长春新碱、更生霉素对发育未成熟睾丸影响的实验研究

目的　探讨长春新碱、更生霉素对幼年期大鼠生精细胞的影响。方法　 3周龄雄性大鼠根据药物种类及剂量随机分为长春新碱组 (VCR)、更生霉素组 (AMD)、长春新碱 +更生霉素组(VDG)、小剂量长春新碱 +更生霉素组 (sVDG)、对照组。应用HE染色、荧光显微镜、TUNEL技术、免疫组化方法检测大鼠用药后生精细胞凋亡及Fas基因的表达。结果　各实验组睾丸曲细精管管腔内及近管腔生精上皮部分精原细胞显示显著凋亡 (P <0 .0 1) ,VDG生精细胞凋亡峰值分别显著高于VG、DG及sVDG(P >0 .0 5 )。睾丸组织Fas蛋白表达水平在实验组无显著上调。结论　长春新碱与更生霉素都将导致未成熟睾丸精原细胞显著凋亡 ,并与用药种类多少、剂量呈正相关 ,其后有可能导致生育能力损害     

9-顺式维甲酸协同γ-干扰素诱导神经母细胞瘤分化、凋亡及生长抑制的实验研究

目的 观察9-顺式维甲酸（9-cis retinoic acid，9-cis RA）联合γ干扰素（gammm interferon，γ-IFN）在体外诱导神经母细胞瘤（neuroblastoma，NB）SK-N-SH细胞分化、凋亡过程中细胞形态、周期的变化，探讨其协同作用的效果及其机制。方法 将细胞分为A（对照组）、B（9-cisRA用药组）、C（γ-IFN用药组）和D（9-cisRA和γ-IFN联合用药组）4组，在处理后适当的时间分别观察细胞形态学变化，进行分化神经节细胞计数，用Hoechst-PI荧光染色法观察细胞的凋亡和死亡，四甲基偶氮唑盐（MTT）法测定NB细胞抑制率，流式细胞仪（flow cytometry，FCM）检测细胞周期分布及其凋亡和死亡。结果 联合用药组细胞形态学分化明显，神经节细胞分化率显著高于其他各组（P〈0．01）；MTT法测定显示联合用药能显著提高NB细胞抑制率并明显优于单独用药；Hoechst-PI荧光染色及FCM钙依赖性磷脂结合蛋白V和碘化丙啶（annexin V／PI）染色法检测一致显示联合用药对于诱导NB细胞早期凋亡和坏死具有显著的协同作用；FCM细胞周期动力学分析发现联合用药组C0-G1期细胞比率增加，S期细胞比率减少。结论 9-cisRA联合γ-IFN在体外对NB细胞的分化、凋亡和生长抑制能产生明显的协同作用，为其联合应用于临床提供理论依据和指导。     

青春期精索静脉曲张对大鼠睾丸的影响与表皮生长因子的变化

目的观察青春期精索静脉曲张对大鼠睾丸的影响和表皮生长因子（EGF）的变化，探讨精索静脉曲张不育的机制。方法雄性wistar大鼠24只建立左侧精索静脉曲张模型作为实验组，18只为假手术组。术后4周、8周和12周采血并取双侧睾丸。光镜观察睾丸组织学变化并对生精功能评分；ELISA法测血清EGF浓度；RT-PCR法检测睾丸EGFmRNA表达。结果实验组睾丸重量减轻，生精功能受损，呈双侧性并进行性加重。术后8周实验组血清EGF水平显著低于假手术组（P〈0．01），4周和12周时差异无显著性意义（P〉0．05）。RT-PCR显示实验组左侧睾丸EGFmR—NA表达量术后4周（P〈0．05）、8周和12周（P〈0．01）均低于假手术组；右侧睾丸表达量术后8周（P〈0．01）和12周（P〈0．05）低于假手术组。结论青春期精索静脉曲张可使睾丸EGFmRNA表达量持续降低，引起睾丸发育障碍。     

肾病综合征患儿血清半胱氨酸蛋白酶抑制剂、巨噬细胞移动抑制因子及尿液足细胞检测的意义

目的 检测原发性肾病综合征（PNS）患儿血清半胱氨酸蛋白酶抑制剂（CysC）、巨噬细胞移动抑制因子（MIF）及尿液足细胞（UPC）水平，探讨三者水平与治疗效应的关系。方法 ELISA测定38例接受治疗的PNS患儿缓解前后血清CysC、MIF水平，间接免疫荧光法检测尿沉渣足细胞特异性标志蛋白Podocalyxin以检测尿液足细胞（UPC）水平；并与15例健康对照组比较。结果 与对照组比较，22例激素敏感型患儿治疗前后MIF均升高（P〈0．05或P〈0．01），CysC及UPC无明显变化（Pa〉0．05）；16例激素耐药型患儿治疗后CysC、MIF及UPC显著升高（Pa〈0．01）。与激素敏感型比较，激素耐药型治疗前CysC、MIF及UPC无明显差异，治疗后均显著升高（Pa〈0．01）。与治疗前比较，激素敏感型治疗后CysC、UPC无显著差异（P。〉0．05），MIF显著降低（P〈0．01）；激素耐药型CysC、UPC显著升高（Pa〈0．05或Pa〈0．01），MIF无显著性差异（P〉0．05）。16例激素耐药型患儿加用环磷酰胺冲击治疗后10例病情缓解，缓解组CysC、MIF、UPC水平均较对照组高（Pa〈0．01），6例未缓解组CysC较缓解组明显升高（P〈0．01），MIF、UPC亦升高（Pa〈0．05）。结论 血清CysC、MIF水平及UPC排泄可作为反映PNS患儿病情、评价治疗效应的预测指标。     

87例肺炎患儿心肌酶谱的临床分析

目的探讨小儿肺炎时心肌酶谱的变化，以指导临床。方法87例肺炎患儿进行心肌酶谱检测，同期门诊40例上呼吸道感染患儿作为对照组进行比较。结果肺炎患儿与上呼吸道感染患儿CK-MB值比较差异有统计学意义（P〈0．05），且小婴儿组差异有显著统计学意义（P〈0．01）。结论肺炎患儿心肌酶谱增高显著，年龄越小心肌酶谱水平越高，心肌损害越明显，对小婴儿肺炎患儿要早期，干预治疗。     

RhoA/Rho激酶在高肺血流肺血管结构重构中的表达及其活性研究

目的探讨RhA／Rho激酶在高肺血流所致肺血管结构重构过程中所起的作用。方法108只4周龄Wistar大鼠随机分到4个分流组、4个对照组,分流组大鼠接受左侧颈总动脉-颈外静脉分流术,对照组大鼠作假手术。在术后1、2、4、8周,分别测量右心室收缩压,作血气分析计算Qp／Qs。肺组织切片HE染色光镜下观察肺血管形态学改变,测量并计算中等血管中膜厚度所占管径百分比（MT％）。免疫组化增殖细胞核抗原（PCNA）染色分析平滑肌细胞增生情况。用TUNEL细胞凋亡标记分析平滑肌细胞凋亡状况。pulldownassay分析RhoA活性,Westernblot检测RhoA、Rho激酶（ROCK2）的蛋白表达以及MYPT1磷酸化程度量化分析Rho激酶活性。结果颈总动脉-颈外静脉分流导致肺循环血流量增加,Qp／Qs平均值为2．26±0．35。分流组RVSP在术后1周和8周较对照组高（t=8．799,t=5．332,P〈0．01）。与对照组相比,分流组以平滑肌细胞过度增生和肥大为特征的中等肺动脉中膜增厚,MT％升高在第4周开始出现,到第8周更为明显（t=9．192,t=11．185,P〈0．01）。分流组PCNA阳性平滑肌细胞在第1周显著升高,第2周达最高值,与对照组相比,差异有统计学意义（t=7．213,P〈0．01）,到第4周下降到显著低于对照组（t=4．183,P〈0．01）,并持续降低,到第8周几乎观察不到增生的平滑肌细胞（t=6．152,P〈0．01）。TUNEL阳性平滑肌细胞在第2周较对照组低（t=2．418,P〈0．05）,并持续降低,到第8周几乎观察不到凋亡的平滑肌细胞（t=4．582,P〈0．01）。RhoA和ROCK2表达在第1周即高于对照组（t=6．056,t=8．411,P〈0．01）,第2周达高峰（t=9．342,t=10．437,P〈0．01）。到第4开始下降,但是到第8周仍然显著高于对照组（t=4．743,t=4．455,P〈0．01）。RhoA和Rho激酶活性在第1周也显著高于对照组（t=10．246,t=19．110,P〈0．01）,到第4周达高峰（t=24．984,t=16．124,P〈0．01）,然后开始降低,但是二者到第8周仍然显著高于对照组水平（t=4．934,t=10．426,P〈0．01）。结论在高肺血流所致的大鼠肺血管收缩和结构重构与RhoA／Rho激酶的表达和活性增强有关。     

Fas、bcl-2与移植血管瘤自然退化的动态研究

目的观察凋亡相关基因Fas和bcl-2与裸鼠移植血管瘤细胞增生、凋亡的关系，探讨其在血管瘤自然退化中的作用。方法采用免疫组化SP法检测裸鼠移植血管瘤不同时期Ki-67及凋亡相关基因Fas和bcl-2的表达。结果增殖期移植血管瘤组织中Fas无表达或低表达，而在移植血管瘤退化期中高表达，其阳性表达率在增殖期和消退期分别是25．0％和100％，两者之间差异有显著性意义（P〈0．001）；增殖期移植血管瘤中bcl-2高表达，而在移植血管瘤消退期中低表达，其阳性表达率在增生期和消退期分别是87．50％和31．25％（P〈0．001）；增殖期移植血管瘤组织中均有Ki-67表达，而在移植血管瘤消退期中表达极弱或无表达，其阳性表达率在增生期和消退期分别是100％和37．50％（P〈0．001）。结论在移植血管瘤增殖消退中存在着逐渐活跃的凋亡现象，细胞凋亡是血管瘤的生物学特征，是血管瘤自然消退的关键因素。Fas和bcl-2参与了血管瘤中细胞凋亡的调节。     

过敏性紫癜患儿治疗前后淋巴细胞凋亡情况分析

目的 探讨细胞凋亡在过敏性紫癜（HSP）发病及病理过程中的作用。方法 对73例HSP患儿治疗前后采用原位末端标记技术进行制片，在光学显微镜下观察，计算出凋亡淋巴细胞的比率。结果 HSP急性期（治疗前）组淋巴细胞凋亡较正常对照组延迟（P〈0．01）；而治疗后组淋巴细胞凋亡较治疗前组明显增多（P〈0．01）。结论 过敏性紫癜（HSP）存在淋巴细胞凋亡延迟，经治疗随着临床症状好转，淋巴细胞凋亡逐渐恢复正常，推测可用促细胞凋亡的药物治疗HSP。     

邻苯二甲酸二(2-乙基)己酯诱导小鼠胚胎Leydig细胞凋亡的体外实验研究

目的研究邻苯二甲酸二（2-乙基）己酯EDi（2-ethylhexyl）phthalate，DEHP]对小鼠胚胎睾丸Leydig细胞凋亡诱导作用。方法DEHP50、100、200ug／ml分别作用于体外培养、纯化的胎龄16d（GD16）小鼠胚胎睾丸Leydig细胞。光镜、电镜观察Leydig细胞形态和超微结构变化，DNA末端原位标记染色法（TUNEL法）和Annexin-V／PI双染法流式细胞仪分别检测kydig细胞凋亡指数、凋亡细胞比例。结果DEHP处理组透射电镜可见凋亡典型的形态学变化，凋亡指数及凋亡细胞比例与对照组比较有显著或非常显著性差异（P〈0．05或P〈0．01）。结论DEHP能诱导小鼠胚胎睾丸Leydig细胞凋亡。     

凋亡调控基因在急性白血病的表达及表其临床意义

研究凋亡调控基因bcl-2和baxmRNA在急性白血病（AL）中的表达、相互关系和临床意义,应用半定量逆转录－聚合酶链反应（RT－PCR）和Southern blot技术检测32例AL患儿bcl-2和baxm RAN水平及其比值,结果显示,AL患儿骨髓细胞中bcl-2mRNA的表达明显高于对照组（P＜0．001）,bclmRNA的表达明显隆低（P＜0．01）,治疗后,bcl-2mRNA在两组间的表达无差异（P＞0．05）,分析bcl-2/baxmRNA表达比值显示,高比值组NR率显著高于低比值组（P＜0．05）,结论：AL的发病与凋亡调控基因bcl-2和bax的异常表达有关,bcl-2和baxmRNA及其比值可作为判断AL预后及指导治疗的主要监测指标。     

二次白消安腹腔注射致小鼠精子再生模型的建立

目的 建立一种小鼠精子再生模型.方法 采用不同剂量白消安(单剂10 mg/kg、20 mg/kg和间隔24 d二剂10 mg/kg)小鼠腹腔注射,分别于给药后第一、二、三、四、六、八周取材,进行光镜和电镜观察.二次给药组采用免疫组化检测生精细胞Ki-67表达,TUNEL标记法检测生精细胞凋亡.结果 间隔24 d二剂10 mg/kg白消安对生精上皮的损伤效应介于单剂10 mg/kg和20 mg/kg之间.第二次给药后2周,生精上皮仅基底部残留以单个型精原细胞(As型)为主精原细胞和支持细胞;第三周As型精原细胞增多;第四周分化精原细胞和精母细胞出现;6～8周出现精子发生,生精上皮结构逐步恢复正常.二次给药后1～2周生精细胞凋亡指数显著高于对照组(P＜0.01),以后逐步下降,至第六～八周恢复至对照组水平(P＞0.05).精原细胞Ki-67阳性率在第一～二周最低,在第三周最高,第四周开始下降,第六～八周与对照组无显著差异.结论 间隔24 d二次白消安(10mg·kg-1·次-1)腹腔注射可建立小鼠精子再生模型,诱导凋亡为白消安致生精细胞损伤的重要机制.二次给药后3周主要为精原干细胞增殖期,4周为分化期,6～8周为精子发生恢复期.     

单侧隐睾大鼠对侧睾丸组织中SCF/c-kit基因表达变化及意义

目的 研究单侧隐睾大鼠对侧睾丸病理变化及SCF/c-kit基因表达,探讨单侧隐睾致对侧睾丸损害的机制.方法 30只SD大鼠随机分为对照组和实验组,实验组复制单侧(左侧)腹腔隐睾模型.3个月后分别取两组右侧睾丸组织进行real-time RT-PCR、Western blot及免疫组化检测干细胞生长因子(SCF)和其受体c-kit基因及其蛋白表达变化,TUNEL法检测细胞凋亡.结果 所有动物均存活,与对照组相比实验组对侧睾丸明显缩小,光镜下观察其曲细精管发生退化,生精上皮变薄,管腔较空,生殖细胞明显减少,细胞凋亡增加.两组凋亡指数分别为14.4±0.63和4.45±0.37,差异有统计学意义(P＜0.05).荧光实时定量PT-PCR检测SCF、c-kit基因mRNA含量,实验组对侧睾丸明显降低,两组相比差异有统计学意义(P＜0.05).Western blot检测SCF及c-kit蛋白表达含量,实验组对侧睾丸同样明显降低,两组相比差异有统计学意义(P＜0.05).免疫组化染色显示各级生精细胞膜均有c-kit表达,SCF主要表达于支持细胞膜表面,但实验组两者的表达均较对照组减弱.相关性检验SCF与AI相关系数r=-0.941,P＜0.01;c-kit与AI相关系数r=-0.908,P＜0.01;SCF与c-kit相关系数r=0.956,P＜0.01,均有统计学意义.结论 单侧隐睾可致对侧睾丸SCF/c-kit基因表达减弱,生精细胞凋亡增加引起不育.     

Maternal vitamin B12 deficiency affects spermatogenesis at the embryonic and immature stages in rats

To evaluate the role of cobalamin (Cbl) on spermatogenesis, the effect of dietary vitamin B(12) deficiency on early spermatogenesis was histologically investigated in male fetuses and newborns in the first filial generation (F(1) males) of rats. There was no difference in the number of gonocytes and supporting cells of Sertoli in the gonad in male fetuses on day 16 of gestation and in the testes in F(1) males at 0 days of age between vitamin B(12)-deficient (VB12-D) and vitamin B(12)-supplemented (VB12-S) groups. However, at 21 days of age, a decreased number of spermatogonia and no spermatocytes were observed in the VB12-D group. Numerous TUNEL positive cells were located among spermatocytes of the spermatogenic epithelium. The ultrastructural features examined using transmission electron microscopy were considered to be indicative of apoptosis. The incidence of seminiferous tubules having apoptotic cells was 51.5% in the VB12-D group. At 60 days of age, aplasia of the spermatids and spermatozoa was detected in the VB12-D group. In the connective tissue between the seminiferous tubules, many interstitial Leydig cells and blood vessels were observed in the VB12-D group, as compared with the VB12-S group. These changes produced by vitamin B(12) deficiency can be reversed by providing a VB12-S diet after weaning at 21 days of age. From these findings, such a vitamin B(12) deficiency during gestation and lactation could affect the germ cells and especially damage spermatocytes in F(1) male rats, which indicates that Cbl may be an essential constituent in the meiosis of spermatogenesis.     

Methotrexate induces germ cell apoptosis and impairs spermatogenesis in a rat

Purpose

The primary toxic effects of methotrexate (MTX) are myelosuppression and/or intestinal mucositis. The objective of the present study is to investigate the effect of MTX on germ cell apoptosis and spermatogenesis in a rat.

Methods

Male Sprague–Dawley rats were divided into three experimental groups: control rats treated with vehicle; MTX-2 rats treated with one dose (20 μg/kg) of MTX given IP and killed on the second day; and MTX rats treated with IP MTX (20 μg/kg) and killed on day 4. Johnsen’s criteria and the number of germinal cell layers in the testes were used to categorize the spermatogenesis. TUNEL assay was used to determine germ cell apoptosis. Western blotting was used to determine Bax and Bcl-2 protein levels. Statistical analysis was performed using the non-parametric Kruskal–Wallis ANOVA test, with p less than 0.05 considered statistically significant.

Results

On day 2, MTX-treated animals demonstrated minimal changes in the histological parameters of spermatogenesis, but germ cell apoptosis increased significantly (threefold increase, p = 0.002) compared to control rats. On day 4, MTX-treated rats demonstrated a trend toward a decrease in germ cell apoptosis, compared to day 2, and showed histological signs of impaired spermatogenesis (decreased number of germ cell layers and Johnsen’s criteria). A significant increase in cell apoptosis in MTX-treated rats was correlated with higher Bax/Bcl-2 protein levels.

Conclusions

MTX induced germ cell apoptosis and impaired spermatogenesis in rat testes.     

Effect of allopurinol on germ cell apoptosis following testicular ischemia–reperfusion injury in a rat

Recent evidence suggests that apoptosis is involved in germ cell loss following testicular ischemia-reperfusion (IR) injury. Allopurinol (Allo) is as a free radical scavenger which prevents tissue damage caused by reperfusion and oxygenation after ischemia; however, its effect on apoptosis in this type of injury has not been studied. To examine the effect of allopurinol on germ cell apoptosis following testicular IR in a rat. Forty rats were divided randomly into 4 experimental groups of 10 rats each: group A (Sham)-Sham operated animals; group B (Sham-Allo)-Sham operated rats treated with allopurinol given PO (by gavage) at a dose of 200 mg/kg, once daily, immediately before and 24 h following operation; group C (IR)-rats underwent 90 min of unilateral testicular ischemia and 48 h of reperfusion; group D (IR-Allo)-rats underwent IR and were treated with allopurinol similar to group B. The ipsilateral and contralateral testes were harvested 48 h following operation. Johnsen's criteria and the number of germinal cell layers were used to categorize spermatogenesis. TUNEL assay was used to determine germ cell apoptosis. Statistical analysis was performed using one-way ANOVA test, with P < 0.05 considered statistically significant. Testicular ischemia in rats led to histological damage in the ipsilateral testis. In the contralateral testis minimal damage was observed. Treatment with allopurinol increased significantly Johnsen's score in both the ischemic (7.3 +/- 0.5 vs 5.6 +/- 0.5, P < 0.05) and contralateral (8.9 +/- 0.1 vs 8.3 +/- 0.2, P < 0.05) testis, compared to IR-animals. Germ cell apoptosis in both the ischemic and the contralateral testis increased significantly after IR. Treatment with allopurinol resulted in a significant decrease in germ cell apoptosis in the ipsilateral testis, expressed as the number of positive tubules per 100 tubules (AI-1, (apoptotic index) threefold decrease, P < 0.005) and the number of apoptotic cells per 100 tubules (AI-2, fivefold decrease, P < 0.005) as well as a significant decrease in germ cell apoptosis in the contralateral testis (AI-1, 3.5-fold decrease, P < 0.05, AI-2- sixfold decrease, P < 0.005) compared to IR animals. In a rat model of testicular IR, treatment with allopurinol decreases germ cell apoptosis in both ischemic and contralateral testes and improves spermatogenesis.     

TSEG-2基因在小鼠睾丸扭转复位模型中的表达特征

目的 探讨睾丸特异表达基因2(testis specific expressed gene 2,TSEG-2)在小鼠睾丸扭转复位模型中的表达特征.方法 昆明小鼠36只,随机分组为对照组(6只)、假手术组(6只)、单侧睾丸扭转复位实验组(24只).实验组分为2组,每组12只,左侧睾丸扭转720°维持2 h,分别于复位后1、7 d取扭转侧睾丸.采用HE染色、原位末端标记技术(TUNEL)观察睾丸组织形态改变;黄嘌呤氧化酶法、硫代巴比妥酸比色法测定超氧化物歧化酶(SOD)、丙二醛(MDA)活性;原位杂交法观测TSEG-2在睾丸生精细胞内的表达定位;实时定量PCR法检测TSEG-2基因在睾丸组织中的表达水平.结果 对照组和假手术组生精上皮排列规则,扭转复位后1、7 d的睾丸组织内生精上皮结构松散,出现生精细胞凋亡,Johnsen's评分分别降低23.4%、64.1%(P<0.01),SOD活性降低11.6%、22.2%(P<0.05),MDA活性升高69.6%、93.2%(P<0.01).TSEG-2基因表达定位于小鼠睾丸生精小管的精原细胞和精母细胞.与对照组比较,扭转复位1、7 d后睾丸组织内TSEG-2表达水平分别上调2.2倍、6.6倍(P<0.01).结论 成功建立小鼠睾丸扭转复位模型,TSEG-2表达上调可能与抗氧化酶活性下降、生精细胞凋亡有关.     

Effects of laparoscopic division of spermatic vessels on histological changes of testes: long-term observation in the model of prepubertal rat

Laparoscopic Fowler-Stephens and Palomo procedures are now commonly performed in children with high positioned intra-abdominal cryptorchidism and varicocele, respectively. During the procedures, the spermatic vessels are ligated and therefore the question of risk related to testicular atrophy is often raised. The long-term follow-up of the histology after the procedures is rare. In this study, we simulated a laparoscopic spermatic vessels clipping and division (SVCD) in a prepubertal rat model, and examined the histological alterations of the testes with regard to spermatogenic arrest between prepuberty and middle age. Thirty-day-old Wistar rats divided randomly into three groups underwent laparoscopic sham operation, unilateral SVCD and unilateral SVCD with additional contralateral orchiectomy, respectively. Histological investigations observed on semithin and paraffin sections were performed at seven different postoperative intervals between day 9 and day 540. We defined partial, most and complete spermatogenic arrest of the seminiferous tubules to correspond with mild, severe spermatogenic arrest and atrophy, respectively. Laparoscopic SVCD induced testicular spermatogenic arrest in a total of 85% of the operated testes with different severity; 27% of operated testes with mild or severe spermatogenic arrest were seen between puberty and middle age (day 45–540 postoperative), and their size was only slightly reduced. Of the operated testes, 51% showed atrophic signs with a striking decrease in size, and their contralateral testes revealed in all cases mild or severe spermatogenic arrest started as early as day 45 postoperatively. Parallel to the spermatogenic arrest, Leydig cell hyperplasia developed frequently in impaired testes, especially in those without contralateral testes, finally reaching a typical adenoma size. Laparoscopic SVCD in prepubertal rats could disturb spermatogenesis with differing severity in most cases. This impairment could persist from peripuberty to middle age, and even involve the contralateral testes, in the case of operated testes and show complete spermatogenic arrest. This study showed that laparoscopic SVCD may have high risk in compromising the operated testis.     

Effect of testicular ischemia-reperfusion on recruitment of neutrophils,E-selectin expression and germ cell apoptosis in the contralateral testis in a rat

Recent evidence suggests that neutrophil recruitment may initiate germ cell apoptosis in the ischemic testis. The purpose of the present study was to examine the relationship between germ cell apoptosis and neutrophil recruitment in the contralateral testis following testicular ischemia-reperfusion (IR) injury in a rat. Adult male Sprague-Dawley rats were divided randomly into two experimental groups: Group A: Sham operated animals; Group B: IR rats underwent 90 min of unilateral testicular ischemia following by 96 h of reperfusion. The rats were sacrificed and testes were harvested. Johnsen's criteria and the number of germinal cell layers were measured to categorize the spermatogenesis. TUNEL assay was used to determine germ cell apoptosis in both the ischemic and contralateral testis. The recruitment of neutrophils was calculated per 100 venules. Expression of E-selectin was determined using immunohistochemical analysis. Statistical analysis was performed using Student's t test, with P less than 0.05 considered statistically significant. Germ cell apoptosis in both the ischemic and the contralateral testis increased significantly after IR. E-selectin expression was significantly greater in ischemic testis from IR rats compared to sham animals. The small increase in E-selectin expression and the concomitant increase in neutrophil recruitment in the contralateral testis of the IR rats (vs. sham animals) were not statistically significant. In conclusion, testicular ischemia causes an increase in germ cell apoptosis in the contralateral testis. Mechanisms other than neutrophil recruitment apparently initiate this process.     

Estimating the testis volume during the fetal period using the stereological method

The purpose of the present study is to assess the development of seminiferous tubule volume, stromal volume and total testis volume in the human fetal testis during the fetal period using the stereological method.In this study, we examined 90 testes of 45 human fetuses with no congenital anomalies and pathologies. They were aged between 12 and 40 weeks and localized between the scrotum and the abdomen. Total testis volume, seminiferous tubule volume, and stromal volume were estimated using Cavalier Principles. The weight and density of the testes were calculated as well.During the fetal period, the testes were firstly found on the right in the 27th week and on the left in the 32nd week in the scrotum. At the end of the third trimester and full term, the migration of the testes into scrotum was completed (98%). When the second trimester, third trimester and full term fetuses were compared, the differences between testis volumes were significant (p<0.001). The density of the testes between the groups was not significant (p>0.05). Testis parameters during the fetal period were not significantly (p>0.05) different between the right and left testes localization. The correlation between the fetal testis parameters was significant (p<0.001).Towards the end of the fetal period, the rate of seminiferous tubule volume to stromal volume changed in the favor of seminiferous tubule volume. It was observed that interstitial tissue became more regular and had a good organized structure with the progress of gestational age. In the third trimester, the lumen in the seminiferous tubules became more regular and clear and the interstitial tissue had a clear appearance.     

Germ-cell degeneration in experimental unilateral cryptorchidism: role of apoptosis

We investigated the possible involvement of apoptosis in the increased germ-cell degeneration in undescended testes (UDT). Experimental unilateral cryptorchidism was induced in 21-day-old rats, and both testes were removed for in-situ TUNEL staining of apoptotic cells at 1, 3, 7, 10, and 14 days postoperation. A gradual increase in the incidence of apoptosis was seen at 21–28 days of age in the control testes, followed by a decrease thereafter. After 10 days postoperation, the weight of the UDT was significantly lower than that of the contralateral descended testis (CDT) and the controls. However, the weight of scrotal testes in each group was similar. UDTs demonstrated a markedly increased incidence of apoptosis. By 7 days postoperation, the percentage of seminiferous tubules containing apoptotic germ cells significantly increased in UDTs compared with that in CDTs and controls (P < 0.001). Moreover, there was a significant difference in the percentage of seminiferous tubules containing apoptotic germ cells between CDTs and controls (P < 0.01). In addition, an increased incidence of seminiferous tubules containing 8–10 and >10 apoptotic germ cells from 7, 10, and 14 days postoperation in UDTs was detected. In-situ TUNEL analysis demonstrated spermatocytes to be the main type of germ cells affected in all groups. These findings suggest that spermatogenesis decreases not only in the␣UDT, but also in the CDT, and that the germ-cell degeneration in cryptorchidism took the form of apoptosis.