Andrology最新文献

Background: Oligoasthenozoospermia is a common cause of male infertility, for which effective treatments are urgently needed. Humanin (HN) is a peptide associated with this condition.

Objectives: To investigate the ameliorative effect of [Gly14]-Humanin (HNG) on oligoasthenozoospermia and the mechanisms.

Materials and methods: Mice were treated with cyclophosphamide (CP) to construct a mice model of oligoasthenozoospermia. The resulting model mice were treated with saline or HNG. Subsequently, the testis weights, organ indices, testicular structure, sperm counts and motilities, litter sizes, and serum testosterone concentrations of the mice were determined. Differential gene expression in testicular tissues was determined by RNA sequencing. TM3, TM4, GC1, and GC2 cells were exposed to erastin to induce ferroptosis, followed by treatment with HNG or HNG + ML385 (a nuclear factor erythroid 2-related factor 2 inhibitor). Levels of reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), and ferrous ions (Fe²⁺) were determined and their expression of ferroptosis-related proteins was determined by immunofluorescence and western blot.

Results: The HNG treatment improved testis and sperm parameters and increased litter size and serum testosterone concentrations in model mice. Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analysis revealed significant differential expression of ferroptosis-related genes. The expression of ferroptosis-related proteins increased in testicular tissues after the HNG treatment. The concentrations of ROS, MDA, and Fe²⁺ decreased and the concentrations of GSH increased in testicular tissues and in TM3 and TM4 cells after HNG treatment. In vitro experiments confirmed that HNG activated the nuclear factor erythroid 2-related factor 2/glutathione peroxidase 4 (Nrf2/GPX4) pathway. However, these effects of HNG were blocked by ML385 treatment.

Discussion and conclusion: HNG demonstrated a therapeutic effect on oligoasthenozoospermia in a mouse model by reducing oxidative stress and ferroptosis. In TM3 and TM4 cells, HNG attenuated cellular oxidative stress and inhibited ferroptosis via the Nrf2/GPX4 pathway.

Background: Common marmosets (Callithrix jacchus) are increasingly recognized as valuable nonhuman primates (NHPs) for biomedical research due to their small size and short reproductive cycle and lifespan relative to other NHP species. Maximizing the utility of captive research marmosets, including genetically manipulated animals, will require the use of assisted reproductive techniques (ART) including manipulation, storage, and sharing of marmoset sperm. Here, we identify characteristics of high-quality semen samples and validate a simple method for selecting high-quality sperm.

Methods: Computer-assisted sperm analysis (CASA) was used to evaluate sperm quality in semen samples collected from 44 marmosets and assessed the use of the swim-up method for the selection of high-quality sperm was also tested in half the samples as a potential means to optimize in vitro fertilization or intrauterine insemination.

Results: For each reference parameter, samples at or below the 5th percentile were categorized as abnormal sperm, while those above the 5th percentile were considered to be normal. Among normal samples, those at or above the 50th percentile were categorized as high-quality. High-quality semen samples exhibited the following characteristics: semen volume ≥ 30 µL; sperm count ≥ 10⁷/ejaculate; total motility ≥ 35%; and normal morphology ≥ 5%. Sperm isolated by swim-up exhibited superior sperm progressive motility (19.7% ± 4.5 vs. 5.6% ± 2.1; P = 0.01) and normal morphology (13.1 ± 1.59 vs. 7.65 ± 1.1; P < 0.001) compared with unselected sperm.

Conclusion: This study defines robust, statistically supported reference values for evaluating marmoset semen samples to assist with the identification of optimal sperm donors and the selection of high-quality sperm samples for assisted reproduction. Ultimately, these reference values combined with a validated selection method will contribute to consistent standards for the international sharing of genetically diverse and/or gene-edited marmoset sperm for research and reproduction.

This paper investigates the complex relationship between the immune system and male reproductive processes, emphasizing how chronic inflammation can adversely affect male reproductive health. The immune system plays a dual role; it protects and regulates reproductive organs and spermatogenesis while maintaining reproductive health through immune privilege in the testes and the activities of various immune cells and cytokines. However, when chronic inflammation persists or intensifies, it can disrupt this balance, leading to immune attacks on reproductive tissues and resulting in infertility.This study provides a detailed analysis of how chronic inflammation can impair sperm production, sperm quality, and the secretion of gonadal hormones both directly and indirectly. It also delves into the critical roles of testicular immune privilege, various immune cells, and cytokines in sustaining reproductive health and examines the impacts of infections, autoimmune diseases, and environmental factors on male fertility.

Background: Imatinib and dasatinib are tyrosine kinase inhibitors (TKIs) increasingly used to treat several diseases in both children and adults at fertile age. We have previously shown that imatinib has adverse effects on developing testis, and imatinib-treated male patients have been reported to have reduced sperm counts. However, the cellular level effects of imatinib and dasatinib on adult male germ cells and germline stem cells (mGSCs) have not been thoroughly investigated.

Objectives: To analyze whether imatinib or dasatinib exposure ex vivo and in vitro is harmful to adult male rodent germ cells and mGSCs.

Materials and methods: Seminiferous tubule segments of adult male mouse or rat were cultured in the presence or the absence of imatinib or dasatinib. Stage-specific effects were monitored by ³H-thymidine incorporation assay (DNA synthesis), immunohistochemistry (cleaved Caspase-3; apoptosis), immunofluorescence (KI67, GFRα1, STRA8, c-KIT, LIN28A; proliferation and spermatogonial differentiation) and flow cytometry (Hoechst). Mouse mGSCs were exposed to imatinib and dasatinib to study proliferation, apoptosis, and differentiation.

Results: Imatinib decreased stage-specific DNA synthesis, and induced apoptosis in cultured rat seminiferous tubule segments. Imatinib also had an adverse effect on mGSC proliferation both in vitro and ex vivo, but did not induce cell death in cultured mGSCs. Imatinib did not impinge on induction of spermatogonial differentiation but suppressed c-KIT expression in nascent differentiating spermatogonia, providing a plausible mechanism for its pro-apoptotic function in spermatogenic cells. Clinically relevant doses of dasatinib did not induce apoptosis in seminiferous tubules but decreased mGSC colony growth in vitro.

Conclusions: Imatinib exposure ex vivo and in vitro impinges on male rodent germ cell proliferation and survival. The plausible mechanism in spermatogenic cells is the inhibition of SCF/c-KIT signaling, and reduced expression of c-KIT. Dasatinib did not show significant adverse effects with clinical doses ex vivo but inhibited mGSC colony growth in vitro.

Aim: To compare clinical outcomes using short and long co-incubation protocols in sibling oocytes based on embryo morphokinetic outcomes measured by time-lapse incubator with stratification based on a woman's age and sperm quality.

Design: Our study included 72 cycles with >6 oocytes retrieved. Sibling oocytes were distributed for two parallel protocols: short (3 h; n = 421) or long (16-20 h; n = 434) insemination, using the same amount of spermatozoa from the same prepared sample. Oocytes were then washed and incubated for 5 days. Time-lapse annotations of embryos were performed by experienced embryologists and artificial intelligence-based Known Implantation Data scores for day 3 and day 5 were calculated with EmbryoScope software.

Results: Short-insemination group exhibited a higher blastulation rate, better morphokinetic indicators, and higher Known Implantation Data scores on day 3 and day 5 of the utilized embryos. However, the fertilization rate and clinical pregnancy rate per embryo transfer did not differ between experimental groups. A higher rate of abnormal fertilization (>2 pronuclei) after long insemination was recorded in women under 35 years old or with a total motile sperm count above 5 million and above 40% motility after preparation. A higher rate of usable embryos was observed after short insemination with a total motile sperm count above 30 million before preparation or 5 million and over 40% motility after preparation.

Conclusions: Our results suggest that a short insemination protocol results in better embryo quality and should be considered as a favorable protocol, especially in young female patients or male patients with high sperm quality.

Background: Infertility affects a significant percentage of couples worldwide, with male infertility contributing substantially in a considerable number of cases. Research indicates that oxidative stress is a critical factor impacting male fertility.

Objective: To explore the relationship between semen static oxidation-reduction potential (sORP), sperm parameters, and validated biomarkers of oxidative stress in infertile men.

Materials and methods: This cross-sectional study involved 202 men diagnosed with idiopathic male factor infertility and male partners from couples with unexplained infertility. Multivariable linear regression to query the associations between sORP, sperm parameters, and oxidative aggression biomarkers (lipid peroxidation, mitochondrial membrane potential, annexin V, and sperm DNA fragmentation).

Results: SORP has no linear association with any semen analysis parameter. Furthermore, its relationship with validated biomarkers of oxidative stress was inconsistent. sORP was inversely related to lipid peroxidation (multivariable linear regression coefficient: -0.64), positively associated with sperm DNA fragmentation (multivariable linear regression coefficient: 3.20), and unrelated to mitochondrial membrane potential or annexin V.

Conclusions: There is no clear or consistent relationship between sORP and validated oxidative aggression biomarkers or sperm parameters. Our findings suggest that sORP is unlikely to be helpful in the evaluation of a male with idiopathic infertility.

Background: Spermatogonial stem cells (SSCs) are essential for adult spermatogenesis. Themolecular mechanisms driving SSC generation are poorly understood.

Objectives: Zou et al. reported that the precursor cells that give rise to SSCs-prospermatogonia (ProSG) require the RNA-binding protein, DDX20, in orderto undergo the obligatory proliferative re-activation step that proceeds SSC formation.

Materials and methods: Literature search.

Results and conclusion: We summarize the authors' discovery that the RNA-binding protein, DDX20, iscritical for driving the proliferative re-activation of ProSG, a key step that proceeds SSC generation in vivo. They provide evidence that DDX20 performs this role through its ability to promote the translation of mRNAs encoding proteins known to be essential for cell-cycle and spermatogonial homeostasis. It remains to be determined whether this role is conserved inhumans. It will also be interesting to elucidate whether other post-transcriptional regulators also have roles in early germ cell development. More broadly, it will be fascinating to determine whether post-transcriptional regulators workin concert with transcriptional regulators to drive germ-cell development.

Background: With the increasing trend ofpopulation aging, erectile dysfunction (ED) among elderly men has emerged as apressing health concern. Despite extensive research on the relationship betweenED and aging, ongoing discoveries and evidence continue to arise.

Objective: Through this comprehensiveanalysis, we aim to provide a more nuanced theoretical framework for thedevelopment of preventive and therapeutic strategies for senile ED, ultimatelyenhancing the quality of life for elderly men.

Methods: This review delves deeper into thecore mechanisms underlying ED in the context of aging and offers acomprehensive overview of published meta-analyses and systematic reviewspertinent to these conditions.

Results and conclusion: Our findings revealthat local structural damage to the penis, vascular dysfunction, neuronalinjury, hormonal alterations, other physiological changes, and psychologicalbarriers all play pivotal roles in the pathogenesis of aging-related ED.Furthermore, more than 20 diseases closely associated with aging have beenimplicated in the occurrence of ED, further compounding the complexity of thisissue.

Background and objective: Studies examining ambulatory blood pressure (BP) response to testosterone replacement therapy are needed owing to inconsistent prior findings across formulations. This study assessed testosterone gel 1.62% and 24-h ambulatory BP.

Materials and methods: Single-arm non-inferiority trial (NCT04274894) conducted at 36 US sites enrolled 246 men with hypogonadism (mean age, 57.6 years; mean office systolic/diastolic BP [SBP/DBP], 129.8/79.5 mm Hg) who were treated for 16 weeks with once-daily testosterone gel treatment (starting dose, 40.5 mg/day; min, max dose, 20.25, 81.0 mg/day) to achieve testosterone concentration of 350-750 ng/dL. Main outcome measures included mean change in 24-h average SBP (primary endpoint) and DBP from baseline to week 16. The non-inferiority threshold was a two-sided 95% confidence interval (CI) upper limit <3.0 mm Hg for 24-h average SBP.

Results: Increase in mean ± SD serum testosterone concentration to a physiologic level (baseline, 244.4 ± 93.9 ng/dL; week 16, 502.5 ± 394.4 ng/dL) was associated with a 1.9-mm Hg mean change in 24-h average SBP observed in the primary analysis (baseline, 123.5 mm Hg; week 16, 125.4 mm Hg; 95% CI, 0.63-3.13 mm Hg; n = 169). As the upper CI limit modestly exceeded the non-inferiority margin (3 mm Hg), study drug effect on SBP could not be ruled out. Non-inferiority was observed in subgroups without hypertension or diabetes (95% CI, upper limit <3.0 mm Hg) and was not observed in those with hypertension or diabetes. Daytime SBP and DBP changes were larger compared with nighttime. No clear cardiovascular adverse events or new safety signals were identified.

Discussion and conclusions: While the effect of testosterone gel 1.62% on 24-h average SBP could not be ruled out based on the study's non-inferiority margin, the clinical relevance of the small-magnitude mean increase of 1.9 mm Hg is anticipated to be minimal considering the results of the TRAVERSE study of testosterone gel 1.62% and major adverse cardiac events.

Clinical trial information: ClinicalTrials.gov identifier: NCT04274894 (registered February 17, 2020).

Background: Histological analysis of the testicular sections is paramount in infertility research but tedious and often requires months of training and practice.

Objectives: Establish an expeditious histopathological analysis of mutant mice testicular sections stained with commonly available hematoxylin and eosin (H&E) via enhanced deep learning model MATERIALS AND METHODS: Automated segmentation and cellular composition analysis on the testes of six mouse reproductive mutants of key reproductive gene family, DAZ and PUMILIO gene family via H&E-stained mouse testicular sections.

Results: We improved the deep learning model with human interaction to achieve better pixel accuracy and reduced annotation time for histologists; revealed distinctive cell composition features consistent with previously published phenotypes for four mutants and novel spermatogenic defects in two newly generated mutants; established a fast spermatogenic defect detection protocol for quantitative and qualitative assessment of testicular defects within 2.5-3 h, requiring as few as 8 H&E-stained testis sections; uncovered novel defects in AcDKO and a meiotic arrest defect in HDBKO, supporting the synergistic interaction of Sertoli Pum1 and Pum2 as well as redundant meiotic function of Dazl and Boule.

Discussion: Our testicular compositional analysis not only could reveal spermatogenic defects from staged seminiferous tubules but also from unstaged seminiferous tubule sections.

Conclusion: Our SCSD-Net model offers a rapid protocol for detecting reproductive defects from H&E-stained testicular sections in as few as 3 h, providing both quantitative and qualitative assessments of spermatogenic defects. Our analysis uncovered evidence supporting the synergistic interaction of Sertoli PUM1 and PUM2 in maintaining average testis size, and redundant roles of DAZ family proteins DAZL and BOULE in meiosis.