Supplementing extender with anandamide enhances quality of low sperm doses during cryopreservation in bulls

The present study explored the effect of anandamide supplementation in the extender on quality of low sperm doses during cryopreservation in Sahiwal bulls. Each fresh semen sample was split into eight aliquots (I, II, III, IV, V, VI, VII and VIII). The aliquots I, II, III and IV were taken as control and diluted to 20, 15, 10 and 5 million spermatozoa/0.25 ml respectively. The aliquots V, VI, VII and VIII were diluted with extender (supplemented with anandamide at 1 µM/ml of extender) to 20, 15, 10 and 5 million spermatozoa/0.25 ml respectively. This was followed by filling of diluted semen into French mini straws, equilibrated at 4°C of 4 hr and cryopreserved. The results revealed that the proportions of motile spermatozoa, live spermatozoa and live acrosome intact spermatozoa were significantly (p < .05) higher in all anandamide-treated sperm doses compared to control. The proportions of moribund spermatozoa, dead acrosome intact spermatozoa and capacitated spermatozoa were significantly (p < .05) reduced in all anandamide-treated sperm doses compared to control, with no difference in proportion of dead acrosome-reacted spermatozoa. In conclusion, anandamide supplementation in the extender increases the post-thaw quality of low sperm doses during cryopreservation in bulls.     

The effect of cancer on semen quality after cryopreservation of sperm

The results of cryopreservation of sperm from cancer patients were retrospectively reviewed in an effort to better understand the relationship between semen quality and the presence of different kinds of cancer. The semen analysis results for 146 patients referred to our infertility center for sperm banking over a 2-year period were examined. These patients were divided into three groups according to their diagnosis: group I, prevasectomy controls; group II, patients with lymphoma and Hodgkin's disease; and group III, patients with testicular cancer, e.g., seminoma, embryonal cell carcinoma, or teratocarcinoma. The seminal parameters assessed included sperm count and prefreeze and postthaw motility and velocity. For these parameters, significant decrease from control values (P less than 0.05 to P less than 0.01) was seen in groups II and III. The specimens from group I patients retained good motility and velocity after thawing. Our results indicate that semen quality is adversely affected by the presence of cancer in the body.     

Panax ginseng improves survival and sperm quality in guinea pigs exposed to 2,3,7,8-tetrachlorodibenzo- p-dioxin

OBJECTIVES: To further assess the effect of Panax ginseng on survival and sperm quality of guinea pigs exposed to 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). MATERIALS AND METHODS: Eighty male guinea pigs were divided into eight equal groups. The normal control (NC) group received vehicle and saline; one dose of 1 micro g/kg body weight TCDD was injected intraperitoneally into the single TCDD-treated (TT) and test groups (P100, P200, C100, C200); G and NC groups received vehicle instead of TCDD. P. ginseng water extract (PG-WE) was injected intraperitoneally at daily doses of 100 (G100, P100, C100) or 200 mg/kg body weight (G200, P200, C200). The PG-WE was administered to the P and G groups for 12 weeks from 1 week before TCDD exposure, and to the C groups for 10 weeks from 1 week after TCDD exposure. After a 4-week discontinuation of PG-WE treatment after the 13th week the surviving males were then tested for fertility by mating them with females. The litter size, death rate, male/female birth ratio and physical abnormalities of the progeny were investigated. After confirming delivery of the offspring, the parent males were killed at 40 weeks, their testes weighed and sperm quality assessed. RESULTS: All TT animals died within 18 days after TCDD exposure, but 40-70% of the PG-WE-treated groups, depending on the group, survived until death at 40 weeks. All the surviving males were fertile regardless of TCDD exposure; there was no difference in litter size between the NC and test groups. Notably the death rate of progeny born to PG-WE-treated groups was lower than that of progeny born to the NC group. The progeny born to TCDD-exposed groups (P200 and C groups) had a preponderance of females. G Group animals had higher sperm quality than that of NCs even long after discontinuing PG-WE. CONCLUSION: P. ginseng improves the survival rate and sperm quality in guinea pigs exposed to TCDD.     

Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.     

Antioxidant effect of hydroxytyrosol on human sperm quality during in vitro incubation

The aim of the study was to evaluate the antioxidant potential of hydroxytyrosol (HT) on human sperm quality during incubation in vitro. Semen samples collected from men attending the Laboratory of Histology‐Embryology of Sfax Faculty of Medicine (Tunisia) for infertility investigations were evaluated for initial sperm parameters. Only normal selected ejaculates (n = 15) were centrifuged and incubated further with or without HT (200ug ml^?1) at room temperature for 45 min. After incubation, sperm motility and viability, DNA oxidation and reactive oxygen species (ROS) production were assessed. The results showed that centrifugation significantly influenced sperm motility and viability. The supplementation of HT in incubating media improved (P = 0.01) significantly sperm viability and decreased sperm DNA oxidation (P < 0.001) and ROS levels (P = 0.03) following centrifugation. It can be concluded that supplementation of HT might be helpful to maintain the human spermatozoon after centrifugation.     

Evaluation of donor semen quality provided by six sperm banks: a retrospective study of 1877 artificial insemination cycles

This study aimed at evaluating the impacts of sperm quality of six national sperm banks on pregnancy rates (PRs) of artificial insemination with donor sperm (AID) in China. A large retrospective analysis was performed on 1877 insemination cycles in 1209 women in a unique setting during a 3.5-year period. Global PRs of 22.1% per cycle and 34.2% per patient were achieved. The PRs of the six banks varied from 15.5% to 29.0% (P = 0.011). Significant differences were observed in the quality of donor semen provided by the six sperm banks. Moreover, in some banks, the poor sperm quality was related to the suboptimal PRs. However, in certain banks, high values of sperm parameters did not result in satisfactory PRs accordingly. These data demonstrated that variability of donor semen quality existed in the different banks. But, sperm parameters after thawing may not be detrimental factors affecting the success rate of AID treatment. Further studies are needed to seek potential molecular markers for predicting fertility potency of donor sperm.     

Study on correlation of sperm quality parameters with antioxidant and oxidant status of buffalo bull semen during various stages of cryopreservation

This investigation was carried out to study the correlation of sperm quality parameters with antioxidant and oxidant status of buffalo bull semen during various stages of cryopreservation. Semen samples were evaluated for sperm parameters (mass motility [MM], concentration [CON], progressive motility [PM], viability [VIB], acrosomal integrity [AI] and hypo‐osmotic swelling [HOS] response), antioxidants (superoxide dismutase [SOD], catalase [CAT], glutathione peroxidase [GPx] and total antioxidant capacity [TAC]) and oxidants (Lipid peroxidation [LPO] and reactive oxygen species [ROS]) at fresh, pre‐freeze and post‐thaw stages. Sperm parameters (PM, VIB, AI and HOS response) and antioxidants (SOD, CAT and TAC) were significantly (p < .05) reduced at fresh stage, and oxidants (LPO and ROS) were significantly (p < .05) increased at pre‐freeze and post‐thaw stages. At fresh stage, MM was negatively correlated with LPO (p < .05), and CON was positively correlated with SOD, TAC and CAT, negatively correlated with LPO and CAT was positively (p < .01) correlated with VIB and HOS response. At pre‐freeze stage, CAT was positively correlated with PM and AI (p < .05), and AI was negatively (p < .05) correlated with ROS. At post‐thaw stage, CAT was positively correlated with PM, VIB, HOS response and AI,, and LPO was negatively correlated with HOS, AI and VIB. The study of correlations of these parameters at different preservation stages with bull fertility may play an important role in developing models for predicting future fertility of bulls in the absence of conception rate data.     

不同来源精子对ICSI治疗后妊娠结局的影响

目的分析不同精子来源和数量对单精子卵胞浆内注射术(ICSI)妊娠结局的影响。方法2000年1月～2005年1月在本中心进行ICSI治疗而妊娠的354例,据精子来源与数量分为精液正常组(47例)、少弱精子组(194例)与手术取精子组(113例)。比较3组的临床妊娠率、流产率、分娩率、妊娠并发症、分娩孕周、新生儿出生体重、畸形、围产期死亡率等组间差异。结果3组患者的年龄、不孕年限、临床妊娠率(41.3%、39.7%、40.4%)、流产率(14.9%、18.6%、15.9%)、分娩率(78.7%、75.8%、79.6%)、双胎率(34.4%、35.4%、37.8%)、妊高症发生率(10.8%、10.2%、11.1%)、前置胎盘发生率(5.4%、6.8%、6.7%)、早产率(27.0%、27.9%、27.8%)、分娩孕周(36.4±5.4)w、(37.6±2.3)w、(37.2±2.8)w、胎儿出生体重(2765.4±624.9)g、(2749.0±523.5)g、(2763.5±684.4)g、先天性畸形发生率(4.5%、5.0%、4.0%)、围产期死亡率(40‰、45‰、48‰),差异均无显著性(P>0.05)。结论不同精子来源与数量不影响ICSI治疗后的妊娠和围产儿结局。     

Cryptorchidism: incidence and sperm quality in infertile men

In a population of 8500 men attending the andrology outpatient clinic, 200 men (2.35%) were recorded as having some disturbances with the descent of the testes into the scrotum. Medical history of the patients revealed that 51 underwent unilateral orchidopexy; 40 bilateral orchidopexy; and 24 were treated with human chorionic gonadotropin in order to induce descent of their testes. In addition, 6 patients reported spontaneous descent of the testes, and 13 others were found to be unilaterally cryptorchid upon physical examination. Results of semen analysis, hormonal profile, testes position, and testicular volume were compared to those of 105 proven fertile men. The major finding of this study shows that post-partum undescended testes suffer from primary Sertoli cell malfunction as reflected by elevated serum follicle stimulating hormone levels. Serum luteinizing hormone and testosterone levels were within the normal range. Surgical descent of the testes did not improve sperm production, proved by low sperm quality of all the study groups, compared to the cryptorchid group. Among the patients who were operated on, no correlation was found between age at operation and semen variables. All groups showed poor sperm quality which can be defined as oligoteratoasthenozoospermia. The degree of spermatogenic damage was in the following order of diagnosis or treatment: bilateral orchidopexy greater than cryptorchid testes greater than hormonal treatment greater than unilateral orchidopexy greater than late spontaneous descent of the testes. Thus, it is advisable to postpone surgical treatment of cryptorchidism and apply this only after a waiting period, and if the hormonal approach has failed to descend the testis.     

二甲双胍对营养性肥胖大鼠附睾精子质量和睾丸抗氧化能力的影响

目的:观察二甲双胍对高脂饮食诱导的肥胖大鼠附睾精子质量和睾丸抗氧化能力的影响。方法:雄性SD大鼠分为正常对照组和造模组,造模组以高脂饲料喂养8周后,取24只肥胖大鼠随机分为模型对照组、二甲双胍组和普通饲料组,除模型对照组继续高脂饲料喂养外,其余3组普通饲料喂养。12周末,所有大鼠均禁食12 h后处死,检测Lee's指数、睾丸和附睾脏器指数,附睾精子浓度、精子活动率和a+b级精子百分率,睾丸组织匀浆中超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)和丙二醛(MDA)含量。结果:Lee's指数模型对照组与其他3组比较显著升高(P<0.01),Lee's指数正常对照组较二甲双胍组升高(P<0.05)。睾丸、附睾、脏器指数模型对照组较其他3组显著降低(P<0.01)。精子浓度、精子活动率和a+b级精子百分率模型对照组与其他3组比较显著降低(P<0.05或P<0.01),正常对照组较二甲双胍组和普通饲料组精子浓度升高(P<0.05)。SOD含量(U/mg prot)模型对照组(90.92±4.06)较正常对照组(101.69±8.49)与二甲双胍组(102.04±10.67)降低(P<0.05)。GSH-Px含量(U)正常对照组(28.32±2.28)较模型对照组(23.49±1.08,P<0.01)、二甲双胍组(25.73±2.14,P<0.05)和普通饲料组(25.77±2.19,P<0.05)升高,模型对照组较二甲双胍组降低(P<0.05)。MDA含量(nmol/mg prot)模型对照组(2.68±0.76)较正常对照组(1.84±0.31,P<0.01)、二甲双胍组(1.88±0.33,P<0.01)和普通饲料组(2.14±0.35,P<0.05)升高。结论:二甲双胍治疗和饮食改善均可提高营养性肥胖大鼠精子质量,改善睾丸组织抗氧化能力。     

Assessment of sperm quality, DNA integrity and cryopreservation protocols in men diagnosed with testicular and systemic malignancies

Men diagnosed with malignancy are often referred for semen banking to preserve their fertility prior to cancer treatment. The chances of cancer patients for achieving future fecundity will be determined by the sperm quality including the integrity of the genomic material in the frozen samples. The objectives of this study were to compare the sperm quality and DNA integrity in men diagnosed with testicular and systemic malignancies before receiving treatment and to identify the optimum cryopreservation protocol for their samples including a remote semen collection option. In comparison with fertile donors, patients with testicular malignancies had significantly lower sperm concentration, while both testicular and systemic malignancy patients had significantly lower sperm motility and cryosurvival rates. In addition, the SCSA defined DNA fragmentation index was significantly higher in patients with testicular and systemic malignancies compared with fertile donors. It was noted that the extent of deterioration in sperm quality and DNA integrity seen in cancer patients did not reach the previously defined statistical threshold for impaired fertility. Freezing spermatozoa with the seminal plasma offers the highest protection against cryo-injury. Nevertheless, remote semen collection can still be used as it yields adequate results.     

禁欲2天对于精子质量的影响

目的 通过对71名人工受精的男性禁欲2d和3～7d的精子质量的分析，探讨禁欲不同天数对于精子质量的影响。方法 第一次人工受精的精液为禁欲3～7d(Ⅰ组)，第二次为禁欲2t(Ⅱ组)，分析2组精液的差异及其精子培养液处理后的差异。结果 Ⅰ组精液的精子密度和A级精子百分比明显高于Ⅱ组，但是按照正常与异常分类，无论是培养液处理前还是处理后均无差异。结论 禁欲2d与3～7d相比，按正常与异常精液划分没有差异，禁欲2d做人工受精，精子质量不受影响。     

Integrity of mitochondrial membrane potential reflects human sperm quality

The aim of this work was to evaluate intracellular reactive oxygen species (ROS) levels, phosphatidylserine (PS) externalisation and mitochondrial membrane potential integrity in the spermatozoa of healthy donors and outpatients who consulted for infertility and to correlate the results with the classic sperm parameters. For the evaluation of intracellular ROS levels, PS externalisation and mitochondrial membrane potential integrity, the fluorescent compounds dihydroethidium, annexin V-FITC and JC-1, respectively, were used and analysed by using flow cytometry. Conventional seminal analysis, including motility, viability, morphology, sperm count and volume, was performed according to the WHO criteria. The mitochondrial membrane potential and ROS results showed significant differences between the spermatozoa of individuals with a normal semen analysis and those of the group presenting abnormality in at least one of the sperm parameters. Mitochondrial membrane potential showed a significant and direct correlation with all the sperm parameters analysed. ROS were inversely correlated with motility, viability and morphology. PS externalisation, however, did not show any differences between the two groups, nor was it correlated with the sperm parameters examined. The evaluation of mitochondrial membrane potential integrity is a test that reflects sperm quality, which makes it highly recommendable to be applied as a complement to routine sperm analyses.     

Poor sperm quality and advancing age are associated with increased sperm DNA damage in infertile men

With increasing evidence for faulty paternal contribution to reproduction, there has been a steady increase in studies highlighting an association between sperm DNA damage, failed/delayed fertilisation and aberrant embryo development. Owing to prevailing ambiguity, the aims of the study were to analyse the genetic integrity of the male gamete and then to understand its association with age, standard semen parameters, lifestyle and occupational factors. The study included 504 subjects, attending university infertility clinic for fertility evaluation and treatment. Semen characteristics were analysed by standard criteria; terminal deoxynucelotidyl transferase-mediated nick end-labelling assay was employed for DNA damage assessment. The average incidence of sperm DNA damage in patients with normozoospermic semen parameters was <10%. Patients with oligozoospermia, severe oligozoospermia, oligoasthenoteratospermia, asthenoteratozoospermia and necrozoospermia had significantly higher level of sperm DNA damage (P < 0.001). Patients above 40 years of age had significantly high levels of DNA damage (P < 0.001) compared with their counterparts. Patients with varicocele and a history of alcohol consumption had higher incidence of spermatozoa with DNA damage (P < 0.01). Poor sperm characteristics in the ejaculate are associated with increased sperm DNA damage. Age-related increase in sperm DNA damage and association of the same with varicocele and alcohol consumption are also demonstrated.     

番茄红素对人精子冷冻损伤的保护作用及机制研究

目的:通过在人精子冷冻保护液中添加不同浓度的番茄红素,研究番茄红素对精子冷冻损伤的可能保护作用及机制。方法:选择吉林省人类精子库捐精者的精液标本25份,每份精液一式4份,3∶1加入冷冻保护液后混匀,不含番茄红素者设为对照组(Ly0),而Ly2、Ly5、Ly10实验组混合液中分别含有2、5、10μmol/L的番茄红素,冷冻复苏精液进行常规分析,采用流式细胞术分析冻融后各组精子的凋亡率;采用硫代巴比妥酸(TBA)比色法检测精子中脂质过氧化产物丙二醛(MDA)浓度。利用JC-1标记法检测线粒体膜电位。结果:冻融后各组精子运动参数较新鲜精液参数均明显下降(P<0.01),Ly5组冻融后精子凋亡率[(25.68±4.36)%]较对照组[(33.26±4.78)%]显著下降(P<0.05);Ly5组线粒体膜电位水平[(66.18±14.23)%]显著高于对照组[(55.24±12.31)%],P<0.05;Ly2、Ly5、Ly10组的MDA水平与对照组比较,无统计学差异(P>0.05)。结论:在精液冷冻保护液中添加一定浓度的番茄红素可以减少线粒体氧化损伤,减轻活性氧对精子质膜的氧化应激损伤,从而提高精子的抗凋亡能力。     

男性血液和精液中微量元素含量与精子活力的相关性研究

目的:探讨男性血液和精液中微量元素的分布及其与男性精液参数的相关性。方法:采用BH-5100五通道原子吸收仪、YY-1001血铅仪和伟力彩色精子自动分析仪,对113例男性血液和精液进行钙、镁、铜、锌、铁、铅的测定及精液常规分析,以了解微量元素和精液参数的相互关系。结果:①血液与精液中铅、铜、锌、铁含量显著相关,其中以铜的相关系数最大;钙、镁没有相关性。②血液和精液中铅、铜、锌、钙、镁、铁6种微量元素分布存在显著性差异。③精液中钙与精子活率、直线运动精子活率、曲线速度、平均移动角度、活跃精子密度、直线速度、平均路径速度、侧摆幅度等指标显著正相关;铅与精子活率、直线运动精子活率、活跃精子密度、活动精子密度、直线运动精子密度等指标呈显著负相关;镁元素与精子活率、直线运动精子活率、直线运动精子密度呈显著负相关;精液酸碱度与钙、镁、铁3种元素呈显著负相关。④血液中的锌与精子活率和直线运动精子活率呈显著负相关,镁与精子活率、直线运动精子活率、平均路径速度呈显著负相关,铅与侧摆幅度呈显著正相关。结论:不同体液中的微量元素分布和精子的运动功能存在不同的相关性。     

Stimulating effects of quercetin on sperm quality and reproductive organs in adult male rats

Aim： To investigate effects of quercetin on weight and histology of testis and accessory sex organs and on sperm quality in adult male rats. Methods： Male Sprague-Dawley rats were injected s.c. with quercetin at the dose of 0, 30, 90, or 270 mg/kg body weight/day （hereafter abbreviated Q0, Q30, Q90 and Q270, respectively）, and each dose was administered for treatment durations of 3, 7 and 14 days. Results： From our study, it was found that the effects of quercetin on reproductive organs and sperm quality depended on the dose and duration of treatment. After Q270 treatment for 14 days, the weights of testes, epididymis and vas deferens were significantly increased, whereas the weights of seminal vesicle and prostate gland were significantly decreased, compared with those of Q0. The histological alteration of those organs was observed after Q270 treatment for 7 days as well as 14 days. The sperm motility, viability and concentration were significantly increased after Q90 and Q270 injections after both of 7 and 14 days. Changes in sperm quality were earlier and greater than those in sex organ histology and weight, respectively. Conclusion： Overall results indicate that quercetin might indirectly affect sperm quality through the stimulation of the sex organs, both at the cellular and organ levels, depending on the dose and the duration of treatment. Therefore, the use of quercetin as an alternative drug for treatment of male infertility should be considered. （Asian J Androl 2008 Mar. 10： 249-258）     

Methamphetamine induces abnormal sperm morphology, low sperm concentration and apoptosis in the testis of male rats

Methamphetamine has been reported to be an important drug in the field of reproductive toxicology. The aim of this study was to investigate the effects of methamphetamine administrations on sperm morphology, sperm concentration and apoptotic activity inside seminiferous tubule in male rats. Rats were administered a dose of 8 mg kg(-1) , intraperitoneally (IP), for acute group and a dose of 4 mg kg(-1) , IP, once daily for 14 days for sub-acute group. Percentage of normal sperm morphology was decreased in acute group when compared with control. Total numbers of sperm count were significantly decreased in acute and sub-acute groups. Apoptotic activities were most abundant in the seminiferous tubules of acute treated animals with a highly significant increase in the number of apoptotic cells per tubule. Those effects of methamphetamine seem to be dose-dependent. The results suggest that methamphetamine not only works as drug of abuse in central nervous system, but also in gametogenesis of males.     

Relationship between donor sperm parameters and pregnancy outcome after intrauterine insemination: analysis of 2821 cycles in 1355 couples

The aim of this study was to investigate whether sperm parameters can affect the pregnancy outcome of artificial intrauterine insemination with cryopreserved donor spermatozoon (AID). A total of 1355 couples received 2821 AID treatment cycles in the Reproductive Medicine Center of the Tongji Medical College between January 2010 and December 2013, and the data were collected and retrospectively analysed. The relationship between pre‐freezing, post‐thawing as well as optimised sperm parameters and AID pregnancy outcome was investigated. Clinical pregnancy rate and cumulated pregnancy rate were also calculated. A total of 728 cycles from 2821 treatment cycles achieved pregnancies, and cumulated pregnancy rate was 25.81%. Pre‐freezing progressive sperm motility in pregnant cycles was higher than that in nonpregnant cycles (P = 0.001); logistic regression analysis also indicated that pre‐freezing progressive sperm motility was the only parameter affecting pregnancy outcome (P = 0.0001). Our study also showed that the cumulated pregnancy rate increased progressively and reached a plateau after the fifth cycle. In conclusion, pre‐freezing progressive sperm motility should be a valuable predictor for AID pregnancy outcome. Female fertility factors should be considered, or IVF/ICSI should be recommended when couples received more than 5 AID cycles without pregnancy.