Subnormal sperm parameters in conventional semen analysis are associated with discrepancies between fertilization and pregnancy rates in in-vitro fertilization and embryo transfer

Three hundred and twenty-eight consecutive treatment cycles in 168 couples were analysed retrospectively in order to examine the influence of conventional semen analysis results on the outcome of in-vitro fertilization and embryo transfer with respect to the occurrence of both fertilizations and pregnancies. All treatments were performed under maximally standardized and controlled conditions. Each of the three main determinants of the spermiogram, namely the concentration, motility and morphology of sperm in seminal plasma, was of significant importance for fertilization and subsequent pregnancy. Best correlations were achieved by counting the number of progressively (a+b) motile sperm and the number of normally formed sperm in seminal plasma. The pregnancy rate was reduced significantly in cases in which the sperm concentration was < 10 x 10(6) ml-1 (P < 0.01), or in which there was < 40% progressively motile sperm (P < 0.001), or < 30% normally formed sperm (P < 0.001). If more than one parameter in the spermiogram was abnormal, the fertilization rate depended mainly on the most disturbed sperm parameter. The implantation rate as well as the pregnancy rate was reduced significantly in patients with low progressive sperm motility and normal morphology rates. The difference could only be attributed partially to the lower number of embryos replaced. In conclusion, subnormal sperm quality seems to interfere with developmental stages beyond the process of fertilization.     

Comparative study of the effects of three semen preparation media on semen analysis,DNA damage and protamine deficiency,and the correlation between DNA integrity and sperm parameters

Semen samples collected from 28 male partners of infertile couples were divided into three equal aliquots and prepared with three selected media,such as PureSperm （Nidacon,Gothenburg,Sweden）,Sil-Select Plus^TM （Fertipro,Beemem,Belgium） and SpermGrad^TM（Vitrolife,Gothenburg,Sweden）. The differences in mean percentages of semen parameters were assessed by repeated measures analysis. Correlations of sperm DNA damage,as measured by terminal deoxynucleotidyl transferase dUTP nick end labeling （TUNEL） assay,and of protamine deficiency,as measured by chromomycin A3 （CMA3） staining with sperm parameters,were determined by Pearson＇s correlation. After preparation with all three media,sperm concentrations decreased （P〈0.05） while percentages of sperm with normal morphology increased （P〈0.05）. Percentages of sperm motility,rapid motility and progressive motile concentration （PMC） increased （P〈0.05） for each ofthese parameters,PureSperm preparation gave the best results （P〈0.05）. The percentage of DNA damage decreased in the PureSperm and Sil-Select Plus preparations （17.9% and 31.3%,respectively,P〈0.05） and increased in the SpermGrad preparation （56.3%,P〈0.05）. Protamine deficiency also decreased in all three kinds of media,59.3%,47.7% and 40.3% for PureSperm,Sil-Select Plus and SpermGrad preparations,respectively （P〈0.05）. The percentage of DNA-damaged sperm was negatively correlated with the percentages of sperm motility,rapid motility and PMC,but was positively correlated with static motility （P〈0.05）. This comparative study and correlation analysis revealed that PureSperm preparation yielded sperm with the best motility and the lowest percentage of protamine deficiency. The Sil-Select Plus preparation yielded sperm with the lowest amount of DNA damage. The SpermGrad preparation had a high percentage of sperm with normal morphology,but also had the highest percentage of sperm with DNA damage. Sperm DNA damage was correlated with percentages of sperm motility,rapid motility,static motility and PMC.     

精子优化处理后孵育时间与夫精宫腔内人工授精临床妊娠率的相关性研究

目的:研究精子优化处理后孵育时间对夫精宫腔内人工授精(AIH-IUI)临床妊娠率的影响。方法:相同促排卵方案的191个AIH-IUI周期,男方精液经密度梯度离心优化处理后,将孵育时间分为0~19、20~39、40~59、60~80 min 4组;根据优化处理后前向运动精子总数(TPMC)分为(0~9)×106、(10~20)×106、(21~30)×106、>30×1064组,分析孵育时间和优化处理后TPMC与临床妊娠率的关系,并对患者年龄、不孕年限、孵育时间、优化处理后TPMC与临床妊娠率进行Logistic多因素回归分析。结果:不同孵育时间临床妊娠率分别为12.7%、18.3%、11.4%、9.1%,孵育20~39 min组显著高于其他3组(P均<0.05);优化处理后4组TPMC临床妊娠率分别为0%、16.7%、11.4%、8.3%,(10~20)×106组显著高于其他3组(P均<0.05);Logistic多因素回归结果显示,女方年龄的增加会导致临床妊娠率显著下降(OR 0.89,95%CI 0.83~0.94),而孵育时间20~39 min(OR 2.11,95%CI 1.34~3.13)和处理后TPMC(10~20)×106(OR 2.06,95%CI 1.32~3.46)均可显著增加临床妊娠率。结论:精子优化处理后的孵育时间是AIH-IUI临床妊娠率的重要影响因素。     

Impact of cigarette‐smoking on sperm DNA methylation and its effect on sperm parameters

DNA methylation is an epigenetic modification of the genome. The purpose of this study was to determine the influence of cigarette‐smoking on sperm DNA methylation from a genomewide survey of sperm samples and to ascertain its effect on sperm parameters. Twenty‐eight sperm DNA samples (from 14 fertile smokers as a case study and 14 proven fertile nonsmokers as controls) were subjected to Infinium 450K BeadChip arrays to identify the changes in the DNA methylation level between the two groups. Then, deep bisulphite sequencing was used to validate five CpGs on 78 samples. The results from the Infinium 450K found that only 11 CpGs showed a significant difference in DNA methylation between the case and the control groups. Five CpGs of the eleven (cg00648582, cg0932376, cg19169023, cg23841288 and cg27391564) underwent deep bisulphite sequencing where cg00648582, related to the PGAM5 gene, and the cg23841288 CpGs, related to the PTPRN2 gene amplicons, showed a significant increase in their DNA methylation level in more than one CpG in the case group. In contrast, a significant decrease was found at cg19169023 and at its various neighbouring CpGs in the TYRO3 gene‐related amplicons. Furthermore, this study demonstrated a significant correlation between the variation in sperm DNA methylation level and standard sperm parameters in the case group.     

全自动精子分析中各运动性参数间相关性研究

目的　考察全自动精子分析中各运动性参数间的相关性。方法　进行了 190份健康受试者精液全自动分析 ,并对其各运动性参数 :直线速度 (vsl)、曲线速度 (vcl)、平均路径速度 (vap)、直线性 (lin)、前向性 (str)、摆动性 (wob)、侧摆幅度 (alh)、平均移动角度 (mad)、鞭打频率 (bcf)、精子密度、精子活动率间进行相关性分析。结果　从相关性分析中可确定运动性参数具有代表意义的是vsl、str、mad。结论　全自动精子分析的众多参数中 ,可通过vsl、str、mad三参数了解精子活动性 ,从而达到简便、快速、准确诊断的目的     

Relationship between donor sperm parameters and pregnancy outcome after intrauterine insemination: analysis of 2821 cycles in 1355 couples

The aim of this study was to investigate whether sperm parameters can affect the pregnancy outcome of artificial intrauterine insemination with cryopreserved donor spermatozoon (AID). A total of 1355 couples received 2821 AID treatment cycles in the Reproductive Medicine Center of the Tongji Medical College between January 2010 and December 2013, and the data were collected and retrospectively analysed. The relationship between pre‐freezing, post‐thawing as well as optimised sperm parameters and AID pregnancy outcome was investigated. Clinical pregnancy rate and cumulated pregnancy rate were also calculated. A total of 728 cycles from 2821 treatment cycles achieved pregnancies, and cumulated pregnancy rate was 25.81%. Pre‐freezing progressive sperm motility in pregnant cycles was higher than that in nonpregnant cycles (P = 0.001); logistic regression analysis also indicated that pre‐freezing progressive sperm motility was the only parameter affecting pregnancy outcome (P = 0.0001). Our study also showed that the cumulated pregnancy rate increased progressively and reached a plateau after the fifth cycle. In conclusion, pre‐freezing progressive sperm motility should be a valuable predictor for AID pregnancy outcome. Female fertility factors should be considered, or IVF/ICSI should be recommended when couples received more than 5 AID cycles without pregnancy.     

反复自然流产与精子DNA完整性的相关性研究

目的 探讨反复自然流产与精子DNA完整性的相关性。方法 按纳入标准将研究对象分成反复自然流产患者丈夫组（n=58）和正常生育男性对照组（n=50），采用吖啶橙实验（AOT）对研究对象精子DNA完整性进行检测，并对研究对象精液进行常规分析。砖果两组研究对象年龄、吸烟史、饮酒史无统计学差异（P〉0．05）；反复自然流产患者丈夫组精液的精子密度、精子活力显著低于正常生育对照组（P〈O．05）；反复自然流产患者丈夫组精子DNA的完整性低于正常生育对照组，差异有统计学意义（P〈O．05）。砖论精子DNA完整性损伤是反复自然流产的可能原因，精子DNA完整性损伤导致反复自然流产的致病机理尚需进一步研究。     

Chronic consumption of alcohol and sperm parameters: our experience and the main evidences

The present article describes the recent clinical experience and the main clinical and experimental evidences on this topic. In the first part, we present retrospective data collected over the last year on the semen quality and hormonal characteristics of the alcohol consumers evaluated in our centre. In the second part, we describe the mechanisms by which chronic alcohol intoxication impairs the testicular function (evidences for an ethanol‐mediated effect at pre‐testicular/testicular and post‐testicular level). In the third part, we present data on ethanol taken a male risk factor of infertility, being present as one among other recreational drugs (also called lifestyle). Finally, is discussed the role of individual susceptibility factors and other variables.     

Effects of the normal sperm morphology rate on the clinical and neonatal outcomes of conventional IVF cycles

The present study assessed the influences of the normal sperm morphology rate on the clinical and neonatal outcomes of conventional in vitro fertilisation cycles. This retrospective study analysed 427 and 2,728 cycles from the normal sperm morphology rate <4% and ≥4% group respectively. The clinical (total fertilisation failure, clinical pregnancy, implantation and abortion) and neonatal (sex, gestational age, preterm birth, birthweight, low birth weight, live births and birth defects of newborns) outcomes were compared. The rate of total fertilisation failure in the normal sperm morphology rate <4% group was significantly higher compared with that in the normal sperm morphology rate ≥4% group (2.8% versus 1.2%, p = .012). Total fertilisation failure was completely resolved by early rescue intracytoplasmic sperm injection. The clinical pregnancy, implantation and abortion rates were not significantly different between the two groups. Additionally, the sex, preterm birth, low birth weight, live births and birth defect rates, gestational age and birthweight of newborns were not significantly different between the two groups. Thus, normal sperm morphology rate <4% significantly increased the total fertilisation failure rate but did not affect the clinical or neonatal outcomes.     

Relationship between age and semen parameters in men with normal sperm concentration: analysis of 6022 semen samples

This study evaluates retrospectively the relationship between age and semen parameters among men with normal sperm concentration. It was based on computerized data and performed in an Academic Fertility and IVF Unit. Six thousand and twenty-two semen samples with sperm concentrations of >or=20 x 10(6) ml(-1) were examined according to WHO criteria and analysed in relation to patients' age. For each age group, mean values +/- SD of semen volume, sperm concentration, percentage of motile spermatozoa, normal morphology, acrosome index, total sperm count/ejaculate, total motile sperm count/ejaculate and sexual abstinence duration were examined. A peak semen volume of 3.51 +/- 1.76 ml(-1) was observed at age >or=30 to <35 years and a lowest volume of 2.21 +/- 1.23 ml(-1) was observed at age >or=55 years (P<0.05). Sperm motility was found to be inversely related to age with peak motility of 44.39 +/- 20.69% at age <25 years and lowest motility of 24.76 +/- 18.27% at age >or=55 years (P<0.05). A reduction of 54% was observed for total motile sperm, between values of 103.34 +/- 107 x 10(6) at age >or=30 to <35 years and 46.68 +/- 53.73 x 10(6) (P<0.05) at age >55 years. A statistically significant and inverse relationship was observed between semen volume, sperm quality and patient age, in spite of prolonged sexual abstinence duration. Top sperm parameters were observed at age >or=30 to <35 years, while the most significant reduction in sperm parameters occurred after the age of 55 years.     

肾移植对精子形态及运动参数的影响

目的 探讨肾移植对人精子形态及运动参数的影响。方法 对15例男性尿毒症患者在肾移植前后分别进行了精液各项指标的测定，并与12名正常男性进行对照。结果 肾移植后患者的精子活力、存活率及精子运动活力参数较术前有明显改善（P〈0．05），而精子密度、精子正常形态率、头部缺陷精子百分率、MAI和SDI及精子运动方式参数与术前相比无明显改善（P〉0．05）。结论 接受肾移植后患者精子运动能力得到了明显改观，但精子形态及密度的改善并不明显。这一结果是否与服用环胞素A引起FSH水平升高有关，需要进一步研究。     

Study on correlation of sperm quality parameters with antioxidant and oxidant status of buffalo bull semen during various stages of cryopreservation

This investigation was carried out to study the correlation of sperm quality parameters with antioxidant and oxidant status of buffalo bull semen during various stages of cryopreservation. Semen samples were evaluated for sperm parameters (mass motility [MM], concentration [CON], progressive motility [PM], viability [VIB], acrosomal integrity [AI] and hypo‐osmotic swelling [HOS] response), antioxidants (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx] and total antioxidant capacity [TAC]) and oxidants (Lipid peroxidation [LPO] and reactive oxygen species [ROS]) at fresh, pre‐freeze and post‐thaw stages. Sperm parameters (PM, VIB, AI and HOS response) and antioxidants (SOD, CAT and TAC) were significantly (p < .05) reduced at fresh stage, and oxidants (LPO and ROS) were significantly (p < .05) increased at pre‐freeze and post‐thaw stages. At fresh stage, MM was negatively correlated with LPO (p < .05), and CON was positively correlated with SOD, TAC and CAT, negatively correlated with LPO and CAT was positively (p < .01) correlated with VIB and HOS response. At pre‐freeze stage, CAT was positively correlated with PM and AI (p < .05), and AI was negatively (p < .05) correlated with ROS. At post‐thaw stage, CAT was positively correlated with PM, VIB, HOS response and AI,, and LPO was negatively correlated with HOS, AI and VIB. The study of correlations of these parameters at different preservation stages with bull fertility may play an important role in developing models for predicting future fertility of bulls in the absence of conception rate data.     

Impact of testicular histopathology as a predictor of sperm retrieval and pregnancy outcome in patients with nonobstructive azoospermia: correlation with clinical and hormonal factors

In this study, our objective was to evaluate the impact of testicular histopathology on the outcome of intracytoplasmic sperm injection (ICSI) cycles of patients with nonobstructive azoospermia and correlate with clinical and hormonal parameters. For this purpose, 271 patients with nonobstructive azospermia (NOA) who underwent testicular sperm extraction (TESE) for ICSI cycles were retrospectively evaluated for sperm retrieval, fertilisation, embryo cleavage, clinical pregnancy and live birth rates among different testicular histology groups. We also correlated hormonal and clinical factors with histological findings. Sperm retrieval and fertilisation rates (FR) were found to be significantly different among all testicular histological groups of NOA except for embryo cleavage, clinical pregnancy and live birth rates. Furthermore, serum follicle stimulating hormone (FSH) level was the most significant variable to predict sperm recovery on TESE. Separate analyses within each testicular histological group revealed that higher FSH was also associated with lower pregnancy rates in only maturation arrest group. In conclusion, testicular histology significantly influences sperm retrieval and FRs but not pregnancy and live birth rates in nonobstructive azoospermia. However, FSH is the best predictor of a successful TESE.     

Human sperm DNA integrity in normal and abnormal semen samples and its correlation with sperm characteristics

Reports indicate an increase in the incidence of DNA fragmentation in male factor infertility and its role in the outcome of assisted reproductive techniques (ART). However, reports are conflicting between the relationships of sperm DNA integrity with conventional semen parameters. We examined the relationship between conventional sperm parameters and DNA integrity using acridine orange (AO) test. The study included 373 patients and 28 fertile volunteers. DNA normality was compared with semen parameters between the patient and donor populations. Significant correlations were noted between DNA normality and sperm concentration ( r  = 0.18, P  = 0.000), motility ( r =  0.21, P  = 0.0001), rapid motility (0.19, P  = 0.000), normal morphology by World Health Organization ( r =  0.15, P  = 0.019) and head defects ( r =  −0.15, P  = 0.023). A significant difference was noted in AO levels between donors and patients with asthenozoospermia ( P  = 0.002) and oligoasthenozoospermia ( P  = 0.001). A significant difference in DNA integrity was noted in samples having <30% and >30% normal morphology. A wide range of % DNA normality was observed in the patient group. Sperm assessment for DNA status using AO is reliable and shows good correlation with sperm count, motility and morphology. Assessment of sperm DNA status with AO staining may be helpful prior to ART.     

Association between total globozoospermia and sperm chromatin defects

Globozoospermia is a severe sperm morphological anomaly leading to primary infertility and low fertilisation following intracytoplasmic sperm injection (ICSI). This phenotype is observed in less than 0.1% of infertile men and is determined by small, round‐headed spermatozoa with absence of an acrosomal cap, acrosome protease and also cytoskeletal proteins. Failure of oocyte activation is considered as the main cause of fertilisation failure in these individuals post‐ICSI. Therefore, artificial oocyte activation (AOA) along with ICSI is commonly implemented. However, based on previous report, fertilisation rate remains low despite implementation of ICSI‐AOA. Therefore, other mechanisms like sperm chromatin packaging and DNA fragmentation may account for low fertilisation and development post‐ICSI‐AOA. Therefore, this study aims to assess and compare the degree of sperm protamine deficiency and DNA fragmentation in large population of infertile men with total globozoospermia (30 globozoospermic men presenting with 100% round‐headed spermatozoa) with 22 fertile individuals using chromomycin A3 and TUNEL assay respectively. Results clearly show that mean of sperm concentration and percentage of sperm motility were significantly lower, while percentage of sperm abnormal morphology, protamine‐deficient and DNA‐fragmented spermatozoa were significantly higher in infertile men with globozoospermia compared to fertile men. Therefore, increased sperm DNA damage in globozoospermia is likely related to defective DNA compaction and antioxidant therapy before ICSI‐AOA could be recommended as an appropriate option before ICSI‐AOA.     

人精子体外获能前后计算机辅助分析参数的比较

目的为临床使用计算机辅助精子分析(CASA)系统的精液分析参数提供参考。方法对48例人类冷冻复温精液和116例临床进行丈夫人工授精(AIH)的新鲜精液体外获能前后CASA检测结果,分组进行配对资料t-检验和独立样本t-检验。结果所有实验组精子体外获能后的曲线运动速度(VCL)、直线运动速度(VSL)、平均路径速度(VAP)、精子头侧摆幅度(ALH)和鞭打频率(BCF)显著提高(P<0．01);除冻存精液精子体外获能后的运动前向性(STR)和头部椭圆度外,其余各组精液精子获能后的STR、直线性(LIN)和头部椭圆度显著下降(P<0,01)。行AIH治疗的精液标本,无论获能前后,已孕组精子VAP、LIN和头部椭圆度显著高于未孕组(P<0．05),而BCF和头部面积低于未孕组(P<0.05);获能前两组的(a b)级精子百分率无显著性差异,获能后差异显著(P<0．05)。结论体外获能处理可显著提高精子的各种运动速度,但精子运动的前向性和直线性降低;精液常规分析结果中的精子运动等级和速度不直接反映精子的受精能力。     

The effect of cysteine and glutamine on human sperm functional parameters during vitrification

Assuming the adverse effects of reactive oxygen species (ROS) on sperm function, this study was conducted to assess the effects of cysteine and glutamine as effective antioxidants on human sperm parameters under vitrification. Twenty normozoospermic samples were used. The samples were subjected to a vitrification process and cysteine (5 and 10 mM) and glutamine (10 and 15 mM). The sperm motility parameters, mitochondrial membrane potential (MMP), plasma membrane integrity (PMI), DNA damage and intracellular ROS damage were assessed for each sample. Statistical analyses showed that motility, mitochondrial membrane potential and DNA damage decreased in the vitrified groups with cysteine 5, 10 mM and glutamine 10, 15 mM separately. Also intracellular ROS increased significantly compared to the fresh group (p < .05). No significant differences were observed for PMI compared with the fresh group (p > .05). Supplementation of cysteine and glutamine in both concentrations separately decreased intracellular ROS and DNA damage of spermatozoa with significant increase in PMI, MMP and progressive motility compared to vitrified control group (p < .05). The results showed no significant effect of a specific concentration in cysteine and glutamine on sperm parameters compared to other concentrations. Both amino acids have the potential to improve the harmful effects of freezing on sperm parameters.     

Antioxidant effect of manganese on the testis structure and sperm parameters of formalin‐treated mice

Manganese inhibits oxidative stress damage. The aim of this study was to investigate the protective role of manganese on testis structure and sperm parameters in adult mice exposed to formaldehyde (FA). Twenty adult male NMRI mice were selected and randomly divided into four groups: (i) control; (ii) sham; (iii) ‘FA’‐exposed group; and (iv) ‘FA and manganese chloride’‐exposed group. The FA‐exposed groups received 10 mg kg^?1 FA daily for 14 days, and manganese chloride was just injected intraperitoneally 5 mg kg^?1 on 2nd weeks. Mice were sacrificed, and spermatozoa were collected from the cauda of the right epididymis and analysed for count, motility, morphology and viability. The other testicular tissues were weighed and prepared for histological examination upon removal. Seminiferous tubules, lumen diameters and epithelium thickness were also measured. The findings revealed that FA significantly reduced the testicular weight, sperm count, motility, viability and normal morphology compared with control group (P ≤ 0.05). In addition, seminiferous tubules atrophied and seminiferous epithelial cells disintegrated in the FA group in comparison with the control group (P ≤ 0.05). However, manganese improved the testicular structure and sperm parameters in FA‐treated mice testes (P ≤ 0.05). According to the results, manganese may improve and protect mice epididymal sperm parameters and testis structure treated with FA respectively.     

Antioxidant supplementations in vitro improve rat sperm parameters and enhance antioxidant enzyme activities against dimethoate-induced sperm damages

Organophosphorus compounds are currently among the most frequently used pesticides worldwide, and therefore, the potential for human exposure to man is considerable. Their toxicity results in negative effects on many organs and systems such as the male reproductive system. So, vitamins that can offer spermatozoa protection are of great importance. This study was designed to investigate (i) the possibility of dimethoate, an organophosphate insecticide, to induce oxidative stress response in rat spermatozoa in vitro and its effect on antioxidant defence system and (ii) the role of vitamin C and vitamin E in alleviating the cytotoxic effects of dimethoate Epididymal spermatozoa were incubated for 3 h at 37 °C with different concentrations of dimethoate (50, 100 and 200 μm) without vitamins or pre-incubated with 20 mm of vitamin C or 2 mm of vitamin E. Sperm parameters, malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) levels were performed. Dimethoate caused a significant induction of oxidative damage in spermatozoa at different concentrations as evidenced by increased MDA levels. However, a significant decrease in sperm mobility, viability and activities SOD, CAT and GPx was observed. Vitamins pre-treated spermatozoa showed a significant protection against the cytotoxic effects induced by dimethoate on studied parameters.     

Seasonality in sperm parameters in normal men and dyspermic patients on medical intervention

This study attempted to investigate the presence of seasonal variations in sperm parameters and to evaluate the season's impact on the response to treatment in men with idiopathic oligozoospermia (IO). To this end, a retrospective analysis of the records of 294 men, who participated in a controlled study, was performed. This sample included IO men ( n  = 106) treated with tamoxifen citrate (10 mg b.i.d.) and testosterone undecanoate (40 mg t.i.d.) or placebo ( n  = 106) and normozoospermic men ( n  = 82) serving as controls. Outcome measures included sperm parameters, functional sperm fraction (FSF) and incidence of pregnancy. Analysis showed a raised frequency of high FSF values and increased area under the response curve (AURC) for FSF mean during autumn-winter seasons in patients on active treatment compared with those in placebo ( P  < 0.05– P  < 0.04). Moreover, receiver operation characteristics (ROC) curves for a >100% FSF rise significantly discriminated autumn-winter from other seasons ( P  < 0.001, all), whereas active treatment showed higher than placebo FSF values particularly during autumn and winter ( P  < 0.001, all). The pregnancy incidence was higher in the autumn in all groups. It is concluded that FSF values showed a better response to active treatment during autumn and winter, indicating that commencement of empirical treatment at this time in IO men may stand a better chance to succeed.