The spermatogenic cycle in the blue fox (Alopex lagopus): relative frequency and absolute duration of the different stages

The spermatogenic cycle of the blue fox was divided into eight distinct stages, based on an analysis of different cell associations of the seminiferous epithelium. The criteria used for classification of the stages were the type of spermatogonia, the occurrence of meiotic figures, and the shape and location of spermatids. The relative frequencies of the stages I to VIII were 25.7, 9.8, 8.7, 5.9, 13.8, 9.9, 10.6 and 15.5%, respectively. The duration of one cycle of the seminiferous epithelium was 12.0 +/- 0.2 days as determined from the progression of 5-bromo-2-deoxyuridine (BrdU)-labelled cells at various time intervals. The absolute duration of stages I to VIII was calculated to be 3.1, 1.2, 1.0, 0.6, 1.7, 1.2, 1.3 and 1.9 days, respectively. The estimated life span of primary spermatocytes was 19.2 days, of secondary spermatocytes less than 0.6 days, of spermatids with round nuclei 9.2 days and of spermatids with elongated nuclei 8.9 days.     

Inhibition kinetics of NAD-linked enzymes by gossypol acetic acid

Effects of gossypol on human spermatozoal enzymes were investigated. This compound was found to be a potent inhibitor of the NAD-linked enzymes, glucelaldehyde-3-phosphate dehydrogenase (GA3PDH), lactate dehydrogenase (LDH), malate dehydrogenase (MDH), and isocitrate dehydrogenase (ICDH). MDH was inhibited by gossypol when the reaction was carried out in malate-oxalacetate (direct) or oxalacetate-malate (reverse) directions. The I50 of gossypol for the direct reaction was 2.9 microM, whereas that for the reverse reaction was 1.2 microM. Reciprocal plots due to Lineweaver-Burk showed that MDH is inhibited in a noncompetitive manner with respect to both reactions. LDH was also inhibited by this compound when pyruvate or alpha-ketobutyrate was used as a substrate. Gossypol was a noncompetitive inhibitor for LDH. The I50 of gossypol for LDH were 9.8 microM and 11.3 microM, when using pyruvate and alpha-ketobutyrate, respectively as substrates. The I50 of gossypol for GA3PDH and ICDH were 110 microM and 2.7 microM, respectively.     

Ultrastructural analysis of rat testes after gossypol acetic acid (GAA) treatment

Summary The present investigations were carried out to show the histological and ultrastructural alterations in rat testes 10 weeks after gossypol acetic acid treatment (dose: 30 mg gossypol acetic acid/kg/day). The morphological findings in the interstitial compartment were compared with the data from studies carried out to investigate the testosterone biosynthesis in gossypol acetic acid treated rats. No morphological changes in the epididymal and vasal epithelia were found; however, the germinal epithelial cells showed vacuolisation, pycnosis, disconnections of junctions, cytolysis and exfoliation of germ cells from the epithelium. The Sertoli cells were affected, too. Gossypol acetic acid seemed to stimulate the physiological activity pathologically; cellular organelles as mitochondria, endoplasmic reticulum, lysosomal vacuoles, pigment granules and nuclei were either enlarged in size and number or malformed in shape. The cellular contact was often restricted to spots or completely disconnected. If gossypol acetic acid was administered for a longer period of time some Sertoli cells were found to be unable to withstand the toxic stimulus, and the cells became necrotic too. In contrast to the toxic process in the germinal and Sertoli cells the Leydig cell compartment did not show any changes in fine structure, and therefore testosterone biosynthesis is presumed to be intact.     

低剂量醋酸棉酚服用前后血钾测定结果分析

于 1993年底至 1996年在山东省进行低剂量醋酸棉酚临床试验 ,服药组 30例 ,对照组 15例。样本于服药前测定血钾 2次 ,服药后第 2月至第 9月每月测定血钾一次 ,停药后再持续测定 4个月。结果 :服药前 2次测定值 (mmol/L)实验组分别为 4.0 4± 0 .40 ,4.6 1± 0 .41;对照组分别为 4.0 3± 0 .32 ,4.5 0± 0 .43。服药后 2～ 9个月实验组血钾在 3 .75～ 4.5 9范围 ;对照组为 3.86～ 4.82范围 ;停药后 4个月血钾值实验组为 3 .92～ 4.2 8,对照组为4.0 4～ 4.0 8范围。经统计学处理两组在各随访月的差异均无显著性。说明男性口服低剂量醋酸棉酚 9个月不会降低血钾 ,停药后 4个月血钾值也在正常值范围内     

An analysis of the direct effects of gossypol on human spermatozoa

The direct effects of gossypol and its acetic acid adduct, on the movement and functional competence of human spermatozoa were investigated employing exposure times of 1, 5 or 15 min and concentrations of 50 μM, 500 μM and 1000 μM. These compounds markedly reduced the motility, velocity, frequency of sperm head rotation and linearity of sperm progression, the most significant effects being observed with gossypol acetic acid on populations of 'capacitated' spermatozoa. Significant direct effects of gossypol on the ability of human spermatozoa to penetrate both cervical mucus and zona-free hamster ova were also observed, which were independent of any effects on motility. These results reinforce the notion that gossypol may serve a contraceptive role in the female as a 'spermicidal' agent, and suggest that this compound may also be of scientific value as a probe for identifying and isolating functionally important components of the human spermatozoon.     

青春期营养性肥胖大鼠睾丸生精细胞周期的变化

目的探讨青春期营养性肥胖大鼠的睾丸生精细胞周期的变化。方法80只新生的雄性清洁级SD大鼠随机分为两组,对照组(n=32只)喂普通饲料,肥胖组(n=48只)喂高脂饲料,分别于喂养后的第3、4、5、6周末观察喂养后体重,计算Lee’s指数,流式细胞分析术检测睾丸生精细胞周期的改变。结果与对照组比较,肥胖组大鼠第3周开始体重有显著性增加(P<0.05),6周内体重持续上升,至第6周末肥胖组大鼠体重超过对照组达26.6%(P<0.01),Lee’s指数随着大鼠肥胖程度的增加而逐渐增高;大鼠G0/G1期细胞在高脂饲料喂养第3周末增多(P<0.05)、肥胖组S期细胞显著下降(P<0.01),此时处于G2/M期细胞的百分数明显增多(P<0.05)。结论青春期时的肥胖可以引起睾丸生精细胞S期细胞百分数逐渐减少,出现G2期细胞阻滞,细胞有丝分裂延迟。     

Inhibition of sialyl transferase activity by gossypol acetic acid in human seminal plasma

Seventeen aliquots of 15 microliters seminal plasma of human origin were incubated with 25 micrograms, 15 micrograms, 5 micrograms and 2.5 micrograms gossypol acetic acid (GAA), dissolved in BWW containing 0.9% sodium chloride and 0.9% benzyl alcohol, at 37 degrees C for 60 minutes. Following incubation, activity of sialyl transferase (S.T.) was determined by a procedure involving incorporation of radioactive sialic acid into asialofetuin. The activity of S.T., expressed as cpm per hour per 15 microliters seminal plasma was compared to controls consisting in incubation of sperm in BWW containing 0.9% benzyl alcohol and 0.9% sodium chloride. The activity of S.T. in controls represented 61.3 +/- 8.0%-67.7 +/- 8.9% of the activity obtained by incubation with BWW only. GAA was found to exert a dose-dependent inhibition of S.T. activity, ranging from 38.3 +/- 20.6% to 53.4 +/- 19.4% (with 25 micrograms) and from 11.3 +/- 14.8% to 21.9 +/- 14.8% (with 2.5 micrograms).     

Effect of Black maca (Lepidium meyenii) on one spermatogenic cycle in rats

Lepidium meyenii (Maca) grows exclusively between 4000 and 4500 m above sea level in the Peruvian central Andes. The hypocotyls of this plant are traditionally used in the Andean region for their supposed fertility-enhancing properties. The hypocotyls have different colours. Of these, Black maca has better effects on spermatogenesis. The present study aimed to test the hypothesis that Black maca has early effects during a spermatogenic cycle (12 days) of male rats. For this, testicular spermatid, epididymal sperm and vas deferens sperm counts were measured after 1, 3, 5, 7 and 12 days of treatment with Black maca. Aqueous extract of Black maca was given orally by daily gavage at a dose of 2 g kg(-1). In a spermatogenic cycle, compared with day 1, daily sperm production (DSP) was lower at day 7 (control), whereas with Black maca, the difference was observed at day 12. Epididymal sperm count was higher in rats treated with Black maca at days 1, 3 and 7, but similar to controls at days 5 and 12; similarly sperm counts in vas deferens was higher in rats treated with Black maca in days 3, 5 and 7, but similar to controls at days 1 and 12. From this, it is suggested that first action of Black maca was at epididymal level increasing sperm count after 1 day of treatment, whereas an increase in sperm count was observed in vas deferens at day 3 of treatment. Finally, an increase in DSP was observed after 7 days of treatment with Black maca. Testicular testosterone was not affected after 7 days treatment with Black maca. In conclusion, Black maca affects sperm count as early as 1 day after beginning of treatment.     

长期给予睾酮致大鼠生精小管伴有生精细胞排列疏松

目的:确定睾酮致大鼠睾丸内睾酮抑制因而精子发生障碍是否伴有生精细胞排列疏松。方法:成年雄性SD大鼠肌注十一酸睾酮[19 mg/(kg.15 d)]130 d后取睾丸组织块,作甲基丙烯酸树脂包埋切片,观察睾丸的组织学变化。结果:除精子形成、精子释放障碍等改变以外,11.5%的生精小管轮廓内生精细胞的排列明显较疏松,成串、成束或成团的生精细胞(主要是精母细胞和精子细胞)之间出现朝向小管腔走行的放射状裂隙。结论:生精细胞排列疏松是大鼠睾丸内睾酮抑制所致重要组织学改变之一。     

高浓度乙酸注射治疗兔VX2肝肿瘤的实验研究

目的利用兔VX2肝移植瘤模型探讨高浓度乙酸注射治疗肝肿瘤的价值,并探索注射治疗肝肿瘤的最适乙酸浓度.方法用移植法建立兔VX2肝肿瘤模型,将40只携瘤兔随机分为A、B、C、D四组.术前以CT测量肿瘤直径.开腹向A、B、C、D组肝肿瘤内分别注入50%、75%、100%乙酸和生理盐水,48 h后重复注射一次,术后一周处死兔子取肝脏观测比较各组肿瘤体积改变情况及坏死灶周围新生灶情况,并取肿瘤组织制作石蜡切片(HE染色),镜下观察肿瘤组织坏死情况.结果注射后一周各实验组肿瘤直径均减小,对照组肿瘤直径明显增大.100%乙酸组10例仅1例在坏死灶周围发现新生灶,而50%乙酸组发现5例,75%乙酸组发现7例.镜下显示100%乙酸组肿瘤中未坏死的肿瘤细胞团较75%和50%乙酸组少见.结论高浓度的乙酸对兔VX2肝肿瘤的灭活效果显著;其中100%的乙酸效果最佳.     

Effect of gossypol on the concentration of sodium and potassium in the rat epididymis

The effects of administration of gossypol acetic acid (7.5 mg/kg daily for 4 weeks) on the concentration of Na+ and K+ in the rat epididymis was assessed. Epididymal fluid samples, collected by micropuncture, from the caput, corpus, proximal cauda and distal cauda epididymis from gossypol-treated and control animals were analysed for Na+ and K+ concentrations. Gossypol-treated males failed to impregnate healthy females, presumably because their sperm were immotile. In gossypol-treated rats, Na+ levels decreased significantly (P less than 0.01) in the caput, corpus, proximal and distal cauda epididymis. In contrast, the K+ concentration was increased significantly (P less than 0.05) only in the caput and corpus epididymis. This altered electrolyte milieu may be responsible, to some extent, for immotility and hence infertility.     

Recovery of sperm production following the cessation of gossypol treatment: a two-centre study in China

A number of variables were investigated in 46 men who had stopped taking gossypol for their predictive association with the degree and time of recovery of spermatogenesis. Thirty-nine (87%) of the men were azoospermic at cessation of gossypol treatment. In those with sperm present the geometric mean concentration and total sperm count were 8.3 X 10(6)/ml and 30.7 X 10(6), respectively. Twenty-eight men (61%) recovered to a defined threshold of spermatogenic function (sperm concentration greater than or equal to 20 X 10(6)/ml), with a median recovery time of 1.1 years. However, 18 men (39%) had not recovered to this degree of spermatogenic function after a median follow-up of 1.9 years and, of these, 10 (22%) remained azoospermic. The influence of individual baseline variables on the time to defined recovery was examined using Kaplan-Meier curves for groups and their joint effect by Cox's regression model. The failure of recovery was strongly associated with longer treatment, greater total dose of gossypol, smaller testicular volume, elevated FSH concentrations and, to a lesser extent, with greater body weight.     

Toxicity of Gossypol at Antifertility Dosages in Male Rats: Statistical Analysis of Lethal Rates and Body Weight Responses

Die Letalität und die Körpergewichtszunahme männlicher Wistarratten während der oralen Verabreichung von Gossypol-Essigsäure (GAA) in Dosierungen zwischen 2.5 mg/kg und 30 mg/kg wurden statistisch ausgewertet. Eine verglichen mit den Placebo-Gruppen signifikante Letalität nach 10wöchiger Behandlung mit niedrigen GAA-Mengen (2.5, 5.0 und 7.5 mg/kg, p < 0.05) und mit hohen GAA-Mengen (15, 20 und 30 mg/kg, p < 0.01) wurde festgestellt. Die bei Placebo-Tieren innerhalb der Versuchsdauer festgestellte deutliche Körpergewichtszunahme blieb in allen mit GAA behandelten Gruppen aus. Die Wirkung hoher GAA-Mengen, welche jeden zweiten Tag gegeben wurden, war mit den Auswirkungen nach täglicher Verabreichung identisch.
Die Antifertilitäts-Wirksamkeit der GAA in diesen Tieren steht in keinem Zusammenhang mit der beobachteten Toxizität dieser Substanz. Aus vorliegender Untersuchung wird auf eine deutliche Toxizität antifertilitärer GAA-Konzentrationen in männlichen Ratten geschlossen.     

锡类散预防酸性灌肠液致兔肠黏膜损伤的实验研究

目的观察锡类散预防酸性灌肠液灌肠所致兔肠黏膜损伤的效果。方法将40只家兔随机分成两组各20只,对照组采用食醋灌肠,观察组采用食醋加锡类散灌肠。观测两组灌肠前后血钾水平变化及粪便涂片肠黏膜细胞脱落情况;将动物腹腔麻醉下取直肠、结肠肠壁制作病理切片,透射电镜观察肠黏膜细胞亚显微结构变化,光镜观察肠黏膜充血、水肿情况。结果对照组灌肠后血钾下降水平较观察组更为显著(P<0.01);病理切片及粪便涂片显示,对照组肠黏膜损伤及肠黏膜细胞脱落情况较观察组严重。结论锡类散可降低酸性灌肠液对兔肠黏膜损伤程度,具有一定的保护作用。     

LM23 is a novel member of the Speedy/Ringo family at the crossroads of life and death of spermatogenic cell

LM23 is a gene specifically expressed in the testis of Rattus norvegicus, as previously reported by our laboratory. The aim of the study is to further investigate the biological function of LM23. Several bioinformatic tools were utilized, including PROSITE and BLAST. To determine the subcellullar localization of LM23, a polyclonal antibody specific for LM23 was generated via the immunization of rabbits. The LM23 gene was cloned from rat testis tissue, and LM23 protein was expressed in Escherichia coli. The biological function of LM23 was analyzed with microarray analysis and immunohistochemistry, using a rat model of LM23 gene knockdown. The results suggested that LM23 belongs to the Speedy/Ringo family. LM23 regulated the G₁/S and G₂/M transitions of the cell cycle during spermatogenesis. Downregulation of the LM23 gene during spermatogenesis could lead to the activation of both the Fas–FasL pathway and the mitochondrial pathway. These novel findings indicate that LM23 has a diverse array of functions that are important in both the life and death of the spermatogenic cell.     

CT透视引导下经皮乙酸消融治疗第二肝门部恶性肿瘤

目的　评价CT透视引导经皮穿刺乙酸注射治疗 (PAI)第二肝门区恶性肿瘤的安全性和效果。方法　搜集西南医院 2 0 0 0年 5月至 2 0 0 3年 10月在实时CT透视导向下经皮穿刺乙酸注射治疗第二肝门区恶性肿瘤 17例 ,肝细胞癌 10例、转移性腺癌 7例 ,共 2 0个病灶。病灶直径 2～ 5cm。根据病灶大小每次注入 5 0 %～ 6 0 %乙酸 2～ 6ml不等 ,乙酸内加入 1ml对比剂混匀 ,以了解乙酸在肿块内的弥散程度和范围。注射后 1周影像检查 ,根据情况重复注射 1～ 2次。结果　在实时CT透视下均能准确穿刺到靶点 ,除 12例轻中度疼痛外无其它并发症。全部病例均经B超或CT随访 ,3个月时 ,其中瘤灶缩小 14个 (70 % ) ,肿瘤大小无变化 4个 (2 0 % ) ,肿瘤增大 2个 (10 % )。 17例随访 3月 ,全部生存 ;12例随访 1年 ,9例生存 ,5例失访 ;6例随访 2年 ,3例生存 ,3例失访。结论　CTF引导下经皮乙酸消融治疗第二肝门部恶性肿瘤是安全有效的     

Superoxide dismutase activity along rat seminiferous epithelial wave: effects of ethane dimethanesulphonate and 3.0 Gy of X—irradiation

Summary. Changes in generation of reactive oxygen species and antioxidant enzyme activities are associated with differentiation processes. The authors have studied the activity of superoxide dismutase (SOD) in sequentially cut stage-defined segments of rat seminiferous tubules. Great variation was observed in SOD activity along the seminiferous epithelial wave. At its highest, fourfold increases were observed in individual tubules. However, these changes showed no clear correlation to the stages of the cycle. To determine the effect of testosterone withdrawal, rats treated with ethane dimethanesulphonate (EDS) were studied. This treatment had no effect on the pattern of SOD activity along the seminiferous epithelial wave. Testes of other rats were exposed to local 3.0 Gy X—irradiation to cause selective loss of germ-cell populations. SOD activity in the seminiferous epithelium was not affected at 30 min or 7 d after X—irradiation. On day 31 post-irradiation, SOD activity increased at stages XIV-VI, peaking at stage III ( P < 0.01 for comparison of stages XIV-VI with the other stages). The data presented here suggest that the activity of SOD in seminiferous epithelium is regulated over a wide range during spermatogenesis. Testosterone plays no major role in the control of seminiferous tubule SOD activity. The loss of spermatocytes and early spermatids by day 31 after X-irradiation revealed a stage-specific increase in SOD activity, which may be associated with the differentiation of elongated spermatids.     

咖啡酸苯乙酯对大肠癌SW480细胞生长和周期的影响

目的　探讨咖啡酸苯乙酯 (CAPE)对体外培养的大肠癌细胞SW 4 80增殖和细胞周期的影响。方法　不同浓度CAPE处理体外培养的大肠癌SW 4 80细胞后 ,采用MTT法检测细胞的增殖活性 ;流式细胞仪检测细胞周期分布。结果　CAPE处理SW 4 80细胞后 ,SW 4 80细胞的生长抑制率明显升高 ,抑制作用表现为剂量依赖性和时间依赖性。流式细胞仪细胞周期分析表明CAPE作用 2 4h后 ,细胞G0 /G1期比例上升 ,S期比例下降。同时发现CAPE作用后 ,细胞出现胞浆混浊 ,胞体缩小、变圆、皱缩 ,核固缩粹裂等凋亡形态学特征。结论　CAPE对大肠癌SW 4 80细胞株具有明显的生长抑制作用 ,其作用机制与阻滞细胞周期G1期和诱导细胞凋亡有关。     

Effect of local testicular heating on spermatogenic cell apoptosis in rats

Aim: To study the effect of testicular local heating on spermatoge-nic cell apoptosis in rats. Methods: Seventy male SD rats were divided into the heat-treated and the control groups. The former was exposed to heat (43 ℃) for 12 hours. Each group was further divided into seven subgroups with respect to the time of observation after heat exposure, i.e., 12 h and 1 days, 3 days, 6 days, 10 days, 50 days and 80 days, respectively. In each subgroup, sper-matogenic cell apoptosis was examined by means of electron microscopy, flow cytometry and terminal deoxynucleotidyl trans-ferase-mediated dUDP-nick end labeling (TUNEL) methods. Results: The percentage of cells with sub-haploid and the percentage of positive TUNEL cells were significantly higher in the heat-treated groups than in the controls (P<0.01). The reaction of cell apoptosis to local heat was highly selective: spermatocytes were the most sensitive, followed by spermatids, spermatozoa and sper-matogonia in a decreasing order. Conclusion: Local testic     

醋酸棉酚对男子垂体－性腺轴功能的影响

报道２２例长期停服棉酚男子性激素的水平及其对ＬＨＲＨ和ｈＣＧ刺激的反应。２２例中，１１例生精功能未恢复者（无精子症组）的ＦＳＨ和ＬＨ基础值及其对ＬＨＲＨ刺激反应均显著高于正常对照组（１１例），而睾酮（Ｔ）基础值和Ｔ／ＬＨ比值及Ｔ对ｈＣＧ刺激反应显著低于正常对照组。生精功能恢复组（１１例）的ＦＳＨ基础值及其对ＬＨＲＨ刺激在应均显著高于正常对照组。但是，ＬＨ和Ｔ基础值及其对ＬＨＲＨ和ｈＣＧ刺激反应，二者差异不显著。这些结果说明，不适当的棉酚治疗所引起的永久无精子症者，全睾丸细胞受到严重损害，垂体－睾丸轴系功能调节发生紊乱；而适量的棉酚所引起的暂时无精子症，生精功能恢复以后，睾丸内分泌一般均正常