Quantitative assessment of the serum-induced stimulation of human sperm motility

In previous studies, sperm populations with high motility and fertilizing potential have been isolated using Percoll gradients and/or addition of serum or caffeine, but no objective measurements of the motility parameters affected have been presented. In addition, the types of sperm populations and the incidence of positive or negative responses to these treatments have never been documented. In this paper, the proportion of samples responding positively or negatively to these treatments and the type of motility stimulation observed was measured using the CellSoft computer-assisted digital image analysis system. Semen, obtained from the Infertility Clinic, was washed on discontinuous Percoll gradients. Sperm motility parameters were measured in the original semen and after washing through a Percoll gradient in the presence or absence of serum or caffeine. Percoll washing permitted a good recovery of essentially pure sperm. The Percoll treatment had little effect on the percentage of motile sperm but caused a decrease in the linearity-related parameters. A high proportion of the washed sperm samples supplemented with serum showed an increase in at least one of the motility parameters when compared to sperm populations in the original semen (68% of cases) or after Percoll washing (96% of cases). These effects were maintained for several hours. The stimulation of motility was concentration-dependent, with an optimum around 25-30% serum. Caffeine (4 mM) increased linear velocity but lowered linearity. In contrast to caffeine, which stimulated sperm motility for less than 1 h, the effects of human serum lasted for more than 16 h.     

Effects of intestinal electrical stimulation on postprandial small-bowel motility and transit in dogs

Background

Intestinal electrical stimulation (IES) with long pulses has been reported to inhibit motility as well as accelerate transit of continuous infusion. However, it is unknown whether there is a correlation between the IES-induced alterations in motility and transit and whether there is a difference in transit during IES between continuous infusion and bolus infusion.

Methods

The study was performed in 2 postprandial sessions (control and stimulation) in dogs with 2 pairs of serosal electrodes and 2 intestinal cannulas. Intestinal motility and transit with and without IES were measured by manometry and phenol red, respectively.

Results

IES significantly decreased intestinal motility and increased transit time. There was a significant correlation between motility index and transit during IES.

Conclusions

IES inhibits both intestinal bolus motility and transit. There is correlation between motility and transit during IES.     

Caffeine- and kallikrein-induced stimulation of human sperm motility: a comparative study

Stimulation of human sperm motility by caffeine and kallikrein was compared in the same semen material. Caffeine, a cyclic nucleotide phosphodiesterase inhibitor, induced an immediate stimulation of sperm motility the intensity being relatively constant during the first two hours of incubation. Kallikrein, a kinin-releasing proteinase, induced a similar enhancement of total sperm motility, but showed a delayed type of reaction with maximum stimulation at 2 hours of incubation. In contrast to the effect of caffeine lasting some hours, enhancement of sperm motility induced by kallikrein was observed 24 hours. Simultaneous addition of serum (kininogen source) and kallikrein to semen samples led to a sitmulation of total sperm motility with higher mean values than those obtained by caffeine of kallikrein alone. However, the ratio of spermatozoa with very good forward progression was highest during caffeine stimulation. Simultaneous addition of caffeine and kallikrein led to a further improvement of sperm motility which was significantly above that produced by caffeine or kallikrein alone. This observation and the finding of a different response of the spermatozoa of two ejaculates towards caffeine of kallikrein indicate that caffeine (cyclic AMP) interferes quite differently in comparison to kallikrein (kinins) in stimulating sperm motility.     

弗司扣林对人精子运动功能的影响

目的 通过研究弗司扣林(forskolin)对体外人精子的运动功能有无影响，了解环磷酸腺苷／蛋白激酶A(cAMP／PKA)信号传导途径是否参与人精子运动功能的调节。方法 10例健康成年男性手淫获得新鲜精液，经上游优化处理后与不同浓度的forskolin一起孵育20、30、60min后，采用计算机辅助的精子分析系统(CASA)检测精子的各项运动参数，并进行对比分析。结果 forskolin在体外能显著提高人精于的活率及前向性运动百分率，而对精子的形态及直线速度(VSL)和曲线速度(VCL)无明显影响。结论 forskolin在体外能提高人精子的运动功能，此结果为证实cAMP/PKA信号传导通道参与调节人精子运动功能提供了一定的实验依据。     

Horsley was the first to use electrical stimulation of the human cerebral cortex intraoperatively

Fedor Krause has previously been credited with being the first surgeon to use electrical stimulation to define epileptic foci in the human cerebral cortex before extirpation. We provide evidence indicating that Krause was probably preceded by Horsley, and Bidwell and Sherrington in England, and by Keen in the United States. Sir Victor Horsley apparently first used electrical stimulation diagnostically in 1884 and slightly later (1886) to define epileptic foci.     

光量子体外照射对精子活力的影响

目的 观察光能对精子活力的影响,探索一条体外不去精浆、提高精子活力的方法。方法 通过对54例因精子活力低下引起男性不育患者的精液,体外应用SB-99型血液处理器进行UV(紫外线)照射,观察同份精液在处理前后活动精子百分率和应用激光光散射测定精子前向运动速率,同时作细菌培养和细菌计数,并作精浆SOD和精浆粘稠度测定。结果 精子速率、活动精子百分率、活力得分、精浆粘稠度及SOD活性处理后较处理前均有明显改善(P<0.01),当UV>500J/cm~3以上,UV照射不同剂量组间精子活力无显著差异(P>0.05);处理最适宜的时间为:3min1000J/cm~3在3～5min时已显示杀菌作用。结论 UV照射引起精浆粘稠度降低、杀灭细菌和SOD活性增加,可能是其提高精子活力的原因。     

Effect of zinc on human sperm motility and the acrosome reaction

This study has assessed the effect of zinc on human sperm motility and the acrosome reaction in vitro. Progressively motile human sperm were selected by swim-up and by glass bead columns and then incubated in a medium in which capacitation happened in an asynchronous way. Different doses of zinc (1, 10, 100 and 1000 microM) were added for periods of 2, 4 or 6 h. Other samples were incubated with zinc (1000 microM), and after 1 h incubation, the zinc was removed. Aliquots of each culture were used to evaluate progressive motility and the acrosome reaction using a triple-stain technique. Sperm motility was reduced when the amount of zinc added was greater than or equal to 100 microM, and these doses also caused a significant reduction in the % of sperm undergoing the acrosome reaction. After removal of zinc and further incubation in zinc-free medium for 1 h, an increase in the percentage of motile and acrosome-reacted sperm was observed. However, the increase in acrosome reaction did not reach the values observed in controls. Results suggest that extracellular zinc acts as an inhibitor of human sperm motility and the acrosome reaction (and/or capacitation and the acrosome reaction). This inhibitory effect is reversible and occurs in a dose-dependent fashion. The probable mechanisms involved are discussed.     

重组人钙调素对精子活动功能的影响

目的：探讨重组人钙调素（ｒｈＣａＭ）对精子活动功能的影响。　方法：用基因工程手段制备并纯化ｒｈＣａＭ。取不育男性精液标本,精子经Ｅａｒｌｅｓ 液处理后,校正精子密度为（５～５０）×１０９／Ｌ,一式３ 份,加入不同浓度的ｒｈＣａＭ,观察不同浓度ｒｈＣａＭ、不同孵育时间对精子运动功能的影响。　结果：２５例活动能力差的精子经孵育１ ｈ,活率未见显著变化（Ｐ＞ ０．０５）,前向运动速度则显著改变（Ｐ＜ ０．０１）,另８ 例活动能力良好的精子,孵育１ ｈ 前向运动速度、活率均无显著性改变（Ｐ＞ ０．０５）。９ 例活动能力差的精子经孵育１ ｈ、２ ｈ 后,活率无显著变化（Ｐ＞ ０．０５）,而前向运动速度、摆幅、鞭打频率改变显著。　结论：外源ｒｈＣａＭ 对精子的活动功能有显著的促进作用,ｒｈＣａＭ 在此过程中可能参与了精子获能前的准备工作。     

The effects of intermittent electrical stimulation on the prevention of deep tissue injury: varying loads and stimulation paradigms

A pressure ulcer is a medical complication that arises in persons with decreased mobility and/or sensation. Deep pressure ulcers starting at the bone-muscle interface are the most dangerous, as they can cause extensive damage before showing any signs at the skin surface. We previously proposed a novel intervention called intermittent electrical stimulation (IES) for the prevention of deep tissue injury (DTI). In this study, we tested the effects of four paradigms of IES and one conventional pressure relief paradigm in preventing the formation of deep pressure ulcers in rats. Loading equivalent to 18, 28, or 38% of the body weight (BW) of each rat was applied to the triceps surae muscle in one hind limb. Treatment groups received IES every 10 min for either (i) 5 or 10 s with moderate or maximal contraction, or (ii) complete pressure removal every 10 min for 10 s (conventional pressure relief). The results showed that conventional pressure relief, emulating a wheelchair push-up every 10 min, was inadequate for the prevention of DTI. In contrast, all IES paradigms were equally effective in significantly reducing the extent of deep muscle damage caused by 28 or 38% BW pressure application. These findings suggest that, in conjunction with existing techniques, IES may be an effective intervention for the prophylactic prevention of DTI.     

Long-term effects of transabdominal electrical stimulation in treating children with slow-transit constipation

Aims

Transcutaneous electrical stimulation (TES) was used to treat children with slow-transit constipation (STC) for 1 to 2 months in a randomized controlled trial during 2006 to 2008. We aimed to determine long-term outcomes, hypothesizing that TES produced sustained improvement.

Methods

Physiotherapists administered 1 to 2 months of TES to 39 children (20 minutes, 3 times a week). Fifteen continued to self-administer TES (30 minutes daily for more than 2 months). Mean long-term follow-up of 30 of 39 patients was conducted using questionnaire review 3.5 years (range 1.9-4.7 years) later. Outcomes were evaluated by confidence intervals or paired t test.

Results

Seventy-three percent of patients perceived improvement, lasting more than 2 years in 33% and less than 6 months in 25% to 33%. Defecation frequency improved in 30%. Stools got wetter in 62% after stimulation and then drier again. Soiling improved in 75% and abdominal pain in 59%. Laxative use stopped in 52%, and 43% with appendicostomies stopped washouts. Soiling/Holschneider continence score improved in 81% (P = .0002). Timed sits switched to urge-initiated defecations in 80% patients. Eighty percent of relapsed patients elected to have home stimulation.

Conclusion

TES holds promise for STC children. Improvement occurred in two thirds of children, lasting more than 2 years in one third, whereas symptoms recurred after 6 months in one third of children.     

睾酮体外对人精子运动参数的影响

目的通过研究睾酮体外对人精子运动参数的影响,探讨睾酮在男性不育症治疗中的作用。方法10例健康生育男性手淫获得精液,经上游优化处理后的精子与不同浓度的睾酮孵育10、30、60min,10例弱精子症患者手淫取精并与睾酮孵育10、60、120min后,采用计算机辅助的精液分析系统(CASA)检测精子的运动参数。结果50ng/dl(1.73nmol/L)睾酮在体外能显著增强正常人精子的直线速度(VSL)、曲线速度(VCL)和平均速度(VAP),而对活率、前向运动百分率无明显影响,而100ng/dl(3.47nmol/L)睾酮使精子活率和前向性运动百分率均有明显下降(P<0.01);1.04nmol/L～1.73nmol/L浓度睾酮能显著增强弱精子症患者精子的活率、前向运动百分率及VSL,并随着浓度的增加,睾酮作用显著增强,而对VCL和VAP无明显影响。结论低浓度(1.04nmol/L～1.73nmol/L)睾酮在体外显著增高弱精症患者精子的运动参数。     

rhGM-CSF体外对人精子运动参数的影响

目的观察体外添加不同浓度的重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)对人精子运动参数的影响,探讨其在精子运动中的作用机制。方法健康生育男性和弱精子症患者各10例手淫取精,经简易上游优化处理后的精子与不同浓度rhGM-CSF溶液孵育10min、30min、60min后,采用计算机辅助的精液分析系统检测精子各项运动参数的变化。结果对精液运动参数正常的标本,1ng/ml和10ng/ml的rhGM-CSF溶液能显著提高精子的活率、前向性运动百分率,在10ng/ml时平均直线运动速度也有显著提高。对弱精子症患者精液标本,1ng/ml和10ng/ml的rhGM-CSF溶液能显著提高精子的前向运动百分率和直线运动速度,10ng/ml的rhGM-CSF对精子活率和平均路径速度也有显著性提高。两类精子的曲线运动速度均无显著性变化。结论1ng/ml～10ng/ml浓度范围的rhGM-CSF体外能显著改善精子的运动功能。     

Clinical elements for the neuromuscular stimulation and functional electrical stimulation protocols in the practice of neurorehabilitation

The physicians and their multidisciplinary teams involved in the clinical practice of neurological rehabilitation have more and more opportunities to apply neuromuscular stimulation (NMS) and functional electrical stimulation (FES) of peripheral nerves as a part of their daily practice. In this article, we outline clinical protocols of NMS and FES in the following clinical conditions of upper motor neuron dysfunction: to prevent consequences of disuse of the neuromuscular system of the upper motor neuron, to facilitate recovery processes of impaired upper motor neuron functions due to acute and/or subacute neurological conditions, to maintain or enhance the trophic state of the muscle, to modify altered control of muscle tone, to modify altered patterns of automatic and volitional functional movements, to enhance functional movement of the single joint muscle group within intact functional multijoint movement, and to modify altered neurocontrol of posture, locomotion, and skillful movements. We emphasize the importance of understanding the motor control alteration while developing clinical protocols and defining the goals. It is very important to be aware that similar clinical findings and due to the same cause can have different features of residual motor control, and therefore potentials for recovery or modification can be very different.     

己酮可可碱对精子活力的体外改善作用

目的体外分析己酮可可碱(Pentoxifylline，PF)对精子活力的改善作用，探讨PF最佳作用浓度和时间，为实施人工授精奠定基础。方法本实验就PF的不同浓度(0．6mmol／L、3．0mmol／L)和不同时间(30min，60min)，恒温下与70例弱精子的体外孵育，并与对照比较，观察精子活力特性的改善情况。结果PF的不同浓度及不同时间对精子活力均有改善作用，而用PF的0．6mmol／L浓度、孵育30min，对体外弱精子的活力有显著增强作用。结论PF对体外弱精子的活力有改善作用，药物浓度和作用时间直接影响孵育效果。     

Transvaginal electrical stimulation in the treatment of urinary incontinence

OBJECTIVE: To determine the effectiveness of transvaginal electrical stimulation (TES) in treating urinary incontinence, and to assess the clinical improvement 6 months after ending the treatment. PATIENTS AND METHODS: In a double-blind randomized controlled clinical trial, 36 women (24 patients and 12 controls) with stress, urge or mixed urinary incontinence were chosen to use TES or placebo (identical equipment but with no electrical current). The patients had their treatment at home twice a day (20-min sessions) for 12 weeks. They completed a voiding diary and had a urodynamic study at the beginning and end of treatment. They were clinically re-evaluated after 6 months. RESULTS: The mean time of use of TES was similar in both groups (approximately 40 h); the treatment group had a significant increase in maximum bladder capacity (P < 0.02), a significant reduction in the total number of voids (over 24 h; P < 0.02), in the number of episodes of voiding urgency (P < 0.001) and, importantly, in the number of episodes of urinary incontinence (P < 0.001). At the first evaluation, after ending the treatment, 88% of the patients had a significant reduction in symptoms or went into remission. At the 6-month re-evaluation, a third of the patients required another therapeutic approach. CONCLUSION: TES is a practical alternative with few side-effects, and is effective for treating the main forms of female urinary incontinence.     

不同阿片类镇痛药对体外人精子活力的影响

目的 探讨阿片类镇痛药芬太尼、阿芬太尼、舒芬太尼、布托啡诺、地佐辛和喷他佐辛对体外人精子活力的影响.方法 男性患者20例,取20份活力正常的精液标本,密度(40～ 80)×106个/ml,接种于培养皿,每皿50μl.采用随机区组设计,分别接受下述处理.对照组:每皿加入等渗盐水50μl;单独用药组:6种阿片类镇痛药分别加入到培养皿中,使其终浓度为10-3、10-4、10-5和10-7mol/L;联合用药组:纳洛酮加入培养皿中,使其终浓度为2× 10-8、1 × 10-7、2×10-7 mol/L,孵育5min后分别加入各浓度的芬太尼、阿芬太尼、舒芬太尼、布托啡诺、地佐辛25μl,各组孵育1h.计算精子活力变化幅度、半数抑制浓度(IC50)和完全抑制浓度(IC100).结果 芬太尼、阿芬太尼和舒芬太尼浓度与精子活力变化幅度的量效曲线均呈直线,精子活力最大降低幅度分别为(56±5)％、(58±7)％和(79±6)％;布托啡诺、地佐辛和喷他佐辛浓度与精子活力变化幅度的量效曲线均呈S型曲线,低浓度均对精子活力无影响,中、高浓度布托啡诺和地佐辛可抑制精子活力,精子活力最大降低幅度均为100％,高浓度喷他佐辛显著增加精子活力,精子活力最大升高幅度(19±6)％.纳洛酮使芬太尼、阿芬太尼、舒芬太尼浓度与精子活力变化幅度的量效曲线右移,最大效应降低,且呈浓度依赖性(P<0.05),使布托啡诺、地佐辛浓度与精子活力变化幅度的量效曲线右移,最大效应不变(P＞0.05),呈浓度依赖性(P<0.05).IC50排列顺序为阿芬太尼≈芬太尼＞地佐辛＞舒芬太尼≈布托啡诺,IC100排列顺序为地佐辛＞布托啡诺.结论 芬太尼、阿芬太尼、舒芬太尼可通过激动精子阿片受体直接抑制精子活力;布托啡诺和地佐辛对精子活力的抑制作用与浓度有关,该作用可能与阿片受体和非阿片受体途径有关;喷他佐辛增强精子活力的作用与浓度有关.     

Laser light-scattering study of the toxic effects of methylmercury on sperm motility

An in vitro study was designed using the laser light-scattering technique to obtain further information on the dose-effect relationship of methylmercury on sperm motility. The technique provided a quantitative evaluation of sperm swimming speed. Semen samples were collected from normal male Macaca fascicularis monkeys by anal electroejaculation. Methylmercury was added to aliquots of sperm suspensions in BWW medium in doses of 10, 5, 2, and 1 ppm. After 3 hours, the relative speed was 35%, 59%, 69%, and 92% of the corresponding controls at doses of 10, 5, 2, and 1 ppm, respectively. The percentage of motile spermatozoa decreased significantly at 10 ppm. By microscopic observation abnormal motility was detected at 5 and 10 ppm, especially after 20 to 40 minutes. Head movement increased from side to side, and many spermatozoa developed coiled tails. The technique proved useful for defining the dose-effect relationship of methylmercury and sperm swimming speed.     

雷米芬太尼和纳洛酮联合体外应用对人精子运动功能的影响

目的:观察雷米芬太尼和纳洛酮联合应用对体外人精子运动功能的影响并探讨其机制。方法:实验选取20例a+b级精子百分率正常的精液标本,每例精液均一式13份,分别经过不同浓度雷米芬太尼或联合不同浓度纳洛酮处理35 min,在计算机辅助的精子分析下观察精子5、10、15、20、35 min时的运动情况。结果:①与对照组相比,0.1～100μg/L雷米芬太尼作用精子5、10 min时a+b级精子百分率降低,并呈浓度依赖性降低,而作用精子15、30 min时a+b级精子百分率无显著性差异;②与对照组相比,0.004～0.04 mg/L纳洛酮作用精子5～35 min时,a+b级精子百分率无显著性差异;0.4～4 mg/L纳洛酮作用精子5～20 min时a+b级精子百分率无显著性差异,而作用精子35 min时a+b级精子百分率显著增加;③与同浓度雷米芬太尼组相比,0.004、0.04、0.4、4 mg/L纳洛酮依次联合0.1、1、10、100μg/L雷米芬太尼共同作用精子5、10 min时a+b级精子百分率显著提高,而作用15、35 min时无显著性差异。结论:雷米芬太尼对精子运动的抑制作用起效和消失快,其抑制作用可被纳洛酮所拮抗,推测可能与μ受体有关。     

Transforming growth factor βreceptor Ⅱ expression in experimental cryptorchidism and apoptosis in spermatogenic cells in rats

Objective: To evaluate the sensitivity of sperm motility assay for detecting the endotoxin effect on human sperm in vitro. Methods: Motile human sperm were separately incubated for up to 24 hours with different concentrations of endotoxin (0.5, 1, 10, 1000, 10 000 and 50 000 ng/mL). Then the sperm motility was determined. The effect of endotoxin on the sperm motility in media without albumin was also determined. In addition, at the endotoxin concentrations of 0.5, 1 and 10 ng/mL, the sensitivity of the assay was compared to those of 1-cell and 2-cell mouse embryo bioassays. Results: At levels of 0.5-1 000 ng/mL endotoxin in media with 2 mg/mL albumin, sperm did not show significant change in motility after 24 h of incubation (P>0.05), while it was significantly inhibited at endotoxin levels of 10 000 and 50 000 ng/mL. In media without albumin, endotoxin levels of 50 000 and 1 000 ng/mL, markedly inhibited the sperm motility after 2 or 8 h of incubation (P<0.01). With media containing 0.5 and 1 ng/mL endo     

Direct effect of alcohol on the motility and morphology of human spermatozoa

Excessive alcohol consumption has been associated with impaired reproductive function by causing the inhibition of penile tumescence and ejaculatory capability. Alcohol intoxication has also been implicated in impaired spermatogenesis and an increase in sperm structural anomalies. The aim of this study was to determine the direct effects of alcohol on sperm motility and morphology in vitro.
Semen samples from 67 subjects were prepared using density centrifugation. Ethanol was added, at concentrations in serum equivalent to social, moderate and heavy drinking, to the medium in which the spermatozoa were cultured. Sperm motility was assessed using computer assisted semen analysis and morphology was assessed by Tygerberg strict criteria after 0, 15, 30, 60, 120, 180 and 240  min exposure. Each concentration of ethanol produced significant decreases in the percentage progressive motility, straight line velocity and curvilinear velocity. The amplitude of lateral head displacement was also depressed by 300 and 500  mg  dL⁻¹ of ethanol. A significant decrease in the number of spermatozoa with normal morphology and an increase in irreversible tail defects were observed after exposure to 300  mg  dL⁻¹ ethanol.
When alcohol is added directly to sperm, at concentrations equivalent to those in serum after moderate and heavy drinking, damaging effects are observed in both sperm motility and morphology.