Differential expression of VASA gene in ejaculated spermatozoa from normozoospermic men and patients with oligozoospermia

Aim： To detect the expression of VASA in human ejaculated spermatozoa, and to compare the expression of VASA between normozoospermic men and patients with oligozoospermia. Methods： Ejaculated spermatozoa were collected from normozoospermic men and patients with oligozoospermia by masturbation, and subsequently segregated through a discontinuous gradient of Percoll to obtain the spermatozoa. Reverse transcription polymerase chain reaction （RT- PCR）, quantitative RT-PCR （QRT-PCR）, immunoflurescence and Western blotting were used to detect the expression of VASA in mRNA and protein levels. Results： VASA mRNA was expressed in the ejaculated spermatozoa. QRT-PCR analysis showed that VASA mRNA level was approximately 5-fold higher in normozoospermic men than that in oligozoospermic men. Immunofluorescence and Western blotting analysis showed that VASA protein was located on the cytoplasmic membrane of heads and tails of spermatozoa, and its expression was significantly decreased in oligozoospermic men, which is similar to the result of QRT-PCR. Conclusion： The expression of VASA mRNA and protein was significantly decreased in the sperm of oligozoospermic men, which suggested the lower expression of the VASA gene might be associated with pathogenesis in some subtypes of male infertility and VASA could be used as a molecular marker for the diagnosis of male infertility.

Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.

Reproductive outcomes of intracytoplasmic sperm injection using testicular sperm and ejaculated sperm in patients with AZFc microdeletions:a systematic review and meta-analysis

Studies have explored the assisted reproductive technology(ART)outcomes of Y-chromosome azoospermia factor c(AZFc)microdeletions,but the effect of sperm source on intracytoplasmic sperm injection(ICSI)remains unknown.To determine the ART results of ICSI using testicular sperm and ejaculated sperm from males with AZFc microdeletions,we searched Embase,Web of Science,and PubMed to conduct a systematic review and meta-analysis.The first meta-analysis results for 106 cycles in five studies showed no significant differences in the live birth rate between the testicular sperm group and the ejaculated sperm group(risk ratio:0.97,95%confidence interval[CI]:0.73-1.28,P=0.82).The second meta-analysis of 106 cycles in five studies showed no difference in the abortion rate between the testicular sperm group and ejaculated sperm group(risk ratio:1.06,95%Cl:0.54-2.06,P=0.87).The third meta-analysis of 386 cycles in seven studies showed no significant difference in clinical pregnancy rates between the testicular sperm group and the ejaculated sperm group(risk ratio:1.24,95%Cl:0.66-2.34,P=0.50).Inevitable heterogeneity weakened our results.However,our results indicated that testicular sperm and ejaculated sperm yield similar ART outcomes,representing a meaningful result for clinical treatment.More properly designed studies are needed to further confirm our conclusions.

Morphometric and kinematic sperm subpopulations in split ejaculates of normozoospermic men

This study was designed to analyze the sperm kinematic and morphometric subpopulations in the different fractions of the ejaculate in normozoospermic men. Ejaculates from eight normozoospermic men were collected by masturbation in three fractions after 3-5 days of sexual abstinence. Analyses of sperm motility by computer-assisted sperm analysis （CASA-Mot）, and of sperm morphometry by computer-assisted sperm morphometry analysis （CASA-Morph） using fluorescence were performed. Clustering and discriminant procedures were performed to identify sperm subpopulations in the kinematic and morphometric data obtained. Clustering procedures resulted in the classification of spermatozoa into three kinematic subpopulations （slow with low ALH [35.6% of all motile spermatozoa], with circular trajectories [32.0%], and rapid with high ALH [32.4%]）, and three morphometric subpopulations （large-round [33.9% of all spermatozoa], elongated [32.0%], and small [34.10%]）. The distribution of kinematic sperm subpopulations was different among ejaculate fractions （P 〈 0.001）, with higher percentages of spermatozoa exhibiting slow movements with low ALH in the second and third portions, and with a more homogeneous distribution of kinematic sperm subpopulations in the first portion. The distribution of morphometric sperm subpopulations was also different among ejaculate fractions （P〈 0.001）, with more elongated spermatozoa in the first, and of small spermatozoa in the third, portion. It is concluded that important variations in the distribution of kinematic and morphometric sperm subpopulations exist between ejaculate fractions, with possible functional implications.

射精功能障碍

在射精功能障碍(ejaculatory dysfunction)中,早泄(premature ejaculation,PE),射精延迟(delayed ejaculation,DE),不射精(anejaculation,AE)和逆行射精(retrograde ejaculation,RE)是最为常见的射精功能障碍类型,其他还包括高潮障碍(orgasmic disorder)、射精疼痛(painful ejaculation)、射精后综合征(postorgasmic illness syndrome)等少见情况[1],根据临床的常见情况,本文重点介绍PE和DE。

Assessment of the Quality of Frozen-Thawed Semen or Epididymal Sperm in Three Native Vietnamese Pig Breeds

Our aim was to evaluate the quality of ejaculated and epididymal frozen-thawed pig sperm of endangered Vietnam native pig breeds. Ejaculated sperm was collected from live boars and epididymal sperm was collected from slaughtered boars of the MuongTe, Kieng Sat and Co BinhThuan breeds and frozen in 0.25 ml straws using a protocol established earlier for modern pig breeds. We evaluated the sperm quality after thawing in terms of motility and rates of viable and abnormal spermatozoa. Our results revealed that the sperm motility and rates of viable and abnormal frozen-thawed sperm were >30%, >44%, and <14%, respectively. The origin of sperm had an effect on the production of pig embryos in vitro. In the Co BinhThuan breed, ejaculated sperm generated higher cleavage, blastocyst and hatching rates than did the epididymal sperm (60.11% vs 56.02%, 17.23% vs 14.31%, 3.78% vs 2.34%, respectively, P < 0.05). Although no difference in cleavage rate, blastocyst formation rate and the average number of cells/blastocysts, the hatching blastocyst rate was different between the breeds (P > 0.05). In the Co BinhThuan breed, the rate of pregnancy of ejaculated groups was similar to that of the epididymal group. In conclusion, the ejaculated and epididymal sperm of native Vietnamese pigs were successfully frozen. We succeeded in creating embryos in vitro and pregnant pigs after artificial insemination from frozen-thawed semen in three native Vietnamese pig breeds for the first time. The use of the ejaculated sperm improved the production of native pig embryos in vitro efficiency.

Effect of Selective Dorsal Neurectomy on Erectile Function in Rats

Objective This study aimed to investigate the effect of penile selective dorsal neurectomy(SDN)on erectile function in rats.Methods Twelve adult male Sprague-Dawley rats(15 weeks old)were divided into three groups(n=4 per group):in control group,rats received no treatment;in sham group,rats underwent a sham operation;in SDN group,rats underwent SDN with half of the dorsal penile nerve severed.The mating test was performed,and the intracavernous pressure(ICP)assessed six weeks after the surgical treatment.Results At postoperative six weeks,the mating test revealed no significant difference in mounting latency and mounting frequency among the three groups(P>0.05),while the ejaculation latency(EL)was significantly longer and ejaculation frequency(EF)lower in the SDN group than in the control and sham groups(P<0.05).There were no significant differences in preoperative and postoperative ICP and ICP/mean arterial blood pressure(MAP)among the three groups(P>0.05).Conclusion SDN does not adversely affect the erectile function and sexual desire of rats,and at the same time it can reduce EL and EF,providing an application basis for SDN in the clinical treatment of premature ejaculation.

Oxytocin in pig seminal plasma is positively related with in vivo fertility of inseminated sows

Background:Identification of relevant in vivo biomarkers for fertility remains a challenge for the livestock industry.Concentrations of the small peptide hormone oxytocin(OXT),involved in male reproductive function and present in the seminal plasma(SP)of several species could be a robust one.This study characterized concentrations of SPOXT in ejaculates from boars used in artificial insemination(AI)programs aiming to evaluate its relationship with sperm quality variables and in vivo fertility of their liquid-stored AI-semen.Seminal OXT concentrations(ng/mL)were measured in 169 ejaculates from 61 boars of the Duroc,Pietrain,Landrace and Large White breeds using a direct competitive immunoassay test based on AlphaLISA®technology.Ejaculate(ejaculate volume,sperm concentration,total sperm count)and sperm parameters(motility,viability,intracellular generation of reactive oxygen species,plasma membrane fluidity)were assessed at 0 h and 72 h in AI-semen samples stored at 17℃.In vivo fertility included only 18 Large White and Landrace boars whose AI-semen was used to inseminated>100 sows and evaluated both farrowing rate and litter size of 3,167 sows.Results:The results showed that SP-OXT differed between boars and between ejaculates within boar(P<0.05)but not between breeds(Duroc,Pietrain,Landrace and Large White).Ejaculates with higher SP-OXT concentration/mL(hierarchically grouped;P<0.001)had larger volume and came from younger boars(P<0.05).Ejaculates of boars showing positive farrowing rate deviation exhibited higher(P<0.05)SP-OXT concentration/mL than those with negative farrowing rate deviation.Conclusion:The SP concentrations of OXT are boar,ejaculate and age dependent,and positively related with ejaculate volume and farrowing rates of liquid-stored semen AI-doses.

Current perspectives on pyospermia： a review

Pyospermia is an abnormal laboratory finding of high concentration of white blood cells in human ejaculates during infertility workup. The role of pyospermia and its impact on fertility is an important consideration in the management of infertility. Etiology, pathogenesis, diagnostic modalities and the management of pyospermia are reviewed in this paper. Current use of antibiotics and the intrinsic production of antioxidants in the management of pyospermia are also discussed in this review.

Cryopreservation of a Small Number of Human Sperm within Empty Zona Pellucidae

Objective To investigate the empty zona pellucida for use in the cryopreservation of human sperm. Materials & Methods Human and hamster zona pellucidae were evacuated and injected with testicular, epididymal and ejaculated sperm. The zona pellucidae with sperm were cryopreserved. Results After thawing, zona pellucidae were easily found, and sperm inside zona pellucidae were also easily observed. There were no differences in post-thaw motility and vitality between ejaculated and epididymal sperm groups (P>0.05), but these two parameters were lowered in testicular sperm group compared to both ejaculated and epididymal sperm (P<0.01). No significant difference was observed in post-thaw motilities among 6%, 7.5%, 9% glycerol concentrations (P>0.05). In addition, obvious differences in post-thaw motilities were not found between human and hamster empty zona pellucidae (P>0.05). Conclusion An evacuated zona pellucida is an ideal vehicle for the cryopreservation of a small number of human sperm.

Viability of Extended Goat Semen Stored at Refrigerated Condition

Due to the small volume of goat semen ejaculate, just a few doses of goat semen were produced when the sperm concentration is 100 × 10^/mL. The study was aimed to determine the viability of extended goat semen at refrigerated condition at 5 ℃ using varying sperm concentrations and evaluated if sperm concentration lower than 100 × 10^6/mL would affect the motility, viability and sperm morphology at refrigerated condition. Using an artificial vagina, ejaculated goat semen was collected from goat semen donor aged 1.5 year. Physical evaluation of the collected semen showed an average volume of 0.54 mL, mean pH of 6.8 and a milky white color with thick consistency indicative of high concentration. Fresh goat semen had an initial average of 76% with an average initial sperm concentration of 128 × 10^/mL. The semen was divided into four treatments： sperm concentration of 100× 10^/mL, 75 × 10^/mL, 50 × 10^/mL and 25 × 10^/mL, and were stored at 5 ℃ for a period of 10 d. The semen evaluation was performed for each of the four treatments every other day. Results showed that the sperm concentration of spermatozoa affected the duration of storage based on the sperm motility percentage, viability and morphology of spermatozoa. The extended goat semen with sperm concentration of 25 and 50 million sperm/mL is optimal for storage within 6 d that gave satisfactory percentage motile, viable and morphologically normal spermatozoa.

Variation among Beef Bulls in the Ratio of X- to Y-Chromosome Bearing Spermatozoa

A study was conducted to evaluate variation in the ratio of X- to Y-bearing sperm of individual ejaculates and to determine any relationship between skewed sex ratio and either routine morphological evaluation or computer assisted sperm analysis (CASA). Semen was collected from bulls weekly for 6 consecutive weeks, sperm DNA was recovered and quantitative, and real-time PCR was used to determine the ratio of X- to Y-bearing sperm in each ejaculate. The overall mean of X-bearing sperm within ejaculates was 54.7% over the 6 weeks of semen collections. The percentages of X-bearing sperm were similar (P > 0.5) across all collections. Between bulls, there were differences (P < 0.05) in the mean percentage of X-bearing sperm. No significant correlations were found between CASA parameters and percentage of X-bearing sperm across bulls, so analysis was done within each bull. Different combinations of CASA and/or morphological parameters were found to correlate with the percentage of X-bearing sperm but the prediction equations were specific for individual bulls and unlikely to be of use across bulls. These results confirm that the ratio of X- to Y-bearing sperm may be skewed in some ejaculates of bull semen. Some sperm parameters measured by CASA or routine morphological evaluation were associated with semen sex ratio.

Selection of High-quality Sperms by the Nanotechnological Method of Magnetic Activation in Brazilian Cervids

Cervids show a high degree of abnormalities in their sperm cells.Thus,this study aimed to select high-quality spermatozoa using magnetic-activated sperm sorting(MASS)compared to density gradient centrifugation(DGC)by assessing the post-selection cell quality.Semen from six Mazama deer was collected by electroejaculation after chemical restraint.The semen was analyzed in four samples:Fresh,DGC,SEMgood-non-apoptotic fraction,and SEMpoor-apoptotic fraction.The material was analyzed for motility and vigor(light microscopy),concentration(Neubauer chamber),semen morphology(phase contrast),and supravital staining test(eosin/nigrosine).The DGC method used 20 x 106 cells in 90%and 45%percoll®gradient.The MASS used 10 x 106 cells with 20μl of iron nanoparticles attached to Annexin V and filtration in a magnetic separation column.Both processing methods(DGC and MASS)were effective in producing high-quality sperm samples,with a marked reduction in abnormalities from 41.83±10.25(fresh)to 14.83±3.17(DGC)and 12±3.01(SEMgood),with 80.3%±2.06 livings cells.These findings suggest that this nanotechnological method,using nanoparticles,effectively produces high-quality semen samples in cervids for use in assisted reproduction.

New insights on premature ejaculation： a review of ：lefinition, classification, prevalence and treatmeni

There are ongoing debates about the definition, classification and prevalence of premature ejaculation （PE）. The first evidence-based definition of PE was limited to heterosexual men with lifelong PE who engage in vaginal intercourse. Unfortunately, many patients with the complaint of PE do not meet these criteria. However, these men can be diagnosed as one of the PE subtypes, namely acquired PE, natural variable PE or premature-like ejaculatory dysfunction. Nevertheless, the validity of these subtypes has not yet been supported by evidence. The absence of a universally accepted PE definition and lack of standards for data acquisition have resulted in prevalence studies that have reported conflicting rates. The very high prevalence of 20%-30% is probably due to the vague terminology used in the definitions at the time when such surveys were conducted. Although many men may complain of PE when questioned for a population-based prevalence study, only a few of them will actively seek treatment for their complaint, even though most of these patients would define symptoms congruent with PE. The complaints of acquired PE patients may be more severe, whereas complaints of patients experiencing premature-like ejaculatory dysfunction seem to be least severe among men with various forms of PE. Although numerous treatment modalities have been proposed for management of PE, only antidepressants and topical anaesthetic creams have currently been proven to be effective. However, as none of the treatment modalities have been approved by the regulatory agencies, further studies must be carried to develop a beneficial treatment strategy for PE.

Sexual function of premature ejaculation patients assayed with Chinese Index of Premature Ejaculation

Aim: To assess the psychometric properties of the Chinese Index of Premature Ejaculation (CIPE). Methods: The sexual function of 167 patients with and 114 normai controls without premature ejaculation (PE) were evaluated with CIPE. All subjects were married and had regular sexual activity. The CIPE has 10 questions, focusing on libido, erectile function, ejaculatory latency, sexual satisfaction and difficulty in delaying ejaculation, self-confi dence and depression. Each question was responded to on a 5 point Likert-type scale. The individual question score and the total scale score were analyzed between the two groups. Results: There were no significant differences between the age, duration of marriage and educational level (P > 0.05) of patients with and vvithout PE and normai controls. The mean latency of patients with PE and normai controls were 1.6±1.2 and 10.2±9.5 minutes, respectively. Significant differences between patients with (26.7±4.6) PE and normai controls (41.9±4.0) were observed on the total score of CIPE (P < 0.01). Using binary logistic regression analysis, PE was significantly related to five questions of the original measure. They are the so-called the CIPE-5 and include: ejaculatory latency, sexual satisfaction of patients and sexual partner, difficulty in delaying ejaculation, anxiety and depression. Receiver Operat ing Characteristic (ROC) curve analysis of CIPE-5 questionnaire indicated that the sensitivity and specificity of CIPE were 97.60 % and 94.74 %, respectively. Employing the total score of CIPE-5, patients with PE could be divided into three groups: mild (>15 point) 19.8 %, moderate (10-14 point) 62.8 % and severe (< 9 point) 16.7 %. Conclusion: The CIPE-5 is a useful method for the evaluation of sexual function of patients with PE and can be used as a clinical endpoint for clinical trials studying the efficacy of pharmacologica] intervention.

Premature ejaculation： current and future treatments

Premature ejaculation （PE） is recognized to be the most common male sexual disorder. PE provides difficulties for professionals who treat this condition because there is neither a universally accepted definition nor a medication approved by the Food and Drug Administration （FDA）. Despite these shortcomings, physicians continue to diagnose their patients with PE according to major guidelines and treat them with either behavioral therapies or off-label medications. This review focuses on current and emerging treatment options and medications for PE. Advantages and limitations of each treatment option are discussed in the light of current published peer-reviewed literature.

Possible association of the 5-HTTLPR serotonin transporter promoter gene polymorphism with premature ejaculation in a Turkish population

We evaluated the genotypes of the serotonin transporter gene （5-HTT） in patients with premature ejaculation （PE） to determine the role of genetic factors in the etiopathogenesis of PE and possibly to identify the patient subgroups. A total of 70 PE patients and 70 controls were included in this study. All men were heterosexual, had no other disorders and were either married or in a stable relationship. PE was defined as ejaculation that occurred within 1 min of vaginal intromission. Genomic DNA from patients and controls was analyzed using polymerase chain reaction, and allelic variations of the promoter region of the serotonin transporter gene （5-HTTLPR） were determined. The 5-HTTLPR （serotonin transporter promoter gene） genotypes in PE patients vs. controls were distributed as follows： L/L 16% vs. 17%, L/S 30% vs. 53% and S/S 54% vs. 28%. We examined the haplotype analysis for three polymorphisms of the 5-HTTLPR gene： LL, LS and SS. The appropriateness of the allele frequencies in the 5-HTTLPR gene was analyzed by the Hardy-Weinberg equilibrium using the Z-test. The short （S） allele of the 5-HTTLPR gene was significantly more frequent in PE patients than in controls （P 〈 0.05）. We suggest that the 5-HTTLPR gene plays a role in the pathophysiology of all primary PE cases. Further studies are needed to evaluate the relationship between 5-HTTLPR gene polymorphism and patient subgroup （such as primary and secondary PE） responses to selective serotonin reuptake inhibitors as well as ethnic differences.

Male sexual dysfunction in Asia

Sex has always been a taboo subject in Asian society. However, over the past few years, awareness in the field of men＇s sexual health has improved, and interest in sexual health research has recently increased. The epidemiology and prevalence of erectile dysfunction, hypogonadism and premature ejaculation in Asia are similar in the West. However, several issues are specific to Asian males, including culture and beliefs, awareness, compliance and the availability of traditional/complementary medicine. In Asia, sexual medicine is still in its infancy, and a concerted effort from the government, relevant societies, physicians and the media is required to propel sexual medicine to the forefront of health care.

HoLEP does not affect the overall sexual function of BPH patients： a prospective study

We aimed to prospectively evaluate the influence of holmium laser enucleation of the prostate （HoLEP） on the overall postoperative sexual function of benign prostatic hyperplasia （BPH） patients with lower urinary tract symptoms （LUTS） and to explore the relationship between sexual function and LUTS. From January 2010 to December 2011, sixty sexually active consecutive patients with BPH who underwent HoLEP were prospectively enrolled in the study. All patients filled out the Male Sexual Health Questionnaire （MSHQ） for evaluation of their overall sexual function and the International Prostatic Symptom Score （IPSS） for pre- and post-operative 6 months evaluation of their voiding symptoms. The LUTS and sexual function changes were statistically analyzed. The preoperative and 6 months postoperative status of the patients was compared using uroflowmetry and IPSS questionnaires. The analysis revealed significant improvements following HoLEP. Among the sub-domains of the MSHQ, postoperative sexual function, including erection, ejaculation, sexual satisfaction, anxiety or sexual desire, did not significantly change after HoLEP （P 〉 0.05）, whereas satisfaction scores decreased slightly due to retrograde ejaculation in 38 patients （63.3%）. Sexual satisfaction improved significantly and was correlated with the improvements of all LUTS and the quality-of-life （QoL） domains in IPSS after surgery （QoL; relative risk [RR]： -0.293; total symptoms, RR： -0.411; P 〈 0.05）. The nocturia score was associated with the erectile function score （odds ratio 0.318, P = 0.029）. The change in ejaculatory scores did not show significant association with IPSS scores. HoLEP did not influence overall sexual function, including erectile function. In addition, sexual satisfaction improved in proportion with the improvement of LUTS.

Vlale sexual dysfunction and infertility associated with neurological disorders

Normal sexual and reproductive functions depend largely on neurological mechanisms. Neurological defects in men can cause infertility through erectile dysfunction, ejaculatory dysfunction and semen abnormalities. Among the major conditions contributing to these symptoms are pelvic and retroperitoneal surgery, diabetes, congenital spinal abnormalities, multiple sclerosis and spinal cord injury, Erectile dysfunction can be managed by an increasingly invasive range of treatments including medications, injection therapy and the surgical insertion of a penile implant. Retrograde ejaculation is managed by medications to reverse the condition in mild cases and in bladder harvest of semen after ejaculation in more severe cases. Anejaculation might also be managed by medication in mild cases while assisted ejaculatory techniques including penile vibratory stimulation and eiectroejaculation are used in more severe cases. If these measures fail, surgical sperm retrieval can be attempted. Ejaculation with penile vibratory stimulation can be done by some spinal cord injured men and their partners at home, followed by in-home insemination if circumstances and sperm quality are adequate. The other options always require assisted reproductive techniques including intrauterine insemination or in vitrofertilization with or without intracytoplasmic sperm injection. The method of choice depends largely on the number of motile sperm in the ejaculate.