Transcriptome analysis of meiotic and post-meiotic spermatogenic cells reveals the potential hub genes of aging on the decline of male fertility.

Genetic and epigenetic changes in sperm caused by male aging may be essential factors affecting semen parameters, but the effects and specific molecular mechanisms of aging on male reproduction have not been fully clarified. In this study, to explore the effect of aging on male fertility and seek the potential molecular etiology, we performed high-throughput RNA-sequencing in isolated spermatogenic cells, including pachytene spermatocytes (marked by the completion of chromosome synapsis) and round spermatids (produced by the separation of sister chromatids) from the elderly and the young men. Functional enrichment analysis of differentially expressed genes (DEGs) in round spermatids between the elderly and young showed that they were significantly enriched in gamete generation, spindle assembly, and cilium movement involved in cell motility. In addition, the expression levels of DEGs in round spermatids (post-meiotic cells) were found to be more susceptible to age. Furthermore, ten genes (AURKA, CCNB1, CDC20, CCNB2, KIF2C, KIAA0101, NR5A1, PLK1, PTTG1, RAD51AP1) were identified to be the hub genes involved in the regulation of sperm quality in the elderly through Protein-Protein Interaction (PPI) network construction and measuring semantic among GO terms and gene products. Our data provide aging-related molecular alterations in meiotic and post-meiotic spermatogenic cells, and the information gained from this study may explain the abnormal aging-related male fertility decline.

H3K36me2 methyltransferase NSD2 orchestrates epigenetic reprogramming during spermatogenesis.

Spermatogenesis is precisely controlled by sophisticated gene expression programs and is driven by epigenetic reprogramming, including histone modification alterations and histone-to-protamine transition. Nuclear receptor binding SET domain protein 2 (Nsd2) is the predominant histone methyltransferase catalyzing H3K36me2 and its role in male germ cell development remains elusive. Here, we report that NSD2 protein is abundant in spermatogenic cells. Conditional loss of Nsd2 in postnatal germ cells impaired fertility owing to apoptosis of spermatocytes and aberrant spermiogenesis. Nsd2 deficiency results in dysregulation of thousands of genes and remarkable reduction of both H3K36me2 and H3K36me3 in spermatogenic cells, with H3K36me2 occupancy correlating positively with expression of germline genes. Nsd2 deficiency leads to H4K16ac elevation in spermatogenic cells, probably through interaction between NSD2 and PSMA8, which regulates acetylated histone degradation. We further reveal that Nsd2 deficiency impairs EP300-induced H4K5/8ac, recognized by BRDT to mediate the eviction of histones. Accordingly, histones are largely retained in Nsd2-deficient spermatozoa. In addition, Nsd2 deficiency enhances expression of protamine genes, leading to increased protamine proteins in Nsd2-deficient spermatozoa. Our findings thus reveal a previously unappreciated role of the Nsd2-dependent chromatin remodeling during spermatogenesis and provide clues to the molecular mechanisms in epigenetic abnormalities impacting male reproductive health.

Machine learning approach identifies meconium metabolites as potential biomarkers of neonatal hyperbilirubinemia.

Background: The gut microbiota plays an important role in the early stages of human life. Our previous study showed that the abundance of intestinal flora involved in galactose metabolism was altered and correlated with increased serum bilirubin levels in children with jaundice. We conducted the present study to systematically evaluate alterations in the meconium metabolome of neonates with jaundice and search for metabolic markers associated with neonatal jaundice. Methods: We included 68 neonates with neonatal hyperbilirubinemia, also known as neonatal jaundice (NJ) and 68 matched healthy controls (HC), collected meconium samples from them at birth, and performed metabolomic analysis via liquid chromatography-mass spectrometry. Results: Gut metabolites enabled clearly distinguishing the neonatal jaundice (NJ) and healthy control (HC) groups. We also identified the compositions of the gut metabolites that differed significantly between the NJ and HC groups; these differentially significant metabolites were enriched in aminyl tRNA biosynthesis; pantothenic acid and coenzyme biosynthesis; and the valine, leucine and isoleucine biosynthesis pathways. Gut branched-chain amino acid (BCAA) levels were positively correlated with serum bilirubin levels, and the area under the receiver operating characteristic curve of the random forest classifier model based on BCAAs, proline, methionine, phenylalanine and total bilirubin reached 96.9%, showing good potential for diagnostic applications. Machine learning-based causal inference analysis revealed the causal effect of BCAAs on serum total bilirubin and NJ. Conclusions: Altered gut metabolites in neonates with jaundice showed that increased BCAAs and total serum bilirubin were positively correlated. BCAAs proline, methionine, phenylalanine are potential biomarkers of NJ.

Diversity of the T cell receptor ß chain complementarity-determining region 3 in peripheral blood of neonates with sepsis: an analysis based on immune repertoire sequencing. / å ç–«ç»„åº“æµ‹åºåˆ†æžæ–°ç”Ÿå„¿è„“æ¯’ç—‡æ‚£è€ å¤–å‘¨è¡€T细胞受体ß链CDR3çš„å¤šæ ·æ€§.

OBJECTIVES: To investigate the diversity of peripheral blood T cell receptor (TCR) ß chain complementarity-determining region 3 (CDR3) based on immune repertoire sequencing in neonates with sepsis and the possible pathogenesis of neonatal sepsis. METHODS: A total of 12 neonates with sepsis were enrolled as the case group, and 9 healthy full-term infants, matched for gestational age, birth weight, and age, were enrolled as the control group. Omega nucleic acid purification kit (SQ blood DNA Kit II) was used to extract DNA from peripheral blood samples, TCR ß chain CDR3 was amplified by multiplex PCR, and then high-throughput sequencing was performed for the products to analyze the diversity of TCR ß chain CDR3 and the difference in expression. RESULTS: The length and type of TCR ß chain CDR3 were similar between the case and control groups, and Gaussian distribution was observed in both groups. With D50 and Shannon-Wiener index as the evaluation indices for diversity, the case group had a significantly lower diversity of TCR ß chain CDR3 than the control group (P<0.05). The frequency of 48 genes in TCR ß chain V segment was compared, and the results showed that compared with the control group, the case group had significantly higher frequencies of TRBV10-3, TRBV2, and TRBV20-1 (P<0.05). The frequency of 13 genes in TCR ß chain J segment were compared, and the results showed that compared with the control group, the case group had significantly higher frequencies of TRBJ2-3, TRBJ2-5, and TRBJ2-7 (P<0.05). CONCLUSIONS: There is a significant change in the diversity of TCR ß chain CDR3 in the peripheral blood of neonates with sepsis, suggesting that it might be associated with the immune pathogenesis of neonatal sepsis.

Value of near-infrared spectroscopy in monitoring intestinal tissue oxygen saturation in preterm infants with hemodynamically significant patent ductus arteriosus: a prospective research. / è¿‘çº¢å¤–å ‰è°±æŠ€æœ¯å¯¹æœ‰è¡€æµåŠ¨åŠ›å­¦æ„ä¹‰çš„åŠ¨è„‰å¯¼ç®¡æœªé—­æ—©äº§å„¿è‚ é“ç»„ç»‡æ°§é¥±å’Œåº¦ç›‘æµ‹ä»·å€¼çš„å‰çž»æ€§ç ”ç©¶.

OBJECTIVES: To study the change in regional oxygen saturation (rSO2) of intestinal tissue in preterm infants with hemodynamically significant patent ductus arteriosus (hsPDA) by near-infrared spectroscopy, and the clinical significance of the change in intestinal oxygen level in preterm infants with hsPDA. METHODS: The preterm infants with patent ductus arteriosus (PDA) who had gestational age <32 weeks and/or birth weight <1 500 g were prospectively enrolled, who were admitted to the Department of Neonatology, Shenzhen Longgang Central Hospital from October 2017 to October 2020.According to the diagnostic criteria for hsPDA, the preterm infants with patent ductus arteriosus (PDA) were divided into two groups: hsPDA and non-hsPDA. According to closure of the ductus arteriosus after oral administration of ibuprofen, the preterm infants in the hsPDA group were subdivided into two groups: hsPDA closure and hsPDA non-closure. Hemodynamic parameters were measured at diagnosis of PDA and after treatment, and the level of intestinal tissue rSO2 was monitored continuously to analyze its change. RESULTS: A total of 241 preterm infants with PDA were enrolled, with 55 infants (22.8%) in the hsPDA group and 186 infants (77.2%) in the non-hsPDA group. There were 36 infants (65%) in the hsPDA closure group and 19 infants (35%) in the hsPDA non-closure group. Compared with the non-hsPDA group, the hsPDA group had a significantly higher left atrial diameter/aortic root diameter ratio and significantly lower left ventricular ejection fraction and fractional shortening (P<0.05). At each time point within 6 hours after diagnosis (1, 2, 4, and 6 hours), the hsPDA group had significantly lower intestinal tissue rSO2 than the non-hsPDA group (P<0.05), and intestinal tissue rSO2 gradually decreased over time in the hsPDA group (P<0.05), with the lowest level of 0.448±0.014 at 6 hours. Compared with the hsPDA non-closure group, the hsPDA closure group had a significantly lower left atrial diameter/aortic root diameter ratio and significantly higher left ventricular ejection fraction and fractional shortening (P<0.05). At each time point within 48-96 hours after treatment (48, 72, and 96 hours), the hsPDA closure group had significantly higher intestinal tissue rSO2 than the hsPDA non-closure group (P<0.05), and intestinal tissue rSO2 gradually increased since 24 hours after treatment in the hsPDA closure group (P<0.05), with the highest level of 0.578±0.031 at 96 hours. CONCLUSIONS: hsPDA has an impact on intestinal tissue oxygenation in preterm infants, and continuous monitoring of intestinal tissue rSO2 by near-infrared spectroscopy can help to guide the clinical management of hsPDA in preterm infants.

Overexpression of MicroRNA-10a in Germ Cells Causes Male Infertility by Targeting Rad51 in Mouse and Human.

Spermatogenesis is a complicated process including spermatogonial stem cells self-renewal and differentiates into mature spermatozoa. MicroRNAs (miRNAs) as a class of small non-coding RNAs play a crucial role during the process of spermatogenesis. However, the function of a plenty of miRNAs on spermatogenesis and the potential mechanisms remain largely unknown. Here, we show that genetically conditional overexpressed miR-10a in germ cells caused complete male sterility, characterized by meiotic arrested in germ cells. Analysis of miR-10a overexpression mouse testes reveals that failure of double strand break (DSB) repairs and aberrant spermatogonial differentiation. Furthermore, we identified Rad51 as a key target of miR-10a in germ cell by bioinformatics prediction and luciferase assay, which may be responsible for the infertility of the miR-10a overexpressed mice and germ cell arrested patients. Our data show that miR-10a dependent genetic regulation of meiotic process is crucial for male germ cell development and spermatogenesis in both mouse and human. These findings facilitate our understanding of the roles of miRNA-10a in spermatogenesis and male fertility.

MicroRNA profile comparison of testicular tissues derived from successful and unsuccessful microdissection testicular sperm extraction retrieval in non-obstructive azoospermia patients.

Non-obstructive azoospermia (NOA) is the most severe clinical diagnosis in cases of male infertility. Although in some cases of NOA spermatozoa can be retrieved by microdissection testicular sperm extraction (micro-TESE) to fertilise eggs through intracytoplasmic sperm injection (ICSI), there remains a lack of potential biomarkers for non-invasive diagnosis before micro-TESE surgery. To determine predictive biomarkers for successful sperm retrieval before micro-TESE, the aim of this study was to explore whether microRNAs (miRNAs) were differentially expressed in testicular tissues in NOA patients in whom sperm retrieval had been successful (SSR) versus those in whom it had been unsuccessful (USR) using next-generation small RNA sequencing (RNA-Seq). In all, 180 miRNAs were identified with significantly altered expression levels between SSR and USR testicular tissues. Of these, the expression of 13 miRNAs was upregulated and that of 167 miRNAs was downregulated in the USR compared with SSR group. Unexpectedly, 86 testicular miRNAs were found to be completely absent in the USR group, but showed high expression in the SSR group, suggesting that these miRNAs may serve as biomarkers for micro-TESE and may also play an essential role in spermatogenesis. Furthermore, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses indicated that the miRNAs that differed significantly between the USR and SSR groups were involved in cell apoptosis, proliferation and differentiation, which are of considerable importance during spermatogenesis. In summary, this study identified a panel of miRNAs highly expressed in testicular tissues of SSR but not USR NOA patients, providing new insights into specific miRNAs that may play important roles in epigenetic regulation during spermatogenesis. The findings provide a basis for further elucidation of the regulatory role of miRNAs in spermatogenesis and clues to identifying useful biomarkers to predict residual spermatogenic loci in NOA patients during treatment with assisted reproductive technologies.

Testicular piRNA profile comparison between successful and unsuccessful micro-TESE retrieval in NOA patients.

PURPOSE: PIWI-interacting RNA (piRNA) is a sub-group of small RNAs about 30 nucleotides length which specifically expressed in mammalian germ cells. Although piRNAs play pivotal roles in spermatogenesis regulation, little is known in the testicular tissues of infertile men. To explore whether piRNA profile could serve as a biomarker for male infertility diagnosis in a clinic, in this study, we systematically investigated the expression profile of piRNAs in testicular tissues from the patients with non-obstructive azoospermia (NOA) between successful and unsuccessful sperm retrieval before micro-dissection testicular sperm extraction (micro-TESE). METHODS: The differential expression levels of piRNAs were evaluated using small RNA-Seq method. Ontologic analyses were performed to determine the presence of enriched biological processes. RESULTS: A total of 18,324 Homo sapiens piRNAs were identified by small RNA-Seq from NOA patient testicular tissues; among them, 959 piRNAs were significantly altered between successful and unsuccessful sperm retrieval groups, of which 951 testicular piRNAs were significantly downregulated and 8 piRNAs were upregulated in NOA patients with unsuccessful sperm retrieval (USR) groups compared to those with successful sperm retrieval (SSR) groups, respectively. Unexpectedly, 553 testicular piRNAs were found completely absent in USR but showing abundant in SSR, which suggests that those piRNAs might serve as a biomarker for micro-TESE application. A total of 20 significantly differential piRNAs (hsa-piR-20830, hsa-piR-4731, hsa-piR-6254, hsa-piR-419, hsa-piR-7152, hsa-piR-7548, hsa-piR-14195, hsa-piR-5026, hsa-piR-11482, hsa-piR-17765, hsa-piR-17102, hsa-piR-4484, hsa-piR-17260, hsa-piR-17098, hsa-piR-20511, hsa-piR-5802, hsa-piR-19121, hsa-piR-2510, hsa-piR-4745, hsa-piR-11873) were selected to further validate the RNA-Seq data by quantitative real-time polymerase chain reaction. In addition, bioinformatic analyses revealed that those altered piRNAs were involved in many important biological pathways, including apoptosis, cell proliferation, and differentiation. CONCLUSIONS: Our results demonstrate that testicular tissues from NOA patients with successful and unsuccessful spermatozoa retrieval exhibit differential piRNA profiles. This study provides a useful resource to further elucidate the regulatory role of piRNAs in spermatogenesis and provides a profound clue to identify useful biomarkers for predicting residual spermatogenic loci in NOA patients during assisted reproductive treatment.

[Ningmitai Capsules combined with doxycycline hydrochloride for Ureaplasma urealyticumpositive chronic prostatitis].

OBJECTIVE: To investigate the effects of Ningmitai Capsules (NMT) combined with doxycycline hydrochloride (DH) on chronic prostatitis induced by Ureaplasma urealyticum (Uu). METHODS: This randomized controlled trial included 240 male patients with Uupositive chronic prostatitis, treated orally with NMT at 4 capsules tid (n= 35), DH at 100 mg bid (n = 78), and NMT+DH at the corresponding doses (n = 127), respectively, all for 2 successive weeks. At 1 week after drug withdrawl, we conducted routine urine analysis, EPS examination, and drug sensitivity test of the cultured Uu. RESULTS: The positivetonegative rate of Uu was significantly higher in the NMT+DH group than in the NMT and DH groups (89.0% ［113/127］ vs 54.3% ［19/35］ and 71.8% ［56/78］, P< 0.05), so were the cure rate (25.2% vs 20.0% and 20.5%, P< 0.05) and total effectiveness rate (89.0% vs 54.3% and 71.8%, P< 0.05). CONCLUSIONS: The combination of Ningmitai Capsules and doxycycline hydrochloride is more effective than either Ningmitai Capsules or doxycycline hydrochloride used alone in the treatment of Uupositive chronic prostatitis.

Long-term reproductive consequences of no-scalpel vasectomy in beagles.

The effects of vasectomy on the reproductive organs in various species are controversial. This study investigated the morphological change and apoptosis of the testis, epididymis, and vas deferens in beagle dogs 12 months after vasectomy. The male beagles were divided into two groups: vasectomized and sham-operated groups (n=5 in each). Histopathological, ultrastructural, and TUNEL evaluation of the changes in the testis, epididymis, and ductus deferens of each animal were conducted 12 months after surgery. The mean lumen diameter, cellular thickness, mean interstitial distance, and lumen area fraction of each seminiferous tubule and ductus epididymis were measured by stereological analysis. The results showed that, compared with the sham-operated group, the seminiferous tubular epithelial cells of the testes in the vasectomized group were disorderly arranged and scattered. Significant atrophy and apoptosis were found in the endothelial cells, and a range of ultrastructural variations were observed in the cells of testes, epididymis, and vas deferens in vasectomized group. It was concluded that complete obstruction of the vas deferens as a traditional contraception method is not absolutely safe in terms of the reversal of fertility in the long run. Techniques of relieving the inner pressure in the vas deferens while maintaining the efficacy of male contraception need to be explored.

Biological evaluation of a novel copper-containing composite for contraception.

OBJECTIVE: To investigate the biocompatibility of a novel copper-containing composite to provide preclinical data for clinical application of intrauterine device (IUD) or intra-vas device (IVD). DESIGN: Prospective experimental study. SETTING: Good laboratory practices laboratories. ANIMALS: Twenty healthy adult mice (SPF grade Kunming white mice, animal code SCXK 2003-0005). INTERVENTION(S): Cytotoxicity tests in vitro were conducted to evaluate the influence of the materials on the morphology, growth, and proliferation of cultured L929 mouse fibroblasts. Acute systemic toxicity tests were conducted to investigate the acute systemic toxic reaction with mice, and then the materials were implanted into the spinal muscle of rabbits (n = 15). The rabbits were sacrificed for pathologic examination at 1, 4, and 12 weeks after surgery. MAIN OUTCOME MEASURE(S): Evaluation of cytotoxicity by MTT assay, cytotoxicity test by direct contact assay, acute systemic toxicity test, and material implantation test. RESULT(S): The cytotoxicity grade of the copper-containing composite was 0-1, suggesting that the material was free of cytotoxicity; no acute systemic toxicity was found in any mice; mild inflammatory reaction was observed in the surrounding tissues of the implanted material in the early implantation stage, which was similar to that of the sham-operated sides. Twelve weeks after implantation, the inflammatory reaction was completely disappeared in the implanted tissue, similarly to the sham-operated sides. The fibrosis membrane surrounding the material became stable gradually over time. CONCLUSION: The copper-containing composite has excellent biocompatibility, which is feasible and safe for the clinical application as a novel contraceptive material.

Initial studies on a novel filtering-type intra-vas device in male dogs.

BACKGROUND: To relieve the side effects induced by the complete obstruction of the vas deferens, we created a filtering-type intra-vas device (IVD) which is made of materials composed of nano-SiO(2)-copper complex cross-linking polymer composites. STUDY DESIGN: Eight male beagle dogs were grouped into nonimplanted control group and IVD-implanted group. We tested the efficacy of the sperm filtering effect of the new IVD material for 12 months and examined the influence of the IVD materials on the cells of the vas deferens, epididymis and testis. RESULTS: The densities of sperm were reduced significantly after the IVD was implanted; no motile sperm were found after the third month. No obvious morphological changes were found in the cells of the vas deferens, epididymis and testis in the IVD-implanted group. CONCLUSIONS: The filtering-type nano-SiO(2)-copper complex/polymer composite IVD is able to filter the sperm of the male dogs, and the IVD material did not cause obvious damage to the cells of the male reproductive organs after 1 year of implantation.

Study on a novel copper-containing composite for contraception.

BACKGROUND: The copper-containing intrauterine devices (Cu-IUDs) are being increasingly used worldwide as an effective contraception for family planning. To avoid abnormal bleeding, pain, partial and complete expulsion, which are associated with the burst release of cupric ions during the first few days, a novel cross-linked composite based on polyvinyl alcohol (PVA) that contained cupric ions, but not metallic copper, was developed by our research team. STUDY DESIGN: As a logical extension of our previous work, the corrosion products and release behavior of this composite after immersing in simulated body fluid (SBF) for 1 year were studied by X-ray fluorescence spectroscopy (XRF), X-ray diffraction (XRD) and atomic absorption spectrophotometry (AAS). RESULTS: No other new elements, such as P, Cl and Ca, appeared on the surface of the composite, and no Cu(2)O was formed after immersing in SBF for 1 year, indicating that the effectiveness of copper can be greatly improved. Furthermore, no significant change on time dependence was found for the release rates of cupric ions in the composite compared with that of metallic copper, suggesting the absent burst release of cupric ions in the composite. CONCLUSION: The present in vitro long-term data suggest that this novel copper-containing composite has potential as a substitute for conventional materials used in the manufacture of IUDs.

Comparison of the release behaviors of cupric ions from metallic copper and a novel composite in simulated body fluid.

The copper-containing intrauterine devices (Cu-IUDs) are being increasingly used worldwide as an effective contraceptive for family planning. To avoid abnormal bleeding, pain, and partial and complete expulsion which are associated with the burst release of copper during the first few days, a novel crosslinked composite based on poly(vinyl alcohol) that contained cupric ions, but not metallic copper, was synthesized. It is hypothesized that the burst release of cupric ions could be avoided and the utility of the cupric ions could be improved by this novel composite. To evaluate these effects of the composite, the corrosion products and the release rate of cupric ions after soaking in simulated body fluid (SBF) for different time spans were studied by environmental scanning electron microscopy, X-ray energy dispersive spectroscopy, X-ray diffraction, and atomic absorption spectrophotometer. In the first week, the release amount of cupric ions in the composite was 0.486 microg/mm(2). In the fifth week, it decreased to 0.0278 microg/mm(2). But for metallic copper, these were 5.93 microg/mm(2) and 0.041 microg/mm(2), respectively. No significant change on time-dependence was found for the release rates of cupric ions in the composite compared with that of metallic copper. Moreover, no other new elements, such as P, Cl, and Ca, appeared on the surface of the composite, and no Cu2O was formed after immersing in SBF for 90 days. All of these results suggested that burst release of cupric ions could be avoided and the effective utility of copper could be improved in this composite. In view of the earlier results, this novel copper-containing composite might serve as a potential substitute for conventional materials of IUDs in the future.

Release behavior of copper ion in a novel contraceptive composite.

PURPOSE: The universally used contraceptive method, the Cu-IUD, an effective contraceptive, is being increasingly used worldwide for family planning. To avoid abnormal bleeding, pain, partial and complete expulsion associated with the burst release of copper during the first few days, a novel cross-linked composite based on poly vinyl alcohol (PVA) that contains copper ions, but not metallic copper, was synthesized. MATERIAL AND METHODS: PVA, well known for its good processability, high strength, long-term temperature and pH stability and biocompatibility, was used as the matrix material. The corrosion products and the release rate of copper ions after soaking in simulated body fluid (SBF) for different time spans were studied by environmental scanning electron microscopy, X-ray energy dispersive spectroscopy, X-ray diffraction and atomic absorption spectrophotometer. RESULTS: No significant change on time dependence for the release rate of copper ions in the composite compared with that of metallic copper was found. Moreover, no other new elements, such as P, Cl and Ca, appeared on the surface of the composite and no Cu(2)O formed after immersing in SBF for 90 days. CONCLUSION: Burst release of copper ions can be avoided by loading copper ions in this polymer material. Release channels would not be obstructed by the deposition of corrosion products and nearly all of the copper loaded in the composites could be an effective contraceptive.

[Establishment of an animal model of azoospermia in male mice].

OBJECTIVE: To establish a stable animal model of azoospermia in male mice. METHODS: Two groups of mice, with 30 in each, were intervened respectively by chemotherapy (intraperitoneal injection of Busulfan and Cyclophosphamide) and subcutaneous injection of Estradiol. At different times after the injection, the microscopic structures of the seminiferous tubules of both groups were observed and compared. The recovery of the germ cells in the seminiferous tubules was also evaluated after the termination of the intervention. RESULTS: A stable animal model was established by chemotherapeutic method with Busulfan and cyclophosphamide, while the model constructed by muscle injection of Estradiol was not stable. CONCLUSION: Compared with estrogen injection, chemotherapeutic intervention is a reliable method for constructing an animal model of azoospermia in male mice.

Effect of sildenafil citrate on penile erection of rhesus macaques.

AIM: To examine the effect of sildenafil citrate on penile erection of male rhesus macaque. METHODS: Twenty Macaca mulatta were divided into the sildenafil treated and the control groups of 10 animals each. The penile size, the corpus cavernosal electromyogram (EMG) and the intra-corpus cavernosal pressure (ICP) were determined. RESULTS: The diameter of penis and the ICP were significantly increased and the corpus cavernosal EMG significantly reduced in the sildenafil group. CONCLUSION: Sildenafil citrate increases the penile size and ICP and reduces the corpus cavernosal EMG in male rhesus macaque.

[Effect of sildenafil ciltrate on the sexual activities of male rats].

OBJECTIVE: To obtain related pharmacodynamic data for the clinical experiment by observing the sexual activities of male rats after using sildenafil ciltrate through stomach irrigation. METHODS: Forty male Sprague-Dawley rats were distributed into 4 groups with different dosages (control with distilled water, low dosage: 0.08%, medium dosage: 0.24% and high dosage: 0.72%). After the male Sprague-Dawlay rats were mated with their female counterparts in pairs, the latent period of chasing, the frequencies of chasing in 60 minutes, the latent period of intercourse and the frequencies of intercourse in 60 minutes were recorded. RESULTS: Compared with the control, the frequencies of chasing were significantly increased and the latent periods of chasing were significantly shortened in both high dosage and medium dosage groups after using sildenafil (P < 0.01); The frequencies of intercourse in 60 minutes were significantly increased and the latent periods of intercourse were significantly shortened in all the groups after the use of sildenafil. CONCLUSIONS: The sexual activities of male rats treated with sildenafil were significantly activated.