Andrologia

Primary ciliary dyskinesia (PCD) is a rare genetic condition characterized by destructive respiratory disease and laterality abnormalities due to randomized left–right body asymmetry. In men, PCD is also often associated with infertility due to immotile sperm owing to a malfunction of the sperm flagella. Pathogenic mutations have been found in more than 50 genes. Nonetheless, not all patients with PCD experience infertility. Therefore, to better understand the impact of PCD-associated mutations on male fertility, it is necessary to clarify the role of these genes in spermatogenesis. The CCDC103 p.His154Pro mutation has a high prevalence in PCD. Here, we present the identification and functional analysis of a biallelic mutation in CCDC103 identified in a familial case of PCD associated with male infertility. The biallelic CCDC103 mutations, NM_213607:c.161_162del(p.His55Serfs9) and NM_213607:c.461A > C (p.His154Pro), were identified by whole-exome sequencing. Sanger sequencing validation was performed on all available family members, and the mutation was recessively separated with an infertility phenotype. The c.161_162del mutation breaks the reading frame of the protein and, therefore, is predicted to produce a nonfunctional protein. The tertiary structure of CCDC103-mutated protein indicated a significant conformational change that likely affected protein function. Transmission electron microscopy of spermatozoa showed that both the mid and principal regions of the flagellum lacked dynein arms, which was confirmed via immunofluorescence staining. Using the method of laser-assisted immotile sperm selection combined with intracytoplasmic sperm injection, the patient’s wife has a successful clinical pregnancy. These results extend the phenotype spectrum of the CCDC103 mutation in PCD.

This study aimed to investigate the potential benefits of frozen testicular sperm and to retrospectively analyze the clinical pregnancy and live birth outcomes following intracytoplasmic injection of fresh or frozen testicular sperm. A total of 468 infertile couples undergoing intracytoplasmic sperm injection cycles utilizing either fresh or frozen testicular sperm extracted between January 2017 and December 2021 were included in this analysis. Participants were categorized into two groups: those utilizing fresh testicular sperm (n = 324) and those utilizing frozen testicular sperm (n = 144). Outcome measures encompassed fertilization rate, embryo development, clinical outcomes, and birth status of infants. Statistical analysis revealed no significant differences in 2PN fertilization rate, Day 3 (D3) available embryo rate, high-quality embryo rate, blastocyst formation rate, clinical pregnancy rate, live birth rate, and preterm birth rate between the two groups. Although the preterm birth rate in the frozen testicular sperm group was lower compared to the fresh testicular sperm group, the difference was not statistically significant (5.6% vs. 15.1%). Notably, there were no discernible distinctions in clinical pregnancy and birth outcomes, as well as infant birth parameters, between the fresh and frozen testicular sperm groups. These findings suggest that frozen testicular sperm holds practical value and warrants consideration for clinical application.

To investigate the correlation between methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism, homocysteine, and male infertility with varicocele in North China. One Hundred infertile males with varicocele (VC; grade II-III, VC group) and 100 healthy males with normal semen parameters and no varicocele (NC group) were recruited for PCR microarray, blood and semen testing. Compared with the CC genotype in the NC group, the TT genotype in the NC group and the CC genotype in the VC group showed no significant changes in sperm motility (; ), sperm density (, ), plasma homocysteine level (; ), and seminal plasma MDA (; ). In contrast, VC patients with the TT genotype had higher plasma homocysteine level and seminal plasma MDA levels (), lower partial pressure of oxygen in seminal pulse (PO₂; ) and poorer sperm quality (), as compared with the CC genotype. This suggests that MTHFR C677C>T may not be a risk factor for male Varicocele in North China. However, this may affect the oxidative stress associated with homocysteine expression, which in turn affects semen parameters in VC patients. Larger studies are needed to validate our findings.

The purpose of this study was to evaluate the differences in sperm function and embryo morphokinetics following sperm cryopreservation by ultra-rapid freezing or rapid freezing methods compared to fresh spermatozoa. Thirty samples of normozoospermia were divided equally into fresh, ultra-rapid freezing, and rapid freezing groups. In the rapid freezing, sperm suspension was placed horizontally on nitrogen vapor. In the ultra-rapid freezing, sperm suspension with a straw-in-straw system was directly immersed in liquid nitrogen. Sperm function was assessed in terms of motility, morphology, viability, mitochondrial membrane potential, sperm DNA fragmentation, and acrosome reaction status. Also, the effects of two cryopreservation methods were assessed on global DNA methylation and DNA methyltransferase activity. Moreover, 730 embryos in three groups were cultured using time-lapse imaging until day 6 for embryo morphokinetics. Progressive motility (38.80 ± 4.21 vs. 34.86 ± 4.19; ) and viability (64.30 ± 6.24 vs. 58.10 ± 8.69; ) in ultra-rapid freezing were significantly higher than rapid group. DNA fragmentation and acrosome reaction were significantly increased in both cryopreserved groups (). However, DNA fragmentation (16.30 ± 1.14 vs. 14.33 ± 2.94; ) was significantly higher in the rapid than the ultra-rapid freezing group. No significant differences were noted in global DNA methylation () and DNA methyltransferases activity () in fresh compared to cryopreservation groups. The kinetic times, including tPB2, tPNa, tPNf, t2, t3, t4, t5, t6, t7, t8, and tM, showed a significant delay in cell divisions in both cryopreservation groups. Furthermore, tPNa, tPNf, and t8 occurred with a significantly higher delay in embryos fertilized by sperm from the rapid freezing compared to the ultra-rapid freezing group. In addition, blastocysts formation was similar in both cryopreservation groups. Ultra-rapid freezing preserved the sperm biological integrity and lead to better embryo morphokinetics compared to the rapid freezing method. However, both methods of sperm cryopreservation were epigenetically safe.

Objective. The presence of infertility in couples not only results in heightened stress but also elevates the risk of developing psychological disorders, further exacerbating their situation. This study is designed to evaluate the relationship between semen analysis and depression, anxiety, and stress, as well as sexual function in infertile men. Method. This descriptive cross-sectional study was carried out at Amir Al-Momenin Hospital of Zabol between March 2019 and April 2020. The couples who were candidates for intracytoplasmic sperm injection according to male factors were recruited for the study. One hundered two men were asked to complete the depression, anxiety, and stress questionnaire (DASS-42) and the International Index of Erectile Function questionnaire (IIEF-15). The findings of the DASS and IIEF questionnaires were compared with the semen analysis test and reported. Results. The frequency of depression, anxiety, and moderate to very severe stress were found in 21 (20.6%), 40 (39.2%), and 23 (22.5%) cases, respectively. Surprisingly, 60 (59%) participants had erectile dysfunction. A significant correlation was found between mood disorders (depression, anxiety, and stress) and some components of the erectile function questionnaire, stress with erectile function (), depression with overall satisfaction (), and anxiety with intercourse satisfaction () and overall satisfaction (). Also, there was a significant correlation between semen analysis parameters such as sperm motility with depression () and anxiety (), normal sperm morphology with stress (), progressive sperm motility with orgasmic function (), and intercourse satisfaction (). Conclusion. The higher the mental health, the higher the erectile function. Improvement of mood disorders leads to improvement of semen parameters and increases the chances of pregnancy. Psychological support may be helpful so, in addition to medical treatment, these people should also receive supportive psychological treatment.

Cold stimulation is the most common stressor in cold regions. Continuous cold stimulation can cause a series of pathophysiological changes in the body, such as aggregated neutrophils, macrophage activation, and increased inflammatory factors, which is also a risk factor for erectile function impairment. In addition, necroptosis is an important form of programmed cell death. However, the mechanisms of necroptosis in erectile function impairment due to cold stimulation have been very poorly studied. Therefore, we explored the mechanism of tumor necrosis factor-α (TNF-α)-mediated receptor interacting protein kinase 1 (RIP1)/receptor interacting protein kinase 3 (RIP3) necroptosis pathway on erectile function among cold-stressed rats. First, we established a cold-stressed rat model using cold stimulation and selected those rats that had developed erectile function impairment. Then, we used Necrostatin-1 (RIP1 specific inhibitor, Nec-1), Etanercept (TNF-α inhibitor, Ent), and Sildenafil (Sil) to intervene for 14 days and subsequently assessed their erectile function by apomorphine test and sexual behavioural test. Lastly, we performed various molecular studies and histopathological analyses of penile tissues collected from these rats after the experiments. We found that erectile function was impaired in cold-stressed rats, with increased penile tissue fibrosis and elevated levels of TNF-α and necroptosis. Contrastingly, intervention with Nec-1 and Ent restored erectile function, reduced penile tissue fibrosis, and decreased TNF-α and necroptosis levels, consistent with the results of intervention with Sil. Based on these results, we confirmed that the TNF-α-mediated RIP1/RIP3 necroptosis pathway was significantly altered in cold-stressed rats. In conclusion, inhibition of the TNF-α-mediated RIP1/RIP3 necroptosis pathway improved erectile function, suggesting that the specific downstream mechanisms need to be further explored.