卡托普利体外对人精子运动参数的影响

目的:探讨卡托普利体外对人精子运动参数的影响。方法:将卡托普利注射液配制成1×10-5、1×10-6及1×10-7mol/L 3组浓度,分别在体外作用于取自正常生育男性并经Percoll梯度离心处理的精子。分别将3组卡托普利溶液各20μl与180μl精子悬液混匀,均于作用5 m in后采用计算机辅助精液分析系统检测精子运动参数。结果:3组不同浓度卡托普利溶液与精子作用5 m in后,精子活率和前向运动精子百分率明显降低(P<0.05);其他运动参数无明显变化。结论:卡托普利能显著降低精子活率和前向运动精子百分率,对其他运动参数无显著影响。     

输精管结扎术时精囊灌注消毒剂杀精效果的比较研究

应用消毒剂新洁尔灭、洗必泰醋酸盐,利凡诺作体外杀精子试验,其最低杀精浓度分别为0.25mg/ml、2.5mg/ml、7.5mg/ml。采用三种药物以最低杀精浓度总量8、10、14和16ml分别进行输精管结扎术时精囊灌注207例。临床观察表明:从精囊灌注后第一次尿中可排出精子3.6～60×10~6个,尿中精子总数在不同灌注量之间差异不明显,各药物组与生理盐水对照组无差异。术后第一次排精和术后1月取精液作常规检查者194例,未发现精子者分别为101例(52％)和169例(87.1％),其余虽有精子,但数量均在4×10~6/ml以内,并且三种药物分别灌注组的精子活力均完全消失。     

白藜芦醇对环磷酰胺诱导的人精子氧化损伤的保护作用乜照燕1,张亚楠2,花蕊2,陈丽君1,李亮1,吴海峰

目的 探讨白藜芦醇(RES)对环磷酰胺诱导的人精子氧化损伤的保护作用。方法 收集2018年1月至4月在河北医科大学第四医院生殖医学中心就诊男性患者的正常精液标本。将用密度梯度离心法处理后的正常精液标本用不同浓度(0、1.0、2.5、5.0、7.5μmol/L)环磷酰胺活性代谢产物4-过氧化氢环磷酰胺(4-HC)对精子进行体外毒性试验,继续培养24h,通过精子活力和精子存活率筛选4-HC作用半抑制浓度(IC50)。将处理后的正常精液用不同浓度RES(5、10、25、50、75μmol/L)预处理2h,再加入IC50浓度4-HC,测定精子活力和精子存活率选择RES最佳保护浓度。选择4-HC和RES浓度后,该实验分为四组:对照组(CON)、环磷酰胺组(4-HC)、环磷酰胺+白藜芦醇组(4-HC+RES)、白藜芦醇组(RES),检测各组精子活力、活率、氧化应激指标以及精子凋亡指标。结果 随4-HC浓度增加,精子活力和存活率降低,呈剂量依赖关系,依据IC50,2.5μmol/L4-HC为后续实验药物浓度。RES预处理后,精子活力和精子活率均得到改善,50μmol/L RES效果最佳,确定为后续实验药物浓度。结果显示:4-HC组较CON组,精子活力和存活率下降,氧化应激水平上升,精子凋亡增加,4-HC+RES组与4-HC组相比精子活力和存活率升高,氧化应激水平降低,精子凋亡降低。结论 RES可以减少化疗药物导致的精子氧化应激水平增加,能预防和缓解化疗药物对人精子造成的损伤。     

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.     

少精子症患者血清、精浆中游离睾酮水平的测定及意义

目的 :通过测定少精子症患者血清、精浆中游离睾酮 (FT)水平 ,分析血清、精浆FT与少精子症的关系。　方法 :正常对照组 (n =4 4 )、少精子症组 (n =4 4 )男性于上午 8:0 0～ 10 :0 0留取血标本 ;正常对照组 (n =30 )、少精子症组 (n =37)同时留取精液。男性精液常规分析判断精子密度 ,放射免疫分析法测定血清、精浆中FT水平。　结果 :少精子症患者血清中FT浓度为 [(94 .88± 4 2 .0 4 )pmol/L],与正常对照组 [(97.5 0± 4 6 .96 )pmol/L]相比差异无显著性 (P >0 .0 5 ) ,但少精子症患者精浆中FT浓度 [(0 .5 2± 0 .4 4 ) pmol/L]显著低于正常对照组 [(2 .0 1±0 .32 )pmol/L],P <0 .0 1。　结论 :精浆中FT的测定较早反映睾丸的功能 ,有利于少精子症患者的早期诊断和治疗。     

阿仑膦酸钠对大鼠成骨细胞增殖的影响

目的观察阿仑膦酸钠对大鼠成骨细胞增殖的影响,探讨局部使用防治人工关节无菌性松动的可行性。方法取2代培养的大鼠成骨细胞在阿仑膦酸钠为1×10-10、1×10-9、1×10-8、1×10-7、1×10-6、1×10-5、1×10-4mol/L浓度的培养液中培养,MTT检测阿仑膦酸钠对成骨细胞增殖的影响。结果10-4、10-5mol/L浓度的阿仑膦酸钠抑制成骨细胞增殖,1×10-9、1×10-10mol/L对成骨细胞的增殖没有影响,1×10-6、1×10-7、1×10-8mol/L促进成骨细胞的增殖,1×10-8mol/L对成骨细胞的增殖作用最强。结论阿仑膦酸钠有利于成骨细胞的增殖功能,并且这种作用具有最适合的浓度。     

黄芪注射液对镉诱导大鼠精子畸形的拮抗作用

目的 :观察黄芪注射液拮抗氯化镉诱导的大鼠精子畸形作用。　方法 :将 30只SD雄性大鼠随机分成 5组 :低浓度黄芪组 (A1)、高浓度黄芪组 (A2 )、环磷酰胺组 (CP)、氯化镉组 (Cd)和对照组 (C)。预先连续 7d分别给A1组和A2 组大鼠腹腔黄芪注射液 5g/ (kg·d)和 10g/ (kg·d) ,Cd组和CP组大鼠分别腹腔注射等量蒸馏水作对照 ,之后连续 2 1dA1组、A2 组和Cd组大鼠分别同时腹腔注射氯化镉溶液 [0 .2mg/ (kg·d) ]。CP组大鼠给予 5 0mg/ (kg·d)腹腔注射染镉后第 2 2d处死大鼠 ,观察睾丸脏器系数、精子头计数、每日精子生成量、附睾尾精子计数和畸形率以及睾丸和附睾病理学。　结果 :A2 组睾丸脏器系数、睾丸精子头计数、每日精子生成量和附睾精子计数 [(5 .6 8±1.19)、(4 9.0 1± 8.78)× 10 6/g、(10 .2 5± 2 .30 )× 10 6/ (g·d)、(4 7.5 1± 2 2 .5 1)× 10 6/ml]明显高于Cd组 [(3.11± 0 .16 )、(37.5 9± 10 .6 3)× 10 6/g、(5 .31± 0 .32 )× 10 6/ (g·d)、(10 .89± 2 .4 5 )× 10 6/ml](P <0 .0 5或P <0 .0 1) ;A2 组大鼠精子畸形率 [(7.0 4± 0 .12 ) % ]明显下降 ,与Cd组 [(17.81± 1.5 5 ) % ]相比 ,差异有极显著性 (P <0 .0 1)。　结论 :黄芪注射液可拮抗氯化镉对生殖系统的损害作用 ,对防护     

瘦素、神经肽Y对离体人卵巢颗粒细胞分泌功能的影响

目的　探讨瘦素、神经肽Y(NPY)对离体人卵巢颗粒细胞雌、孕激素生成的影响。　方法　将来自体外受精 胚胎移植的 2 5例离体人卵巢颗粒细胞纯化后 ,在不同浓度瘦素 (1、10、10 0 μg/L)、NPY(1× 10 -8、1× 10 -7、1× 10 -6mol/L)单独或与人绒毛膜促性腺激素 (hCG ,0、2 0IU/L)联合作用下体外培养 ,采用放射免疫方法测定培养液中雌二醇 (E2 )、孕酮(P)的水平。　结果　 (1)无论单独或与hCG联合不同浓度瘦素组E2 、P水平均无显著性差异 (P >0 .0 5 ) ;(2 )NPY浓度 1× 10 -7mol/L时 ,E2 水平显著低于空白对照组 (P <0 .0 5 ) ,但与hCG联合组显著高于单独组 (P <0 .0 5 ) ;(3)NPY浓度 1× 10 -7mol/L、瘦素浓度 10 μg/L组 ,E2 水平明显高于相应浓度NPY单独作用组 (P <0 .0 5 )。　结论　 (1)瘦素不直接影响体外培养颗粒细胞E2 、P的分泌 ,可能通过影响其它因子的作用而间接调节卵巢功能 ;(2 )NPY单独作用时 ,对颗粒细胞E2 的分泌有一定抑制作用 ,但当hCG存在时作用消失 ,说明在正常月经周期中NPY对黄体的生成和维持可能无明显不利影响 ;(3)瘦素可能阻断NPY对颗粒细胞分泌E2 的抑制作用 ,表明二者可能对卵巢功能有相互协调作用。     

精子顶体素ELISA检测法的建立与临床应用

从人精子中提取顶体素，制备兔抗人精子顶体素抗体，建立定量检测顶体素的ELISA方法。本法线性范围为（0．35～3．0）×10－3g／L，批内、批间cV分别为7．8％～9．6％和8．4％～10．8％。临床检测结果表明，在精子密度为50×109／L，生育组（n=15）顶体素含量（X±S）为（1．04±0．10）×10－3g／L；不育组（n=30）顶体素含量为（0．81±0．18）×10－3g／L，两组之间差异显著（P＜0．01），顶体素含量与精子密度、精子顶体完整率之间呈正相关（r=0．987、r=0．821），提示精子顶体素含量的降低是引起男性不育的原因之一。     

活性氧致精子运动参数变化的观察

我们采用次黄嘌呤 黄嘌呤氧化酶反应体系产生活性氧 (ＲＯＳ) ,在体外与具有正常生理功能的精子作用 ,观察精子运动参数的变化 ,报告如下。材料与方法　正常生育男性及本院男科门诊不育男性 68例。精子密度 >60× 1 0 6 /ｍｌ,ａ +ｂ级精子 >50 % ,ＷＢＣ<1× 1 0 6 /ｍｌ。禁欲 3～ 5ｄ后 ,手淫法留取精液于洁净玻璃容器内。按文献方法制备正常精子[1 ] 。将制备好的正常精子悬液 1式 4份 ,每份 0 .5ｍｌ分四组 :ａ组 ,精子悬液 ;ｂ组 ,精子悬液 +次黄嘌呤 (终浓度 1ｍｍｏｌ/Ｌ) ;ｃ组 ,精子悬液 +黄嘌呤氧化酶 (终浓度 50ｍＵ/ｍｌ) ;ｄ…     

体外rhTNF-a对人精子运动、线粒体功能影响的实验研究

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白念珠菌体外感染对人精子运动和超微结构的影响

目的: 体外研究白念珠菌(Ca)对人精子的运动功能有无直接影响并观察精子超微结构变化,对其影响机制作初步探讨。　方法: 将从念珠菌性阴道炎患者的分泌物中分离纯化的Ca制成活菌悬液,对 10例健康青年男性手淫获得并经上游优化处理的精子进行体外感染。按细菌与精子比例不同分成A组 ( 1∶1 )、B组 ( 1∶10 )、C组(1∶100)、D组(1∶1 000)、E组(1∶10 000)、F组(空白对照)分别孵育,于 0、1、2、4h取样,计算机辅助精子分析系统(CASA)检测人精子运动参数(前向性运动百分率、直线速度、曲线速度、平均路径速度、头部侧摆幅度 )、精子存活率、精子形态及凝集情况。透射电镜观察孵育 4h后的各组精子的超微结构变化。　结果: Ca与精子体外孵育后,前向运动百分率所受影响最大,且与菌体密度及时间密切相关;其他参数与对照组相比也相继出现显著性差异。观察到精子黏附于菌体和凝集现象。精子超微结构产生变化:精子核空泡增多,顶体破裂,质膜破损,线粒体排列紊乱。　结论: Ca体外可显著降低精子存活率和抑制精子运动功能,其机制可能与Ca对人精子的黏附作用和超微结构的损伤有关。     

Evaluation of Hamilton-Thorn automated semen analysis system.

Sperm concentration and percentage motility values generated by the HT M-2030 system (CASA) were compared with those obtained by subjective semen analysis (SSA). Three calibrations arbitrarily designated as A, M, and D for gates and discriminators and three calibrations arbitrarily designated LI, MI, and HI for default pixel count (DPC) and default intensity (DI) were evaluated. The best correlation between CASA and SSA was observed utilizing M calibration (n = 114) with which sperm concentration was +0.3 x 10(6)/mL (r = .96) and motility was -6.3% (r = .89) compared to the values obtained by SSA. It was found that 35.9% of sperm concentration values and 34.2% of sperm motility values were within 10% of the values obtained by SSA. When sperm concentration was between 50 and 100 x 10(6)/mL the difference in motility was reduced (-3.2%) while the difference in sperm concentration was reversed (-2.6 x 10(6)/mL). LI calibration (DPC = 4, DI = 86) gave acceptable results with M calibration for sperm concentration (+2.1 x 10(6)/mL) and motility (-6.9%) compared to the values obtained by SSA. In the presence of sperm clumping, the difference between CASA and SSA was reversed for sperm concentration (+0.56 x 10(6)/mL for normal samples vs. -2.2 x 10(6)/mL for samples with clumping) and was reduced for sperm motility (-7.14% vs. -4.55%, respectively). HT M-2030 under proper calibration can be used as a rapid, objective, and reliable alternative to conventional semen analysis in routine and for research purposes.     

Predictive factors associated with successful varicocele repair a study of 139 infertile men with valicocele

PURPOSE: To determine pretreatment parameters which predict improvements following varicocele repaire in semen quality. MATERIALS AND METHODS: We retrospectively evaluated a total of 139 infertile patients who underwent varicocelectomy from February 1995 to March 2000. A logistic regression analysis was performed to identify parameters associated with improvements in semen quality. Parameters evaluated included varicocele grade, age, testicular volume, serum testosterone, liteinizing hormone (LH), follicle-stimulating hormone (FSH), preoperative sperm density and sperm motility. RESULTS: Of 139 patients 71 (51.0%) improved sperm concentration and 59 (42.4%) improved sperm motility postoperatively. Overall, median sperm density significantly increased from 10 x 10(6)/ml preoperatively to 30 x 10(6)/ml postoperatively. Sperm motility also significantly increased 33% to 45%. In logistic regression analysis, varicocele grade (odds ratio [OR] = 5.7; 95% confidential interval [CI : 1.9-17), FSH level ([OR] = 0.76; [CI]: 0.60-0.96) and sperm motility ([OR] = 1.03; [CI]: 1.0-1.1) were independent predictive factors for improvement in sperm concentration. CONCLUSION: Varicocelectomy improves sperm concentration and motility. Our data suggest that patients with grade 3 varicocele, low serum FSH level and high sperm motility are more likely to benefit from varicocele repair in sperm concentration.     

Effects of exposure to new inhalational anesthetics on spermatogenesis and sperm morphology in rabbits

The effects of chronic exposure to new inhalation anesthetics (sevoflurane and isoflurane) on spermatogenesis and sperm morphology were examined in 23 rabbits, randomly divided in 3 groups. Rabbits received 20 exposure hours (four hours/day x 5 days), as follows: group I: 2.3% (1.2 MAC) sevoflurane + 2L/min oxygen, group II: 1.3% (1.2 MAC) isoflurane + 2L/min oxygen, and group III (control): 2L/min oxygen. Semen was collected on the 12th, 19th, 26th, 33rd, and 41st days of exposure. Sperm concentration, motility and morphological changes were evaluated. On the 41st day, testicular biopsies were taken and observed with light microscopy. Sperm concentration and motility significantly decreased in the sevoflurane and the isoflurane groups, compared to control. There were no significant changes in the control group. It is concluded that chronic exposure to the new inhalational anesthetics had negative effects on spermatogenesis and sperm morphology.     

硝苯地平对人精子膜功能完整性及压力敏感性Ca~(2+)内流影响的实验研究

目的:探讨模拟正常血药浓度下硝苯地平体外对人精子膜功能完整性及压力敏感性Ca2+内流的影响。方法:将10例生育男性的精液经上游优化处理后,分别与20、100、20×103ng/ml的硝苯地平在体外共同孵育60min,同时设立对照组进行低渗肿胀(HOS)试验及测定压力导致的精子细胞内钙离子浓度的变化。结果:20×103ng/ml组的精子HOS百分率明显低于对照组,差异具有显著性(P<0.01)。20、100、20×103ng/ml组的精子细胞内Ca2+荧光差值均明显低于对照组,差异具有显著性(P<0.01)。结论:硝苯地平在体外可以改变精子膜功能完整性,抑制压力敏感性Ca2+内流。治疗剂量的硝苯地平在体外可影响精子质量。     

Free radical scavenger effect of rebamipide in sperm processing and cryopreservation

Aim: To study the effect of rebamipide added to semen samples and cryoprotectant on reactive oxygen species (ROS) production. Methods: Semen samples from 30 fertile and healthy volunteers were collected by masturbation after 2 days - 3 days of abstinence. After liquefaction, the specimens were diluted with sperm wash media to a uniform density of 20×106/mL. Rebamipide was added to semen samples and cryoprotectant to a final concentration of 10 μmol/L, 30 μmol/L, 100 μmol/L or 300 μmol/L. Specimens were incubated at 37 ℃ in a 0.5 % CO2 incubator for 1 h or cryopreserved at -196 ℃ LN2 for 3 days. The sperm motility and viability and the levels of ROS and lipid peroxidation of sperm membrance were assessed before and after incubation and cryopreservation by means of computer assisted semen analyzer, eosin-nigrosin stain, chemiluminescence and thiobarbituric acid assay, respectively. Results: The sperm motility was significantly increased after incubation with 100 μmol/L and 300 μmol/L rebamipide (P     

Secular and seasonal changes in semen quality among young Danish men: a statistical analysis of semen samples from 1927 donor candidates during 1977-1995

The objective of this study was to investigate whether semen quality has changed during the years 1977-1995 in a group of unselected semen donor candidates, and to determine whether semen quality is subject to seasonal variation, by analysis of time- and season-related changes in semen quality using multiple regression and ANOVA. The study was based on analysis of the first semen sample delivered by 1927 semen donor candidates in Copenhagen during the period 1977-1995, with determination of semen volume, sperm concentration, total sperm count, percentage motile spermatozoa, and a semiquantitative sperm motility score. Multiple linear regression analysis with year, sexual abstinence and season as covariates showed a significant increase in mean sperm concentration from 53.0 x 10(6)/mL in 1977 to 72.7 x 10(6)/mL in 1995 (p < 0.0001) and in mean total sperm count from 166.0 x 10(6) to 227.6 x 10(6) (p < 0.0001). Mean semen volume and percentage motile spermatozoa did not change. Sperm motility deteriorated, as the spermatozoa in 74.2% of the samples were of excellent motility in 1977-1980 compared to only 41.9% in 1993-1995 (chi 2 = 130.0, p < 0.0001). Analysis of variance showed significant variation between seasons regarding sperm concentration (p < 0.0001) and total sperm count (p < 0.0001). Highest sperm counts were found in spring, with a mean concentration (95% C.I.) of 77.6 x 10(6)/mL (71.9-83.7), and lowest in summer, with a mean of 57.5 x 10(6)/mL (50.1-65.4). No other semen parameter varied with season. It is concluded that sperm counts increased, whereas sperm motility decreased, in a group of Danish semen donor candidates, from 1977 to 1995. Due to the retrospective design and the anonymity of the donors, we were unable to control for variation in donor age, and we cannot exclude the possibility that some donor candidates were selected by being accepted as donors by other semen donor services in Copenhagen. With these limitations in mind, we suggest our results should be interpreted cautiously and regarded as a contribution to the ongoing dispute on whether or not there is a continuous decrease in sperm quality. The seasonal variations found in sperm concentration and total sperm count were pronounced and were not attributable to seasonal differences in the length of sexual abstinence. Additionally, the same seasonal pattern was observed in five successive year-intervals. These findings strongly indicate that human testicular function is influenced by season, a phenomenon well known in many lower mammals.     

Sperm count and motility influence the results of human fertilization in vitro

In order to select sperm characteristics that can predict the outcome of in-vitro fertilization-embryo transfer (IVF-ET), semen samples delivered in conjunction with this treatment were studied carefully. We have analysed these data retrospectively in relation to the outcome of treatment. Ninety-one couples were treated for tubal infertility by IVF-ET. Fifteen women became pregnant. Sperm were isolated from semen using a swim-up technique and in most cases 40-80 x 10(3) (range 20-120 x 10(3)) motile sperm per ovum were used for insemination. The couples were divided into three groups: group A who achieved pregnancies, group B who achieved cleaved ova but no pregnancies, and group C who achieved no ova that were cleaved 48 h after oocyte recovery. Comparisons between these groups showed that some characteristics of the native semen samples and the swim-up preparations were significantly different: the sperm concentration (P = 0.001) and total sperm count (P = 0.003) in the native sample, the number of sperm recovered during 30 min of swim-up (P = 0.001), and the specific progressive motility of sperm in the swim-up preparation, both at the time of insemination and on each day, up to 5 days thereafter (P = 0.002-0.028). No pregnancy was achieved with a sperm concentration below 26 x 10(6) ml-1 in the native sample. Some of the sperm characteristics studied in this paper may be of value in the pretreatment evaluation for IVF treatment.