐up methods. While improvements in intermediate outcomes are consistently reported, the evidence for improved live birth rates remains less robust, with some studies failing to reach statistical significance. This underscores the need for larger, randomized clinical trials to validate the clinical utility of these techniques and establish standardized protocols for their use.
BackgroundAndrogen from the testis is critical for maintaining immune environmental homeostasis and male spermatogenesis. Z. mioga Roscoe flower buds (ZM-FB) are edible plants belonging to the ginger family and popular among consumers for their high nutritional and medicinal value. However, the antiandrogen effect of ZM-FB and, subsequently, the reproductive outcome has not been evaluated.AimThis study aimed to explore the effect of ZM-FB on the reproductive health of male mice and its potential mechanism.MethodsThe body weight, sperm concentration, testosterone level in serum, abundance of androgen synthesis-related genes, and testicular transcriptome were detected in mice from the control and experimental groups, which were fed 125 or 250 mg/kg fresh or frozen ZM-FB powder daily for 3-5 weeks.ResultsCompared with the control group, mice fed 125 or 250 mg/kg fresh ZM-FB for 5 weeks exhibited no significant alterations in reproductive and immune indices. In contrast, administration of 250 mg/kg frozen ZM-FB powder for 3 weeks significantly reduced the testis/body weight ratio and sperm concentration and markedly altered the testicular transcriptomic profile in mice. Additionally, RNA-Seq analysis identified 480 differentially expressed genes (DEGs) in mouse testicular tissue, which were mainly enriched in pathways related to androgen synthesis and immune function. These DEGs included notably downregulated genes (Scd1, Star, cytochrome P450 family 11 subfamily A1 [Cyp11a1], Hsd3b6, Cyp17a1, Hsd17b3, luteinizing hormone receptor [Lhcgr]) and upregulated MHC class II marker genes. These transcriptomic findings were further validated by qRT-PCR.ConlusionZM-FB is a typical medicinal and edible ingredient. Fresh ZM-FB has no impact on male animal spermatogenesis and androgen synthesis, while frozen ZM-FB exerts adverse effects on spermatogenesis. This study provides reliable theoretical guidance for the rational dietary application of ZM-FB.
Background: An unhealthy lifestyle has been linked to declined semen quality; however, most previous studies have focused only on a single lifestyle factor, and the combined effect of an unhealthy lifestyle on semen quality remains unclear. Methods: We conducted a longitudinal study with repeated semen measurements among 3785 sperm donation volunteers at the Human Sperm Bank of Guangdong Province from 2020 to 2023. An unhealthy lifestyle score was used to assess participants' unhealthy lifestyle levels and comprised six indicators, including body mass index (BMI), cigarette smoking, alcohol consumption, physical activity, sleep quality, and diet diversity. A higher score indicated a more unhealthy lifestyle. Semen analysis was conducted according to the 5(th) edition of the World Health Organization laboratory manual for the examination and processing of human semen. Linear mixed models and linear regression models were used to evaluate the association between lifestyle and semen quality. Results: Each 1-point increase in the unhealthy lifestyle score was significantly associated with a reduction of 0.1 mL (95% CI: 0.04, 0.1; adjusted p = 0.003) in semen volume, 6.5 & times; 10(6) (2.1, 10.9; adjusted p = 0.016) in total sperm number, 3.9 & times; 10(6) (1.1, 6.8; adjusted p = 0.016) in total motile sperm number, and 3.8 & times; 10(6) (1.0, 6.6; adjusted p = 0.016) in total progressive sperm number. Stratified analyses showed significantly poorer semen quality among individuals aged <25 years and those with high perceived stress levels. Conclusions: We provide evidence that an unhealthy lifestyle is associated with lower semen quality. Adopting a fully healthy lifestyle has positive implications for semen quality and male reproductive health.
This study aimed to develop and validate a multivariable prediction model for benign prostatic hyperplasia (BPH) among Chinese middle‐aged and elderly men using pooled cross‐sectional data from the 2011, 2013, and 2015 waves of the China Health and Retirement Longitudinal Study (CHARLS). A total of 8637 male participants aged ≥45 years were included and randomly divided into training ( n = 6048) and validation ( n = 2589) cohorts. Through univariate and multivariate logistic regression analyses, 12 independent predictors were identified: Frailty index (FI35), age, education level, urban/rural residence, smoking status, body mass index (BMI), sleep duration, instrumental activities of daily living (IADL), hypertension, diabetes, geographic region (North/South), and nap duration. A nomogram was constructed and demonstrated moderate predictive performance, with area under the curve (AUC) values of 0.709 (95% CI: 0.690–0.727) in the training cohort and 0.702 (95% CI: 0.673–0.731) in the validation cohort. The model exhibited good calibration and clinical utility, offering a practical tool for early BPH risk assessment in community‐based primary care settings.
Erectile dysfunction (ED) is a common sexual disorder worldwide significantly affects men’s quality of life. However, there remains a lack of comprehensive bibliometric analysis of ED treatment. This study aims to utilize bibliometric methods to assess the research progress in ED treatment and identify the research hot spots and emerging trends in this field. The publications associated with the treatment of ED from 1993 to 2023 were obtained from the Web of Science Core Collection (WOSCC) database. Articles and review articles published in English from 1993 to 2023 were selected and screened manually. The bibliometric software, CiteSpace, was used to conduct cooperative network analysis, keyword analysis, and reference co‐citation analysis. A total of 5627 publications including 4696 articles and 931 review articles from 861 journals were extracted for analysis. The publication counts and citations reached its peak of 337 in 2023 and 10,556 in 2021, respectively. The United States accounted for the largest number of publications (32.2%), as well as the highest betweenness centrality (BC) among countries/regions (0.18). The most productive institution was the University of California System ( N = 264). Furthermore, the Journal of Sexual Medicine published the most papers related to ED ( N = 750). ED treatments were summarized by analyzing the keywords and co‐cited references, including oral phosphodiesterase type 5 inhibitors (PDE5Is), testosterone supplements, intracavernosal injection, intraurethral suppository, wave therapy, and stem cell treatment. This study showed an overview of the treatment of ED from a fresh perspective. Wave therapy, stem cell treatment were emerging trends in future research in this field. An analysis of the intellectual bases, research hot spots, and emerging trends in the field will help to shape research strategies and treatment approaches in the future, offering potential benefits for patients with ED.
BackgroundMale infertility is a complex condition with significant genetic heterogeneity. Globozoospermia, a rare form of teratozoospermia characterized by round-headed sperm, is frequently associated with mutations in the DPY19L2 gene. While the link between DPY19L2 and infertility is established, the co-occurrence of male infertility and congenital heart defects (CHDs) in offspring within consanguineous families highlights the necessity of investigating compounded genetic risks and independent segregation of recessive traits.MethodsWe recruited a consanguineous family comprising a male proband with infertility and a history of adverse reproductive outcomes, including an offspring with transposition of the great arteries (TGA). Semen analysis was performed according to WHO guidelines (2010, 5th Edition). Genetic investigation included karyotyping and whole-exome sequencing (WES) for the proband, his spouse, and family members. Sanger sequencing was used to validate identified variants.ResultsSemen analysis of the proband revealed 100% round-headed sperm (globozoospermia) with normal volume and density but reduced motility. WES identified a homozygous mutation in the DPY19L2 gene (c.63dupG; p.Arg22Alafs & lowast;66). This mutation was also confirmed in the proband's affected brother. Pedigree analysis suggested an autosomal recessive inheritance pattern resulting from consanguinity. The specific mutation explains the fertilization failure in previous intracytoplasmic sperm injection (ICSI) cycles and highlights the necessity for artificial oocyte activation (AOA).ConclusionThis study confirms the pathogenic role of the DPY19L2 c.63dupG mutation in familial globozoospermia. Furthermore, it highlights the critical importance of comprehensive genetic screening in consanguineous couples presenting with infertility and congenital anomalies. Early genetic diagnosis can guide appropriate assisted reproductive strategies, such as ICSI with AOA, and inform risk assessment for future pregnancies.
Objectives: Matrix metalloproteinases (MMPs) are implicated in tissue remodeling and inflammation, yet their causal roles in prostate diseases remain unclear. This study employed two-sample Mendelian randomization (MR) to investigate genetic evidence for causal relationships between MMPs and prostate diseases. Materials and Methods: First, univariable MR (UVMR) was used to assess the causal effects of ten genetically predicted MMPs on four prostate diseases. Genetic instruments for MMPs were derived from genome-wide association study (GWAS) catalog data, while prostate disease outcomes were sourced from FinnGen R12. Inverse variance weighted (IVW), MR-Egger, weighted median, and weighted mode methods were applied. For robust associations, multivariable MR (MVMR) was performed to investigate the mediating effects of key inflammatory markers. Sensitivity analyses included Cochran's Q, MR-PRESSO, and leave-one-out tests. Results: In the UVMR analysis, genetically predicted higher levels of MMP9 were causally associated with a modest, but statistically significant, increased risk of benign prostatic hyperplasia (BPH; odds ratio [OR] = 1.052, 95% confidence interval [CI]: 1.019-1.085, p = 0.0015). This was the only association to survive false discovery rate (FDR) correction (q < 0.10). Nominal associations were observed for MMP10 with prostatitis and MMP12 with malignant prostate neoplasm. Critically, in the MVMR analysis, the causal effect of MMP9 on BPH was substantially attenuated and became nonsignificant after adjusting for genetically predicted levels of C-reactive protein (OR = 1.046, 95% CI: 0.983-1.114, p = 0.152) and other inflammatory mediators. Conclusion: This study provides robust genetic evidence supporting a causal relationship between elevated MMP9 levels and the risk of BPH, which appears to be largely mediated by inflammatory pathways.
Introduction Moringa oleifera (M. oleifera) is a medicinal plant rich in dietary antioxidants that may protect reproductive cells from oxidative stress-induced damage and male fertility problems. This study investigated M. oleifera aqueous leaf extracts (ALE) effects on TM3 Leydig cells exposed to hydrogen peroxide (H2O2). Methods M. oleifera ALE and ascorbic acid (10-1000 & micro;g/mL) were assessed for antioxidant activity. TM3 Leydig cells were exposed to various concentrations of M. oleifera ALE (0-1000 & micro;g/mL) or alpha-tocopherol (50 & micro;M) for 24 h, alongside 200 & micro;M H2O2, with or without hCG (4 ng/mL). Cell viability, testosterone production, mitochondrial membrane potential (MMP), total protein and oxidative stress parameters (reactive oxygen species [ROS], superoxide dismutase [SOD], reduced glutathione [GSH], total antioxidant capacity [TAC] and lipid peroxidation [MDA]) were assessed. Results Ascorbic acid demonstrated superior DPPH radical scavenging and FRAP activity compared to M. oleifera ALE at most concentrations (p < 0.0001). H2O2 (200 & micro;M) exposure significantly reduced cell viability, MMP, SOD, GSH and TAC levels, while increasing ROS and MDA levels (p < 0.05). Treatment with M. oleifera ALE or alpha-tocopherol significantly increased cell viability, total protein, SOD activity and GSH levels while reducing ROS and MDA compared to H2O2 control (p < 0.05). However, testosterone levels remained unchanged (p > 0.05). Conclusions M. oleifera ALE effectively protects the TM3 Leydig cells from H2O2-induced oxidative stress by restoring the redox balance, though unchanged testosterone levels suggest limitations in using TM3 Leydig cells for assessing testosterone synthesis.
Essential oils are well-known for their strong antioxidant properties and are valuable supplements for enhancing the quality of thawed sperm. Among these, citrus fruits such as Citrus sinensis, Citrus limon, and Punica granatum are particularly rich in antioxidants, especially in their peels, which exhibit the highest antioxidant activity. In this study, essential oils were extracted from the peels of Citrus sinensis, Citrus limon, and Punica granatum and added to diluted Sanjabi ram semen at different concentrations (0.25%, 0.5%, 1%, and 2% v/v) during the freezing process. The effects of these three essential oils were evaluated using various qualitative parameters after thawing, including sperm total motility, viability, membrane functionality, DNA integrity (DI), total abnormality rates, and malondialdehyde (MDA) levels. The analysis was conducted with seven replicates, and the resulting data were assessed using SPSS software. Duncan's test was applied to compare the means at a significance level of 5%. Incorporating 0.25% v/v Punica granatum peel oil into the freezing medium for ram sperm significantly enhanced viability (70.19 +/- 2.03; p < 0.01), total motility (66.38 +/- 1.12; p < 0.01), and membrane integrity (67.53 +/- 2.29; p < 0.01) after the freeze-thaw process compared to other treatment groups. Furthermore, both 0.25% v/v Punica granatum (16.02 +/- 0.39) and Citrus sinensis peel (16.40 +/- 0.51) essential oils in the freezing medium resulted in lower MDA levels (p < 0.03) and improved DI (89.57 +/- 0.1 and 91.44 +/- 1.03, respectively; p < 0.0065) after freeze-thaw when compared to other treatments. Additionally, the inclusion of 0.25% v/v Citrus limon essential oil led to a reduction in total sperm abnormalities following freeze-thaw. In contrast, higher concentrations of 1% and 2% v/v of both Citrus limon and Punica granatum peel essential oils significantly decreased viability (p < 0.01), total motility (p < 0.01), and both membrane (p < 0.01) and DI (p < 0.01) parameters after freeze-thaw compared to the control group. Moreover, the 2% v/v concentrations of Citrus limon (28.63 +/- 0.6) and Punica granatum peel (28.41 +/- 0.58) essential oils resulted in increased MDA levels (p < 0.05) compared to the control. Overall, adding appropriate concentrations of essential oils from Citrus sinensis, Citrus limon, and Punica granatum peels, known for their antioxidant properties, significantly improved sperm quality parameters after thawing.
Renovascular hypertension (RVH) is associated with oxidative stress and impaired reproductive function. Galangin, a polyphenolic compound with antioxidative properties, may mitigate these adverse effects. This study aimed to evaluate the therapeutic effects of galangin on sperm quality in an RVH rat model. Male Sprague-Dawley rats were induced using a two-kidney, one-clip (2K-1C) model by clipping the renal artery. The mice were divided into sham (control), 2K-1C, 2K-1C + galangin (30 and 60 mg/kg body weight), and 10 mg/kg losartan groups. Sperm motility and concentration, and body and organ weights were measured. Testicular angiotensin II type 1 receptor (AT1R) and NADPH oxidase 2 (NOX2) protein expression and plasma and testicular malondialdehyde (MDA) levels were determined to evaluate oxidative stress. 2K-1C rats had significantly increased testicular MDA levels and reduced sperm motility and concentration. The 2K-1C rats also showed upregulated testicular AT1R and NOX2 expressions. Galangin reduced oxidative stress and downregulated AT1R and NOX2 expression. It also improved sperm quality, as evaluated by sperm motility and concentration, compared to that in the untreated 2K-1C group. Galangin ameliorates sperm dysfunction in 2K-1C rats by reducing oxidative stress and modulating the renin-angiotensin system. These findings suggest a potential therapeutic role for galangin in patients with male infertility with renal artery stenosis.
BackgroundQinghuayin is a traditional Chinese medicine (TCM) formulation created by Professor Dong Shangpu, a renowned TCM expert from Hebei Province. This clinical prescription is developed through years of research and practice. This study establishes chronic nonbacterial prostatitis (CNP) rat and prostate epithelia RWPE-1 cell pyroptosis models both in vivo and in vitro for further exploration of its potential mechanisms.Methods and ContentThe quality control and quantitative analysis of QHY were studied by high-performance liquid chromatography (HPLC). The inhibitory effect of Qinghuayin on CNP was assessed via HE staining, Masson's staining, immunohistochemistry (IHC), and the determination of inflammatory factors with enzyme-linked immunosorbent assay (ELISA). The morphological changes within the prostate epithelia RWPE-1 cells were observed using a microscope. Cell viability assessment was conducted using multiple approaches: lactate dehydrogenase (LDH) toxicity release quantification to measure membrane damage and Calcein/propidium iodide (PI) dual staining to evaluate membrane integrity, while pyroptotic cell death was detected through specific immunofluorescence labeling techniques. The expression levels of pyroptosis-related proteins were ascertained by Western blot.ResultsThe study revealed that Qinghuayin markedly decreased the presence of inflammatory cells within the prostate tissue of CNP-induced rat models while simultaneously improving the tissue's pathological alterations. The combination of LPS and ATP decreased the viability of prostate epithelial RWPE-1 cells, resulting in cell swelling, pyroptotic vacuole formation, membrane integrity impairment, an increase in LDH toxicity release, and a rise in PI-positive cells. Experimental results indicated that Qinghuayin effectively alleviated pyroptosis changes while decreasing the levels of inflammatory mediators IL-1 beta, IL-18, and LDH toxicity release in vitro. The expression of P-NF-kappa B protein, nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, and gasdermin D (GSDMD) was inhibited in vivo and in vitro.ConclusionThis study demonstrates that the combination of LPS and ATP is capable of promoting the nuclear translocation of nuclear transcription factor kappa B (NF-kappa B), activating the NLRP3 inflammasome, and inducing pyroptosis in prostate epithelial cells. Qinghuayin can reduce CNP symptoms by inhibiting NF-kappa B regulation of NLRP3 inflammasome-mediated pyrodeath.
In this article, we firstly integrated scRNA-sequencing and bulk RNA-sequencing data to mine hub biomarkers and to clarify the microenvironment or mechanisms of diabetic erectile dysfunction (DMED) from the perspective of genomics. Bulk RNA-sequencing and scRNA-sequencing data for DMED were acquired from the GSE2457 and GSE206528 datasets, respectively. Hub biomarkers were identified by the machine learning method, and their potential mechanisms were also revealed in DMED. Based on these differently expressed genes in DMED, four different kinds of machine learning methods were simultaneously applied and compared. Therein, random forest (RF) was the optimal machine learning method, and its top five genes (Slco1a6, Angptl4, Mepe, Cidea, and Ubd) were regarded as hub biomarkers in DMED. These five hub biomarkers' PPI networks, chemical-gene-ED networks, TF-gene-ED networks, RBP-gene-ED networks, and LncRNA/miRNA/mRNA networks, were also displayed by us for their potential mechanisms in DMED. By scRNA-sequencing data analysis, our results showed that Angptl4 and Cidea were highly expressed in DMED compared to the control group. Moreover, Angptl4 was expressed in five annotated cell clusters, while Cidea was mainly expressed in the fibroblast (FB) cluster and the endothelial cell (EC) cluster. Further experimental results confirmed that Angptl4, Cidea, Mepe, and Slco1a6 were consistent in both the bulk RNA-seq data analysis of GSE2457 and our qRT-PCR results (all p < 0.05).In conclusions, Angptl4, Cidea, Mepe, and Slco1a6 could be hub biomarkers, providing novel targets for DMED in the future, and their involved mechanisms could provide novel insights into the DMED microenvironment.
Background Oligoasthenoteratospermia (OAT) represents a major cause of male infertility. The molecular mechanisms underlying sperm morphological defects, particularly their potential interplay with environmental exposure-associated molecular alterations, remain poorly understood and warrant exploratory investigation. Methods Semen and plasma samples were collected from normal controls (NC, n = 6), mild morphological defect (MMD, n = 6), and severe morphological defect (SMD, n = 6) subjects in this exploratory pilot study designed to detect large-effect signals. Integrated proteometabolomic profiling was performed using 4D proteomics and LC-MS. Bioinformatic analysis focused on screening differential expression, functional enrichment, exploring potential interactions between exposure-associated metabolites and proteins, and identifying coherent patterns to potentially differentiate groups. Results Comparative analysis identified biphasic dysregulation patterns in key cytoskeletal proteins. The prominent hub ACTN4 exhibited depletion in MMD sperm while displaying pathological spatial dislocation with 83% showing midpiece signal loss and fragmented tail fluorescence, accompanied by extracellular vesicle release in SMD sperm. Quantitative immunofluorescence validation confirmed these divergent phenotypes, that is a 56% systemic reduction in MMD compared to controls (p < 0.0001), with SMD sperm also showing 41% absolute depletion versus NC (p < 0.0001) despite proteomic upregulation, which indicates functional loss through vesicular shedding. Plasma metabolomics revealed distinct profiles between severity groups, quantifying 357 differentially abundant metabolites (including prioritized selenometabolite-bile acid interactions) between SMD and MMD. Exploratory correlation analysis prioritized inverse associations including arsenocholine (an established environmental arsenic exposure biomarker) and ACTN4 (rho = -0.867, p = 0.0025), glycodeoxycholic acid versus PPP1CC (rho = -0.917, p < 0.001), and Se-adenosylselenomethionine-PGK1 (rho = -0.867, p = 0.0025), reflecting ultrastructural damage patterns. Conclusion This severity-stratified analysis demonstrates that sperm morphological defects in OAT associate with compartment-specific ACTN4 depletion patterns, that is quantitative loss systemically impairing head-tail coupling in MMD versus spatial dislocation with pathological vesiculation causing functional nullification in SMD. The exploratory findings suggest potential associations with multisystem exposure effects, particularly indicated by the arsenocholine-ACTN4 correlation, though causal relationships cannot be established from this observational study. In addition, exposure-protein associations involving PPP1CC and PGK1 also support the hypothesis of environmental perturbation affecting sperm integrity, though these observations require validation in independent cohorts.
Background The impact of partial azoospermia factor c (AZFc) deletions and deleted in azoospermia (DAZ) deletions located on the long arm of the Y chromosome (Yq) on the risk of oligozoospermia and azoospermia remains a subject of debate. Objective This study aims to investigate the prevalence of partial AZFc deletions and DAZ deletions in infertile males without AZF microdeletions in Guangxi province, China, and to analyze their association with clinical phenotypes. Materials and Methods Karyotyping was performed on patients with abnormal sperm concentrations. Multiplex polymerase chain reaction (PCR) was utilized to analyze AZF microdeletions among patients with a normal karyotype and healthy controls. Partial AZFc deletions were also assessed using multiplex PCR. Additionally, PCR-restriction fragment length polymorphism (PCR-RFLP) analysis was employed to evaluate DAZ deletions in patients lacking AZF microdeletions compared to healthy controls. Results Among 934 patients with a normal karyotype, AZFc deletions were the most prevalent, occurring in 3.64% of patients, closely followed by sY1192-independent microdeletion at 3.43%. Among 888 patients without AZF microdeletions, 10.47% carried partial AZFc deletions, with gr/gr deletions being the most prevalent (7.66%). Among patients with gr/gr deletions, 60 had gr/gr+DAZ1/DAZ2 deletions, whereas eight had gr/gr+DAZ3/DAZ4 deletions. The AZFc deletion rate in patients with nonobstructive azoospermia (NOA) was significantly higher than that in those with severe oligozoospermia, oligozoospermia, and healthy controls (p < 0.05). Similarly, the sY1192-independent microdeletion rate was higher in patients with NOA and severe oligozoospermia than in healthy controls (p < 0.05). The gr/gr deletion rate was significantly higher in patients with NOA or severe oligozoospermia compared to those with oligozoospermia or healthy controls (p < 0.05). Furthermore, the DAZ1/DAZ2 deletion rate in patients with NOA was higher than that in those with oligozoospermia or healthy controls (p < 0.05). Conclusions Our findings indicate that the sY1192-independent microdeletion, gr/gr deletions, and DAZ1/DAZ2 deletions are linked to disrupted spermatogenesis in infertile males.
Background Smoking remains a leading modifiable risk factor for male infertility, impairing semen quality through oxidative stress from reactive oxygen species (ROS) in tobacco smoke, alongside toxins like nicotine and heavy metals that damage sperm membranes and DNA. While cigarettes are well-studied, emerging data on water pipe (shisha/hookah) and vaping highlight regional gaps, especially in high-prevalence areas like Jordan, where a 2025 national survey reported 51.6% adult tobacco use (71.2% males), exceeding WHO projections. Infertility affects 15% of couples globally, with male factors in 20%-50% of cases; in Jordan, cultural stigma often overlooks male contributions despite semen analysis as the cornerstone diagnostic tool. Methods This cross-sectional study enrolled 299 men aged 20-50 years undergoing infertility evaluation at four Jordanian fertility centers (King Abdullah University Hospital, Istiqlal Hospital, Islamic Hospital, and Finlandi Hospital) from January to December 2025, using convenience sampling. Participants self-reported smoking type/intensity: cigarettes (light: <10/day; moderate: 10-20/day; and heavy: >20/day), water pipe (sessions/week, duration), and vaping (nicotine strength). Exclusions encompassed chronic illnesses (e.g., diabetes, varicocele), substance use, medications affecting fertility, and hormonal therapy. Data via supervised questionnaires included demographics and smoking history; semen analysis followed WHO 6th edition protocols (volume >= 1.4 mL, concentration >= 16 & times; 106/mL, total motility >= 42%, progressive >= 30%, and morphology >= 4%). Analysis used Jamovi: descriptive stats, one-way ANOVA (continuous parameters), chi-square (categorical vs. WHO thresholds), and p < 0.05 significant. Ethical approval from Yarmouk University ensured informed consent and confidentiality. Results Participants comprised non-smokers (NOs) (n = 76), light (n = 72), moderate (n = 87), and heavy smokers (n = 64; primarily cigarettes, with subsets for shisha/vaping). Means showed no significant ANOVA differences: volume (3.0 +/- 1.6 mL overall, F = 0.25, p = 0.858), concentration (34.5 +/- 33.3 & times; 10(6)/mL, F = 1.56, p = 0.202), total motility (38.9 +/- 18.0%, F = 0.04, p = 0.990), progressive motility (25.4 +/- 18.2%, F = 1.88, p = 0.136), and morphology (6.9 +/- 15.0%, F = 0.30, p = 0.822). Chi-square revealed one significant link: motility categories (progressive >32%, immotile <32%; chi(2) = 14.5, df = 6, p = 0.0246), with NOs favoring >32% and smokers more <32%/immotile. No differences in volume (chi(2) = 2.58, p = 0.461), concentration (chi(2) = 5.10, p = 0.165), total count (chi(2) = 4.39, p = 0.222), pH (chi(2) = 2.56, p = 0.862), or morphology. Conclusion Smoking shows selective motility associations without broad semen impacts, urging expanded studies for mechanistic insights and public health interventions in high-burden settings like Jordan.
Background: Male infertility is an increasing global health concern. In African traditional medicine, red palm oil is used alone or with herbs or honey to enhance male fertility by improving sperm quality and libido. Its rich bioactive composition supports its therapeutic potential, yet its oily consistency may limit absorption and bioavailability. Aim of the Study: This study evaluated the effects of crude palm oil (EGCMO) and its emulsified form (EGEMO) on male reproductive parameters in Wistar rats to validate their ethnomedicinal relevance and improve pharmacological applicability. Materials and Methods: The antioxidant capacities (1,1-diphenyl-2-picrylhydrazyl [DPPH], 2,2 '-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) [ABTS], and ferric reducing antioxidant power [FRAP]) and phenolic contents of EGCMO and EGEMO were determined in vitro. Male Wistar rats (10-12 weeks) received 1 or 2 mL/100 g body weight of EGCMO or EGEMO. Controls received distilled water, while a reference group received testosterone enanthate (4 mg/kg, intramuscular, every 3 days). Treatments lasted 30 or 60 days, after which reproductive organs and blood were analyzed for biochemical, hormonal, and histological parameters. Results: EGCMO showed higher FRAP and EGEMO greater ABTS activity. EGCMO (2 mL/100 g) and both EGEMO doses significantly increased daily and total sperm production (p < 0.05) and improved sperm motility compared to controls and testosterone-treated rats. EGEMO enhanced catalase (CAT) activity after 30 days, while EGCMO did so after 60 days, with no adverse lipid effects. Conclusions: Both EGCMO and EGEMO improved male reproductive function, supporting the ethnomedicinal use of palm oil against infertility. The emulsified form enhanced efficacy and bioavailability, bridging traditional African remedies with modern nutraceutical innovation.
