人类精子低渗试验超微结构分析

目的：观察低渗试验精子的超微结构，以探讨低渗试验的机制。方法：采用透射电镜观察分析两种不同渗量低渗试验中精子的超微结构变化。结果：活精子头部顶体膜间隙增宽，界限清晰部分明显肿胀破裂。“ｇ”型精子尾部向４条周致密纤维一侧卷曲，中段线粒体鞘损坏，尾部卷曲试验精子尾肿胀明显，部分外周致密纤维扭曲移位。     

不同浓度低渗肿胀试验结合伊红染色对精子质量的评估

本文应用不同浓度精子低渗肿胀试验九红染色法，对８例生育男性和１０例不育男性的精子膜功能进行评估。结果表明：１、活精子尾部低渗肿胀率均达到９０％以上，生育组与不育组无显著性差异；２、在１５０ｍＯｓｍ低渗溶液的稀释倍数为３－７倍之间，精子头部膜的损害程度在生育组与不育组之间存在显著性差异；３、在不同浓度的低渗溶液中，精子头部膜与精子尾部膜对低渗溶液的顺应性并不一致。实验证明不同浓度低渗肿胀试验结合伊红     

不同浓度低渗肿胀试验结合伊红染色对精子质量的评估

本文应用不同浓度精子低渗肿胀试验结合伊红染色法，对８例生育男性和１０例不育男性的精子膜功能进行评估。结果表明：１．活精子尾部低渗肿胀率均达到９０％以上，生育组与不育组无显著性差异；２．在１５０ｍＯｓｍ低渗溶液的稀释倍数为３～７倍之间，精子头部膜的损害程度在生育组与不育组之间存在显著性差异；３．在不同浓度的低渗溶液中，精子头部膜与精子尾部膜对低渗溶液的顺应性并不一致。实验证明不同浓度低渗肿胀试验结合伊红染色可正确评价精子膜的整体功能。     

精子尾部卷曲试验与精子低渗肿胀试验的共性与特性

精子尾部卷曲试验与精子低渗肿胀试验的共性与特性施云飞，汤文葳，茅拥军关键词精子，低渗肿胀试验，男性不育中图法分类号Ｒ３２１．１自１９８４年Ｊｅｙｅｎｄｒａｎ等［１］报告采用精子尾部低渗肿胀试验评估精子膜完整性及授精能力以来，ＷＨＯ于１９９２年推荐了该...     

精子尾部低渗肿胀试验在评价男子生育方面的应用

本文应用精子尾部低渗肿胀试验和常规精液分析,对120例有生育力男子和693例不育男子进行了包括精子膜功能在内的综合精液质量评定。结果发现:生育组男子精子尾部低渗肿胀率明显高于不育组男子的精子肿胀率(P<0.01)。同时还发现生育组精子尾部低渗肿胀率与常规精液分析中精子活动率、精子存活率、正常形态精子百分率间呈显著正相关(r=0.3164,0.5306,0.3034,P<0.0005,<0.0005,<0.005)。实验证明,精子低渗尾部肿胀试验是一种有价值的评价精子功能的方法。     

正常男性附睾中精子功能的初步研究

目的　检测正常成年男性附睾中不同部位的精子指标 ,为男性附睾精子功能的研究提供实验资料。方法　将附睾切断分成附睾头、附睾尾和附睾体 ,分别纵向剖开 ,置于含有 10 %小牛血清的F10 溶液中 ,37℃孵育 30min ,孵育液作精子常规分析、低渗肿胀试验 (HOS)、精子 -宫颈粘液穿透试验。结果　附睾不同部位精子的形态和功能存在明显差异 ,尾部和体部精子的活率、活动力、HOS试验和CM穿透试验结果明显高于头部精子 ,差异有显著性 (P <0 .0 5或 0 .0 1) ,以尾部精子的各项指标最高 ,与正常排出体外的精子无显著差异 (P >0 .0 5 )。头部附睾未成熟精子、畸形精子比例明显高于尾部和体部附睾的精子 (P <0 .0 1和 0 .0 5 )。结论　附睾精子可应用于辅助生育技术 ,作为治疗梗阻性无精子症的最好选择。     

精子尾部卷曲试验影响因素的探讨

本文介绍了精子尾部卷曲试验的影响因素。３０例正常生育男性和４３例不育男性精子尾部郑曲与精子活率具有高度的相关性，与精子密度无相关性，生育与不育男性之间精子尾部卷曲率具有显著性差异。     

人类精子低渗试验指数分析

目的：探讨精子低渗试验指数（ＳＨＯＴＩ）对评估男性生育力的临床价值。 方法：采用两种不同渗量的低渗液结合活体染色技术，对１２９例男性精液进行精子低渗试验。 结果：生育组与不育组比较精子卷尾率、ｇ型肿胀精子率和精子低渗试验指数差异均显著（Ｐ〈０．０１），生育组ＳＨＯＴＩ分布范围为：０．０９６￣０．３３６；不育组ＳＨＯＴＩ分布范围为０．０２５￣０．２１５。 结论：ＳＨＯＴＩ综合反映了精子的整体膜功能及     

局部振动对雄性免生殖功能的影响

目的通过对雄性新西兰兔精液中精子质量及受精能力的检测，观察睾丸大体形态及精子细胞超微结构的改变，探讨局部振动对雄性兔的生殖功能的影响。方法将新西兰兔随机分为A组（接振强度为3．02m／s^2）、B组（接振强度为6．13m／s^2）、C组（接振强度为12．26m／s^2）和1个对照组。于接振后30d测定精浆中精子的密度、活动力、活动率、畸形率，通过去透明带仓鼠卵．精子穿透试验和精子低渗膨胀试验评价精子的受精能力，光镜下观察睾丸组织学变化，电镜下观察睾丸精子细胞超微结构的改变。结果接振试验后与对照组相比较，A、B、C3个试验组新西兰兔精液精子活动率、活动力明显降低（P〈0．01）；精子的畸形率明显升高（P〈0．05）。精子的受精率及尾部肿胀的精子比率明显降低（P〈0．05）。睾丸生精细胞数量减少，精子细胞的部分细胞器破坏、裂解、坏死。结论后肢接振试验可降低精子的活动力、活动率及卵子受精率及低渗肿胀率，增加精子的畸形率，睾丸组织中生精细胞减少，精子细胞的部分细胞器破坏、裂解、坏死，从而抑制精子的成熟，影响生殖功能。     

用有效活精指数评价精液质量

同时应用有效活精指数及精子尾部低渗肿胀试验、精子前向运动级别、毛细管穿透试验，精子顶体染色，对９０例正常男性和１２９例不育男性精液进行检测。结果显示：有效活精指数与精子总肿胀率（Ｔ），ｇ形精子率（Ｇ），前后运动级别（Ｐ），穿透力评分（Ｓ），顶体完整率（Ａ）之间，生育组呈显著正相关，不育组呈非常显著正相关。以上各项试验参数在两组间均有非常显著差异。不育男性结果表明，有效活精指数与Ｔ、Ｇ、Ｐ、Ａ的异常     

Review on the screening of semen by hypo‐osmotic swelling test

The hypo‐osmotic swelling test (HOST) is widely used as a valuable test for determining sperm quality by evaluating the membrane integrity of spermatozoa of various domestic animals including cattle, horses and swine. The HOST has also been used as an indicator of the fertilising capacity of spermatozoa. This test is based on the swelling ability when functional spermatozoa submitted to hypo‐osmotic solutions. This test is commonly used as an important parameter for the evaluation of semen due to its strong correlation with semen evaluation parameters. The objective of this review was to analyse its significance in semen evaluation, swelling of spermatozoa under various osmolarities and variations in swelling percentage under different seasons.     

In situ viability detection assays induce heat‐shock protein 70 expression in spermatozoa without affecting the chromatin integrity

To differentiate dead spermatozoa from viable but immotile spermatozoa, several techniques are being used during ICSI. As processed spermatozoa from poor‐quality ejaculate are confronted with a higher risk of experiencing stress on exposure to altered osmotic conditions or chemicals, this study was undertaken to determine the expression of stress response gene Hsp70 and chromatin integrity in spermatozoa subjected to in situ viability assays such as hypo‐osmotic swelling (HOS) test, modified hypo‐osmotic swelling (M‐HOS) test and pentoxifylline in 25 fresh and frozen–thawed asthenozoospermic ejaculates. RT‐PCR and immunofluorescence detection of Hsp70 were performed to elucidate the expression and localisation of Hsp70 in spermatozoa, whereas DNA fragmentation analysis was performed by sperm chromatin dispersion assay. Exposure of fresh and frozen–thawed asthenozoospermic spermatozoa to M‐HOS and pentoxifylline significantly increased Hsp70 expression as evidenced by increased RNA expression and immunolocalisation of Hsp70 protein in sperm head (P < 0.05–0.001). However, chromatin integrity was not significantly affected in any groups until 6 h of post‐exposure time period. Our results suggest that conventional HOS may be preferred for the in situ detection of the viability as there was no immediate stress response and chromatin instability in the exposed spermatozoa.     

Hypo‐osmotic test in cat spermatozoa

The hypo‐osmotic (HOS) test has been used in other species as an indicator of the fertilising capacity of spermatozoa. The aims of this study were to assess the response of domestic cat spermatozoa to the hypo‐osmotic test, to determine the type of solution, concentration and time of incubation needed to obtain a maximum percentage of swelling, to correlate the selected combination with the percentages of progressive motility and to evaluate whether dilution of the ejaculate alters the results. Incubation for 30 and 45 min in solutions of fructose and of citrate of 50 and 100 mOsmol kg^?1 was evaluated. The highest percentage of swelling was obtained using the 50 mOsmol kg^?1 solution, and no significant differences were observed between the times of exposure to the solutions. A positive correlation was observed between the percentage of individual progressive motility and the percentage of sperm swelling in a 50 mOsmol kg^?1 fructose solution, with no significant differences being observed between raw and diluted semen samples. The results of this study suggest that the HOS test could be useful for evaluating membrane function in domestic cat spermatozoa, both in raw semen and in samples diluted in the EZ Mixin^® commercial extender, and thus could be incorporated into routine semen evaluation protocols.     

Normal live birth after vitrified/warmed oocytes intracytoplasmic sperm injection with immotile spermatozoa in a patient with Kartagener's syndrome

The present article is a report on two cases of male Kartagener's syndrome enrolled in intraconjugal IVF programme due to akinetospermia. Viable spermatozoa were selected using a hypo‐osmotic swelling test (HOST) and pentoxifylline activation and subsequently microinjected into vitrified/warmed oocytes. The treatment enabled one of these two couples to achieve a pregnancy and to give birth to a healthy baby girl.     

香烟烟雾对大鼠精子核DNA成熟度和精子膜功能的影响

目的 :探讨吸烟对精子核DNA成熟度和精子膜功能的影响。　方法 :自制吸烟机将大鼠制成被动吸烟模型 ,取其精子应用精子核吖啶橙荧光染色法和精子低渗肿胀试验 ,观察吸烟大鼠精子核DNA成熟度和精子膜功能。结果 :大鼠被动吸烟 8周后其双链DNA精子百分率为 (5 2 0± 5 6 ) % ,明显低于相应的对照组 [(6 8 5± 3 3) % ](P <0 0 1) ;精子低渗肿胀率 [(2 0 7± 4 3) % ]也较正常对照组 [(5 2 5± 3 8) % ],明显降低 (P <0 0 1)。　结论 :吸烟可致大鼠精子核DNA成熟度降低并使大鼠精子膜更脆弱 ,降低大鼠精子受精能力。     

Sperm plasma membrane integrity measurement: a combined method

Sperm plasma membrane characteristics were measured by a combined method consisting of the hypo-osmotic swelling test and staining with either the eosine Y (HOS-eosine test) or propidium iodide dye (HOS-propidium test). Sperm samples were washed and resuspended in BWW medium (fraction I). Aliquots of the washed spermatozoa were treated by a swim-up technique to select motile spermatozoa (fraction II). After separation of motile cells, residual sperm pellets were treated separately (fraction III). These three fractions were subjected to the hypo-osmotic swelling test, lipid peroxidation measurement, and the HOS-eosine and HOS-propidium tests. The HOS-eosine test makes it possible to distinguish 4 types of spermatozoa: type 1: HOS+/eosine-; type 2: HOS-/eosine-; type 3: HOS-/eosine+ and type 4: HOS+/eosine+ (Fig. 1). HOS-propidium test shows equal results as HOS-eosine test. Fraction I spermatozoa showed 55.2 +/- 3.6% type 1; 12.6 +/- 1.0 type 2; 28.0 +/- 2.9 type 3; and 4.2 +/- 0.6 type 4 cells. Fraction II spermatozoa were characterized by high percentages of type 1 cells, low percentages of types 3 and 4, and very low values of lipid peroxidation (5 times smaller than fraction I). Fraction III showed a low percentage of type 1, a high percentages of the other types, and an enhanced value of lipid peroxidation (2 times higher than fraction I). The prognostic value of the HOS-eosine test was evaluated in an IVF programme. Preliminary results show that a high incidence of types 2 and 4 spermatozoa is often associated with fertilization failure.     

人类精子低渗试验指数分析

目的：探讨精子低渗试验指数（ＳＨＯＴＩ）对评估男性生育力的临床价值。方法：采用两种不同渗量的低渗液结合活体染色技术，对１２９例男性精液进行精子低渗试验。结果：生育组与不育组比较精子卷尾率、ｇ型肿胀精子率和精子低渗试验指数差异均显著（Ｐ＜０．０１），生育组ＳＨＯＴＩ分布范围为：０．０９６～０．３３６；不育组ＳＨＯＴＩ分布范围为０．０２５～０．２１５。结论：ＳＨＯＴＩ综合反映了精子的整体膜功能及受精能力，可作为评估男性生育力的一个新的指标。初步确定临床参考值为ＳＨＯＴＩ＜０．０９６，提示精子功能不良，不能生育；在０．０９６～０．２１５之间精子功能部分正常，有机会生育；在０．２１５～０．３４０之间预示精子功能正常，生育力良好。     

The effect of colloid osmotic pressure in human spermatozoa exposed to hypoosmotic conditions

The use of a protein source such as serum and albumin had been extensively employed as supplements of culture media for handling and culture of gametes and embryos. Protein molecules behave as colloids in solution and contribute to the osmotic pressure of fluids. The interaction of proteins in solution and spermatozoa needs to be assessed in order to determine their possible role in osmoregulation. The aim of this study was to assess possible osmoregulatory mechanisms of protein supplementation against exposure to hypoosmotic conditions by assessing the sperm's response to those environments. A stock hypoosmotic solution (HOS) was prepared by using a mixture of fructose and sodium citrate and adjusted to an osmotic pressure of 150 mOsm l-1. Another stock solution was prepared by diluting a preparation of synthetic serum supplement [SSS; 6% (v/v) total protein] with distilled water to obtain an osmotic pressure of 150 mOsm l-1 (hypoosmotic SSS or H-SSS). Three additional solutions were prepared by mixing the stock HOS and H-SSS solutions in the following proportions (v/v): (i) 75% H-SSS/25% HOS, (ii) 50% H-SSS/50% HOS and (iii) 25% H-SSS/75% HOS. Aliquots of washed spermatozoa from 18 men were diluted 1 : 10 (v/v) with each of the testing solutions and incubated for 60 min. Specimens were assessed on wet mounts for total and specific swelling patterns. Swelling patterns were classified as maximal (>50% tail length swollen) and minimal (<50% tail length swollen) swelling with or without associated sperm motility. The major finding of this study was that increasing the concentration of protein supplementation resulted in a decrease in the proportion of maximal sperm tail swelling patterns and an increase in the proportion of minimal tail swelling patterns. A proportion of spermatozoa which exhibited minimal swelling patterns were still motile in all solutions tested, and the percentage of those spermatozoa increased as the protein supplementation was also increased in the testing solutions. Incorporation of protein supplementation as described in this study delays the effect of sperm swelling in hypoosmotic conditions.     

Intracytoplasmic sperm injection with motile and immotile frozen-thawed testicular spermatozoa (the Hungarian experience)

The authors summarize their experience in 75 in vitro fertilization cycles, where frozen-thawed testicular spermatozoa were used for intracytoplasmic sperm injection. In 32 cases, motile spermatozoa could be observed in the frozen-thawed sample. In 34 cases, motility could be induced by pentoxifylline and in nine cases immotile spermatozoa, selected with hypoosmotic swelling test, were used for fertilization. The fertilization rates obtained with motile and immotile spermatozoa (66.1% versus 52.3%) were not significantly different. Our data demonstrate that freezing of testicular spermatozoa opened new possibilities for the treatment of azoospermic men. The clinical pregnancy rate per embryo transfer (ET) (21.87%) was comparable with previous results use of fresh testicular spermatozoa (27.7%). The quality and number of transferred embryos had the most significant impact on the pregnancy rate. The fertilization rate and frequency distribution of good-quality embryos were lower in the case of immotile spermatozoa, and pregnancies were only achieved when motile spermatozoa had been used.     

Association of foreign DNA with porcine spermatozoa.

The aim of this project was to establish if DNA molecules bind to porcine spermatozoa after incubation in vitro. In an initial experiment, the DNA was mixed with porcine spermatozoa to test for the presence of DNases. From bacterial transformation and gel electrophoresis studies, it was established that the presence of DNases within the sperm or in the sperm suspension was negligible. By the addition of radiolabeled DNA, spanning a large size range, it was demonstrated that the DNA molecules interacted with the sperm during a 15 to 20 min period of incubation. These samples were centrifuged and washed extensively to confirm that the DNA was closely associated with the sperm. Under the experimental conditions used, it was calculated that approximately 3.8 x 10(2) DNA molecules were associated with each spermatozoon. Percoll gradient experiments, which allowed differentiation between motile and nonmotile sperm, showed that motile sperm were more efficient at capturing DNA molecules than nonmotile sperm. In situ hybridization studies revealed that approximately 30% of the motile sperm carried the DNA on their surface, and the associated DNA was attached only to the sperm head with no attachment to the sperm tails. Heterologous foreign DNA can become closely associated in a nonrandom manner with porcine spermatozoa.