Bovine mucus penetration test and routine semen analysis of fresh and cryopreserved human spermatozoa

Semen samples from 18 fertile men and 18 infertile men were evaluated both fresh and after freezing and thawing using traditional semen parameter, the hypoosmotic swelling test (HOS-test) and the bovine cervical mucus penetration test (BMP-test). Major differences in the semen analysis between the fertile and infertile men were observed for motility, morphology (head and tail defects), the bovine mucus penetration test and the hypoosmotic swelling test. Cryopreservation resulted in a decrease regarding motility, the BMP-test and the HOS-test. Percentagewise the losses due to cryopreservation were higher in the infertile group than in the fertile group. The results of this study show that the bovine mucus penetration test as well as the HOS-test are valuable additions to the routine semen analysis of fresh and cryopreserved spermatozoa.     

Multivariate discriminate analysis of the relationship between the hypo-osmotic swelling test and the in-vitro fertilizing capacity of human sperm

Multivariate discriminant analysis was used to evaluate the usefulness of routine semen parameters and the hypo-osmotic swelling test (HOST) as predictors of the in-vitro fertilizing capacity of human sperm as assessed by the zona-free hamster egg penetration assay (HEPA). Eighty-eight semen samples from untreated patients attending an infertility clinic were analysed. Semen samples were classified into the following three groups before statistical analysis: group 1--positive sperm penetration (greater than or equal to 10%, n = 39); group 2--borderline penetration rates for HEPA (greater than 0% but less than 10%, n = 39) and group 3--negative sperm penetration (0%, n = 10). The percentage of sperm with normal morphology and sperm count were found to be significant in discriminating between semen samples exhibiting different in-vitro fertilizing capacity. These two discriminating variables in combination gave an overall correct classification rate of 45.5%. The multivariate discriminant analysis was also performed after excluding the data of group 2 semen samples (n = 39), which exhibited borderline sperm penetration rates. As a result, three discriminating variables including semen volume, sperm count and the percentage of sperm with normal morphology were selected. These three variables in combination could accurately predict whether a semen sample would exhibit positive sperm penetration (group 1) or negative sperm penetration (group 3) with an overall accuracy of 75.5%. The percentage of swollen sperm after hypo-osmotic treatment was not related to the HEPA result, as determined by linear correlation and multiple regression analyses, and did not give additional information about the in-vitro fertilizing capacity of sperm as evaluated by multivariate discriminant analysis.     

Seminal lead and copper in fertile and infertile men: Blei und Kupfer im Spermaplasma bei fertilen und infertilen Männern

Lead and copper concentrations were determined by atomic absorption spectroscopy in semen from 18 fertile and 172 infertile men. Significant correlations between copper concentrations in semen and sperm concentration (r = 0.32, P less than 0.001), percentage progressive motility (r = 0.23, P less than 0.005) and normal morphology (r = 0.22, P less than 0.005) were observed, while no such correlation existed for lead. However, semen copper concentrations of infertile men (194.99 +/- 5.70 micrograms l-1) and fertile men (183.39 +/- 14.37 micrograms l-1) did not differ significantly. Mean lead concentration in semen of fertile men was 11.18 +/- 0.62 micrograms l-1 and significantly higher than lead concentration in semen of fertile men (5.61 +/- 0.53 micrograms l-1, P less than 0.006). Reinvestigation of 18 infertile men after 2 years showed a significant drop of lead concentrations in semen from 17.31 +/- 1.41 to 6.94 +/- 1.32 micrograms l-1 (P less than 0.0002), which might be related to the decreasing use of leaded petrol in the Federal Republic of Germany.     

Comparison of the penetration ability of human spermatozoa into bovine cervical mucus and zona-free hamster eggs

In vitro bovine cervical mucus (BCM) penetration tests, sperm penetration assays (SPA) using zona-free hamster eggs, and routine semen analyses were performed on a total of 136 freshly collected semen samples from men who were seen at an infertility clinic. The correlations between bovine cervical mucus penetration and other semen parameters were the percent motile spermatozoa (r = 0.48), progressive motility grade (r = 0.44), sperm count (X 10(6)/ml) (r = 0.47), the percent normal morphology (r = 0.32) and the percent eggs penetrated (r = 0.46) (P less than 0.0001 for each correlation coefficient). When known fertile (n = 32) and infertile (n = 18) groups were tested, positive mucus penetration was associated 75% correctly and positive egg penetration was associated 90% correctly to clinical status. The mucus test had no false-negative results and the SPA had no false-positive results in these groups. It appears, then, that the mucus test and sperm penetration assay, although contributing different elements of data to an infertility evaluation, are both useful adjuncts to a semen analysis.     

Relationship between hypoosmotic swelling test, semen analysis, and zona-free hamster ovum test

The hypoosmotic swelling test is a simple test for measuring the functional competence of the human sperm membrane. Fifty-four patients with idiopathic infertility were assessed by hypoosmotic swelling test and the results were compared with those of routine semen analysis and zona-free hamster ovum human sperm penetration test (ZSPT). Semen samples with abnormal semen parameters had lower percentage of swollen sperm in comparison with those with normal semen parameters. A positive correlation was observed between sperm concentration and sperm swelling (r = .50, p less than .05). A strong positive correlation was observed between the percentage of sperm motility and the percentage of swollen sperm (r = 0.60, p less than .01), and between motile sperm concentration and sperm swelling (r = .62, p less than .01). On the other hand, sperm swelling correlated only weakly with the percentage of sperm penetration. The results indicate that the hypoosmotic swelling test appears to evaluate different functional qualities of sperm than ZSPT.     

Effect of freeze–thawing procedure on chromatin stability, morphological alteration and membrane integrity of human spermatozoa in fertile and subfertile men

Cryopreservation is known to impair sperm motility and decrease the fertilization rate by detrimental effects on acrosomal structure and acrosin activity. However, the consequences of cryopreservation on the integrity of the sperm nucleus, chromatin stability and centrosome are less clear. The present study was designed to determine the effect of the freeze-thawing procedure on chromatin condensation (aniline blue staining) and the morphology (strict criteria) and membrane integrity of human spermatozoa. The structural and functional characteristics of the sperm plasma membrane were measured by the eosin-test and hypo-osmotic swelling test which were done separately. Sperm cryopreservation was performed on semen samples from two groups of men classified as fertile (n = 20) and subfertile (n = 72), based on their reproductive history and semen analysis according to WHO guidelines. The mean percentage of condensed chromatin, morphologically normal spermatozoa and membrane integrity in all semen samples investigated (n = 92) decreased significantly (p = 0.0001) after freeze-thawing, in comparison to the value observed prior to freezing. By comparing the semen samples between fertile and subfertile patients, significantly (p = 0.0009) greater damage was demonstrated in the subfertile than in the fertile group. Furthermore, no significant difference was observed between the two groups with regard to the morphological alteration and structural as well as functional damage of the sperm membrane. In conclusion, the freeze-thawing procedure significantly affects chromatin structure and sperm morphology, especially in the head and the tail regions, and this may explain the lower fertilization rate and IVF/ICSI outcome when frozen-thawed spermatozoa are used. In addition, this study demonstrates that chromatin condensation is a sensitive parameter for the evaluation of cryodamage of semen samples from fertile and subfertile patients, though subfertile patients with very poor semen characteristics have yet to be studied. It is therefore recommended that chromatin condensation be used as an additional parameter for the assessment of sperm quality after freeze-thawing.     

Relationship between the human sperm hypo-osmotic swelling test and sperm penetration assay

The hypo-osmotic swelling (HOS) test has been proposed as a useful assay in the diagnosis of the infertile male. A good correlation between the HOS test and the sperm penetration assay (SPA) in fertile and normal semen samples was initially found, but subsequently, no significant correlation was demonstrated with fertile and infertile patients. To validate the potential clinical usefulness of the HOS test, we evaluated 92 ejaculates using the HOS test, SPA, and traditional semen parameters. The methodology originally described by Jeyendran et al (1984) was used for the HOS test. The SPA was performed by the original procedure using an 18-hour preincubation period, and for 28 ejaculates, a modified procedure using TEST-yolk buffer was performed. Values of 60% or more for the HOS and 1% or more for the SPA were considered positive, and less than 60% for HOS and 0% for SPA were considered negative when the standard SPA was performed. For the TEST-yolk buffered SPA, values of 20% or more were considered positive. The sensitivity of the HOS test was 87%, but the specificity was 36%. The association of the two tests over and above that expected by chance (Kappa) was only 0.23. Using logistic regression, both sperm count (P less than 0.001) and morphology (P less than 0.025) were significant predictors of the SPA classification, but the HOS test did not improve the predictive results (P greater than 0.50).(ABSTRACT TRUNCATED AT 250 WORDS)     

Oocyte Penetration and Acrosome Reactions of Human Sperms II: Correlation with other Seminal Parameters

Washed sperm suspensions of fertile men and men consulting for infertility were evaluated for their ability to penetrate zona-free hamster eggs and for their ability to exhibit an acrosome reaction in vitro. Furthermore, the swelling of the spermatozoa under hypoosmotic conditions as indication of their membrane integrity was determined. In the group of fertile men and in the group of patients with normal spermiogram, significantly more acrosome reactions were observed than in the group of infertile men with abnormal spermiogram parameters (p less than 0.05). This difference was still more significant when men with a positive hamster penetration test (H.O.P. test) and men with a negative H.O.P. test were compared (p less than 0.005). However, within the groups the level of acrosome reactions after incubation appeared to be highly variable. In a second series of experiments, working with semen obtained during our in vitro fertilization program, we found that the fertilization of human ova does not seem to be dependent on a strong progress of the acrosome reaction. Finally, swelling of the spermatozoa in a hypoosmotic medium was weakly correlated (r = 0.70, p less than 0.01; n = 73) with trypan blue exclusion. No significant correlations with other semen parameters, hamster ova test included, were found.     

Correlation of the hypo-osmotic swelling test, semen score and the zona-free hamster egg penetration assay in humans

The correlation between the hypo-osmotic swelling test, the modified Eliasson score for human semen analysis and the hamster egg penetration assay was examined. The results showed a weak but significant correlation between the group of subjects with swelling rates below 50% and the above 80% group. These groups showed hamster egg penetration assay rates of 9 +/- 14% and 39 +/- 29%, respectively (P less than 0.035). A significant correlation was also found between modified Eliasson scores (17 +/- 10 and 5 +/- 7, respectively) and swelling rates below 50% and above 70% (P less than 0.001). These results suggest that spermatozoa showing a low swelling rate have a poor fertilizing capacity. Therefore, the swelling tests and analysis of the patients semen can be used to discriminate sperm quality.     

An evaluation of the hypo-osmotic sperm swelling test as a predictor of fertilizing capacity in vitro

The ability of sperm to swell in hypo-osmotic conditions was examined in 211 semen samples from the partners of patients about to undergo oocyte retrieval for in-vitro fertilization (IVF). The test was performed using aliquots of semen, the remainder of which was then prepared for IVF. No significant difference was found, in either the percentage of swollen sperm or the type of swelling response, between samples that achieved fertilization in vitro and those that did not, or between any of the diagnostic categories of infertility (tubal damage, unexplained infertility, oligospermia). In samples which achieved fertilization in vitro there were correlations between sperm swelling and sperm motility (r = -0.51) and abnormal morphology (r = 0.33), but no such correlations were demonstrated in samples that failed to achieve fertilization. Moreover, there was no significant difference between the percentage of swollen sperm in semen (mean motility 64%), in samples immediately after preparation for IVF (mean motility 96%) or in capacitated sperm 24 h after preparation (mean motility 91%). These results demonstrate that the hypo-osmotic sperm swelling test does not assist in the prediction of the fertilizing capacity of human sperm in vitro.     

Sperm preparation: DNA damage by comet assay in normo- and teratozoospermics

The present study was carried out on semen samples of human fertile and infertile subjects, teratozoospermics (TZs) and idiopathics (IDs), with neat semen and sperm prepared by swim up or Percoll density gradient centrifugation procedures. Sperm morphology analysis revealed that only head and midpiece defects in TZs and IDs were significantly (P < 0.001) higher compared to fertile subjects. Infertile subjects indicated significantly higher (P < 0.001) sperm DNA damage compared to fertile subjects. Fertile subjects with sperm prepared from neat and Percoll density gradient centrifugation exhibited a comet tail DNA percentage of 20% and 15%, respectively. The TZs and IDs infertile subjects had higher levels of comet tail DNA of 33% and 25% and 25% and 19%, respectively. A significant (F = 24.01; P = 0.0059) decrease in mean comet head DNA percentage or sperm DNA integrity was observed in neat samples from fertile and infertile subjects by Repeated Measures ANOVA. In Percoll prepared samples from fertile, TZs, and IDs, there was a significant increase in sperm DNA integrity. Similarily, there was a decrease in abnormal sperm morphology in swim up and Percoll prepared sperm compared to neat samples. The Percoll density gradient centrifugation procedure yields sperm with an increase in sperm DNA integrity relative to swim up. Sperm DNA damage of TZs with both sperm preparation methods was significantly (P < 0.01) higher when compared to fertile and IDs. But the level of DNA damage was higher in IDs compared to fertile subjects. Compared to the other methods tested, the Percoll method yielded sperm with improved DNA integrity. In conjunction with semen analysis, the assessment of nuclear integrity improves the characterization of the semen sample and may be used as a tool for allocating the patients to specific assisted reproductive treatments.     

Sperm dysfunction in subfertile patients with varicocele and marginal semen analysis

A comparative analysis of potentially functional spermatozoa per ejaculate, progressive motility, hypo-osmotic swelling test, acrosome integrity and sperm viability (24 and 48 h) was carried out in a group of 40 subfertile patients with varicocele and marginal semen analysis and 40 fertile subjects, in order to identify subclinical abnormalities that may explain subfertility. Patients with varicocele had lower numbers of potentially functional spermatozoa per ejaculate, progressive motility, acrosome and membrane integrity and sperm viability. These abnormalities were not related to the grade of varicocele, testicular volume or serum FSH concentration. A positive correlation between the hypo-osmotic swelling test and progressive motility (r = 0.71) and between potentially functional spermatozoa and the hypo-osmotic swelling test (r = 0.69) was found in patients with varicocele. These data suggest that some of the deleterious effects produced by the varicocele might be related to sperm migration and viability in the female genital tract and others to sperm-zona interaction and/or sperm-egg fusion.     

Analysis of spermatozoa from the proximal vas deferens of vasectomized men

This study assessed the condition of spermatozoa from the proximal vas deferens of men after vasectomy. The fluids of both proximal vas deferens were collected from 67 vasectomized men by cannulating the vas deferens at the time of vasectomy reversal. Selected sperm parameters were analysed after incubation of the spermatozoa for 30 min at 37°C. Spera concentration in the proximal vas from vasectomized men (16 312 ± 21 496 million per ml, geometric mean: 7948 ± 398 million per ml) was significantly higher than that of fertile men and was maintained at a constant level independent of the duration of vas obstruction. The means of sperm motility (36.2 ± 26.2%), spermatozoa with normal morphology (50.7 ± 21.7%), sperm viability (53.0 ± 25.3%) and hypo-osmotic swelling test (HOS-test, 53.9 ± 21.7%) were statistically lower than the respective values for normal fertile men. There was no significant correlation between the duration of vas obstruction and the above semen parameters. In 46.4% of vas fluids all spermatozoa were immotile and this condition was more common after 3 years of vasectomy. Immotile spermatozoa in the proximal vas fluids at the time of vasectomy reversal may be an important factor for predicting semen quality and fertilizing ability after vasovasostomy. There were no significant differences in the results of sperm-cervical mucus penetration test (CMPT) between spermatozoa fiom vasectomized and fertile men. Antisperm antibodies on the surface of spermatozoa from the vas of vasectomized men were determined by the immunobead test (IBT; 78.6% for IgG, 32.1% for IgA) and sperm cervical mucus contact test (SCMC, 36.4%). The presence of antisperm antibodies on the spermatozoa from the vas of vasectomized men may explain, in part, the lower pregnancy rate after vasovasostomy. These parameters of spermatozoa from the proximal vas of vasectomized men may closely reflect those in the cauda epididymis after vasectomy.     

''Water-test'': a simple method to assess sperm-membrane integrity

The functional integrity of the membrane of sperm from semen samples collected from 10 fertile men (Group A) and from 50 infertile men (Group B) was assessed by studying the swelling reaction of sperm when suspended in a medium of distilled water ('Water-test'). The results were correlated with routine semen-analysis and with the results of Eosin-Y staining. The mean values for the 'Water-test' were significantly different (P less than 0.01) between Groups A and B (84.8 +/- 3.5 versus 70.9 +/- 13.3, respectively). No significant correlations were observed in either Group A or B, between values for the 'Water-test' and values for the sperm count, the percentage of sperm with normal morphology and the percentage of motile sperm. There was a good correlation in both Group A (r = 0.86, P less than 0.01) and Group B (r = 0.92, P less than 0.01) between values for the 'Water-test' and those for the Eosin-Y test. These results indicate that the 'Water-test' is a simple and reliable test for evaluating sperm membrane integrity.     

Prediction of spontaneous conception based on semen parameters

Accurate prognosis of male fertility based on semen measurements is still not straightforward. This study was designed to identify the best predictors of fertility and to develop a multiple regression model predicting fertility using selected parameters of semen analysis. The predictive value of standard semen parameters and selected functional tests were studied in 113 fertile men and in 109 subfertile men whose spouses had a normal infertility workup. Individual semen parameters were evaluated using the receiver operating characteristic curve. Logistic regression based on linear functions of analysed sperm parameters was used to predict the chance of spontaneous conception. Logistic regression modelling revealed that the best prediction of spontaneous conception was obtained using 12 semen parameters: sperm concentration, total progressive motility (A + B), motility grade C or D, normal sperm morphology, defects of: head, acrosome, midpiece and tail, spontaneous acrosome reaction, hypo-osmotic swelling (HOS) test and acid aniline blue test. This mathematical model reached 90.3% accuracy in predicting in vivo conception and 90.8% for its lack. A satisfactory prediction of male fertility was also obtained using only four semen measurements: sperm concentration, total progressive motility (grade A + B), normal morphology, and HOS test; this model correctly identified as fertile 84.1% of those who conceived and identified as subfertile 88.1% of those who did not achieve pregnancy. In conclusion, basic semen analysis and selected functional tests of sperm provide important information regarding male fertility status.     

Testicular function in potential sperm donors: normal ranges and the effects of smoking and varicocele

Testicular exocrine (semen analysis) and endocrine (plasma LH, FSH, prolactin and testosterone) function was assessed in 119 consecutive healthy men presenting for screening as potential sperm donors. Since these volunteers were unbiased with respect to their fertility status, this sample of the general male population was suitable to determine normal ranges and the influence of a variety of physical (height, weight, standardized body weight, varicocele) and demographic (age, marital and fertility status, tobacco and alcohol consumption) factors on normal human testicular endocrine and exocrine function, without the confounding effects of bias in selection of subjects. The statistical distribution of all seminal parameters was non-gaussian, but cube-root transformation of the data normalized the distribution, allowing for parametric statistical analysis. The median (and 95% confidence limits) for the various semen parameters was 73.0 (10.6-235.3) million sperm per ml; 189.0 (12.6-868) million sperm per ejaculate; 50.4 (5.9-181.9) million motile sperm per ml; 133.0 (6.9-661.7) million motile sperm per ejaculate; 54.0 (7.0-172.9) million morphologically normal sperm per ml and 138.5 (7.5-672) million morphologically normal sperm per ejaculate. Testicular volume was correlated positively with measures of physique such as standardized body weight (r = 0.25, P less than 0.01) and body surface area (r = 0.30, P less than 0.002), and negatively with plasma levels of FSH (r = -0.31), P less than 0.002) but not LH. Sperm output was positively correlated with testicular volume (r = 0.28, P less than 0.005) and negatively correlated with plasma FSH (r = -0.31, P less than 0.002) and plasma LH (r = -0.31, P less than 0.002). Smoking was associated with a highly significant reduction in sperm output and motility. Men with varicocele (25%) were significantly taller, had slightly lower haemoglobin levels and moderate left (but not right) testicular atrophy, but neither seminal nor hormonal parameters were different from men without varicocele. There was no difference in any markers of human testicular function between men according to marital or fertility status, grades of moderate alcohol consumption or the presence of low titres of sperm antibodies.     

TEST-egg yolk buffer storage increases the capacity of human sperm to penetrate hamster eggs in vitro

A total of 85 semen samples from infertility clinic patients were examined to study the effect of storage at 4 degrees C in TES-Tris (TEST)-egg yolk buffer for 24 h on the penetrating capacity of sperm in the zona-free hamster egg penetration assay (HEPA). The mean sperm penetration rate and the fertilization index increased significantly after storage in TEST-egg yolk buffer. Only five out of the 85 samples (5.9%) failed to show any improvement in sperm penetration rate after cold storage. The sperm penetration rate before cold storage showed no significant correlations with routine semen characteristics, semen ATP concentration or the functional integrity of sperm membranes as measured by the hypo-osmotic swelling technique. Significant but low correlations were observed between sperm penetration rate after cold storage and the following semen parameters: sperm count, % motility, total number of motile sperm, % normal sperm morphology, total number of normal sperm, semen ATP concentration and sperm penetration rate before cold storage. Partial correlation analysis revealed that the positive correlation between semen ATP concentration and sperm penetration rate after cold storage was not a direct relationship but was due to the correlation with sperm count. The combination of sperm penetration rate before cold storage, sperm count and % normal sperm morphology accounted for 26.2% of the variation in sperm penetration rate after cold storage by stepwise multiple regression analysis, while sperm penetration rate before cold storage alone explained 13.5% of the variation. The results indicate that TEST-egg yolk buffer treatment can enhance sperm penetration rate in vitro and may be useful in the treatment of impaired sperm fertility.(ABSTRACT TRUNCATED AT 250 WORDS)     

Evaluation of computerized analysis of sperm movement characteristics and differential sperm tail swelling patterns in predicting human sperm in vitro fertilizing capacity

Multivariate discriminant analysis was used to evaluate the usefulness of computer image analysis of sperm movement characteristics and differential patterns of sperm tail swelling after hypoosmotic treatment for predicting the human sperm in vitro fertilizing capacity assessed by the zona-free hamster egg penetration assay. Fifty-five semen samples, mostly normospermic, from untreated infertility clinic patients were analyzed. The % normal sperm morphology, linearity of seminal sperm movement, seminal sperm head beat frequency, mean and maximum amplitudes of lateral head displacement, and hypoosmotic sperm tail swelling patterns c, d and f were selected by multivariate discriminant analysis to be capable of discriminating the samples exhibiting the presence or the absence of sperm in vitro fertilizing capacity. The % total sperm tail swelling did not give additional information about in vitro fertilizing capacity. These preliminary data suggest that computer image analysis of sperm movement characteristics and differential evaluation of hypoosmotic sperm tail swelling might be useful for the prediction of human sperm fertility. Further prospective studies are necessary to validate their predictive functions.     

Reduced penetration of zona-free hamster ova by cryopreserved human spermatozoa

The effect of cryopreservation on human sperm fertilizing potential was assessed by using the human sperm penetration into zona-free hamster ovum test. Semen samples from 12 fertile men were compared before and after cryopreservation for motility, sperm penetration rate, and number of sperm cells incorporated per oocyte. In fresh samples sperm concentration was 102 +/- 51 X 10(6) cells/ml, motility 66 +/- 14%, penetration rate 77.8 +/- 19%, and sperm incorporation 4.3 +/- 3.9 sperm per ovum. Frozen-thawed sperm cells showed a marked reduction of 61 +/- 21% (p less than 0.001) in motility. Penetration rate was reduced by 53 +/- 34% (p less than 0.01), and sperm incorporation dropped by 50 +/- 28% (p less than 0.05). Despite this substantial reduction in all three parameters, 75% of the samples maintained a penetration rate exceeding 14%, which is the lower limit for fertile semen. For the individual subject the decrease in sperm motility did not reflect actual fertility potential as expressed by its ability to penetrate zona-free hamster ova. These findings are related to morphological and biochemical changes in frozen-thawed semen and apparently are correlated with the decrease in pregnancy rates after cryopreservation. This test may be a valuable supplement to routine microscopic semen analysis for semen cryopreservation candidates.     

精浆游离L-肉毒碱水平及其与精子密度、活动率及活力的相关性研究

目的:研究正常生育及不育男性精浆中游离L-肉毒碱水平差异及其与精子密度、活动率(a+b+c级精子百分率)及活力(a+b级精子百分率)之间的相关性,探讨精浆中游离L-肉毒碱水平对男性生育力的影响及其在不育症检查和治疗中的作用。方法:分别采用高效液相色谱法和计算机辅助精液分析系统,测定了230例不育症患者(精子密度正常117例,少精子症81例,无精子症32例)和30例正常生育男性精浆中游离L-肉毒碱水平及精子密度、活动率、活力等参数。根据检查结果对不育症患者分组后,以SPSS12.0软件包进行统计学分析,比较各组间游离L-肉毒碱水平的差异以及游离L-肉毒碱水平与精子密度、活动率、活力之间的相关性。结果:正常生育组精浆游离L-肉毒碱水平明显高于不育组(P<0.01)。精液中精子密度越低、活力越弱,这种差异性越显著。相关性分析结果显示,精浆游离L-肉毒碱水平与精子密度呈显著正相关关系(r=0.521,P<0.01),与精子活动率和活力之间也具有正相关关系(r=0.319,P<0.01;r=0.251,P<0.01)。结论:精浆L-肉毒碱水平与精子密度、活动率和活力之间密切相关,其含量测定作为一项有用的生化指标,可为男性不育症检查及临床诊治和进行有关男性生殖功能机制研究提供参考。