Sperm Density Research Articles

Determination of the effects of in-ovo chrysin addition to fertile quail eggs on testicular histology, oxidative stress and semen quality.

In this study, the effects of in-ovo injection of Chrysin (CR) into Japanese quail eggs on testicular histology, oxidant status and epididymal sperm quality were investigated. 720 Japanese quail eggs were divided into 4 groups and 0.1 mL saline was given to the control group, 0.25 mg CR to the 0.25 group, 0.50 mg CR to the 0.50 group and 0.75 mg CR to the 0.75 group. On the 60th day after the laying of eggs, 8 Japanese quails were randomly selected from each group and sacrificed under mild sevoflurane anesthesia. The abdominal cavity was opened and both testicular tissues and epididymal parts were removed. The right testicular tissue was used for histopathological examinations and the left one was used for biochemical analyses. The epididymal part at the tip of the vas deferens at the end of the right testis was trimmed in 100 μL saline at 37°C. Histopathological examinations showed that histological scoring was higher in the CR 50 and CR 75 groups. A decrease in oxidant status was observed in all CR groups compared to the control group. Spermatozoa density was higher in CR groups compared to the control group. Total motility value was statistically significantly higher in CR 50 and CR 75 groups compared to control and CR 25 groups. There was no statistical difference between the groups in terms of dead and abnormal spermatozoa. In-ovo CR injection at doses of 0.50 and 0.75 mg/egg improved testicular histological score, decreased oxidative stress, and increased epididymal sperm quality.

Effect of sea buckthorn (Hippophae rhamnoides) on kidney and testicular damage, sperm quality and expression of Irisin and Asprosin in Streptozotocin-Induced diabetic rats

In recent years, many antioxidants have been used against hyperglycemia and oxidative damage in diabetes. The purpose of this study is to investigate the protective effects of Sea buckthorn (Hippophae rhamnoides) against adverse effects of diabetes on testicular and renal tissues. A total of 39 male Sprague-Dawley rats were assigned to 5 groups as follows: control, citrate, SBT, diabetes, and diabetes+SBT. Diabetes induction was made by streptozotocin (50 mg/kg intraperitonally) to diabetes and diabetes+SBT group. SBT oil was administered to SBT and diabetes+SBT group (50mg/ kg/48h by oral gavage). At t e end of study, testicular and kidney samples for histochemical and immunohistochemical examinations, serum samples for biochemical examinations and sperm samples for spermatogenic examinations were collected. The results of the analysis showed that SBT reduced body weight loss and lowered blood glucose levels by reducing the harmful effects of diabetes-induced oxidative stress. When serum TAS and TOS levels were evaluated, it was determined that TAS level was the highest in the SBT group and TAS level increased in the diabetes + SBT group compared to diabetes and the other groups. While TOS level increased in the diabetes group, it decreased in the diabetes+SBT group. SBT also increased sperm density and motility and reduced total abnormality (head-tail) in diabetic rats. In SBT-treated diabetic rats, histopathological changes during the course of diabetes significantly reduced. In addition, decreased irisin expression in renal tissue and decreased irisin and asprosin expression in testicular tissue in the diabetes group were significantly normalized in the diabetes+SBT group. In this study, it was found that the application of SBT oil significantly prevented hyperglycemia and oxidative stress in diabetes, and protected testicular and renal tissues from the functional and histopathological changes in these organs caused by hyperglycemia and oxidative stress in diabetic animals. These results showed that SBT oil was an effective nutritional supplement that can be used to protect against the adverse effects of diabetes.

P-401 Predicting fertility in young boys after gonadotoxic treatments: recovery of spermatogenesis

Abstract Study question Can the cyclophosphamide equivalent dose (CED) given to young boys be used to predict future infertility? Summary answer Our data suggests that CED alone is not a good predictor of subsequent infertility. What is known already Currently young males are referred for fertility preservation based on the estimation of risk of infertility after gonadotoxic treatment utilizing the calculation of the cyclophosphamide equivalent dosage (CED). It has been generally assumed that, in young males, a CED of 3-4 g/m2 results in a risk level of 50 to 80% for future infertility and that ≥ 6 g/m2 results in a higher risk of infertility. However, data regarding the relative CED and subsequent adult recovery of spermatogenesis for these males is sparse. Study design, size, duration Testicular Tissue Cryopreservation (TTC) is offered at our centre1 to families of young boys with childhood cancer or immunologic disorders who will receive gonadotoxic treatment with >3 g/ m2 CED. A number of these boys are now adults and our aim was to assess the impact of this chemotherapy on their subsequent fertility as assessed by the presence of sperm in ejaculates. Participants/materials, setting, methods TTC has been performed for prepubertal or peripubertal boys (total =255), of whom 67 are now aged ≥ 18 years. 23 adult males have returned to produce a semen sample. Semen was provided a minimum of 5 years after cessation of treatment in all cases. Data is available for 11 boys who had testicular tissue frozen at age ≤13 years and 12 who had sperm observed during dissection of tissue (age 13-18 years). Main results and the role of chance In those who were pre /peri pubertal at the time of TTC, 9/11 adult males had some viable sperm and 5/11 had a normal density (>20 million/ml). The lowest CED at which no sperm were detected was 6.5 g/m2. However, 2 adult males had recovery to normal density following high CED (14 and 16 g/m2). In adult males who had some sperm frozen during TTC as post pubertal males, 10/12 had some viable sperm and 4/12 had normal densities. The lowest CED at which there were no sperm detected was 12 g/m2 but recovery to normal range also took place following 12 g/m2 in 1 male. The results showed wide variability in sperm density with CED, with a trend to lower sperm density at higher CED, and a rapid decline in sperm density at around 4 g/m2 CED in both groups. Limitations, reasons for caution The number of males who have returned to produce a semen sample for evaluation is low and needs to be increased to draw more reliable conclusions. Wider implications of the findings Discussion regarding chemotherapy related future infertility should be had with all parents / boys having a CED > 4 g/m2 and options provided to preserve testicular tissue and /or sperm. Reference: 1. Ho et al., (2017) Clinical Endocrinology. 87(3): p. 279-285 Trial registration number No

Chlorophenols suppress gametogenesis by disrupting sex hormone signaling through DNA methylation in zebrafish.

Chlorophenols suppress gametogenesis by disrupting sex hormone signaling through DNA methylation in zebrafish.

Testicular function after immune-checkpoint inhibitors treatment.

e24096 Background: Immune-checkpoint inhibitors (ICIs) have revolutionized cancer treatment, demonstrating durable survival benefits across a wide range of malignancies, from advanced to early-stage disease. While endocrine toxicities of ICIs are well-known, their impact on male reproductive function remains poorly understood. This prospective study aimed to evaluate the effects of ICIs on testicular function in men with melanoma, and no prior gonadotoxic treatments. Methods: This observational, mixed longitudinal and cross-sectional study enrolled men of reproductive age with melanoma requiring systemic therapy at two Australian centres. Baseline andrological assessments included medical history, physical examination, and sperm cryopreservation. Semen analyses (volume, density, motility, morphology) and reproductive hormone measurements (serum LH, FSH, testosterone, SHBG) were performed at baseline and during follow-up. Patients with prior gonadotoxic treatments such as pelvic radiotherapy were excluded. Changes in reproductive parameters were analysed using mixed-effects models for repeated measures with adjustment for potential confounders. Results: Among 29 men with melanoma (22 stage II/III, 7 stage IV), aged 25 ± 1 years, 26 were included in the analysis after excluding 3 who received gonadotoxic treatments. Treatment regimens included anti-PD1 in 24 patients (nine alone, seven with anti-CTLA4, four with anti-LAG3, and six with BREF/MEK inhibitors). Over a median follow-up of 2.8 years (IQR 1.7, 4.5 years), there was a marginally significant reduction in sperm output (34% reduction from baseline median of 193 to 127 million; p = 0.053) and sperm density (30% reduction from 61 to 43 million/mL; p = 0.075), with no changes in motility or morphology. One 28-year-old patient developed isolated azoospermia (persisting three years post-treatment) accompanied by increased serum FSH, while serum testosterone and LH levels remained stable. No other medications were administered, and no immune-related endocrinopathies were identified. Sensitivity analyses excluding this patient showed no significant reductions in sperm parameters. Conclusions: ICIs, widely used across multiple cancers, have minimal impact on male fertility, with stable testicular endocrine function. The observed mild impact on sperm parameters was primarily driven by a single case of persistent azoospermia ( < 5%). These findings highlight the importance of sperm cryopreservation prior to ICI initiation and the need for systematic reproductive health monitoring during and after treatment. Further larger studies are required to confirm these findings, explore potential mechanisms, and develop protective strategies to preserve fertility in male cancer patients undergoing immunotherapy.

Long-term exposure to environmentally relevant concentrations of tri-m-cresyl phosphate induces fecundity decline and gonadal dysplasia by disrupting reproductive endocrine homeostasis.

Long-term exposure to environmentally relevant concentrations of tri-m-cresyl phosphate induces fecundity decline and gonadal dysplasia by disrupting reproductive endocrine homeostasis.

P-233 Does paternal age affect live birth outcomes after frozen oocyte donation? the impact of semen preparation, use of antioxidants on culture media, and insemination methods

Abstract Study question Does advanced paternal age affect clinical outcomes after egg donation treatment and does the method of culture, preparation or insemination play a role? Summary answer Advanced paternal age negatively impacts clinical outcomes in egg donation, but caution is necessary since this association with may reflect maternal factors. What is known already Although female age is well-established as a key factor in embryo development and genetic constitution, a decline in male reproductive function with age may also be significant. While miscarriages are often linked to chromosomal abnormalities inherited from the mother, recent research has increasingly highlighted the role of advanced paternal age in influencing pregnancy outcomes, particularly miscarriage rates warranting further attention in clinical and research contexts. This shift in focus calls for a more comprehensive understanding of paternal age effects. Study design, size, duration This is a retrospective analysis of 3216 patients who underwent treatment with frozen oocyte donation over 11 years (2014-2023) at a single clinic. The aim was to identify whether paternal age was a significant predictor for clinical outcomes. Secondary analyses addressed whether the method of semen preparation, the difference in embryo culture media usage, the addition of antioxidants and the method of oocyte insemination contributed to the outcomes. Participants/materials, setting, methods Sperm preparation was performed with either microfluidics sperm sorting or density gradient centrifugation (DCG). Warmed oocytes were injected either by ICSI or PICSI. Embryos were cultured using two types of culture media one of which had the addition of antioxidants. Combinations of the above treatments were analysed to see difference in effect. Logistic regression and matched analysis performed to confirm statistical significance. Main results and the role of chance While fertilization and blastocyst formation showed no significant difference across paternal age groups (<40, 40-45, 46-50, 51+), both CPR and LBR declined with advancing paternal age. Specifically, LBR per embryo transferred decreased from 45% in the <40 group to 33% in the 51+ group (p = 0.04). Multivariate regression analysis addressed paternal age, maternal age, egg donor age and number of eggs thawed. While paternal age was a significant predictor of live birth outcomes (OR 0.9 CI: 0.68-0.99),when recipient age was added to the model, neither paternal age nor recipient age remained significant, suggesting they may be collinear factors related to overall parental age. The study also explored sperm preparation techniques and culture media options. Using microfluidics sperm sorting for sperm preparation resulted in a significant improvement in LBR (44% vs 53%, p = 0.01) compared to DCG. Using antioxidants in the culture media improved CPR (43% vs 49%, p = 0.01). The use of PICSI also showed a significant CPR difference (44% vs 54%, p = 0.001), though the sample size was small. The above findings are very interesting as they might guide us for tailored treatment in advanced paternal age or patients with high DNA fragmentation. Limitations, reasons for caution Challenges such as the collinearity between paternal and maternal age, along with the need for fertility treatment, introduce underlying clinical factors that can skew outcomes. Additionally, the retrospective design of the study introduces the potential for treatment biases, which should be carefully considered when interpreting results and drawing conclusions. Wider implications of the findings The study suggests that advanced paternal age may negatively impact clinical outcomes after egg donation , highlighting the need for tailored treatments. It indicates the potential value of further research into paternal age and sperm quality, and may prompt personalized treatment approaches, informing counselling, and updated guidelines in assisted reproduction. Trial registration number No

P-049 Female reproductive tract-on-a-chip for selecting sperm with ultra-low DNA fragmentation index

Abstract Study question How to reduce DNA fragmentation in spermatozoa through in vitro sperm selection? Summary answer We designed and fabricated a female reproductive tract (FRT)-on-a-chip (FRToC) platform to select sperm with ultra-low DFI. What is known already High DNA fragmentation index (DFI) in sperm is a critical determinant of infertility, impacting successful embryonic development and the health and well-being of the offspring.In assisted reproductive technology procedures, the sperm selection methods, the swim-up or density gradient centrifugation (DGC) method, is commonly employed to select motile sperm from semen. However, prolonged contact between motile sperm, immature sperm, and cellular debris may induce the production of reactive oxygen species (ROS), leading to DNA fragmentation in motile sperm. However, selecting sperm with low DFI remains a tremendous challenge. Study design, size, duration We recruited eight males with high DNA fragmentation index (exceeding 27%) from June 2023 to August 2024. Participants/materials, setting, methods We used FRToC device, membrane-based sperm sorter (MSS) method and swim-up methods to select sperm. Then, the DFI was analyzed using sperm chromatin structure assay (SCSA). The sperm motility parameters were analyzed using a computer-assisted sperm analysis (CASA) system. Finally, the proteomics analysis of sperm selected by various methods. Main results and the role of chance Our organ-level selection strategy that isolates sperm with ultra-low DFI, reducing it to as low as 0.13%, compared to 34.57% in original samples. We design and fabricate a female reproductive tract (FRT)-on-a-chip (FRToC) device that mimics the entire physiological conditions of in vivo sperm selection. The FRToC selects sperm with ultra-low DFI (mean: 0.71%) from eight clinical sperm samples with high DFI (mean: 41.93%). Sperm selected by the FRToC exhibit not only exceptionally low DFI but also superior motility and intact acrosome, indicating a heightened potential to enhance fertilization success. Additionally, proteomic analysis has some common pathways when compared with both the swim-up and MSS groups, including hydrogen peroxide metabolic process/reactive oxygen species metabolic process, regulation of hydrolase activity and regulation of proteolysis. Limitations, reasons for caution Clinical research is needed to investigate the effectiveness of this spermatozoa selection method. Wider implications of the findings Our organ-scale selection method underscores the potential of the FRToC to select high-quality sperm, offering a promising improvement for assisted reproductive technologies (ARTs). Trial registration number No

P-036 Sperm Selection Using Microfluidic Techniques significantly improves Sperm DNA Fragmentation (SDF) and Reproductive Outcomes: a systematic review and metanalysis

Abstract Study question Do microfluidic sperm selection improve semen quality parameters and reproductive outcomes versus standard sperm selection methods, swim-up or density gradients? Summary answer Microfluidics improves sperm quality and reproductive outcomes, particularly for patients with high sperm DNA fragmentation, although limited evidence hinders strong recommendations for routine clinical practice. What is known already Despite remarkable technological advances and increasing success rates in recent decades, the success of assisted reproduction techniques remains limited, frequently needing multiple treatments before achieving healthy offspring. Sperm selection is one of the key aspects to be improved, since each single sperm is genetically unique, and its proper selection could enhance the proportion and quality of available embryos, thereby improving success rates per microinjected oocytes, and IVF/ICSI treatments. Microfluidic sperm selection has emerged as a promising sperm selection tool, based on sperm shape and motility behavior through microscopic channels, yet robust evidence is still lacking to determine its actual effectiveness. Study design, size, duration An automated search was conducted in the PubMed and SCOPUS databases, as well as a manual search of the references from the studies identified through the automated search. The search strategy used in all cases was the same: “sperm” AND “microfluidic” AND “human” AND (“reproductive results” OR “DNA fragmentation”). These searches were conducted between October 2023 and October 2024. Participants/materials, setting, methods A total of 196 studies were found, with 115 publications excluded based on title or abstract. Of the remaining 81, 21 were duplicates. After reviewing the full text, 39 studies (articles, abstracts, and conference poster) were included. Data of interest were extracted from the selected publications to create a database used for statistical analysis (ReviewManager5). Mantel-Haenszel model was applied, and Forest plots and Funnel plots were generated, with significant differences considered at a p-value<0.05. Main results and the role of chance The analysis revealed that sperm capacitation using microfluidics enables lower sperm DNA fragmentation (MD=-9.98 [-13.19, -6.76], p < 0.00001), increased progressive motility (MD = 14.50 [7.84, 21.71], p = 0.04), total motility (MD = 10.68 [6.04, 15.31], p < 0.00001) and morphology (MD = 1.41 [0.67, 2.16], p = 0.0002). Significant differences were also found in fertilization rate (OR = 1.22 [1.01, 1.46], p = 0.04), implantation rate (OR = 4.51 [1.42, 14.37],p=0.01), clinical pregnancy (OR = 1.73 [1.22, 2.45],p=0.002), ongoing pregnancy (OR = 1.99 [1.03, 3.83], p = 0.04), live birth rate/first cycle (OR = 1.59 [1.12, 2.24],p=0.009) and per all embryo transfer (OR = 1.65 [1.06,2.55], p = 0.03). No significant differences were found in embryo euploidy (OR = 1.34 [0.88,2.04], p = 0.77), biochemical pregnancy (OR = 1.23 [0.84,1.80],p=0.29), miscarriage rate/cycle (OR = 0.84 [0.54, 1.31], p = 0.35) and per pregnancy (OR = 0.71 [0.50,1.02], p = 0.07), live birth rate/first embryo transfer (OR = 1.60 [0.80,3.22], p = 0.18) and per concluded cycle (OR = 1.03 [0.53,2.00], p = 0.92). Limitations, reasons for caution The study’s limitations such as data heterogeneity and moderate to severe risk of bias in retrospective studies highlight the need for better-designed, randomized controlled studies to confirm these findings. Wider implications of the findings Microfluidic sperm selection involves better reproductive outcomes, and its incorporation into clinical practice should be considered, as improved basic semen parameters and DNA fragmentation lead to enhanced reproductive outcomes. However, specific follow-up of results, especially in particular infertility groups, is advised to determine its real cost/benefit ratio. Trial registration number No

Yi-Jing Decoction Ameliorates Oligoasthenozoospermia by Inhibiting the Oxidative Stress-p38 Mitogen-Activated Protein Kinase-Mediated Mitochondrial Apoptosis Pathway in Leydig and Sertoli Cells

Background: To explore the therapeutic effects of Yi-jing decoction (YJD) on oxidative stress-induced oligoasthenozoospermia (OAZS) in mice, with a focus on its role in regulating mitochondrial apoptosis and testosterone synthesis. Methods: OAZS was induced in Bagg’s albino (BALB)/C mice by intraperitoneal injection of cyclophosphamide for 5 days, followed by oral administration of YJD at low (0.525 g/mL), medium (1.05 g/mL), and high (2.1 g/mL) doses for 5 weeks. A total of 75 mice were randomly divided into five groups: control, model, low-dose YJD, medium-dose YJD, and high-dose YJD. Sperm motility and count and sex hormone levels were measured, and testicular and epididymal tissues were analyzed using hematoxylin and eosin staining, transmission electron microscopy, and Western blotting to evaluate mitochondrial function, oxidative stress markers, and apoptosis-related protein expression. TM3 Leydig cells and primary Sertoli cells were used to assess the role of p38 mitogen-activated protein kinase (p38MAPK) in oxidative stress-induced apoptosis. Results: YJD significantly improved sperm motility and density and serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testosterone (P < 0.05). Histological and ultrastructural analyses showed reduced mitochondrial damage and improved morphology of the seminiferous tubules in the YJD-treated groups. YJD downregulated the expression of apoptosis-related proteins (BCL2-Associated X [BAX], cytochrome complex [Cyt-C], caspase-3) and upregulated B-cell lymphoma 2 (Bcl-2), reducing the BAX/Bcl-2 ratio (P < 0.05). Mitochondrial membrane potential was restored in the YJD groups. In TM3 and Sertoli cells, YJD inhibited the p38MAPK signaling pathway, preventing oxidative stress-induced mitochondrial apoptosis and enhancing the expression of the testosterone-synthesis enzymes steroidogenic acute regulatory protein (StAR) and P450scc. Conclusion: YJD effectively mitigates oxidative stress-induced spermatogenic dysfunction in OAZS by modulating the p38MAPK mitochondrial apoptosis pathway, improving mitochondrial function, and enhancing testosterone synthesis and utilization. These findings suggest that YJD is a potential therapeutic strategy for OAZS.

Safranal Alleviates Cyclophosphamide Induced Testicular Toxicity in Rats.

Safranal, the principal component of Crocus sativus essential oil, is primarily responsible for the characteristic aroma and distinct odor of saffron. Cyclophosphamide (CP), a chemotherapeutic agent commonly used in the treatment of both malignant and non-malignant conditions, is known to induce cytotoxicity in various tissues, particularly within the male reproductive system. This study aimed to investigate the protective effects of safranal against CP-induced reproductive toxicity in Wistar albino rats. CP was administered orally at a dose of 15 mg/kg once per week for 56 days to establish a model of testicular toxicity. In parallel, the treatment group received safranal via oral gavage at a daily dose of 200 mg/kg for the same duration. At the end of the treatment period, spermatological, biochemical, and histological analyses were performed on collected tissue samples. CP administration led to increased dead/live and abnormal sperm ratios, elevated levels of NF-κB, IL-6, TNF-α, and MDA, and a reduction in sperm motility and density, Nrf-2 expression, as well as GSH and GSH-Px activity. In contrast, safranal treatment significantly ameliorated these detrimental effects. In conclusion, safranal demonstrated protective and therapeutic effects against CP-induced reproductive toxicity, suggesting its potential as a supportive agent during chemotherapy.

P-042 The impact of microfluidic sperm sorting device and density gradient centrifugation on sperm DNA fragmentation index: A blinded controlled analysis

Abstract Study question In men undergoing fertility treatment, is microfluidic device a superior processing option compared to Density Gradient Centrifugation (DGC) in terms of DNA Fragmentation Index (DFI)? Summary answer Microfluidic sperm sorting devices appear to provide sperm populations with reduced DNA fragmentation compared to DGC. What is known already DFI has been proposed as an additional predictor of male fertility, as elevated sperm DNA fragmentation has been associated with poor clinical outcomes in Assisted Reproductive Technology (ART). So far, numerous studies have investigated the potential adverse effects of DGC on sperm motility, viability and DNA integrity. Microfluidic devices (MFD) address this challenge by sorting a smaller population of progressively motile and morphologically normal spermatozoa with the required DNA integrity. At the moment, there is no consensus on the most appropriate sperm processing method for optimizing outcomes. Study design, size, duration This blinded, controlled observational clinical study included 34 men undergoing ART and was conducted at Embryolab Fertility Clinic, Thessaloniki, Greece, from June 2023 to December 2024. Each semen sample was evaluated for DFI in its unprocessed state, after DGC and after processing with MFD. DFI was assessed in 500 spermatozoa per sample, using the same sperm chromatin dispersion test. The evaluations were performed by three experienced embryologists, and were blinded to the processing method. Participants/materials, setting, methods Sperm samples were collected via masturbation and were allowed to liquefy at room temperature. Basic semen analysis was conducted according to World Health Organization (WHO) guidelines. The mean age was: 41.5±8.07. DFI was assessed and interpreted as the percentage of spermatozoa with fragmented DNA. Exclusion criteria included retrograde ejaculation, surgically retrieved sperm, severe oligo-astheno-teratospermia, and azoospermia. The cut off limit for DFI was 30%. Main results and the role of chance The statistical software R was utilized for data analysis (age and by-subject variability were accounted for). A p-value below 0.05 was taken to indicate that the difference observed reached statistical significance. Both processing methods demonstrated statistically significant differences compared to the DFI of unprocessed semen samples. The mean DFI of the raw samples was found to be 20.88±15.39%. The DFI of sperm processed using a microfluidic device was 8.29±7.42% (p < 0.05), while the DFI of sperm processed with DGC was 43.09±23.77% (p < 0.05). The difference in DFI between the two processing methods (MFD versus DGC) was 34.79±24.9% (p < 0.05). Subsequently, the samples were divided into two groups based on their raw sample DFI: Group A with normal DFI (24 samples) and Group B with abnormal DFI (10 samples). Using the microfluidic device revealed a statistically significant decrease of DFI in both groups (Group A, p < 0.05 and Group B, p < 0.05) while a similar statistically significant increase in DFI was observed with DGC (Group A, p < 0.05 and Group B, p = 0.0025). Comparing the two sperm processing methods (MFD and DGC) in both groups (A and B) the difference in DFI was 28.71±22.34% in Group A and 49.4±24.33% in Group B. Limitations, reasons for caution The study was performed on a small sample size and did not include clinical outcomes. Wider implications of the findings The outcome of the study indicates that the use of the MFD yields spermatozoa with lower DFI, compared to DGC, improving overall patient prognosis by counterbalancing paternal factor. This finding was consistent to all samples, independent of the DFI in the unprocessed samples. Trial registration number No

Evaluating the Potential Adverse Effects of Favipiravir on Biochemical, Histopathological, and Spermatological Parameters in Male Rats' Testicular Tissue.

Favipiravir is a selective RNA polymerase inhibitor and a broad-spectrum antiviral drug. Favipiravir reduces cell proliferation by inhibiting RNA transcription, particularly in rapidly proliferating cells such as spermatogonia. The aim of this study was to investigate the effects and mechanism of action of favipiravir (T-705) on sperm quality and testicular tissue in rats. A total of 60 Sprague-Dawley rats, 30 in each group, were used in our study. Rats were randomly divided into two groups, control and experimental. Rats were killed on Day 14, Day 21, and Day 50 to observe short- and long-term effects. Oxidative stress, apoptosis, proliferation, aromatase activity, inflammation, histopathological changes, and epididymal sperm quality were examined in the testicular tissue of rats. Favipiravir administration decreased SOD activity and GSH levels and increased MDA levels and 8-OHdG levels in the testes of rats. It increased the levels of Caspase-3 and NF-ĸB, which are apoptotic markers, and decreased the levels of NRF2, PI3K, and Bcl-2, which play a role in the regulation of apoptosis. Favipiravir led to disruption of the seminiferous tubules and disturbances in the structure of cells in the testis. In spermatological analysis, total motility value and epididymal spermatozoa density decreased. On Day 50, the favipiravir groups had higher rates of abnormal spermatozoa, DNA damage, and acrosome damage. In conclusion, favipiravir administration induced oxidative stress by increasing MDA levels and decreasing SOD activity and GSH levels in the testicular tissue of rats. It also affects the release of reproductive hormones by altering the hypothalamic-pituitary axis. Favipiravir administration decreases the expression of genes that induce sperm capacitation and acrosome reaction and decreases sperm quality by causing changes in testicular histoarchitecture. Study results reveal that favipiravir treatment negatively affects testes and semen quality.

Mechanism of Shu-Mu Brain-Kidney Acupoint in Treating Oligoasthenospermia in Rats via the Hypothalamic-Pituitary-Testicular Axis.

Oligoasthenospermia is a common cause of male infertility. The hypothalamic-pituitary-testicular (HPT) axis regulates gonadal differentiation and maturation through reproductive hormone synthesis and release, playing a vital role in male fertility. Disrupting HPT axis stability impairs sperm production, reducing semen quality. Investigating electroacupuncture's effect on HPT axis regulation may provide insights into treating oligoasthenospermia. Fifty 8-week-old male SD rats were randomly divided into blank, model, Shu-Mu Brain-Kidney, non-acupoint, and L-carnitine groups (n = 10 per group). Except for the blank group, rats received adenine intragastrically for 28 days to establish the model. Post-modeling, the Shu-Mu Brain-Kidney group underwent electroacupuncture at designated acupoints, while the non-acupoint group received sham treatment for 30 min daily. The L-carnitine group received L-carnitine (10 mL/kg) intragastrically once daily. Treatments continued for 28 days. General conditions, organ coefficients, and semen quality were assessed. HE staining analyzed tissue morphology, and ELISA detected serum hormone changes. Compared to the model group, the Shu-Mu Brain-Kidney, and L-carnitine groups exhibited significant improvements in spirit, diet, and bowel movements, with increased body weight, while the non-acupoint group showed no significant change. Renal organ coefficients decreased significantly in the Shu-Mu Brain-Kidney and L-carnitine groups but remained unchanged in the non-acupoint group. Testicular organ coefficients showed no significant differences among treatment groups. Sperm count, density, survival, and motility rates improved significantly in the Shu-Mu Brain-Kidney and L-carnitine groups, but not in the non-acupoint group. H&E staining showed ameliorated kidney and testicular tissue damage in the Shu-Mu Brain-Kidney and L-carnitine groups. ELISA revealed increased T, GnRH, and INHB and decreased LH, FSH, E2, and PRL levels in these groups (p < 0.001), with no significant changes in the non-acupoint group. These findings indicate that Shu-Mu Brain-Kidney acupoint therapy improves sperm quality by regulating the HPT axis, offering a potential treatment for oligoasthenospermia.

Research progress on the adverse effects of high-altitude environment to the male reproductive system: a review study.

An increasing number of people are being exposed to high-altitude environments as they become more important in economic development, resource exploitation, and other areas. This review is focused on the impact of the high-altitude environment on the male reproductive system. In high-altitude areas, the unique conditions lead to complex and significant changes in male reproductive hormone levels. The secretion of GnRH is inhibited, which in turn affects the levels of FSH and LH, ultimately influencing testosterone synthesis and secretion, thus disrupting the normal endocrine regulatory network. Testicular tissue also shows marked morphological changes. The seminiferous tubule structure becomes disordered, and the number and function of spermatogenic and interstitial cells are damaged. These alterations have a direct impact on sperm quality, resulting in a decrease in sperm density and motility, an increase in the deformity rate, and damage to genetic material integrity. Additionally, sexual function is affected, with symptoms such as decreased libido and erectile dysfunction being common. The underlying mechanisms involve oxidative stress damage, an abnormal increase in apoptosis, and enhanced autophagy. Nevertheless, current research, especially human-based studies, is restricted by small sample sizes and insufficiently in-depth exploration of these mechanisms.

The effects of cigarette smoke on the epididymal tissues in adult male albino rats and the ameliorative effect of the Sidr honey.

The present study investigated the effects of cigarette smoke (CS) on the epididymal tissues and the protective effect of Sidr honey in adult male rats. 24 adult male rats were divided into four groups: Group 1: normal control group; Group 2:rats received Sidr honey orally (100 mg/kg b.w./d.) for 4 weeks; Group 3: rats were exposed to five lit cigarettes per day (5 times per day) for 4 weeks; and Group 4: rats received Sidr honey orally (100 mg/kg b.w./d.) for 2 weeks, then the rats were treated with cigarette smoke generated by a machine after taking the Sidr honey for 4 weeks. The results presented the gross morphology of the vas L deferens of rats in the group 3was smaller than that of the other groups. Moreover, the testes in group 3 were bigger as compared to all other groups. In addition, results demonstrate the rats in group 3 showed many histopathological changes in epididymal tissues when compared with the group 1. While the epididymal tissues from the rats in the group 4displayednearly normal histological structure of the tubules with normal sperm density as compared to the group 3. This study indicates that Sidr honey has a protective effect against CS-induced epididymal damage in adult male albino rats.

Effects of Dendrobium nobile on antioxidant capacity, hormone levels, testicular metabolism, and reproductive performance of aged roosters.

Oxidative stress is a major cause of semen quality decline in old roosters. Dendrobium nobile Lindl (DNL), a Chinese herbal medicine, exhibits excellent antioxidant activity. Therefore, the present study aimed to investigate the effects of dietary DNL supplementation on semen quality, antioxidant capacity, reproductive hormone levels, and testicular tissue structure in aged roosters. This study further aimed to elucidate the potential mechanism for improving reproductive performance. Thus, the expression of antioxidant defense system-related genes was verified, and metabolomic analysis was performed. Twenty 56-week-old recessive, white-feathered roosters were randomly assigned into two groups. The DNL group was fed a basal diet supplemented with 2500 mg/kg DNL for 60 days, whereas the control group was fed a basal diet. Here, DNL improved the semen quality (sperm density and motility) and antioxidant capacity of aged roosters, increased the expression of genes in the Nrf2 pathway, increased serum hormone levels, and delayed testicular tissue degradation. Seventy-six differential metabolites that are mainly enriched in amino acid biosynthesis pathways, taurine and hypotaurine metabolism, and cysteine and methionine metabolism were identified. DL-serine, DL-cysteine, and α-ketoglutarate were related to improved testicular antioxidant capacity. In this study, dietary supplementation with 2500 mg/kg DNL delayed the decline in reproductive performance by improving the antioxidant capacity of aging roosters. These findings could facilitate the use of DNL as a feed additive to improve the reproductive performance of aged roosters.

Comparison of reproductive performance and functional analysis of spermatogenesis factors between domestic yak and semi-wild blood yak

This study investigates differences in reproductive performance, testicular histology, and transcriptomic profiles between male Subei (SB; semi-wild) yaks and two domestic yaks, Gannan (GN) and Qinghai (QH). Key metrics including mating age, utilization time, breeding capacity, morphometric traits, and testicular indices were analyzed. SB yaks exhibited superior reproductive metrics, including earlier sexual maturity, prolonged utilization periods, and enhanced breeding capacity compared to GN and QH (P < 0.05). Morphologically, SB yaks demonstrated significantly greater body weight, and testicular dimensions. Compared with GN and QH yaks, the seminiferous tubules of SB yaks exhibited significantly larger spermatogenic cells and luminal cavities, along with a notably higher sperm density within the luminal cavity. Transcriptomic analysis identified 2,403 and 4,428 differentially expressed genes (DEGs) in GN vs. SB and QH vs. SB comparisons, respectively. Eight key genes (TPPP3, SMAD3, PAFAH1B3, BMP7, ARSA, CTNNB1, SMAD4, STAT3) and three pathways (Hippo, pluripotency regulation, TGF-β) were implicated in testicular development and spermatogenesis. These findings underscore the genetic and physiological advantages of SB yaks, offering insights for enhancing male yak reproductive performance.

Evaluation of the acute effects of single‐dose TPN171 on semen quality in healthy Chinese male volunteers

AimsTPN171 is a novel phosphodiesterase type 5 inhibitor for the treatment of erectile dysfunction. This study aimed to assess the immediate impact of a single dose of TPN171 (10 mg) on semen quality in healthy male Chinese volunteers. Additionally, the study investigated the exposure of TPN171 in seminal plasma and observed the safety of TPN171 in the participants.MethodsEighteen healthy male volunteers were enrolled in this double‐blind, randomized, placebo‐controlled, 2‐period, 2‐sequence, crossover study. Semen samples were collected to compare semen parameters between the TPN171 and placebo groups. The concentrations of TPN171 were measured in seminal plasma and blood plasma to calculate the ratio of TPN171 concentration in seminal plasma to that in blood plasma, along with the exposure level and percentage of TPN171 in the seminal plasma.ResultsThe administration of a single 10‐mg dose of TPN171 did not result in statistically significant effects on sperm motility, count, density, morphology, viscosity or volume in healthy male Chinese volunteers. The ratio of TPN171 concentration in seminal plasma to that in blood plasma was 0.72, with the amount of TPN171 in the seminal plasma measured at 78.5 ng. The percentage of exposure to the administered dose (10 mg) was 0.00085%. No adverse events were reported in the TPN171 group.ConclusionThe findings indicate that a single oral dose of 10 mg TPN171 did not induce acute effects on semen quality in healthy male Chinese volunteers. The exposure of seminal plasma to TPN171 was &lt;0.001% of the administered dose.Clinical Trial Registration NumberNCT05585931.

Efficacy and mechanism of Cistanches Herba extract in treating reproductive dysfunction in rats with kidney-Yang deficiency based on metabolomics

This study investigates the reproductive protective effect and potential mechanism of Cistanches Herba extract(CHE) on a rat model of kidney-Yang deficiency induced by adenine. Rats were randomly divided into five groups: normal, model, low-dose CHE(0.6 g·kg~(-1)·d~(-1)), high-dose CHE(1.2 g·kg~(-1)·d~(-1)), and L-carnitine(100 mg·kg~(-1)·d~(-1)). The rats were administered adenine(200 mg·kg~(-1)·d~(-1)) by gavage for the first 14 days to induce kidney-Yang deficiency, while simultaneously receiving drug treatment. After 14 days, the modeling was discontinued, but drug treatment continued to 49 days. The content of components in CHE was analyzed by high-performance liquid chromatography. The adenine-induced kidney-Yang deficiency model was assessed through symptom characterization and measurement of testosterone(T) levels using an enzyme-linked immunosorbent assay kit. Pathological damage to the testis and epididymis was evaluated based on the wet weight and performing hematoxylin-eosin staining. Sperm density and motility were measured using computer-aided sperm analysis, and sperm viability was assessed using live/dead sperm staining kits, and sperm morphology was evaluated using eosin staining, thereby determining rat sperm quality. Metabolomics was used to analyze changes in serum metabolites, enrich related metabolic pathways, and explore the mechanism of CHE in improving reproductive function damage in rats with kidney-Yang deficiency syndrome. Compared to the normal group, the model group exhibited significant kidney-Yang deficiency symptoms, reduced T levels, decreased testicular and epididymal wet weights, and significant pathological damage to the testis and epididymis. The sperm density, motility, and viability decreased, with an increased rate of sperm abnormalities. In contrast, rats treated with CHE showed marked improvements in kidney-Yang deficiency symptoms, restored T levels, alleviated pathological damage to the testis and epididymis, and improved various sperm parameters. Metabolomics results revealed 286 differential metabolites between the normal and model groups(191 upregulated and 95 downregulated). Seventy-five differential metabolites were identified between the model and low-dose CHE groups(21 upregulated and 54 downregulated). A total of 24 common differential metabolites were identified across the three groups, with 22 of these metabolites exhibiting opposite regulation trends between the two comparison groups. These metabolites were primarily involved in linoleic acid metabolism, ether lipid metabolism, and pantothenic acid and coenzyme A biosynthesis, as well as metabolites including 13-hydroperoxylinoleic acid, lysophosphatidylcholine, and pantethine. CHE can improve kidney-Yang deficiency symptoms in rats, alleviate reproductive organ damage, and enhance sperm quality. The regulation of lipid metabolism may be a potential mechanism through which CHE improves reproductive function in rats with kidney-Yang deficiency. The potential bioactive compounds of CHE include echinacoside, verbascoside, salidroside, betaine, and cistanoside A.