Porcine sperm vitrification I: cryoloops method

The aims of this study were to evaluate porcine sperm vitrification in cryoloops, with and without two different cryoprotectants and assess two warming procedures. Extended (n = 3; r = 4) and raw (n = 5; r = 2) semen was diluted in media without and with cryoprotectants (4% dimethylformamide and 4% glycerol) to a final concentration of 20 × 10⁶ spermatozoa ml^?1 and vitrified using the cryoloops method. Two warming procedures were evaluated: rapid method (30 s at 37°C) and an ultra‐rapid method (7 s at 75°C, followed by 30 s at 37°C). Total motility (phase contrast), sperm viability (6‐carboxifluorescein diacetate and propidium iodide stain), membrane function (hypo‐osmotic swelling test), acrosome integrity (phase contrast), chromatin condensation (toluidine blue stain) and chromatin susceptibility to acid denaturation (acridine orange stain) were evaluated before and after vitrification and analysed using Friedman's test. In all media, the only seminal parameters that were maintained after vitrification were chromatin condensation and integrity. Vitrification of porcine spermatozoon using cryoloops, both in the presence or absence of cryoprotectants and independent of the warming procedure used, permits conservation of sperm chromatin condensation and integrity. It would be interesting to further verify this by producing porcine embryos using vitrified spermatozoon with intracytoplasmic sperm injection.     

Processes involved in assisted reproduction technologies significantly increase sperm DNA fragmentation and phosphatidylserine translocation

Sperm preparation techniques in assisted reproduction technologies (ART) are potential generators of exogenous stresses that cause additional DNA damage. DNA fragmentation tests, such as the sperm chromatin structure assay, involve freezing sperm samples in the absence of cryoprotectant. Thermal, oxidative stress (OS) and freezing are detrimental to sperm DNA fragmentation and phosphatidylserine (PS) translocation. The primary aim of this study was to subject mature sperm to environmental insults that normally occur during ART. We tested the hypotheses that OS, thermal stress and freeze‐thawing caused sperm nuclear and membrane damage and that a positive correlation exists between PS translocation and DNA fragmentation. Sperm DNA integrity deteriorates in semen samples from men with advancing age and a sperm concentration of <15 m  ml^?1. The significant increase in sperm DNA fragmentation at 37 °C after merely 1 h is important clinically as semen liquefaction and short‐term sperm storage in an ART cycle involve incubating samples at this temperature. Freezing without a cryoprotectant significantly increases the level of sperm nuclear damage, so it is important not to freeze neat semen prior to DNA fragmentation testing. This study highlights the importance of minimising the production of exogenous stresses during sperm preparation in ART.     

Correlation of sperm parameters with apoptosis assessed by dual fluorescence DNA integrity assay

Failed fertilization after intracytoplasmic sperm injection or miscarriages occurs in cases involving apoptotic and necrotic sperm. Identifying normal sperm is important for successful assisted reproductive technologies (ART) procedures. The study was conducted to correlate sperm parameters with intact sperm with normal DNA assessed by the dual stain assay in 118 separate individuals. The results showed differences in percent DNA intact sperm in individuals with normal W.H.O. sperm features (62 +/- 1.1; mean +/- S.E.M.) compared with oligoasthenoteratozoospermia patients (38 +/- 5.3). Individuals whose sperm had fertilizing capacity had higher percentages of intact DNA (60 +/- 1.3 versus 47 +/- 2.4). The percentages of intact DNA sperm were significantly correlated to total motility in semen (R = 0.7), post-wash motility (R = 0.6), rapid progression (R = 0.6), intact acrosome (R = 0.5), and strict morphology (R = 0.5). There were no correlations with the remaining parameters. The dual stain assay identified sperm with normal physiology and fertilizing capacity. The dual stain assay measures DNA integrity and is a promising method to select normal sperm for ART.     

Effect of leukocytospermia on sperm DNA integrity: a negative effect in abnormal semen samples

Controversy exists over levels of DNA integrity in the sperm of fertile and infertile men. In addition, the effect of leukocytospermia on sperm DNA in these 2 groups is unclear. We decided to address these questions by collecting semen samples from men known or presumed to be fertile and men from infertile couples. Samples were analyzed and assessed for sperm concentration, motility, and morphology. Samples failing to meet World Health Organization (WHO) standards in one or more of these parameters were judged abnormal. Samples were then arbitrarily assigned normalized scores in each of the above parameters, and scores were summed to give a normalized value for overall sperm quality. DNA abnormality was determined by an in situ DNA denaturation test with acridine orange and expressed as a percentage of cells with abnormal DNA integrity (ADI). Assessment of 187 samples revealed a moderate inverse correlation between ADI and sperm quality (r =.58), although a large degree of ADI dispersion was observed in abnormal semen samples. The average ADI for normal and abnormal semen samples was 18% +/- 2.8% and 36% +/- 5.8%, respectively, with the threshold of 95% probability set at 30%. When sorted for leukocytospermia, the difference in ADI between normal and abnormal semen groups without leukocytospermia was much smaller (17% +/- 2.2% and 22% +/- 4.6%; P =.023). Leukocytospermia had no significant effect on ADI in the normal semen group (P = .46); however, ADI was more than double the ADI in the abnormal semen group (18% +/- 2.4% and 50% +/- 11%; P < .001). The results of our analysis show that at least 3 factors affect net DNA integrity in leukocytospermic samples that fail to meet WHO standards: 1) primary DNA damage, which is moderately inverse to sperm quality, in particular to sperm concentration; 2) effect of leukocytes increasing primary or provoking potential DNA damage in a cascade-like manner, particularly in sperm with poor morphology and motility; and 3) a decreasing proportion of cells with damaged DNA in semen with the worst quality.     

Male age is more critical to sperm DNA integrity than routine semen parameters in Chinese infertile males

This retrospective study evaluated the correlation between the sperm DNA integrity results and infertile male age or sperm motility in 654 infertile men undergoing infertility evaluations from 2013 to 2016. The correlation between the results of sperm DNA integrity and male age was positive, while a negative correlation was detected between sperm DNA integrity and sperm motility in all subjects. According to age (≤30, 30–35 and ≥35), men with normozoospermia or abnormal semen parameters were, respectively, divided into groups 1, 2 and 3, or groups A, B and C. The sperm DNA fragmentation index (DFI) and DFI abnormality rates in groups 3 and C were highest among their respective cohorts. But they were not significantly different between groups within the same age range. Statistically significant differences were found in male age, progressive motility, as well as total motility between patients with normal DFIs and those with abnormal DFIs in group C, but not in group 3. Older (≥35 years) infertile men have increased sperm DNA fragmentation, independent of conventional semen parameters. Male age is more critical to sperm DNA integrity than routine semen parameters.     

The association of seminal leucocytes,interleukin‐6 and interleukin‐8 with sperm DNA fragmentation: A prospective study

The effect of seminal leucocytes on sperm DNA integrity has been discussed controversially in literatures. Moreover, the studies investigating the in vivo effect of pro‐inflammatory cytokines interleukin‐6 and interleukin‐8 on sperm DNA fragmentation are scarce and inconsistent. The association of standard sperm parameters with sperm DNA fragmentation is also a matter of ongoing discussion. Hence, the aims of this study were, first, to evaluate the effect of seminal leucocytes, interleukin‐6 and interleukin‐8 on sperm DNA integrity and, second, to examine whether standard semen parameters are associated with sperm DNA fragmentation. Seminal leucocytes, interleukin‐6, interleukin‐8 and standard semen parameters, including total sperm number, sperm concentration, progressive motility, nonprogressive motility, immotility and normal morphology, were determined in 134 consecutive men. The concentrations of seminal leucocytes, interleukin‐6 and interleukin‐8, did not correlate with sperm DNA fragmentation. In contrast, total sperm number, sperm concentration, progressive motility, nonprogressive motility and normal morphology exhibited significant inverse correlations with sperm DNA fragmentation. Immotile spermatozoa were directly correlated with sperm DNA fragmentation. In conclusion, seminal leucocytes, interleukin‐6 and interleukin‐8, are not associated with sperm DNA fragmentation. Poor standard semen parameters are significantly related to the high levels of sperm DNA fragmentation.     

Pressure flow pattern of varicocele veins and its correlation with testicular blood flow and semen parameters

The pressure pattern in varicocele veins of infertile patients and its correlation with semen quality and testicular blood flow was determined. Consecutive patients at andro‐urology clinic of a teaching hospital undergoing microsurgical varicocelectomy were included. Their semen quality and testicular blood flow were determined. Peak systolic velocity (PSV) and resistive index (RI) of subcapsular and intraparenchymal branches of testicular artery were noted by colour Doppler ultrasonography. During surgery before ligation of varicocele veins, intravenous pressures of internal spermatic (ISV) and external spermatic (ESV) veins were determined at baseline and after Valsalva manoeuvre. Thirty patients, 20–45 years old, were evaluated. Baseline pressure for maximum dilated ISV (A), less dilated ISV (B) and ESV was 15.93 ± 6.34, 12.38 ± 4.60 and 12.92 ± 5.65 mm. Hg, respectively, which increased after Valsalva by 104.4%, 116.2% and 38.22% respectively. Correlation (r = ?.71; p < .05) was appreciated between percentage increase in pressure of ISV B with PSV of intraparenchymal testicular arteries and progressive motility (r = ?.759; p < .05), nonprogressive motility (r = ?.738; p < .05) and morphology (r = ?.653; p = .07) of spermatozoa. In conclusion, ISV develops higher pressure on Valsalva as compared to ESV and has correlation with semen quality and testicular blood flow.     

Relationship between conventional sperm parameters and motile sperm organelle morphology examination (MSOME)

With the motile sperm organelle morphology examination (MSOME), spermatozoa morphology may be assessed directly on motile spermatozoa at high magnification (up to 6600×). This procedure describes more precisely spermatozoa abnormalities, especially head vacuoles. However, no consensus has been established concerning normal or abnormal MSOME criteria. The aim of our study was to define MSOME vacuole criteria assessed objectively with a digital imaging system software to establish a potential relationship between conventional semen parameters. A total of 440 semen samples were obtained from males consulting in Rouen University Hospital Reproductive Biology Laboratory. Conventional semen analysis (volume, sperm concentration, progressive motility, vitality and morphology) and MSOME assessment {sperm head length, width and area as well as vacuole number, vacuole area and relative vacuole area to sperm head [RVA (%) = [vacuole area (μm(2))/head area (μm(2))] × 100)]} were performed for each semen sample. Among our 440 males, 109 presented normal conventional semen parameters and 331 abnormal ones. Sperm head vacuoles were significantly larger in abnormal semen samples (p < 0.0001). RVA was the most discriminative MSOME criterion between normal and abnormal semen samples according to ROC curves analysis, and was negatively correlated with poor sperm morphology (r = -0.53, p < 0.0001). We concluded to (i) the normal occurrence of vacuoles in sperm head whatever the normality or abnormality of semen parameters, (ii) the discriminative function of the RVA to distinguish semen samples with normal and abnormal parameters, and (iii) the strong correlation between high RVA and poor sperm morphology.     

Sperm DNA damage and its clinical relevance in assessing reproductive outcome

The routine examination of semen, which assesses sperm concentration, percentage motility and morphology,does not identify subtle defects in sperm chromatin architecture. The focus on the genomic integrity of the male gamete has intensified recently due to the growing concern that genetic diseases may be transmitted via assisted reproductive techniques (ART). Accordingly, the intent of this review is to describe the details of the informationpertaining to mitochondfial/nuclear sperm DNA damage with an emphasis on its clinical significance and its relationship with male infertility. Assessment of sperm DNA damage appears to be a potential tool for evaluating semen samples prior to their use in ART. Testing DNA integrity may help select spermatozoa with intact DNA or with the least amount of DNA damage for use in assisted conception. In turn, this may alleviate the financial, social and emotional problems associated with failed ART attempts.     

Famotidine does not affect human sperm fertility characteristics (motility,viability and DNA integrity) in vitro

Famotidine, a histamine‐2 receptor antagonist, is commonly used to relieve the acid‐related gastrointestinal diseases; however, its effect on human sperm parameters, and hence on sperm function, is still undetermined. Here, we intended to measure human sperm motility, viability, and DNA integrity of ejaculated human sperm in the presence of famotidine at 0, 0.1, 1 and 10 mM concentrations in vitro. Forty‐nine semen samples of normal count, motility, and morphology were included in this study. Sperm motility was assessed using Makler counting chamber and a phase contrast optics (200× magnification), whereas sperm viability was assessed using eosin–nigrosin staining procedure. The effect of famotidine on sperm DNA integrity was measured using flow cytometry. Famotidine at 0.1, 1 or 10 mM had insignificant effect on human sperm motility (progressive, p = .9594; and total, p = .8420), sperm viability (p = .6471), and content of DNA breaks in sperm (p > .05) compared with the control. In conclusion, famotidine at 0.1, 1 or 10 mM did not alter human sperm motility, viability or DNA integrity in vitro. Although, these findings indicate safety of famotidine in human sperm, further in vivo studies are required to establish the drug's safety.     

Factors affecting fecundity among sperm donors: a multivariate analysis

The study was aimed at identifying the predictors of the male fertility potential among sperm donors. Fifty anonymous donors undergoing 683 intracervical insemination (ICI) cycles between January 2002 and December 2006 were retrospectively evaluated according to semen characteristics in terms of reproduction rate (RR). We used RR as a parameter to determine the fertility potential among sperm donors. The overall RR was 26.79%. There were no significant differences among low, mean and high RR groups with regard to most sperm routine parameters. However, the RR was notably higher in the sperm morphology of ≥18% than in the <18% group (26.2% versus 19.4% respectively; P < 0.01). Both post-thaw total motility and progressive motility were proportional to RR (P < 0.01). Differences in RR were seen when the percentage of propidium iodide-negative spermatozoa was ≥45% (26.2% versus 16.4% respectively; P < 0.01) and DNA fragmentation index (DFI) was <8% (37.5% versus 17.9% respectively; P < 0.01) in post-thaw samples. Using stepwise linear regression analysis, the percentage of normal morphology, post-thaw progressive motility, PI-negative spermatozoa, DFI had the maximum power to predict the donor fecundity in ICIs. Conclusion: Both the integrity of plasma membrane and DNA in spermatozoa are crucial factors affecting the fecundity of sperm donors. Therefore, the addition of some of these new tests to routine semen analysis could significantly improve the recruitment of sperm donors and the clinical pregnancy rate of anonymous donors.     

Relationship between seminal ascorbic acid and sperm DNA integrity in infertile men

Ascorbic acid has recently been reported to protect sperm DNA from the damage induced by exogenous oxidative stress in vitro. But, there is no report on seminal ascorbic acid and sperm DNA fragmentation in infertile men. In this study, we asked whether sperm DNA damage correlates with seminal ascorbic acid levels. Sperm DNA fragmentation index (DFI) was analysed in 75 men by flow cytometry after acridine orange staining. We also measured the levels of seminal plasma ascorbic acid and total antioxidant capacity. Abnormal sperm DNA integrity (DFI >or= 30%) was observed in 12% of the patients with normal semen parameters and in 52% of the patients with abnormal semen parameters. There were significant correlations between the level of DFI and conventional semen parameters including sperm count, motility and morphology (r = -0.29, -0.55 and -0.53 respectively; p < 0.05). Seminal ascorbic acid level was significantly lower in the patients with leucospermia than the patient with normal semen parameters. Interestingly, a significantly greater percentage of men with abnormal DFI were observed in the patients with low levels of seminal ascorbic acid compared with those with normal or high levels of ascorbic acid (59% vs. 33%, p < 0.05). Men with insufficient seminal ascorbic acid frequently have sperm DNA damage.     

Canthaxanthin protects human sperm parameters during cryopreservation

Different antioxidants have been introduced to reduce oxidative stress during the cryopreservation. The main goal of this study was to evaluate the effects of canthaxanthin on human sperm parameters during the freeze‐thaw process. This study was performed on 25 normozoospermic semen samples dividing into five groups including 0, 0.1, 1, 10, and 25 µM of canthaxanthin. The prepared spermatozoa were cryopreserved by rapid freezing technique. Sperm motility, viability (eosin‐nigrosin), morphology (Papanicolaou), acrosome reaction (double staining), DNA denaturation (acridine orange), chromatin packaging (aniline blue and toluidine blue), and DNA fragmentation (sperm chromatin dispersion test) were evaluated before freezing and after thawing. All sperm parameters after thawing significantly were decreased compared to before freezing. Twenty‐five micromolar canthaxanthin could significantly improve the progressive and total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Ten micromolar canthaxanthin significantly improved total motility, viability, normal morphology, chromatin packaging, acrosome integrity and DNA denaturation and fragmentation. Whereas, in 1 µM group, there were significant differences only in improvement of acrosome integrity, chromatin packaging (toluidine blue) and DNA denaturation and fragmentation. But, in 0.1 µM group, there were no significant differences in any of measured parameters. It seems that canthaxanthin ameliorates detrimental effects of cryopreservation on human sperm parameters.     

Prognostic value of male diagnostic profiles in intracytoplasmic sperm injection (ICSI)

Possible correlations between male hormone and semen parameters with pregnancy and oocyte fertilization rates following intracytoplasmic sperm injection (ICSI) were investigated. The study is based on 290 couples who underwent ICSI therapy for the first time. The parameters evaluated were male age, serum levels of follicle stimulating hormone (FSH) and testosterone, sperm concentration, sperm motility, normal sperm morphology, index of teratozoospermia (TZI) and sperm vitality. A marginal, barely significant association was found between the fertilization rate and serum FSH levels in the male partner ( p =  0.046). There was no relevant association between male parameters and pregnancy rates. The study confirms that male hormonal and semen parameters are of low prognostic value for the outcome of ICSI.     

精子DNA完整性与体外受精-胚胎移植临床结局的相关性研究

目的 探讨精子DNA完整性与精液参数及体外受精-胚胎移植（IVF-ET）/卵胞浆内单精子注射（ICSI）临床结局的关系.方法 选择2008年6月～2009年6月在解放军105医院生殖医学中心接受IVF/ICSI治疗的179对不育夫妇作为研究对象,采用吖啶橙试验（AOT）对116例实施IVF和63例实施ICSI治疗的男性患者进行精子DNA完整性分析,根据精子DNA碎片指数（DFI）将患者分为DFI≤30%组和DFI＞30%组,比较两组间精液参数、受精率、卵裂率、优胚率、胚胎冷冻率、着床率和临床妊娠率.结果 DFI ＞30%组精子畸形率显著高于≤30%组（P＜0.01）,但两组间精子密度、活动率、前向运动精子（a＋b）均无显著性差异（P＞0.05）;DFI＞30%组IVF和ICSI的优胚率、ICSI的胚胎着床率和临床妊娠率均显著低于DFI≤30%组（P＜0.01,P＜0.05）.结论 精子DNA完整性与精子形态密切相关,精子DNA损伤在IVF/ICSI过程中对胚胎质量有负面影响,并显著影响ICSI的胚胎着床率和妊娠率,建议行ICSI前应对精子DNA完整性进行评估.     

Sperm motility and DNA integrity affected by different g‐forces in the preparation of sperm in urine specimens

Retrograde ejaculation, a common type of anejaculation, is attributable to many causes, some of which can be treated with medication and some of which cannot. For infertility treatment, sperm must be collected from the urine of the patients. Our study attempts to ascertain the effects of different g‐forces on sperm motility, morphology and DNA integrity in sperm preparation by the Sil‐Select? density gradient method of isolating sperm from urine specimens. Forty‐seven semen samples with normal semen analyses according to World Health Organisation (WHO) 1999 criteria were included in this study. Semen samples of 1 ml were mixed with 20 ml alkalinised normal urine and then divided equally into tubes A and B. The two samples were prepared by the Sil‐Select? density gradient centrifugation method at 350 g (tube A) and at 700 g (tube B). Total motile sperm after centrifugation at 700 g was significantly higher than after centrifugation at 350 g [6.7 (0.4–23.0) million versus 3.1 (0.1–13.7) million] (P < 0.001). There was no significant difference between the either the percentage of sperm with normal morphology or with DNA damage between centrifugation at 350 g and 700 g (P > 0.05), although centrifugation at 700 g achieves a higher number of total motile sperm compared with Sil‐Select? sperm preparation at 350 g centrifugation.     

Effect of repeated sequential ejaculation on sperm DNA integrity in subfertile males with asthenozoospermia

The aim of this work was to study the possible beneficial effect of repeated sequential ejaculation on sperm DNA integrity in subfertile males and its possible implementation in assisted reproduction. The study included 20 infertile males with idiopathic asthenozoospermia or oligoasthenozoospermia. They underwent detailed history taking, complete clinical assessment and hormonal assessment. Patients were asked to bring two semen samples (taken within 1-3 h). Two consecutive samples were assessed with regard to semen volume, sperm count, motility grading, and morphology and sperm DNA integrity using the comet assay. There was a significant improvement in the sperm motility pattern and DNA integrity in the second sample in comparison with the first sample. Therefore, it is concluded that due to its positive impact on sperm motility and DNA integrity, repeated sequential ejaculation is recommended in subfertile males with idiopathic asthenozoospermia who pursue assisted reproduction.     

Comparative analysis of tests used to assess sperm chromatin integrity and DNA fragmentation

Male infertility has a complex etiology, and many times, the cause is unknown. While routine semen analysis provides an overview of basic semen parameters, such as sperm concentration, motility, viability and morphology, a significant overlap of these parameters has been reported in fertile and infertile men. Moreover, conventional semen parameters do not reveal the cellular or molecular mechanisms of sperm dysfunctions leading to infertility. Therefore, sperm functional parameters, including sperm chromatin integrity, are evaluated to provide information on subtle sperm defects that are not routinely identified. Incomplete or defective sperm chromatin condensation increases the susceptibility of the sperm DNA to oxidative damage or other factors. To evaluate sperm chromatin integrity, different methods with varying degrees of diagnostic and prognostic capabilities are available. Among these assays, SCSA, TUNEL and SCD assays are most commonly used. While these assays rather evaluate the DNA directly for damages, the aniline blue and chromomycin A3 stains test for the quality of chromatin condensation. Thus, this review discusses and compares different methods used to evaluate sperm chromatin integrity and condensation, and their inclusion in the routine evaluation of the male infertility.     

Impact of body mass index values on sperm quantity and quality

Body mass index (BMI) has been demonstrated to affect female fertility; however, little information is available on the impact of BMI on male fertility or semen parameters. Therefore, the study objective was to determine the relationship between BMI and semen parameters, including sperm chromatin integrity. We analyzed data on semen samples from 520 men who were grouped based upon calculated BMI values (normal, 20-24 kg/m(2); overweight, 25-30 kg/m(2); obese, >30 kg/m(2)). The data collected included patient height and weight, semen volume, sperm concentration, percent sperm motility, percent sperm morphology (normal forms), and sperm chromatin integrity (DNA fragmentation index [DFI]). Data were analyzed by regression analysis and analysis of variance (ANOVA) with Tukey's test for multiple pairwise comparisons. The overall BMI mean (+/-SEM) was 27.5 (+/-0.49) kg/m(2). Linear regression revealed a significant (P < .05) and negative relationship between BMI and the total number of normal-motile sperm cells. ANOVA revealed a significant difference (P < .05) in the total number of normal-motile sperm cells among the different BMI groups. The number of normal-motile sperm cells per BMI group was as follows: normal, 18.6 x 10(6); overweight, 3.6 x 10(6); and obese, (0.7) x 10(6). All multiple pairwise comparisons were found to be significantly (P < .05) different. The overall DFI mean (+/-SEM) was 24.7 (+/-2.57). Linear regression revealed a significant (P < .05) and positive relation between BMI and DFI. Men presenting with a BMI greater than 25 kg/m(2) have fewer chromatin-intact normal-motile sperm cells per ejaculate. Therefore, to ensure maximum fertility potential, patients may be advised to reduce body weight.     

养精赞育颗粒对弱精子症患者精子DNA完整性的影响

目的探讨养精赞育颗粒治疗弱精子症患者对精子DNA完整性的影响。方法选择弱精子症患者86例,口服养精赞育颗粒3个月。通过吖啶橙试验（AOT）与计算机辅助精液分析（CASA）检测DFI（精子DNA断裂指标）和精液常规参数。再将86例患者按照DFI值（大于10%或小于10%）进行分层研究并使用SPSS11.5软件进行数据分析。结果治疗3个月后,a级精子百分比和（a＋b）精子百分比与治疗前相比差异均有统计学意义（P〈0.01）;所有86例患者DFI绝对值（6.89&#177;4.08）与治疗前（11.31&#177;7.21）相比差异有统计学意义（P〈0.01）;DFI正常组DFI绝对值（4.71&#177;1.87）与治疗前（5.25&#177;2.61）相比差异无统计学意义（P〉0.05）;DFI异常组DFI绝对值（8.61&#177;4.53）与治疗前（16.11&#177;5.94）相比差异有统计学意义（P〈0.01）。结论应用养精赞育颗粒治疗男性不育可显著改善精子的活力和精子DNA的完整性;补脾益肾是提高精子DNA完整性的有效方法。