Impact of chronological ageing on semen parameters in southern Indian men visiting infertility centre:A retrospective study

Objective:To investigate the association between age and semen parameters among male partners of subfertile couples.Methods:This retrospective study analyzed the semen of 1523 infertile men aged 26 to 50 years.Data were extracted from GarbhaGudi IVF Centre database from January 2019 to September 2020.The basic semen parameters were interpreted according to the WHO manual 2021,6th edition.Semen parameters in different age groups were compared.Results:Total and progressive motile sperms were significantly higher in the age group of 26-30 years compared to other age groups(P<0.05).Normal sperm count was significantly higher in the age group of 26-30 years compared to the age groups of 41-45 years and>46 years(P=0.001).However,sperm head defects,neck and midpiece defects,tail defects,and cytoplasmic droplets showed statistically insignificant difference in all the age groups(P>0.05).Semen viscosity showed no statistical difference in all the age groups compared to the reference age group of 26 to 30 years.Conclusions:Higher age can lead to a significant decrease in normal sperms and motility in subfertile men.Hence,male partner age should be considered as one of the major determining factors for reproductive outcomes.

Abnormal expression of cen-trosome protein (centrin) in spermatozoa of male human infertility

To study the relations between male infertility and centrosome protein (centrin) and the functions of centrin in spermatogenesis, the matured spermatozoa of 10 normal male people and 18 male infertility patients were stained by immunofluorescence labeling antibody against centrin. The results showed that two fluorescence signal dots appeared in the normal male spermatozoa and were located at the base of flagellum. They are proximal centriole and distal centriole. However, in some spermatozoa of the male infertility, centrin protein was located abnormally at the base of flagellum and its staining signals were spread, the normal proximal and distal centrioles were confused and could not be recognized separately. These results suggest that abnormality of centrosome protein may be related to male infertility. This discovery may be used as a marker of abnormal sperm and male infertility.

The Influence of Sperm DNA Damage and Semen Homocysteine on Male Infertility

Background:To explore the relationship of sperm DNA fragmentation index(DFI),serum and seminal plasma homocysteine(Hcy),and semen parameters in patients with severe spermatogenetic dysfunction.Methods:A total of 77 infertile males treated in our hospital for severe spermatogenetic dysfunction from January 2016 to November 2017 were recruited.The involved patients were divided into two groups:oligozoospermia(SOM group,35 cases)and asthenozoospermia(OAT group,42 cases).The control group(NM group)contained 31 healthy males without reproductive dysfunctions.All the participants involved were tested in the items below:spermatozoa parameters,spermatozoa DFI,serum Hcy level and seminal plasma Hcy level,concentration of seminal plasma malondialdehyde(MDA),and total antioxidant capacity(TAC).Results:Between the SOM group and NM group,there were significantly difference in sperm concentration,motility and vitality,concentration of MDA,and TAC.The spermatozoa DFI and Hcy levels in SOM group were significantly higher than those of the NM group.Sperm DFI was positively correlated with serum Hcy level(r=0.083,P<0.05).Serum Hcy level was negatively correlated with sperm concentration(r=−0.186,P<0.05)and sperm vitality(r=−0.216,P<0.05).The serum Hcy level was not correlated with sperm Hcy level(r=0.103,P>0.05).Conclusions:The elevated Hcy level and spermatozoa DFI may be important factors of the severe spermatogenetic dysfunction,which can be used as semen index to evaluate sperm quality and male fertility.

Proteomic signatures of infertile men with clinical varicocele and their validation studies reveal mitochondrial dysfunction leading to infertility

To study the major differences in the distribution of spermatozoa proteins in infertile men with varicocele by comparative proteomics and validation of their level of expression. The study-specific estimates for each varicocele outcome were combined to identify the proteins involved in varicocele-associated infertility in men irrespective of stage and laterality of their clinical varicocele. Expression levels of 5 key proteins （PKARIA, AK7, CCT6B, HSPA2, and ODF2） involved in stress response and sperm function including molecular chaperones were validated by Western blotting. Ninety-nine proteins were differentially expressed in the varicocele Eroup. Over 87% of the DEP involved in major energy metabolism and key sperm functions were underexpressed in the varicocele group. Key protein functions affected in the varicocele group were spermatogenesis, sperm motility, and mitochondrial dysfunction, which were further validated by Western blotting, corroborating the proteomics analysis. Varicocele is essentially a state of energy deprivation, hypoxia, and hyperthermia due to impaired blood supply, which is corroborated by down-regulation of lipid metabolism, mitochondrial electron transport chain, and Krebs cycle enzymes. To corroborate the proteomic analysis, expression of the 5 identified proteins of interest was validated by Western blotting. This study contributes toward establishing a biomarker ＂fingerprint＂ to assess sperm quality on the basis of molecular parameters.

Acupuncture-moxibustion Therapy for Infertility due to Sperm Abnormality

Objective： To observe the clinical efficacy of acupuncture-moxibustion therapy for infertility due to sperm abnormality （SAI）. Methods： We treated a series of 35 cases of SAI with electroacupuncture combined with herb cake-partitioned moxibustion, observing the variation before and after treatment in symptom integral, sperm status, sex hormone and prostate function. Results： After treatment, the patients were remarkably improved in symptom integral, sperm status, sex hormone and prostate function as compared with those prior to treatment. Conclusion： Acupuncture-moxibustion is an effective therapy for SAI.

Effect of Acupuncture-moxibustion Therapy on Sperm Quality in Infertility Patients with Sperm Abnormality

Objective： Working on seminal plasma acid phosphatase, to explore the mechanism by which acupuncture improves sperm quality （concentration, viability and motility） in infertility patients. Methods： A total of 118 patients received acupuncture-moxibustion treatment. Before and 3, and 6 months after the treatment were detected their seminal plasma acid phosphatase, sperm concentration, sperm viability and sperm motility were measured. Results： The differences between before and after treatments in sperm motility and seminal plasma acid phosphatase levels were statistically significant （P〈0.01）. Conclusion： Acupuncture-moxibustion can improve seminal plasma acid phosphatase levels in infertility patients.

Varicocele repair in improving spermatozoa,follicle-stimulating hormone,and luteinizing hormone parameters in infertile males with azoospermia:a systematic review and meta-analysis

Patients with azoospermia show a prevalence of varicocele of 10.9%and a 14.8%contribution to male infertility.Patients with azoospermia are thought to produce high-quality semen following varicocele treatment.Advising varicocelectomy prior to sperm retrieval in a reproductive program is still debated.This study reviewed the impact of varicocele repair on male infertility using several factors.A literature search was conducted using Scopus,PubMed,Embase,the Wiley Online Library,and Cochrane databases.Sperm concentration,sperm progression,overall sperm motility,sperm morphology,and follicle-stimulating hormone(FSH)and luteinizing hormone(LH)levels were also compared.Outcomes were compared between those who received treatment for varicocele and those who did not.The data from the pooled analysis were presented as standardized mean difference(SMD)along with a 95%confidence interval(CI).Heterogeneity was evaluated using I2.Additionally,we conducted analyses for publication bias,sensitivity,and subgroup analysis as appropriate.Nine studies were included after screening relevant literature.Statistical analysis revealed a significant improvement in sperm concentration(SMD:1.81,95%CI:0.84–2.77,P<0.001),progressive sperm motility(SMD:4.28,95%CI:2.34–6.22,P<0.001),and sperm morphology(SMD:3.59,95%CI:2.27–4.92,P<0.001).Total sperm motility showed no significant difference following varicocele repair(SMD:0.81,95%CI:−0.61–2.22,P=0.26).No significant differences were seen in serum FSH(SMD:0.01,95%CI:−0.16–0.19,P=0.87)and LH(SMD:0.19,95%CI:−0.01–0.40,P=0.07)levels as well.This study supports varicocele repair in infertile men with clinical varicocele,as reflected by the improvement in sperm parameters after varicocelectomy compared with no treatment.There were no significant improvements in serum FSH and LH levels.

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.

Whither must spermatozoa wander？ The future of laboratory seminology

This commentary celebrates the publication of the 5th for the Examination and Processing of Human Semen edition of the World Health Organization Laboratory Manual This is the most complete text to date on the creation of a conventional semen profile and includes invaluable reference limits for specific aspects of semen quality based on the analysis of over 1 900 recent fathers. The new edition of the manual also includes detailed protocols for monitoring different aspects of sperm function and new chapters on the preparation of spermatozoa for assisted conception and cryopreservation. Given that this publication is the definitive statement on how to perform a descriptive semen analysis, we might speculate on the future of this field and the sorts of tests that might feature in future editions of the manual. Cell biologists are currently being empowered by the ＇omics revolution, which is placing at their disposal technologies of unprecedented power to examine the biochemical composition of cells such as spermatozoa. Indeed, spermatozoa are perfect vehicles for this kind of analysis because they can be obtained as extremely pure suspensions, exist naturally in isolation and can be induced to express their capacity for fertilization and the initiation of embryonic development in vitro. The application of ＇omics technologies to these cells, in concert with detailed assessments of their functional competence, should provide insights into the biochemical basis of defective semen quality. This information will then help us understand the causes of male infertility and to develop rational methods for its treatment and possible prevention.

The usefulness and significance of assessing rapidly progressive spermatozoa

It is possible and clinically relevant to distinguish between slow and rapid progressive spermatozoa in basic semen analysis. This is discussed in light of the different purposes of semen analysis for the subfertile couple and the male patient. The two groups of progressive spermatozoa should be distinguished to help ensure that pertinent information available in the semen sample is not neglected.

Preparation and incubation conditions affect the DNA integrity of ejaculated human spermatozoa

Appropriate semen processing and assessment are critical for successful infertility treatment. We investigated whether laboratory procedures including semen preparation and incubation affect sperm DNA integrity. A total of 153 infertile men were involved. Conventional semen parameters and sperm chromatin structure assay （SCSA） parameters, that is, DNA fragmentation index （%DFI） and high DNA stainability （%HDS）, were assessed on the flesh ejaculated semen samples, which were treated and incubated under different conditions. Negative correlations were identified between the %DFI and sperm concentration, motility, progressive motility and morphology. A lower percentage of DFI was detected in spermatozoa when density gradient centrifugation （DGC） was followed by swimup treatment in comparison with DGC alone （P 〈 0.01）. Although the %DFI increased in a time-dependent manner with incubation both at room temperature （RT） and at 37℃ in air, the %DFI after 24 h at RT was significantly lower than that at 37℃ （P 〈 0.05）. Incubation with 5% CO2 was effective in maintaining sperm motility （P 〈 0.01）; however, it induced further elevation of %DFI （P 〈 0.001）. Thus, sperm DNA damage was associated with longer incubation periods. Interestingly, common culture conditions, such as maintaining pH and temperature, compromised the sperm DNA integrity.

Chlamydia trachomatis and sperm lipid peroxidation in infertile men

<abstract>Aim: To relate the presence of anti-Chlamydial trachomatis IgA in semen with sperm lipid membrane peroxidation and changes in seminal parameters. Methods: Semen samples of the male partners of 52 couples assessed for undiagnosed infertility were examined for the presence of IgA antibody against C. trachomatis. The level of sperm membrane lipid peroxidation was estimated by determining the malondialdehyde (MDA) formation. Results: Sperm membrane of infertile males with positive IgA antibodies against C. trachomatis showed a higher level of lipid peroxidation than that of infertile males with negative IgA antibody (P<0.05). There was a positive correlation (P< 0.01) between the level of C. trachomatis antibody and the magnitude of sperm membrane lipid peroxidation. All the other tested semen parameters were found to be similar in the two groups. Conclusion: The activation of immune system by C. trachomatis may promote lipid peroxidation of the sperm membrane. This could be the way by which C. trachomatis affects fertility.

Compound heterozygous mutations in PMFBP1 cause acephalic spermatozoa syndrome:A case report

BACKGROUND Acephalic spermatozoa syndrome(ASS)is an extremely rare form of severe teratozoospermia,where in most of the sperm either appear to lack heads or have disconnected or poorly connected heads and tails.CASE SUMMARY We reported the case of a male patient with secondary infertility whose sperm showed typical ASS upon morphological analysis.Whole-exome sequencing was performed on the patient’s peripheral blood,which revealed two heterozygous variants of the PMFBP1 gene:PMFBP1c.414+1G>T(p.?)and PMFBP1c.393del(p.C132Afs*3).CONCLUSION It is speculated that the compound homozygous mutation of PMFBP1 may be the cause of ASS.We conducted a literature review in order to provide the basis for genetic counseling and clinical diagnosis of patients with ASS.

Relationship between serum vitamin D concentration and parameters of gonadal function in infertile male patients

Background Vitamin D(vitD)deficiency could affect male reproductive function.Our objective was to investigate the relationship between serum vitD concentrations and hormonal and seminal parameters in infertile patients and to compare the results with those in healthy controls.Materials and methods Infertile patients(n=29)and normozoospermic healthy donors(n=27)were recruited for the study.Serum concentrations of vitD,total testosterone,estradiol,and sex hormone-binding globulin were determined using chemiluminescence assays,and free testosterone concentration was determined by radioimmunoassay.Semen analysis was performed as suggested by the World Health Organization.Statistical analysis was conducted using Student’s t test,contingency tables,and linear regression studies.Results VitD concentrations were lower in patients than in controls(p<0.001).A significant association(p<0.001)was observed between vitD concentrations<20ng/mL and infertility.In the control group,significant correlations were reported between vitD concentrations>30 ng/mL and the concentrations of testosterone(p<0.05),free testosterone(p<0.01),and estradiol(p<0.05).A direct correlation was found between vitD concentration and percentage of sperm vitality(p=0.01).VitD also positively correlated with the percentage of progressive sperm motility(p<0.05)and sex hormone-binding globulin concentrations(p<0.01).Conclusions VitD may affect male reproductive parameters,and its deficiency could be associated with infertility.

男性不育症患者精子DNA碎片指数检测意义

目的分析男性不育症患者精子DNA碎片指数(DFI)检测意义。方法选定本院2021年5月至2023年5月就诊的150例男性不育症患者设为观察组,另选取同期本院150例孕前体检并于随访1年内女方受孕者设为对照组,采集精液标本,检测、比较2组DFI、精液参数,根据DFI将男性不育症患者分为3组,DFI≤15%的36例患者设为A组,DFI在>15%~30%的64例患者设为B组,DFI>30%的50例患者设为C组,比较3组精液参数,Pearson分析DFI与精液参数的相关性。结果观察组DFI高于对照组(P<0.05)。观察组前向运动精子百分率(PR)、精子浓度、精子正常形态率、精子存活率均低于对照组(P<0.05)。C组PR、精子浓度、精子正常形态率、精子存活率均低于B组、A组(P<0.05),B组PR、精子浓度、精子正常形态率、精子存活率均低于A组(P<0.05)。DFI与PR、精子浓度、精子正常形态率、精子存活率均呈负相关性(r=-0.403、-0.381、-0.376、-0.396,P均<0.001)。结论男性不育症患者DFI较高,DFI与精液参数存在一定的相关性,可作为评估患者病情的重要指标。

精索静脉曲张患者血清生殖激素、精子形态、精子活率的变化

目的探讨精索静脉曲张(VC)对生殖激素及精子形态、精子活率的影响。方法检测81例VC所致男性不育患者和18例健康生育男性精液及血清标本,其中不育患者按VC临床分度分为Ⅰ°、Ⅱ°和Ⅲ°组,用放射免疫法测定血清催乳素(PRL)、卵泡刺激素(FSH)、黄体生成素(LH)、睾酮(T)、雌二醇(E2)水平;采用计算机辅助精液分析仪测定精子活率。结果Ⅱ°和Ⅲ°组FSH、LH低于对照组(P<0.05);Ⅰ°、Ⅱ°和Ⅲ°组的T、PRL和E2与对照组相比差异无统计学意义(P>0.05);Ⅱ°和Ⅲ°组正常形态精子百分率低于对照组(P<0.05);VC各组精子活率均低于对照组(P<0.05),且Ⅲ°组精子活率低于Ⅰ°和Ⅱ°组(P<0.05)。结论VC可引起血清生殖激素FSH、LH变化,但VC临床分度对血清生殖激素水平的影响关系不明显。VC能够导致正常形态精子百分率和精子活率下降,VC程度越重,对精液正常形态精子率与活率的影响程度越大。

男性不育患者精浆中游离睾酮、游离左旋肉碱水平与精液参数的相关性分析

目的分析男性不育患者精浆中游离睾酮和游离左旋肉碱水平与精子密度、活动率和活力参数间的相关性,探讨二者对男性生育力的影响及在不育症检查和治疗中的作用。方法分别采用化学发光免疫分析法、液相色谱-质谱/质谱联用技术和计算机辅助精液分析系统,测定192例不育男性(正常精液组36例,少精组48例,弱精组60例,少弱精组48例)精浆中游离睾酮、游离左旋肉碱水平及精子密度、活动率、活力等参数。采用SPSS13.0软件包进行统计处理,分析各组游离睾酮和游离左旋肉碱水平的差异以及游离睾酮和游离左旋肉碱水平与精子密度、活动率、活力间的相关性。结果正常精液组游离睾酮和游离左旋肉碱水平高于其他3组(P<0.05)。相关性分析结果显示:精浆中游离睾酮水平与精子密度存在正相关关系(r=0.427,P<0.05),精浆游离左旋肉碱水平与精子密度存在正相关关系(r=0.318,P<0.01),精浆中游离左旋肉碱水平与精子活动率及活力间呈现正相关关系(r=0.641,P<0.01;r=0.568,P<0.01)。结论精浆中游离睾酮水平与精子密度正相关,游离左旋肉碱水平与精子密度、活动率、活力正相关,二者可作为一项有效的生化指标,为男性不育症临床诊治提供参考。

精子DNA碎片率及顶体酶活性对男性不育的诊断价值

【目的】探讨精子DNA碎片率与顶体酶活性检测对男性不育的诊断价值。【方法】回顾性分析近三年前来本院就诊的902例男性患者的精液常规参数及精子DNA碎片率和顶体酶活性水平。【结果】按DNA碎片率水平将所有患者分为4组(≤10%,11~20%,21~30%,≥31%),各组患者的年龄、精子浓度、精子总活力、前向运动率以及顶体酶活性存在统计学差异;按精子顶体酶活性正常与否将患者分为两组后,两组间精子浓度、精子总数、精子总活力、前向运动率、正常形态率、畸形精子指数、DNA碎片率均有统计学差异。相关性分析显示,DNA碎片率与精子浓度、精子活力、前向运动率、精子总数及顶体酶活性呈负相关关系,与畸形精子指数呈正相关关系;顶体酶与精子浓度、精子活力、前向运动率、精子总数、正常形态率呈正相关关系,与头部异常率、畸形精子指数、DNA碎片率负相关。【结论】精子DNA碎片率及顶体酶活性水平均可以在一定程度上反映精子参数水平,其中DFI更能反映精子活力水平,顶体酶活性则与精子形态更为相关。两者都是精子质量评估更为客观而全面的指标。

少精子症、弱精子症和畸形精子症患者精子凋亡率比较

目的:比较少精子症、弱精子症和畸形精子症患者精子凋亡率。方法:应用计算机辅助精液分析(CASA)对生育组(n=14)、不育组[(n=48),少精子症(精子密度<10×106·mL-1)11例、弱精子症(a+b级精子百分率<50%)25例、畸形精子症(形态正常精子百分率<14%)12例]进行精液参数检测,瑞-姬染色检测精子凋亡率。结果:不育组及不育组中的少精子症组精子凋亡率均明显高于生育组(P<0·05);弱、畸形精子症组精子凋亡率与生育组比较差异均无显著性(P>0·05)。密度异常组(精子密度<20×106·mL-1)精子凋亡率明显高于密度正常组(精子密度≥20×106·mL-1)(P<0·05);a+b级异常组(a+b<50%)精子凋亡率与正常组比较差异无显著性(P>0·05);精子形态异常组(形态正常精子<14%)凋亡率与正常组比较差异无显著性(P>0·05)。结论:不育组精子凋亡率高于生育组,尤其是少精子症患者精子凋亡率增加更明显。

男性不育患者精子形态与精浆锌和精子顶体酶活性关系的研究

目的：研究男性不育患者精子形态与精浆锌和精子顶体酶活性关系。方法455例男性不育患者分别根据年龄和精子正常形态百分率分成5组和8组，采用 Diff-Quik 染色法、改良 PRA 法和改良 Kennedy 法分别进行精子形态、精浆锌和精子顶体酶活性的检测。结果各年龄组的正常形态精子百分率和精浆锌浓度差异无统计学意义（P ＞0．05），精子顶体酶活性在36～50岁年龄段显著降低（P ＜0．05），其他年龄段精子顶体酶活性差异无统计学意义（P ＞0．05）。在精子正常形态百分率不同的8组中，年龄差异无统计学意义，精浆锌总量差异无统计学意义，精子顶体酶活性随着精子正常形态百分率降低而降低（r ＝0．93，P ＜0．01）。结论精子正常形态百分率不受年龄和精浆锌的影响，与精子顶体酶活性呈正相关。