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.     

Analysis of spermatozoa from the proximal vas deferens of fertile men

Fluids from the left and right proximal vas deferens were collected from 105 normal fertile men by cannulating the vas deferens during vasectomy, and sperm parameters analysed. Sperm motility (73.1 k 13.3Y0), normal sperm morphology (75.2 k 11.1"/o), sperm viability (72.7 k 18.8%) and the hypo-osmotic swelling test (73.3 k 19.2%) were in the normal range, compared with that of ejaculated spermatozoa. However, sperm Concentration in the proximal vas deferens (6274.6 k 5103.8 × 10" ml^-' was higher than that in semen. Sperm concentration in the right vas deferens was significantly higher (P<0.05) than that in the left and the percentage of spermatozoa showing abnormal cervical mucus penetration was Significantly higher (47%) for the left than for the right (18%). There were no anti-sperm antibodies on the surface of spermatozoa from the vas deferens as determined by the sperm cervical mucus contact test and immuno bead test. These parameters of spermatozoa from the proximal vas may reflect those of spermatozoa from the human cauda epididymis.     

Sites of residual spermatozoa after irrigation of the distal vas deferens using normal saline solution during vasectomy in a rat model

We have previously reported that irrigation of the distal vas deferens using a normal saline solution (NSS) is successful in removing a large amount of spermatozoa from the tract. However, this technique does not completely remove all the motile spermatozoa from the ejaculate. The aim of the present study is to evaluate the location of the residual spermatozoa after distally irrigating the vas deferens. Twenty male Sprague-Dawley rats (400-450 g) constitute our study population. The animals were divided into two groups: group 1, control group (n = 10), rats that undergo only vasectomy and group 2, experimented group (n = 10), rats that undergo vasectomy and distal irrigation of the vas deferens using 3 mL of NSS. In both groups, the middle and terminal parts of the vas deferens including the seminal vesicles are removed and sent for spermatozoa count. The post-vasectomy urine samples containing spermatozoa are obtained by mid-ventral cystocentesis and the concentration is determined using a haemocytometer. More spermatozoa was found in the urine samples of the experimented group than the control (21.3 +/- 10.61 vs. 0.2 +/- 0.20 million/ml, p-value = 0.068), and lesser residual sperms reside at both the middle and the terminal parts of the vas deferens (0.5 +/- 0.31 vs. 3.0 +/- 0.00; p = 0.008 and 1.1 +/- 0.99 vs. 2.0 +/- 0.00; p = 0.036 respectively). No sperms were present in the seminal vesicles of the control group, but two of 10 rats in the experimented group had few to moderate amount of sperms in their seminal vesicles (p = 0.180). After the distal irrigation of the vas deferens using NSS, some residual sperms resided in the middle and more at the distal part of the vas with a few that escaped into the seminal vesicles.     

Comparison of spermatogenic damage induced at 6 months after ligation of the vas deferens at proximal and distal locations in the rabbit

Recent study in rabbits demonstrated that vasectomy via the inguinal canal did not result in any spermatogenic damage 3 months postoperation; this study aimed to determine whether the damage would occur in a longer term. The left or right vas deferens was ligated near the epididymal head (unilateral proximal vasectomy, 12 animals) or via the inguinal canal (unilateral distal vasectomy, 11 animals) in adult male rabbits, with a sham operation being performed on the contralateral side. Six months postoperation, testes, epididymides and vasa deferentia were removed and methacrylate resin-embedded sections prepared to evaluate spermatogenesis by histological (qualitative) and stereological (quantitative) studies. The juxta-epididymal segment of the occluded vas deferens was severely distended (filled with sperm) in 10 of the 11 animals with distal vasectomy and moderately or slightly distended in nine of the 12 animals with proximal vasectomy. Severe spermatogenic damage occurred in seven animals with proximal vasectomy (the juxta-epididymal vas moderately or slightly distended), in only one animal with distal vasectomy (the vas not severely distended). In conclusion, spermatogenic damage occurred at 6 months postvasectomy in some animals, especially those with proximal vasectomy and therefore shorter occluded reproductive tract for sperm storage; the damage was probably intra-tract pressure mediated.     

Microrecanalization after vasectomy in man

Previously spermatozoa in the semen of vasectomized men were reported in 62 of 63 specimens from 24 men 2 to 31 years postvasectomy (Freund and Couture, 1982). A morphologic basis and term, "microrecanalization," was proposed for this observation. Serial sections (5 mu at 200-mu intervals) of 40 specimens removed at vasovasostomy from 20 men (2 to 14 years postvasectomy) were examined and microcanals (small epithelial-lined channels) were demonstrated in 27 specimens from 18 men. In nine of the 27 specimens, spermatozoa or sperm heads were found within the microcanals. Microcanals occurred in smooth muscle, connective tissue and scar tissue, in each segment, testicular, central and abdominal, in the presence or absence of the vas deferens. Microcanal continuity was traced for 200 to 1140 microns by computerized image analysis. Microrecanalization is characterized by the absence of inflammation or sperm extravasation and is histologically distinct from vasitis nodes or sperm granuloma. Microrecanalization provides morphologic and physiologic bases for the protection of the testis and maintenance of spermatogenesis in man after vasectomy.     

Intravasal "toothpaste" in men with obstructive azoospermia is derived from vasal epithelium, not sperm

PURPOSE: Men undergoing vasectomy reversal many years after vasectomy are at increased risk for secondary epididymal obstruction. When this occurs the intravasal fluid is often a thick, white, toothpaste-like material devoid of sperm. In this study we characterize the vasal fluid found in men with a newly described entity, segmental dysplasia of the vas deferens, in which at least 2 distinct sites of vasal obstruction are present. We determine the significance of this fluid in men with obstructive azoospermia. MATERIALS AND METHODS: Three men who underwent scrotal exploration for obstructive azoospermia due to segmental dysplasia of the vas deferens were evaluated. Each underwent scrotal exploration including bilateral vasotomy and testicular biopsy. Intravasal fluid was collected, evaluated microscopically and sent for cytopathological evaluation. RESULTS: All men had isolated segments of the vas 2 to 5 cm in length that were not connected to the epididymis or ejaculatory ducts. We have named this condition segmental dysplasia of the vas deferens. Vasotomy was performed between aplastic segments, revealing thick, white, toothpaste-like material identical to that seen in men with secondary epididymal obstruction undergoing vasectomy reversal. Cytopathological evaluation of this fluid revealed proteinaceous concretions and rare clusters of degenerated columnar epithelial cells, but no sperm or sperm products. CONCLUSIONS: Thick, white, toothpaste-like material is produced between 2 obstructed segments as seen in men with segmental dysplasia of the vas deferens and with secondary epididymal obstruction. Our findings in men with segmental dysplasia of the vas deferens indicate that vasal "toothpaste" must be derived from vasal epithelium, not sperm.     

输精管滤过装置节育术的临床研究

本文报告51例输精管腔植入由作者研制的输精管滤过装置与40例钳穿法输精管结扎术经过1至3年的临床比较研究。其结果:两组不同术式的节育有效率均为100％,滤过装置组术后1～3年仍有51.6％(16/31)测出中性α-糖苷酶活性,至今仅有12例附睾轻度肿胀和近附睾段输精管轻度增粗;而结扎组一年后均有不同程度的附睾肿胀和近附睾段输精管增粗,并有1例并发附睾淤积症。两组间有非常显著的统计学差异(P<0.005)。结果提示:输精管滤过装置不仅能限制精子通过而达到节育的目的,又不完全影响附睾液的排出,较好地避免或减少了附睾淤积的发生。因此,有可能作为一种新型的、非阻塞性的男性节育方法用于临床。     

无缝合非灼烧式输精管上皮刮除术：人类实验性研究

输精管上皮刮除术可能是一种安全有效的男性绝育方法。本实验探究了输精管上皮刮除术实现绝育的有效性。实验中我们使用了一种称为Vas—X的新型刮匙,对12名要求绝育的正常男性的输精管上皮进行刮除。手术后连续6个月每月进行一次精液分析,同时测量疼痛状况。术后3个月,所有男性的精子浓度均降到20万／毫升以下,其中7名男性精子浓度为0,术后疼痛微弱。最终9名男性实现并保持不育,但术后4到6个月期间,3名男性的精子浓度增加,需要重新行刮除术。输精管显微检查发现这三名男性的精子浓度升高是由输精管再通造成。本实验表明：输精管刮除术可以有效的实现绝育,但是有1／4的手术对象由于输精管再通而不能成功实现绝育。本研究表明,输精管刮除术还不够作为切实可行的绝育手段,还有待于进一步完善。     

输精管结扎术四种不同残端处理与节育效果的比较性研究

８１６例自愿接受输精管绝育术男子按照节育手术常规施行输精管结扎术，受术者随机均分为４个队列，分别施行不同的输精管残端处理方法：（１）单纯两断端结扎；（２）结扎加精索筋膜隔离；（３）结扎加苯酚涂灼；（４）电灼。术中以１：３０００新洁尔灭溶液每侧输精管５ｍｌ行精囊灌注，术后３年４种方法精子消失率分别为９０．０％（１８０／２００），９４．１％（１７５／１８６），９３．７％（１７９／１９１）及９４．５％（１７２／１８２），术后并发症为０．４％（３／８１６）。在此对输精管结扎术后自发再通问题进行了讨论，并认为１：３０００新洁尔灭溶液行精囊灌注是安全可行的。     

Assessment of seminal plasma laminin in fertile and infertile men

Aim： To assess laminin levels in the seminal plasma of infertile and fertile men, and to analyze the correlation of laminin levels with sperm count, age, sperm motility and semen volume. Methods： One hundred and twenty-five recruited men were equally divided into five groups according to their sperm concentration and clinical examination： fertile normozoospermia, oligoasthenozoospermia, non-obstructive azoospermia （NOA）, obstructive azoospermia （OA） and congenital bilateral absent vas deferens （CBAVD）. The patients＇ medical history was investigated and patients underwent clinical examination, conventional semen analysis and estimation of seminal plasma laminin by radioimmunoassay. Results： Seminal plasma laminin levels of successive groups were： 2.82 ± 0.62, 2.49 ± 0.44, 1.77 ± 0.56, 1.72 ± 0.76, 1.35 ± 0.63 U/mL, respectively. The fertile normozoospermic group showed the highest concentration compared to all infertile groups with significant differences compared to azoospermic groups （P 〈 0.05）. Testicular contribution was estimated to be approximately one-third of the seminal laminin. Seminal plasma laminin demonstrated significant correlation with sperm concentration （r = 0.460, P 〈 0.001） and nonsignificant correlation with age （r = 0.021, P = 0.940）, sperm motility percentage （r = 0.142, P = 0.615） and semen volume （r = 0.035, P = 0.087）. Conelusion： Seminal plasma laminin is derived mostly from prostatic and testicular portions and minimally from the seminal vesicle and vas deferens. Estimating seminal laminin alone is not conclusive in diagnosing different cases of male infertility.     

影响显微外科输精管吻合术后复孕的多因素研究

通过采用作者改良的显微外科输精管吻合法和总结查找近睾端输精管液中精子的方法，对56例输精管结扎术后要求复有者进行了输精管管吻合术．术后2年随访50例，复通率达100％，复孕率达60％、以抗精子抗体、精子穿卵试验、输精管结节精子肉芽肿、精液质量等21个指标对孕组和未孕组进行了对照现实，经过Logistic回归等方法进行统计学处理，筛选出术后SIT、精子存活率、精子密度、IBT结合IgA和IgG、MAR、TAT和精子活动率等8个影响吻合术后受孕的主要因素，其中前2个最为主要。     

Reconstructive microsurgery of the vas deferens. Experimental study of suture materials

Microsurgery is the procedure of choice for vasectomy reversal. The aim of this study was two compare two different suture materials for vasovasectomy - a nonresorbable material (nylon 10/0 with a BV 6 needle), which is widely used, and a resorbable material (polyglycolic acid, also with a BV 6 needle), which has not yet been evaluated for this use. 28 Sprague-Dawley rats were operated on under microscope. Two groups were then compared, group A with nylon (n = 14) and group B with polyglycolic acid (n = 14). In each group, 8 animals had a vas deferens section and 6 had a previous vasectomy by ligature. Ten days postoperatively, the patency rate of the anastomosis was evaluated by the presence of sperm on both sides of the suture line. The contractility was assessed by mechanical stimulation. The existence of a sperm granuloma was considered as indicative of a non functional anastomosis. Three days later a fertility test was performed, lasting three months, and the number of litters was checked. The rats were sacrificed after three months, and each vas deferens was examined histologically or by electron microscope. The macroscopic results were: 57% patent anastomoses in group A and 77% in group B. 16% patent anastomoses after ligature in group A (n = 6) and 75% in group B (n = 6). The pregnancy rate was 54% in group A and 77% in group B. After previous ligation, the corresponding figures were 20% and 83% respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

The ratio of second to fourth digit length in azoospermic males undergoing surgical sperm retrieval: predictive value for sperm retrieval and on wubsequent fertilization and pregnancy rates in IVF/ICSI cycles

The differentiation of the urogenital system and the appendicular skeleton in vertebrates is under the control of Homeobox (Hox) genes. It has been shown that this common control of digit and gonad differentiation has connected the pattern of digit formation to spermatogenesis and prenatal hormone concentrations in males. We wished to establish whether digit patterns, particularly the ratio between the lengths of the second and fourth digit in males (2D : 4D), was related to spermatogenesis and, more specifically, the presence of spermatozoa in testicular biopsies from azoospermic men undergoing surgical sperm retrieval. Forty-four men were recruited, of whom 16 were diagnosed with nonobstructive azoospermia and 4 with congenital bilateral absence of the vas deferens, and 24 previously fertile men were azoospermic after previous vasectomy. Our results show that men with previous fertility or of an acquired form of azoospermia had significantly lower 2D : 4D ratios than men with nonobstructive azoospermia. In nonobstructive azoospermia, there was a significantly lower 2D : 4D ratio on the left side in men who had successful retrieval than those with unsuccessful retrieval. For these men who had a successful retrieval, none had a 2D : 4D ratio more than 1 on the left side, whereas 4 of 7 men in whom sperm was not found had a 2D : 4D ratio greater than 1. On successful sperm retrieval, subsequent fertilization and clinical pregnancy rates were unaffected by 2D : 4D ratios.     

Some vasovasostomized men are characterized by low levels of P34H, an epididymal sperm protein

During epididymal transit, sperm surface proteins involved in the fertilization process can be added or modified. P34H, a human epididymal-sperm protein, is proposed to be involved in the interactions between spermatozoa and the zona pellucida. We have previously demonstrated that P34H is present in men of proven fertility and is absent in 50% of men presenting with idiopathic infertility. Spermatozoa with a low amount of P34H exhibit a dramatic reduction in their ability to interact with zona pellucida. Even if the surgical success of vasectomy reversal is high, fertility is not always reestablished, possibly due to epididymal damage caused by vasectomy. In this study, western blot analyses were performed to determine the level of P34H present on spermatozoa of men who underwent vasectomy reversal. Spermatozoa obtained from different semen samples from a given individual had similar P34H levels; however, samples from different men were highly variable. When quantified by densitometric scanning, P34H levels from vasovasostomized men varied between 1.5% and 149% compared with that from a fertile donor who represented 100%. Eighteen of 25 vasovasostomized men had a P34H level lower than 30% of the normal value, while the remaining 7 males were in the normal range. Furthermore, the population of vasovasostomized men with P34H levels lower than 30% was significantly different from the control group of 19 fertile men. The high variation of P34H levels observed in vasovasostomized men did not correlate with the spermiogram values (P > 0.05). An important factor in determining sperm P34H level appears to be the period of time elapsed between the vasectomy and vasovasostomy. In summary, our results show that the P34H level varied from one man to another and that low levels of the epididymal sperm protein is associated with vasectomy reversal.     

Effect of vasectomy via inguinal canal on spermatogenesis in rabbits

Aim： To determine whether vasectomy away from the epididymal tail （via the inguinal canal） in rabbits can reduce the early postoperative effects on spermatogenesis. Methods： Twenty-nine normal male Japanese white rabbits （aged 4- 6 months） were subjected to unilateral close-ended （conventional） or open-ended （the cut end of the juxta-epididymal vas deferens not ligated） vasectomy via the inguinal canal. Ten days and 3 months after operation, testes, epididymides and vasa deferentia were removed and methacrylate resin-embedded sections prepared. The histology of the testis, epididymis and vas deferens was examined under light microscope, and the volume and diameter of the seminiferous tubules were quantitatively studied using stereological methods. Results： Neither of the methods of vasectomy led to apparent damage to spermatogenesis on the vasectomized side in comparison with the contralateral shamoperated side, but the juxta-epididymal vas deferens on the vasectomized side was highly distended and contained numerous sperm 3 months after operation. Conclusion： Vasectomy away from the cauda epididymis has no significant early postoperative effects on spermatogenesis in rabbits.     

Localization of intrinsic and extrinsic SVS II immunoreactivity in rat spermatozoa.

A glycoprotein, designated SVS II, is secreted in an androgen-dependent manner from lateral prostate and seminal vesicles of the rat. The pI of the protein is 10.5 and it has a molecular mass of 49 kDa. F-actin isolated from skeletal and heart muscle is precipitated at a ratio of 2:1 by SVS II. Using a polyclonal rabbit antibody against SVS II, we found an immunoreaction at the head region of rat spermatozoa removed from the vas deferens. In addition, immunoreactive material was observed in the principal piece of the sperm tail in those spermatozoa. We have studied the distribution of SVS II-immunoreactive material in spermatozoa isolated from seminiferous tubules, proximal (efferent ductules), and distal (caudal epididymal duct) epididymis, both in sexually active and inactive rats and found a differential reactivity pattern. Immunoreactivity observed in the principal piece of the sperm tail develops only immediately before spermiation and does not change during the epididymal transit of the spermatozoa. Immunolabelling seen in the head portion is first observed in spermatozoa from proximal epididymis. Simultaneous with its appearance, rhodamine-labelled phalloidin, indicating the presence of F-actin, no longer binds to that region. While the immunoreaction of the sperm head is attributed to extrinsic SVS II, added to the sperm head in proximal epididymis, the immunoreactivity of the sperm tail seems to result from a cross reactive intrinsic sperm tail protein that achieves its final structure briefly prior to the onset of spermiation.(ABSTRACT TRUNCATED AT 250 WORDS)     

Irrigation of the distal vas deferens during vasectomy: does it accelerate the post-vasectomy sperm-free rate?

A prospective, non-randomized, partially blinded, controlled trial was conducted to evaluate the efficacy of irrigation with normal saline solution (NSS) during no-scalpel vasectomy (NSV) compared with NSV alone in 62 men. In the NSS irrigation group, an Angiocath 24-gauge needle was inserted into the distal vas lumen, and 20 mL NSS solution was used to irrigate the vas manually on both sides. Post-operative follow-up included urine samples collected immediately and semen samples for sperm count at 2, 6 and 12 weeks post-vasectomy. The difference in the number of spermatozoa appearing in the post-vasectomy urine samples and the mean urine sperm count in both groups were significantly different ( p < 0.0001 and p < 0.01, respectively). The numbers of post-operative ejaculations, the mean sperm concentration, and the number of patients who achieved sterility (defined as no motile spermatozoa in the ejaculate) in both groups at 2, 6 and 12 weeks were similar ( p > 0.05). It is concluded that although irrigation of the distal vas with NSS was successful in removing a large number of spermatozoa from the tract, this procedure did not significantly accelerate the rate of achieving absence of motile spermatozoa in the ejaculate.     

Sperm function tests after vasovasostomy

Aim: To evaluate the sperm function after vasovasostomy. Methods: Semen samples from 42 subjects after vasovasostomy (Group A: 1 -6 months, Group B: 6 - 12 months; Group C: 12 - 18 months after vasectomy reversal) were investigated. Semen from 34 normal fertile men was used as controls. Sperm function tests, including hyposmotic swelling test (HOST), acridine orange (AO) fluorescence, acrosome reaction (triple-stain), cervical mucus penetration test (CMPT), etc were done. Results: After vasectomy reversal, the percentage of HOST was significantly lower than that of the normal fertile men. In regard to AO, there were no significant differences between the three vasovasostomy groups and between these 3 groups and the controls. With triple-stain, the percentage of normal acrosome reaction was significantly lower in Group A as compared with the controls, but not in Groups B and C. There were no significant differences in the results of CMPT between the vasovasostomy groups and the controls. However, the number of “poor“ type was significantly higher in Groups A and C than in the controls; the percentage of “negafive“ type were higher in Groups A and B than in the controls. Conclusion: After vasovasostomy a lower level of HOST remained for one year and gradually recovered after one year. Six months after vasectomy reversal, the percentage of acrosome reaction could be changed from lower level to normal range. The data of AO indicated that the genetic material (double-stranded DNA) in spermatozoa was not affected by vasovasostomy. To evaluate the result of CMPT after vasectomy reversal, not only the normal results but also the abnormal results (“poor“ and “negative“ types) should also be considered.     

The effects of vasectomy on the autonomic innervation of the human vas deferens

Neurohistochemical and fine structural techniques have been employed to examine the intramural autonomic innervation of the human vas deferens following surgical division of the duct one to 15 years previously. Samples from sites on the distal (testicular) and proximal (urethral) aspects of the original vasectomy have been compared with control specimens obtained at vasectomy as to the arrangement and distribution of autonomic nerves. In contrast with tissue from the proximal part and from controls, the distal samples revealed a marked reduction in the noradrenergic innervation of the muscle coat. In addition acetylcholinesterase-containing nerves associated with the basal aspect of the epithelium were usually absent from the distal portion of the vas deferens. These findings have been considered in relation to the contractile and secretory activities of the organ following vasovasostomy and may be of importance to the maturation and fertility of spermatozoa.     

Seminal human papillomavirus originates from the body surface and is not a frequent aetiological factor in azoospermia

Human papillomavirus (HPV) DNA has been detected in the testis tissue of 6.5% of 185 men with non‐obstructive azoospermia (NOA). Others have suggested that seminal HPV originates from contamination from the genital skin and mucosa. One hundred unselected azoospermic men and 43 normal men undergoing vasectomy were recruited. Testicular biopsies for HPV examination were collected from all the men. Additionally, the normal men undergoing vasectomy delivered a semen sample and had a swab for HPV examination taken from the genital skin before vasectomy. A piece of each Vas deferens obtained during the vasectomy was examined for the presence of HPV. Two of the primarily azoospermic men were shown to have cryptozoospermia. It was not possible to detect HPV in the testis tissue of any of the included 98 azoospermic men or the 43 proven fertile men. In the proven fertile men, HPV DNA was detected in the semen of 15 men (35%), on the genital skin of 28 men (65%), and in the Vas deferens in three cases (7%). In 13 (87%) men with HPV‐positive semen samples, HPV DNA was also detected in the skin swabs, and in 11 men (73%), identical HPV genotypes were found in the two locations.