人精浆抗精子抗体与顶体酶活性关系的探讨

目的 :研究精浆抗精子抗体 (AsAb)对精子顶体酶活力的影响。　方法 :用间接血凝法测定 34 32例不育男性和 6 5例生育男性精浆的AsAb ,并对其中的 2 882例不育男性精子用比色法进行了顶体酶活性的测定。　结果 :34 32例不育者精浆AsAb阳性率为 10 .2 0 %,2 882例进行了顶体酶活性测定的不育者精浆AsAb阳性率为 9.37%,对照组精子顶体酶活性明显高于各不育组 (P <0 .0 0 1)。不育组AsAb阳性组与AsAb阴性组比较顶体酶活性没有明显差异 (P >0 .0 5 ) ,顶体酶活性正常与异常组AsAb阳性率比较差异无显著性 (P >0 .0 5 )。　结论 :精浆AsAb对精子顶体酶活性没有影响。     

抗精子抗体对精子顶体酶活性的影响

本文旨在观察抗精子抗体对精子顶体酶活性的影响。选择男性不育者50例与正常生育者20例，采用固相酶染色法测抗精子抗体，以固定明胶薄膜法测精子顶体酶活性。结果发现50例不育者抗精子抗体阳性率为52％。不育者精子顶体酶活性明显低于生育者；抗精子抗体阳性者须体酶活性明显低于阴性者，表明抗精子抗体可降低精子顶体酶活性。     

Problems in evaluating male fertility: valuable factors in evaluating male fertility and normal values of seminal parameters

To determine the valuable factor for evaluating male fertility, a comparative study was done as to various seminal parameters between fertile and infertile groups. The fertile group consists of 57 proven fertile males and the infertile group consists of randomly chosen 67 infertile patients. Seminal parameters assessed were sperm concentration, motility, mean velocity, total sperm output, total motile sperm output, sperm morphology, acrosin activity and sperm penetration rate on zona-free hamster egg penetration assay (SPA). The infertile group was significantly different from the fertile group in every parameter except acrosin activity. However, the range of each parameter in the two groups overlapped each other. The diagnostic rate of each parameter, which is the percentage of an infertile male correctly diagnosed as infertile, was calculated by using 95% specificity threshold value of fertile males. The 95% specificity threshold values of sperm concentration, motility and % normal shaped sperm were 24.9 x 10(6)/ml, 34.9% and 55%, respectively, and they could be acceptable for the normal limit of seminal parameters. The diagnostic rate was highest in penetration rate (72.4%). In other words, penetration rate is the most valuable factor in various parameters for making a distinction between fertile and infertile males. Sperm motility and mean velocity showed the next highest diagnostic rate. On the other hand, sperm concentration showed a poor diagnostic rate (36.8%). In addition, there was no significant correlation between penetration rate and any other seminal parameters. These results suggest that the SPA will be an essential test for evaluating male fertility and penetration rate may be a marker of male fertility in the treatment of male infertility.     

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.     

Biochemical and Genetic Investigation of Round-headed Spermatozoa in Infertile Men including two Brothers and their Father

Acrosin and the outer acrosomal membrane (OAM) were studied in the spermatozoa of 9 infertile patients who differed in the number of round-headed spermatozoa between 14 and 71% in their ejaculates. These sperm components were also investigated in two infertile brothers who exhibited exclusively round-headed spermatozoa in their ejaculates, and in their fertile father. It turned out that round-headed spermatozoa lack both acrosin and the OAM as studied by indirect immunofluorescent and immunoperoxidase staining technique, gelatinolysis tests and by acrosin activity measurements. The normally shaped spermatozoa of 6 of the 9 infertile patients were found to be positive for acrosin and the OAM as expected, but in the remaining three patients even these spermatozoa were abnormal; in one patient they were unstainable for acrosin and in two patients they were unstainable both for acrosin and the OAM. These results have been confirmed by studies with the gelatinolysis test. The father of the two brothers with exclusively acrosomeless spermatozoa had more than 94% of normally shaped spermatozoa. However, only 10% of these spermatozoa were acrosin positive and only 30% were positive for the OAM. On the basis of these results we postulate that the mode of inheritance of the round-headed spermatozoa syndrome is polygenic rather than monogenic as suggested by previous authors.     

Sperm chromatin heterogeneity as an infertility factor

Semen samples from husbands with a history of unexplained infertility (n = 33), of women with habitual abortion (n = 36), or normal fertile donors (n = 20) were subjected to conventional semen analysis (SA), Acridine orange test (AOT), and zona-free hamster egg penetration test (HEPT). The three tests operate independently. The most discriminatory test was AOT (p = 0.0001) followed by HEPT (p = 0.019). The frequency of sperm chromatin heterogeneity as detected by AOT red fluorescence was highest in habitual abortion (39.4%), followed by unexplained infertility (16.4%), and, last, donors (9.4%). However the percentage of penetration was highest in habitual abortion (50.7%), followed by donors (43.1%), and least in unexplained infertility (33.9%). Conventional semen parameters (sperm density, motility, abnormality, and vitality) were the least to discriminate between the three groups. The presence of abnormal sperm chromatin may lead to infertility as a result of early pregnancy loss.     

217例男性不育症精子顶体酶检测与分析

目的　研究人精子顶体酶与男性不育之间的关系。　方法　 2 1 7例不育男性 ,年龄 2 4～ 43岁 ,根据精液常规检查结果分为不育 A、B两组 ;46例生育男性为对照组。手淫留取精液标本 ,常规洗涤后孵育 ,采用固定明胶底膜法检测精子顶体酶。　结果　正常生育组的精子顶体酶活性阳性率为 86.5% ,酶活性亮区直径为 42 .9μm。不育 A组酶活性阳性率为 57.0 % ,酶活性亮区直径为 3 8.5μm。不育 B组酶活性阳性率为3 5.2 % ,酶活性亮区直径为 2 7.5μm。经统计学处理 ,生育组与不育组之间精子顶体酶阳性率差异有显著性 ( P<0 .0 1 )。　结论　顶体酶阳性率和活力低下是男性不育的一个重要原因 ,精子顶体酶是受精过程中的关键酶     

Contribution of the male factor to unexplained infertility: a review

The more exhaustive the evaluation of couples with unexplained infertility, the more likely is the opportunity for detecting the aetiological factor responsible for infertility. Transport of spermatozoa through the upper genital tract and their ability to fertilize the oocyte are two obscure areas for the conventional evaluation of infertility. Although research in the former area is limited, there is indirect evidence that impaired sperm transport could be one of the causes of infertility in some couples with otherwise unexplained infertility. On the other hand, the availability of sperm function tests and the correlation of their results with in-vitro fertilization rates have allowed the detection of a previously hidden male factor in couples with unexplained infertility. It has been demonstrated that couples suffering unexplained infertility have significantly lower in-vitro fertilization rates in comparison with patients with tubal problems. These results can be explained because several case control studies in patients with unexplained infertility have reported defects in capacitation and sperm motion characteristics, binding of the spermatozoa to the zona pellucida, acrosome reaction, acrosin activity of the spermatozoa, and the ability of the spermatozoa to penetrate zona-free hamster cocytes. These observations suggest that methods for assessing the fertilizing capacity of the spermatozoa have to be incorporated in the evaluation of couples with unexplained infertility in order to amplify the scope of the workup and to better decide the appropiate treatment for these couples.     

人精子顶体反应的检测及其与穿卵试验的相关性

选择正常生育男子（生育组）、输精管结扎再吻合（吻合组）、不育症患者（不育组）各２５例的精液进行人精子顶体蛋白酶及酸性磷酸酶测定,顶体组织化学三色染色和人精子穿透去透明带金黄仓鼠卵试验,比较不同生育条件下精子功能和生育能力的相关性。结果显示,精子获能后顶体反应率明显高于获能前（Ｐ＜０．０１）。生育组顶体反应率和穿透试验高于吻合组和不育组（Ｐ＜０．０１）。结果表明,精子穿卵试验结合精子顶体反应测定可作为客观评价人类精子受精能力的依据     

Influence of the Concentration of Motile Sperm Inseminated on the Ovum Penetration Assay Results: Toward a Standardized Method

The influence of the concentration of motile sperm inseminated on the zona-free hamster egg penetration rate has been studied. In 3 fertile men, the higher the concentration of motile sperm inseminated, the higher the ovum penetration rate. This correlation emphasizes the usefulness of standardizing the technical conditions under which the ovum penetration assay is performed. It is also suggested to refer to a standard curve for results analysis. In the 22 patients studied, the outcome of the heterologous ovum assay was in most cases related to the initial sperm parameters. However, some disagreement was also observed. In the absence of a correlation between the initial sperm parameters and the ovum penetration rates, further investigation is needed in order to either more precisely determine the aetiology of unexplained couple infertility, or, in case of intrauterine insemination or in vitro fertilization, to better understand the conditions under which spermatozoa are capacitated in vitro.     

Boar Proacrosin Correlation between Total Proacrosin Content and Sperm Fertilizing Capacity

A method of acidic extraction at pH-1.3 was used for quantitative estimation of the proacrosin and acrosin content in boar spermatozoa after various times of storage in four different sperm dilutors. The proacrosin and acrosin contents were correlated with the results of biological tests for sperm penetration in zona-free hamster eggs and sperm survival. The results have shown that proacrosin quantitation should be a convenient biochemical test of sperm fertilizing capacity.     

Assessment of sperm functions in infertile patients with varicoceles

Acrosin activity, acrosome reaction and nuclear chromatin condensation were studied in 24 infertile patients with varicoceles and 26 fertile men with or without varicocele. Chromatin condensation, assessed by aniline blue staining, and acrosin activity, evaluated by gelatinolysis technique, were significantly affected in the group of infertile patients. Defective chromatin condensation and defective acrosin activity were detected in 67% and 50% of the infertile patients, respectively. No significant difference was found between the two groups regarding the acrosome reaction, which was assessed by the triple staining technique after exposure of spermatozoa to low temperature (4 degrees C). This study identified a subgroup of infertile patients with normal standard semen parameters but impaired sperm functions. Results of the sperm function tests and standard semen parameters were not correlated. Therefore, it is concluded from this study that important sperm functions are impaired in patients with varicocele and that the gelatinolysis technique and aniline blue staining are effective tools for assessment of the fertilization potential of varicocele patients.     

A prospective study of the relationship between semen quality and fertility in cases of unexplained infertility

The male partners of 68 couples exhibiting 5.1 +/- 0.3 (SEM) years of unexplained infertility were assessed using the conventional criteria of semen quality, the movement characteristics of the spermatozoa and the outcome of the zona-free hamster egg penetration test. After a follow-up period of 2.3 +/- 0.06 (SEM) years, 25 (37%) of these patients were found to have initiated a pregnancy, thereby permitting an analysis of those aspects of semen quality which most accurately predicted their subsequent fertility. A multivariate discriminant analysis revealed that the conventional semen profile, per se, was not of significant value in discriminating the incidence of pregnancy. However, significant discrimination (P = 0.0173) was obtained when the postcapacitation movement characteristics of the spermatozoa were incorporated into the analysis. The accuracy of this prognosis was further increased if either the duration of infertility or the outcome of the zona-free hamster egg penetration test was taken into consideration. Overall classifications of fertility were then 76.3% and 76.5% accurate, respectively. These results suggest that in vitro assessments of human sperm function are of significant value in evaluating male fertility.     

Zona-free hamster egg-sperm penetration assay: II. Correlative study with routine semen analysis

This work studied the effect of variations in the different parameters of routine semen analysis on the penetration rate of the spermatozoa into the zona-free hamster eggs, using semen samples from 21 fertile donors, 57 infertile patients with normal spermiograms, 63 infertile patients with subnormal spermiograms, and 19 frozen semen samples. The results of the hamster test do not correlate with the percentage of motile spermatozoa, the percentage of live spermatozoa, or the percentage of spermatozoa with normal morphology in the semen samples. A positive correlation was found between the results of the hamster test and three other parameters of the semen analysis: the sperm concentration, the progressive type of motility, and a pH range of 7.2-7.7, but these positive correlations were statistically insignificant.     

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.     

Adenosine 5''-Triphosphate (ATP) Concentration and Acrosin Activity in Human Spermatozoa. Their Relation with the Sperm Penetration Assay (SPA)

Fertility of men depends on the quality of semen. The aim of the present paper is to determine both the acrosin activity by radioimmunoassay and ATP concentration by bioluminescence in human spermatozoa, and evaluate these results in those samples with normal or low sperm penetration according to SPA test. Ejaculates obtained from 42 untreated men, were studied one hour after the obtention. These materials were divided into two groups:20 human semen samples with "in vitro" potentiality to penetrate zone-free hamster ova, between 15% to 98% and 22 human semen with SPA test between 0% to 14%. When we compare the group with normal penetration response vs that group with low or absent penetration ones, a significant decrease of ATP and acrosin concentrations was observed (P less than 0.001). Nevertheless no significant difference was observed in relation with percentage of motility, volume (ml), sperm concentration (10(6)/ml), percent of quick progressive spermatozoa and number of gametes capable of migrating into the medium layer (10(6)), between the group with low or absent penetration test against that one with normal zona-free hamster egg test.     

Diagnostic value of sperm function tests and routine semen analyses in fertile and infertile men

The results of routine semen analyses, the zona-free hamster oocyte penetration test, the hypoosmotic swelling test, and semen adenosine triphosphate levels were studied in 66 fertile and 130 infertile men. Multivariate discriminant analysis demonstrated that routine semen parameters including semen volume, sperm count, percent sperm motility, and percent normal spermatozoa in combination could predict the fertility of these patients with 70.4% accuracy. Of the three sperm function tests evaluated, the zona-free hamster oocyte penetration test and the hypoosmotic swelling test were selected by the multivariate discriminant analysis as variables capable of providing significant information on the fertility status of the patients. However, the addition of the results of these two tests to the routine semen analysis did not significantly improve the predictability of fertility. The overall correct prediction rate was 77.6% after incorporation of the results of these two sperm function tests. In this group of subjects, the presently available sperm function tests did not predict the fertility status of a patient with a high degree of accuracy.     

Effects of white blood cells on the in vitro penetration of zona-free hamster eggs by human spermatozoa

The presence of white blood cells in semen has been associated with male infertility. Previous studies indicate that pyospermia occurs in conjunction with decreases in sperm motility, number of normal sperm forms, and penetration rates in the zona-free hamster egg sperm penetration assay. We have evaluated the relationship of seminal white blood cells and sperm function, as reflected in the zona-free hamster egg penetration assay, and have investigated the possible mode of action of the white cells. Egg penetration rates decreased when white blood cells from fertile or potentially fertile donors were added to their sperm suspensions prior to preincubation and at insemination in the in vitro assay. Zona-free hamster egg penetration assay results were also inhibited when the supernatant from white blood cells incubated in Biggers, Whitten, and Whittingham (BWW) medium overnight were introduced to sperm-oocyte suspensions at insemination. Conversely, egg penetration rates were enhanced in samples from hypofertile individuals when white blood cell concentrations in the semen or WBC/sperm ratios were reduced, either by physical removal or as a result of antibiotic therapy. The physical presence of leukocytes, and possibly, the extracellular release of lysosomal enzymes may be responsible for the inhibitory effects in vitro. Although the mechanism(s) by which white blood cells interfere with the fertilizing capacity of spermatozoa are not clear, it is quite obvious that their presence in the in vitro environment is undesirable and can mask an individual's actual fertilizing potential.     

Acrosin activity in patients with idiopathic infertility

Acrosin is a sperm acrosomal enzyme that is involved in the acrosome reaction, the binding of spermatozoa to the zona pellucida, and fertilization. This study was designed to determine whether sperm acrosin measurements can identify subpopulations of infertile or subfertile patients that are not recognized by routine semen analyses. We measured the total acrosin activity of ejaculates in a group of 19 men (15 suspected subfertile patients and 4 fertile donors). The acrosin activity was measured in liquefied semen specimens using the methodology described by Kennedy et al. [1989) J Urol 10:221-231). Ten patients in the suspected subfertile group had a mean acrosin value of 7.8 microIU acrosin/million sperm, which is clearly in the abnormal range (less than 14 microIU/10(6) sperm). Three patients had a mean acrosin value of 20.1 microIU/10(6) sperm, which is in the indeterminate range. Two other patients and four proven fertile donors had acrosin values in the normal range (greater than 25 microIU/10(6) sperm) (Agarwal A, Loughlin KR (1990): 2d International Meeting of Andrology, Como, Italy; Abstr 22). The normal fertile controls had a mean acrosin value of 32.5 microIU/10(6) sperm.     

Penetration of sperm from teratospermic men into zona-free hamster eggs

The ability of sperm samples from male partners of infertile couples, with isolated teratospermia (ITS), to penetrate zona-free hamster eggs was examined. The ITS patients had a significantly lower proportion of normal spermatozoa than did a control group of men of proven fertility (23.6% and 47.8%, respectively; P less than 0.001), while the mean total sperm count and the % motility did not differ between the two groups. The mean hamster egg penetration rate of sperm from ITS patients was 2.64% compared to 31.1% in the control group. A significant negative correlation was found in the ITS group between the proportion of a) abnormal forms and the ability to penetrate zona-free hamster eggs, and b) pyriform-shaped sperm and the penetration rate. Of the sperm parameters which were examined, only the morphology could be correlated with the rate of penetration.