大鼠精子在附睾成熟中精子膜变化的研究

我们运用Ｐｅｒｃｏｌｌ离心技术对ＳＤ大鼠附睾头、体、尾各段的精子分离纯化后，再用硫代巴比妥酸法、酶法、ＳＤＳ－ＰＡＧＬ电泳技术等对精手膜依次进行唾液酸、甘油－３－磷酸胆碱（ＧＰＣ）及蛋白质含量变化的检测。结果显示：精子膜唾液酸、ＧＰＣ量不断降低且有显著统计学意义（Ｐ＜０．０１）。附睾头、体、尾各段精子膜唾液酸和ＧＰＣ量分别为１０．１８±２．８２、８．４２±３．０７、７．８３±２．７９μｇ／１０８精子；１１２．３１±２８．１４、１０９．３３±３７．１６、７４．５０±２５．１３ｎｍｏｌ／１０８精子（－ｘ±ｓ）。膜蛋白变化主要是由大分子蛋白转变成较小分工的蛋白。大鼠精子附睾转运中精子膜的变化与精子成熟具有十分密切的关系。     

精索静脉曲张不育患者精子核成熟性研究

利用苯胺兰细胞化学染色法和ＳＤＳ、ＳＤＳ一ＥＤＴＡ处理精子研究精子核的成熟性。结果发现，２１例精索静脉曲张不育患者精子核蛋白存在明显的转型缺陷（不育组２７．８２±９．７３，对照组４０．６２±１１．７３，Ｐ＜０．０１）；其精子核抗解聚能力明显下降，Ｐ＜０．０１（ＳＤＳ）和＜０．０５（ＳＤＳ一ＥＤＴＡ）；α一糖苷酶活性越低，核蓝染精子百分数越高（ｒ＝－０．８１，Ｐ＜０．０１），而核稳定精子百分数越低（ＳＤＳ组ｒ＝０．７５；ＳＤＳ一ＥＤＴＡ组ｒ＝０．７２，Ｐ＜０．０１）．结果说明，精子核的成熟性异常与精索静脉曲张不育有密切的联系。附睾功能的异常可能是精索静脉曲张导致不育的机制之一。     

输精管吻合术后睾丸cAMP含量与精子密度的关系

日本大耳白雄兔３２只，随机分为输精管结扎组（ＶＧ）、输精管结扎－吻合组（ＶＡＧ）和假手术组（ＳＯＧ）。结果显示：ＶＡＧ的精子密度明显低于ＳＯＧ（Ｐ＜０．０１），并且与睾丸组织ｃＡＭＰ含量呈明显正相关（ｒ＝０．８４１，Ｐ＜０．０１）。血清睾酮水平３组无显著差异。ＶＧ，ＶＡＧ的睾丸重量均低于ＳＯＧ（Ｐ＜０．０１，Ｐ＜０．０５），但ＶＡＧ与ＶＧ比较有显著差异（Ｐ＜０．０５），且ＶＡＧ和ＳＯＧ的睾丸重量与精子密度也呈明显正相关（ｒ＝０．６９９，Ｐ＜０．０１）．说明输精管结扎及吻合后一段时间内睾丸生精功能处于相对抑制状态，且与对垂体促性腺激素反应性下降有关。     

切除颌下腺对大鼠睾酮和精子顶体酶活力的影响

本实验对大鼠切除颌下腺３０和４８天时，睾丸和附睾重量、血清睾酮水平及精子顶体酶活力进行了观察。结果显示：（１）切除颌下腺的实验组，睾丸和附睾重量明显低于对照组（Ｐ＜０．０５，Ｐ＜０．０１），睾丸脏器系数也明显降低（Ｐ＜０．０１），但附睾脏器系数无明显降低（Ｐ＞０．０５）；（２）血清睾酮水平３０天时实验组略低于对照组，４８天时为２．４１ｎｍｏｌ／Ｌ，明显低于对照组的４．４９ｎｍｏｌ／Ｌ（Ｐ＜０．０１）；（３）实验组的精子顶体酶活力分别为７．１９和６．００ｍＵ／ｍｌ明显低于对照组（Ｐ＜０．０１）分别降低了５５．１％和６１．１％。结果表明表皮生长因子（ＥＧＦ）参与了调节睾酮的分泌，并能影响精子顶体酶活力以及睾丸和附睾的重量。     

男性不育症病人的精液分析与性激素检测的探讨

本文测定了４０例正常生育男性和５０例不育男性的精液分析，以及血清ＦＳＨ、ＬＨ、ＰＲＬ的浓度，按照精子密度分为正常组、少精症组和无精症组。我们观察到正常组、少精症组、无精症组之间的精液液化时间、ｐＨ、量的差异均无显著性（Ｐ＞０．０５）．而精子的活动率与正常形态精子却有明显的不同，精子的活动率差，其畸形精子的百分率就高，且呈非常显著的关系。少精症组与无精症组中的ＦＳＨ、ＬＨ、ＰＲＬ的浓度明显高于正常组（Ｐ＜０．０５），而无精症组的ＦＳＨ、ＬＨ、ＰＲＬ值又明显高于少精症组（Ｐ＜０．０５）。当ＦＳＨ、ＬＨ明显升高时，精子的活动率、密度的百分率明显下降（Ｐ＜０．０１）。本文结果是无精症组血清ＰＲＬ与正常组有显著性差异（Ｐ＜０．０５），而少精症组则无明显差异（Ｐ＞０．０５）。     

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。     

大鼠附睾精子运动调节的有关因子研究

精子在附睾成熟中其运动能力的获得和发展过程十分复杂，受众多因素的影响和调节，包括附睾内精子本身因素、附睾内环境及一些附睾内调节因子对精子的作用。我们用附睾微穿刺等技术获得附睾液，并分别应用生物化学、生物发光等方法分段研究了大鼠附睾精子成熟过程中精子ＡＴＰ、钙调蛋白和附睾液肉毒碱含量及其活性变化规律。结果发现大鼠附睾液内肉毒碱含量随附睾头、体、尾移行而逐渐增加，均值（－χ±Ｓ－χ）分别为４８．７±３．６４．２±３．６、９０．２±５．３ｍｍｏｌ／Ｌ；附睾精子ＡＴＰ含量（×１０－１０ｍｏｌ／ｌ０６精子）也呈此种变化，分别为１．５±０．２、２．８±０．４、４．６±２．５；附睾精子钙调蛋白活性（×ｌ０－４ｎｍｏｌ／ｌ０８精子）分别为２．６±０．７、３３±０．７、１．４±０．２，其中以体部精子最高，尾部精子最低。并针对该结果及这些调节因子对精子附睾成熟中运动能力的调节作用及其机理作了一些探讨。     

大鼠附睾精子成熟过程中膜流动性变化的研究

精子在附睾内的运行过程中，结构和功能发生变化，最终达到成熟，并贮存于附睾尾部。本文用５－Ｄｏｘｙｌ－ｓｔｅａｒｉｃａｃｉｄ（５ＤＳＡ）和１６－Ｄｏｘｙｌ－ｓｔｅａｒｉｃａｃｉｄ（１６ＤＳＡ）为标记，在电子自旋共振（ＥＳＲ）波谱仪上研究成年ＳＤ雄鼠附睾头、体、尾各段的精子膜流动性的变化，并用低渗膨胀（ＨＯＳ）试验测定精子膜对渗透压的反应能力。结果表明，从附睾头部到尾部，精子膜膜脂的流动性逐步降低，特别是从体部到尾部更为明显（Ｐ＜０．０１），而精子膜对渗透压的反应能力也逐渐降低，主要表现在从头部移行到体部的精子（Ｐ＜０．０１）。精子成熟过程中的脂质过氧化反应，可能是降低膜脂流动性的原因之一。     

昆明山海棠提取物TH5对雄性大鼠的抗生育活性

为了开发避孕新药，作者对昆明山海棠提取物ＴＨ５的抗生育活性进行了研究。５批Ｗｉｓｔａｒ成年雄性大鼠灌服ＴＨ５（１１６ｍｇ／ｋｇ）３０天后的雄性抗生育有效率平均为９７％（８６／８９）。停服ＴＨ５２０天后的大鼠附睾尾部三项精子参数统计值明显低于对照组（Ｐ＜０．０５或Ｐ＜０．０１）；停服ＴＨ５５０天后，９０％的受试大鼠恢复生育，其附睾尾部的三项精子参数值与对照组相比无显著差异（Ｐ＞０．０５）。ＴＨ５对大鼠体重、睾丸重量与大小等无影响。认为ＴＨ５具有发展为男用避孕药的良好前景。     

5例正常人胆管壁的组织构筑研究

对５例正常人胆管壁的组织构筑及其平滑肌（ＳＭＣ）、弹力纤维（ＥＦ）、胶原纤维（ＣＦ）进行了定量研究。结果发现：从胆总管胰后段至左右肝管汇合部，平滑肌的面积百分数从１３．３５±０．６５降到３．３４±０．３２；弹力纤维的含量以肝总管和左右肝管汇合部为高，与胆总管胰后段和十二指肠上段比较，差别有显著性（Ｐ＜０．０５）；胶原纤维的含量，肝外胆管各段及肝内胆管各段差别无显著性（Ｐ＞０．０５），但肝内、外胆管各段比较，差别有显著性（Ｐ＜０．０５）。对胆管壁的组织学观察发现，肝内与肝外胆管壁的组织构筑存在一定的差异，说明胆管不同部位的生理功能不同。     

Free and Bound Sialic Acid in Rat and Hamster Epididymal Fluid/Freie und gebundene Sialinsäure in der Nebenhodenflüssigkeit der Ratte und des Hamsters

Total, free and bound sialic acid concentrations were determined in sperm-free luminal fluid removed by micropuncture from different regions of the rat and hamster epididymides. In the rat, total sialic acid concentrations (mean +/- s.e.m.) in the proximal caput, the mid corpus and the proximal cauda were, respectively, 25.7 +/- 1.3, 23.9 +/- 1.7 and 28.8 +/- 1.9 mM compared to 4.4 +/- 0.1 mM in blood plasma. In the hamster, total sialic acid concentrations in the distal corpus, the proximal cauda and the distal cauda were, respectively, 32.9 +/- 3.8, 26.4 +/- 1.4 and 26.6 +/- 3.2 mM compared to 4.7 +/- 0.5 mM in blood plasma. Free sialic acid accounted for approximately 70-80% of total sialic acid present in the epididymal plasma of the rat. Similarly, 82% of sialic acid in the rat blood plasma was in free form. The levels of free and bound sialic acid were not changed in different regions of the rat epididymis. Unilateral ligation of the rat efferent ducts had no effect on total, free or bound sialic acid concentrations in all regions except in the proximal cauda in which a transient increase (P less than 0.01) in free sialic acid was observed on Day-3 after EDL.     

Alteration of epididymal function and its relation to maturation of spermatozoa

Alteration of epididymal function and its relation to maturation of spermatozoa was studied in 54 adult male albino rats. Levels of free and bound sialic acid in the spermatozoa and luminal contents of the epididymis and vas deferens were determined. A group of 10 received rabbit antiserum to ovine luteinizing hormone (LHAS) sc .2 ml/day for 5 days. 2 groups of 8 animals each received 2.5 mg cyproterone acetate twice daily for either 15 or 30 days. 16 animals served as intact controls and 12 animals served as castrate controls. Epididymis and vas deferens sperm counts were not affected by LHAS for 5 days or by cyproterone acetate for 15 days; however, sperm counts were decreased in the corpus (p less than .02), cauda (p less than .05), epididymidis and vas deferens (p less than .01) when rats were treated with cyproterone acetate for 30 days. Castration resulted in a marked reduction in all regions within 5 days. In the intact rats spermatozoa sialic acid decreased in the cauda epididymidis (p less than .01) and increased in the vas deferens (p less than .001). Sialic acid concentration was similar in those treated with either LHAS or cyproterone acetate for 30 days. Bound sialic acid in the epididymal fluid increased (p less than .02) to a maximum in the corpus and cauda and decreased in the vas deferens (p less than .05). LHAS or cyproterone acetate caused a reduction in bound sialic acid in the fluid of the epididymis and vas deferens.     

大鼠睾丸扭转复位后附睾唾液酸含量变化及意义

目的 :探讨大鼠睾丸扭转 2h和 4h复位后 2 4h附睾唾液酸含量的变化和意义。　方法 :用 2 4只雄性SD大鼠建立左侧睾丸扭转复位模型 ,分为对照组、扭转 2h组和 4h组 ,每组 8只。 5 甲基苯二酚法检测扭转侧附睾唾液酸的含量。　结果 :睾丸扭转 2h复位后 2 4h扭转侧附睾唾液酸含量 [(2 3.385± 9.2 2 0 )mg/mgprot]改变不明显 ;睾丸扭转 4h复位后 2 4h附睾唾液酸含量 [(13.72 5± 7.80 1)mg/mgprot]下降明显 (P <0 .0 5 )。　结论 :睾丸扭转 2h复位后 2 4h附睾分泌唾液酸功能不受影响 ,扭转 4h复位后 2 4h附睾分泌唾液酸功能下降 ;附睾耐受缺血再灌注损伤的时间可能较长。     

Compartmentalization of lysosomal enzymes in cauda epididymis of normal and castrated rats

The mammalian epididymis is an organ particularly rich in acid hydrolases, consistent with a developed lysosomal apparatus. However, some of these enzymes could also play a role in an extracellular environment, since they are actively secreted by the epithelium. In this study the authors measured the activity of five acid hydrolases distributed between the epithelium, fluid, small vesicles, and spermatozoa of the rat cauda epididymis in adult rats, and compared with that distribution under conditions of deprivation of luminal testosterone and testicular compounds (hemicastration). Lysosomal enzymes are differently compartmentalized in rat cauda epididymis. Most of beta-galactosidase (beta-GAL) and aryl sulfatase (approximately 70%) were found in soluble form within the fluid. Some 60% of N-acetyl-beta-D-glucosaminidase (beta-NAG) and alpha-mannosidase (alpha-MAN) become transiently bound to sperm, and beta-glucuronidase (beta-GLU) was mostly concentrated in the epithelium. After remotion of testis this distribution changed, as the retention of alpha-MAN, beta-GAL, beta-GLU, and beta-NAG by the epididiymal tissue increased. The increase of beta-GLU followed an increase of synthesis of the enzyme. The distribution of enzymes in the epididymis from the contralateral side was similar to that in normal rats. The different roles for each enzyme in the epididymis are discussed.     

Antispermatogenic/antiandrogenic properties of solasodine (C27H43O2N) obtained from solanum xanthocarpum berries on the male genital tract of dog (Canis-familiaris). A Histophysiological approach

Chronic administration of solasodine (20 mg/kg alt. day for 30 days) caused testicular lesions resulting in a severe impairment of spermatogenic elements. The epididymides were devoid of spermatozoa. Total protein, sialic acid and glycogen contents of the testis and epididymis were reduced significantly whereas the testicular cholesterol was elevated. Acid Phosphatase enzyme activity of the testes was low after solasodine treatment. Serum enzymes (SGPT, alkaline phosphatase) serum protein, triglycerides, non esterified fatty acid levels were in normal range when compared with their own controls. Cholesterol and phospholipid levels were elevated after solasodine treatment to intact dogs. Reduced androgen production was reflected in low levels of sialic acid in the testes and epididymides and reduced Leydig cell nuclei. Castration alone brought about reduction in size of the epididymis. Castration followed by solasodine treatment caused epididymal degeneration. Simultaneous administration of TP to solasodine treated castrated dogs failed to stimulate the epididymal growth. Antispermatogenic/antiandrogenic activity of the compound solasodine is discussed. Solasodine administration in dogs definitely rendered the male infertile as evidenced by the absence of sperms in the cauda epididymis and ductus deferens.     

Effect of papaya seed extract on microenvironment of cauda epididymis

Aim: To evaluate the effect of aqueous Carica papaya seed extract on microenvironment of cauda epididymis. Methods: Adult male albino rats were intramuscularly administered with 0 (control) or 0.5 mg papaya seed extract/kg body weight for 7 days. Cauda epididymal tubular content was collected by micropuncture technique; epididymal luminal fluid and sperm pellets were separately analyzed. Results: The results revealed that the extract treatment caused significant reduction, as compared with control, in total protein and sialic acid contents in both epididymal fluid and sperm pellet. As compared with control, significantly lowered acid phosphatase activity was recorded in sperm pellet but was higher in epididymal fluid after the treatment. The extract treatment also caused significant reduction in level of inorganic phosphorus in the epididymal fluid. Conclusion: It is concluded that the aqueous papaya seed extract alters cauda epididymal microenvironment.     

Effect of piperine on the epididymis of adult male rats

Aim： To study the effect of piperine on the epididymal antioxidant system of adult male rats. Methods： Adult male rats were orally administered piperine at doses of 1 mg/kg, 10 mg/kg and 100 mg/kg body weight each day for 30 consecutive days. Twenty-four hours after the last treatment, the rats were weighed and killed with ether and the epididymis was dissected from the bodies. Sperm collected from the cauda region of the epididymis was used for the assessment of its count, motility and viability. Caput, corpus and cauda regions of the epididymis were separated and homogenized separately to obtain 10 % homogenates. The supernatants were used for the assays of sialic acid, superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, lipid peroxidation and hydrogen peroxide generation. Results： Body weight of the piperine-treated rats remained unchanged. The weights of the caput, corpus and cauda regions of the epididymis significantly decreased at dose of 100 mg/kg. Epididymal sperm count and motility decreased at 10 mg/kg and 100 mg/kg, and sperm viability decreased significantly at 100 mg/kg. Sialic acid levels in the epididymis decreased significantly at 100 mg/kg while significant decrease in the cauda region alone was observed at 10 mg/kg. A significant decline in the activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione reductase, along with an increase in hydrogen peroxide generation and lipid peroxidation were observed at 10 mg/kg and 100 mg/kg. Conclusion： Piperine caused a decrease in the activity of antioxidant enzymes and sialic acid levels in the epididymis and thereby increased reactive oxygen species levels that could damage the epididymal environment and sperm function.     

热休克蛋白70-2基因沉默对大鼠精子附睾成熟的影响

目的 观察热休克蛋白(HSP)70-2基因沉默对大鼠附睾中精子成熟的影响.方法 构建HSP70-2特异的短发夹RNA(shRNA)质粒载体,注入大鼠附睾中,1周后处死大鼠,无菌切取附睾,用逆转录-聚合酶链反应(RT-PCR)方法 检测附睾中HSP70-2 mRNA表达,Western blot观察HSP70-2蛋白表达,应用苏木素-伊红(HE)染色检测附睾管内精子密度.结果 空白对照组HSP70-2蛋白表达量为1.31±0.10,附睾管内成熟精子密度为(63.46±14.43)%.基因沉默组HSP70-2蛋白表达量为0.93±0.13,附睾管内成熟精子密度为(31.51±10.67)%,与对照组比较均显著减少,差异有统计学意义(P＜0.01).结论 HSP70-2基因沉默影响附睾中精子成熟.     

大鼠精索静脉曲张后附睾上皮细胞凋亡及管腔α-1,4-葡糖苷酶、唾液酸含量观察

目的:探讨大鼠左侧精索静脉曲张后同侧附睾上皮细胞凋亡与附睾管腔中-α1,4-葡糖苷酶和唾液酸含量改变的关系及意义。方法:16只雄性SD大鼠建立左侧精索静脉曲张模型,分为对照组和实验组,每组8只。缺口末端转移酶标记技术检测附睾上皮细胞凋亡;硝基酚法和5-甲基苯二酚法分别检测附睾管腔液-α1,4-葡糖苷酶和唾液酸含量。结果:大鼠左侧精索静脉曲张模型建立后的第7 d,实验组同侧附睾上皮细胞凋亡指数显著高于对照组(P<0.001);实验组附睾管腔液-α1,4-葡糖苷酶、唾液酸含量显著低于对照组(P<0.05,P<0.005)。结论:大鼠左侧精索静脉曲张可导致同侧附睾上皮细胞凋亡增加,进而影响该侧附睾上皮细胞的合成和分泌功能。     

A comparative study of experimental and human pancreatic carcinoma with special reference to histochemical findings

This study was undertaken in order to histochemically investigate the mucoid substances (acid mucopolysaccharide and sialic acid) in experimental and human pancreatic carcinoma. The degree of alcian blue staining in the ductal and interstitial portions of the pancreas increased, especially in hamster tubular adenocarcinoma which had been induced by N-nitrobis (2-hydroxypropyl) amine. The pancreatic carcinoma of the rats, induced by di-methyl benz (alpha) anthracene, was usually poorly differentiated carcinoma, in which alcian blue and toluidine blue staining was high in the interstitial portion when compared with that of the hamster pancreatic carcinoma, which was usually well differentiated. The rates of enzyme digestion of acid mucopolysaccharide and sialic acid showed more prominently in the rat pancreatic carcinoma than in the hamster pancreatic carcinoma. Mucoid substances in the ductal portion of human pancreatic carcinoma revealed properties similar to those of hamster pancreatic carcinoma whereas those in the interstitial portion were similar to those of rat pancreatic carcinoma. These results suggested that mucoid substances, especially acid mucopolysaccharide, both in experimental and human pancreatic carcinomas, may play an important role in tumor growth.