不同日龄隐睾复位大鼠睾丸组织结构观察

目的 观察不同13龄隐睾复位大鼠睾丸组织结构的变化.方法 72只21 d雄性SD大鼠随机分为单侧隐睾组、双侧隐睾组、假手术对照组各24只.建立单、双侧隐睾动物模型.2周后行隐睾大鼠睾丸下降固定术,于日龄40、60 d处死取睾丸,采用苏木素.伊红染色光镜下观察各组大鼠精曲小管生育力指数(TFI)和平均精曲小管直径(MTD);生物素-dUTP/酶标亲和素法(TUNEL法)检测睾丸生殖细胞凋亡情况.结果 隐睾侧睾丸MTD、TFI显著低于阴囊内睾丸,而隐睾生殖细胞凋亡指数(AI)明显增高于阴囊内睾丸(P＜0.05);单侧隐睾组阴囊内睾丸TFI低于相应日龄的假手术对照组,但无统计学意义(P＞0.05).40 d时单侧隐睾组隐睾侧睾丸生殖细胞AI较双侧隐睾组低(P＜0.05),日龄60 d,各组隐睾侧睾丸AI较40 d时明显降低(P＜0.05),但单侧隐睾和双侧隐睾AI比较无统计学差异(P＞0.05).结论 实验隐睾复位大鼠睾丸AI升高,同时单侧隐睾鼠对侧睾丸组织存在不同程度的损害.随着复位时间的延长,隐睾组织的病理损害有恢复的趋势.     

人绒毛膜促性腺激素对单侧隐睾大鼠睾丸生殖细胞凋亡的影响

目的探讨人绒毛膜促性腺激素（HCG）对单侧隐睾大鼠睾丸生殖细胞凋亡的影响。方法将sD雄性大鼠40只随机分为单侧隐睾组和假手术组，各20只，于日龄21d制备单侧隐睾模型。单侧隐睾组和假手术组各又分为HCG治疗组和未治疗组。日龄22d时HCG治疗组开始肌肉注射HCG20U，隔日1次，共7次。日龄35、60d时处死大鼠，取其睾丸，采用生物素-dUTP／酶标亲和素法（TUNEL法）检测其生殖细胞凋亡水平。结果单侧隐睾组隐睾睾丸生殖细胞凋亡指数（AI）高于假手术组，但无统计学差异（P〈0．05）；单侧隐睾各组对侧睾丸生殖细胞AI高于假手术未治疗组（P〉0．05）。假手术和单侧隐睾模型HCG治疗组阴囊内睾丸生殖细胞AI高于相应未治疗组，且35d假手术组治疗组与未治疗组间差异有统计学意义（P〈0．05）。60d单侧隐睾HCG治疗组大鼠隐睾侧睾丸生殖细胞AI高于相应未治疗组，但无统计学差异（P〉0．05）。结论单侧隐睾时不仅患侧睾丸生殖细胞凋亡增加，而且隐睾对侧阴囊内睾丸生殖细胞凋亡也增加；HCG的应用可加重睾丸生殖细胞凋亡，且停用后仍存在着一些不可逆的病理损害，故临床应用HCG要慎再，应尽早手术治疗。     

冷诱导RNA结合蛋白在小鼠隐睾中的表达及与生精细胞损害的关系

目的 探讨冷诱导RNA结合蛋白(cold inducible RNA-binding protein,CIRP)在小鼠隐睾睾丸中的表达,及其与隐睾生精细胞损害的关系.方法 雄性BALB/c小鼠20只用手术的方法建立左侧隐睾模型,分别于术后第7天和第10天取双侧睾丸称重,光镜下观察睾丸组织学变化,RTPCR法检测睾丸CIRP mRNA的表达水平,Western-blot检测睾丸CIRP蛋白的表达水平,并用Annexin V-FITC/PI双染流式细胞仪检测生精细胞凋亡.结果 CIRP强表达于正常小鼠睾丸中,隐睾模型建立后,隐睾睾丸CIRP mRNA和蛋白的表达水平均明显降低(P<0.05),术后第10天隐睾睾丸CIRP表达水平明显低于第7天(P<0.05).同时隐睾侧睾丸重量较对侧睾丸明显减轻(P<0.05),术后第10天对侧睾丸重量明显重于第7天(0.102±0.006)g比(0.080±0.005)g(P<0.05);而术后第7天和第10天隐睾睾丸重量相比,(0.072±0.007)g比(0.062±0.004)g(P>0.05),差异无统计学意义.另外生精细胞的凋亡明显增加(P<0.05),术后第10天隐睾侧睾丸的生精细胞凋亡率明显高于第7天(29.2%±1.3%)比(20.2%±1.6%)(P<0.05);而对侧睾丸生精细胞凋亡率无明显差异(5.8%±1.5%)比(5.6%±1.8%)(P>0.05).组织切片显示隐睾侧睾丸生精上皮变薄,生精细胞排列紊乱.结论 小鼠隐睾中CIRP的表达明显降低,CIRP表达的降低可能在隐睾生精细胞损害中起着重要的作用.     

The influences of Di-2-ethyithexyl phthalate and its' metabolites (single-ethylhexyl phthalate, MEHP) on spermatogenic epithelium in young wistar rats

目的 了解环境内分泌干扰因子邻苯二甲酸二乙基已酯(Di-2 ethyhhexyl phthalate,DEHP)及代谢产物邻苯二酸-单2乙基己酯(single-ethylhexyl phthalate,MEHP)对幼鼠生精上皮有否损害.方法 生后2周龄的Wistar雄性幼鼠96只,随机分8组,每组12只.随机选择1组行生理盐水(0.2 ml·kg-1·d-1,喂养3周)灌胃,作为正常对照组(NC组);再选1组行环磷酰胺(CTX 100mg·kg-1·d-1,喂养1周)灌胃,作为阳性对照组(PC组);余各组分别用DEHP、MEHP按低剂量(100mg·kg-1·d-1,喂养3周)、中剂量(200mg·kg-1·d-1,喂养2周)、高剂量(300 mg· kg 1·d-1,喂养1周)灌胃制作动物模型.观察不同时期、不同剂量下睾丸生精上皮的变化,并应用TUNEL法检查生精上皮凋亡,免疫组织化学法检测睾丸组织中c kit、PI3K的表达,放射免疫法检测血清性激素(卵泡刺激素FSH、黄体生成素LH以及睾酮T)水平.结果 睾丸生精上皮的变化:用药组生精上皮萎缩、发育落后,数量少、线粒体肿胀、空泡化等,但在用药的高剂量组短时间内损害与低剂量组长时间的损害一致;①睾丸组织中生精上皮凋亡:与NC组(凋亡指数AI:0.015±0.002)比较,用药组生精上皮凋亡(DEHP和MEHP组的AI分别为:0.051±0.004、0.073±0.005)显著增多(P＜0.01),但在高剂量短时间喂养组与低剂量长时间喂养组间比较,差异无统计学意义(P＞0.05);②睾丸组织中c-kit、PI3K的表达:与NC组(c-kit:0.389±0.014、PI3K:0.457±0.011)比较,用药组睾丸组织中c-kit(DEHP和MEHP组分别为:0.185±0.018、0.165±0.014)及其下游信号分子PI3K (DEHP和MEHP组分别为:0.205±0.019、0.189±0.017)的表达显著减少(P＜0.01),但高剂量短时间喂养组与低剂量长时间喂养组间比较,差异也无统计学意义(P＞0.05).血清性激素水平:与NC组比较,用药组FSH、LH水平升高,T水平下降(P＜0.05);但用药的高剂量组与低剂量组间比较差异无统计学意义(P＞0.05).结论 DEHP及代谢产物MEHP对幼鼠生精上皮有明显损害,推测可能对小儿的生精功能也有损害,小儿应避免过多的长期接触含DEHP及代谢产物MEHP的物质.     

大鼠隐睾、隐睾下降固定对生殖细胞发育、凋亡影响的实验研究

目的　探讨大鼠隐睾、隐睾下降固定对生殖细胞发育、凋亡的影响。方法　 6 8支SD雄性大鼠 ,分为三组 :隐睾组 (5 2 ) ;隐睾下降固定组 (8)及对照组 (8)。于日龄 2 2d时复制单侧隐睾模型 ,一部分于单侧隐睾模型术后 14d复制隐睾下降固定模型。采用生物素 dUTP/酶标亲和素测定法检测睾丸生殖细胞凋亡。结果　 1.单侧隐睾模型术后第 13d内隐睾侧睾丸生殖细胞凋亡指数随时间的延长而增加 ;从术后第 4天起 ,与自身对侧正常睾丸 (4.8± 0 .9) %相比 ,隐睾侧睾丸 (11.2±3.6 ) %发生凋亡的生殖细胞数显著增加 (P <0 .0 1)。 2 .与隐睾组 (47.5± 8.6 ) %相比 ,隐睾下降固定组 (6 .2± 1.8) %发生凋亡的生殖细胞数显著降低 (P <0 .0 1)。结论　 1.大鼠隐睾模型建立术后13d内隐睾侧睾丸生殖细胞凋亡指数随时间的延长而增加 ;2 .在日龄 36d时隐睾下降固定能使大鼠隐睾恢复正常的生精功能 ;3.隐睾患儿应及早施行睾丸下降固定术 ,可以恢复其生育能力。     

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.     

单侧隐睾对侧睾丸损害机制的实验研究

目的 神经分子水平方面探讨单侧隐睾鼠对侧睾丸损害的机制。方法 30只SD雄性大鼠,分为对照组(A组)、隐睾组(B组)、隐睾加生殖股神经(GFN)切断组(C组)。每组各10只。免疫组化技术观察降钙素基因相关肽(CGRP)阳性细胞的神经分布;生物素-dUTP/酶标亲和素测定法检测睾丸生殖细胞凋亡;透射电镜观察Sertoli细胞的超微结构;化学比色法测定丙二醛(MDA)的含量。结果 A组睾丸中含有大量CGRP的神经细胞。术后90d,与A组比较,B组对侧睾丸CGRP明显降低,MDA升高,细胞凋亡增加(P<0.01),Sertoli细胞有缺血性改变。C组的这种损害减轻(P<0.01)。结论 单侧隐睾鼠对侧睾丸的损害是通过隐睾GFN传入到交感神经,反射性引起对侧睾丸CGRP降低,睾丸缺血缺氧,氧自由基升高,生殖细胞凋亡增加所引起的。     

褪黑激素对大鼠睾丸扭转缺血和再灌注损伤的保护作用

目的 探讨褪黑激素对大鼠睾丸扭转的治疗作用.方法 选取青春期雄性SD大鼠48只,随机分为3组:空白对照组(A组);扭转复位组(B组);扭转复位+褪黑激素组(C组).B组和C组大鼠建立睾丸扭转复位模型,对照组不扭转.扭转4h后复位睾丸,复位前15 min B组腹腔注射生理盐水1 ml:C组腹腔注射褪黑激素1ml(17 mg/kg).复位后4h处死所有动物取睾丸待测.以原位缺口末端标记法(TUNEL)检测生精细胞凋亡指数;化学比色法测定睾丸组织内总抗氧化能力(T-AOC).结果 B组T-A0C( 20.31±2.55)U/mg比A组(33.62±3.29) U/mg明显降低,差异有统计学意义(P＜0.01).而C组T-AOC(30.05±2.08)U/mg较B组明显升高(P＜0.05).B组凋亡指数(42.2±3.21)％明显高于A组(5 7±0.67)％(P＜0.01),而C组凋亡指数(12.2±1.34)％较B组显著下降(P＜0.05).结论 褪黑激素具有明显对抗睾丸扭转复位后的氧化损伤,对因睾丸扭转导致的缺血再灌注损伤具有保护作用.     

人绒毛膜促性腺激素对内分泌型双侧隐睾大鼠生精细胞凋亡的影响

目的探讨人绒毛膜促性腺激素(HCG)对内分泌型双侧隐睾大鼠生精细胞凋亡的影响。方法取60只SD大鼠,随机抽取20只作为正常对照组,余40只采用皮下注射17-β雌二醇方法制成双侧隐睾模型,随机分为隐睾+HCG组、隐睾+9 g.L-1盐水组(隐睾+NS组)。隐睾+HCG组自日龄26 d起隔日肌注HCG 20 U,共10次;隐睾+NS组自日龄26 d起隔日肌注9 g.L-1盐水1 mL,共10次。于日龄45 d、60 d每组各抽10只采集血清,取其睾丸组织后处死。采用ELISA间接法测定其血清抗精子抗体(AsAb)水平,放射免疫方法测其血清雌二醇(E2)、睾酮(T)水平,生物素-脱氧尿嘧啶核苷三磷酸(dUTP)/酶标亲和素(TUNEL)法检测其生殖细胞凋亡指数(AI)。结果日龄45 d和60 d时,隐睾+HCG组双侧睾丸生殖细胞AI均明显高于同日龄其他各组(Pa<0.01);隐睾+HCG组血清E2水平较同日龄其余各组均显著增高(Pa<0.01),而血清T水平均降低(Pa<0.01);隐睾+HCG组AsAb水平均高于同日龄其他组(Pa<0.01)。结论 HCG注射治疗双侧隐睾大鼠不仅能增加AsAb产生,且能加重睾丸生殖细胞的凋亡。     

胚胎期氟他胺暴露对睾丸Connexin43基因表达的影响

目的 探讨胚胎期雄激素受体阻滞剂氟他胺(flutamide,Flu)暴露对睾丸间隙连接蛋白43 (Connexin43,Cox43)基因表达的影响.方法 在胚胎期12～21 d,给予SD孕鼠氟他胺25 mg·kg-1·d-1皮下注射.应用免疫组织化学法和Real-time PCR技术,分析出生后13、15、17、20 d SD幼鼠睾丸Cox43 mRNA表达及蛋白质定位分布情况.结果 出生后(postnatal day,PD) 13、15d,Flu暴露组睾丸Cox43 mRNA和蛋白表达均明显低于正常对照组及玉米油对照组(P＜0.01);PD17 Flu暴露组与正常对照组及玉米油对照组相比,睾丸Cox43 mRNA表达差异无统计学意义(P＞0.05),而生精小管基底膜精原细胞层有Cox43蛋白异常表达分布;PD20 Flu暴露隐睾组Cox43 mRNA表达明显低于正常对照组(P＜0.05)、玉米油对照组(P＜0.05)和非隐睾组(P＜0.01),并且PD20 Flu暴露各组生精小管基底膜精原细胞层均有Cox43蛋白异常分布.结论 胚胎期Flu暴露导致睾丸Cox43mRNA转录下降,生精小管内Cox43蛋白表达分布异常.     

Effect of diclofenac on germ cell apoptosis following testicular ischemia-reperfusion injury in a rat

Recent evidence suggests that enhanced cell apoptosis is responsible for germ cell loss following testicular ischemia-reperfusion (IR) injury. A nonsteroidal anti-inflammatory drug diclofenac sodium (Voltaren) is a prostaglandin-synthesis inhibitor, which is widely used in many testicular disorders. The purpose of the present study was to examine the effect of diclofenac (DIC) on germ cell apoptosis in the ischemic and contralateral testes following testicular IR in a rat. Forty rats were divided randomly into four experimental groups of ten rats each: group A (Sham)—Sham operated animals; group B (Sham-DIC)—Sham operated rats that were treated with DIC given subcutaneously at a dose of 10 mg/kg, once daily, 24, 48 and 72 h following operation; group C (IR) underwent 90 min of unilateral testicular IR; group D (IR-DIC)—rats underwent 90 min of unilateral testicular IR and were treated with DIC similarly to group B. Ninety-six hours following operation, the rats were sacrificed and testes were harvested. Johnsen’s criteria and the number of germinal cell layers were used to categorize the spermatogenesis. TUNEL assay was used to determine germ cell apoptosis in both the ischemic and contralateral testes. Statistical analysis was performed using the non-parametric Kruskal–Wallis ANOVA test, with P less than 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. Germ cell apoptosis in both the ischemic and the contralateral testes increased significantly after IR. Treatment with DIC did not change histologic parameters of spermatogenesis in both the ischemic and contralateral testes, but decreased germ cell apoptosis in both testes following testicular IR. We conclude that testicular ischemia causes an increase in germ cell apoptosis in the contralateral testis. Diclofenac may be beneficial for spermatogenesis following testicular IR by decreasing germ cell apoptosis.     

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.     

Cryptorchidism: a morphological study of 670 biopsies

Among a series of 512 boys with an empty scrotum, 495 (96.7%) were found to have cryptorchidism, 4 had ectopia and 13 unilateral anorchia. Cryptorchidism was bilateral in 106 boys (21.4%). The only anomaly consistently associated with cryptorchidism was a detached epididymis, present in 31 patients. A total of 670 biopsies were studied, 441 of which came from cryptorchid and 229 from scrotal testes. Spermatogonial counts, performed according to Mancini's method, showed the germ cell population to be diminished in nearly all cryptorchid testes. The seven boys who still had a well preserved germ cell population were found in a group of 51 patients operated before age three; four of the seven boys with normal counts were below age one. No difference in the mean spermatogonial counts was found between uni- und bilateral cryptorchidism and ectopia, with the exception of bilaterally intraabdominal testes whose spermatogonial cell loss was particularly severe. Mean counts remained constant during childhood, no gradual increase with age having been observed. The scrotal testes in unilateral cryptorchidism showed cell loss in 30.1% of the cases, the germ cell depletion being severe in one out of every six cases. In the remaining scrotal testes, the counts were in the low normal range with a significantly lower mean than that found in scrotal testes associated with anorchia. Control biopsies were performed several months or years after orchidopexy in 18 boys with unilateral and in 24 boys with bilateral cryptorchidism. Orchidopexy does not improve the number of germ cells in either originally cryptorchid or in scrotal testes, the only postoperative change being an increase in tubular diameter. A search for malignant tumours which could have developed in this series has remained negative. According to our data, no optimal time for orchidopexy can be proposed. The damage to germ cells, once established, seems to remain unchanged during childhood at least after age three, and does not warrant special timing for operative correction of cryptorchidism.     

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.     

卡维地洛对免疫性心肌炎小鼠心肌凋亡调控蛋白影响

目的 观察卡维地洛对免疫性心肌炎小鼠心肌凋亡蛋白Bcl-2、Bax及Fas的影响,探讨其在免疫性心肌炎中的作用.方法 60只4～5周龄近交系雄性BALB/C小鼠,随机将小鼠分为3组:免疫性心肌炎模型组(模型组),卡维地洛干预组(干预组),空白对照组(对照组),每组20只.眼球取血后处死,留取心脏及血液标本.光镜观察各组小鼠心肌组织病理学改变,电镜观察心肌细胞超微结构,化学发光免疫法检测血清心肌肌钙蛋白I(cTnI)水平;免疫组化法检测心肌Bcl-2、Bax及Fas蛋白表达的含量;原位缺口末端标记(TUNEL)法检测心肌细胞凋亡情况.结果 模型组小鼠心肌细胞光镜下见大量炎症细胞浸润,电镜下可见细胞核固缩、染色质边集,对照组小鼠心肌细胞光镜下未见炎症细胞浸润,电镜下染色质边集不明显.模型组Bcl-2、Bax及Fas蛋白表达较对照组高(23.48±2.24 vs 6.64±1.60,26.15±2.02 vs 5.09±0.85,21.22±3.62 vs 5.86±1.37,P＜0.01);干预组与模型组比较,小鼠心肌组织炎症积分轻(2.60±0.3l vs 2.02±0.26,P＜0.05),电镜下细胞核固缩轻,心肌细胞Bcl-2、Bax及Fas蛋白表达低(17.13±1.94 vs 23.48±2.24,17.66±2.62 vs26.15±2.02,16.79±2.83 vs 21.22±3.62,P＜0.05),凋亡指数低[(16.61±4.67)%vs(24.51±4.70)%,P＜0.05],cTn-I水平低[(1.878±0.48)ng/ml vs(1.102±0.23)ng/ml,P＜0.05].结论 卡维地洛可减轻免疫性心肌炎小鼠心肌细胞凋亡,对免疫性心肌炎起一定的保护作用.

Abstract：

Objective To observe the effects of carvedilol on the expression of Bcl-2, Bax and Fas in autoimmune myocarditis (AM).Methods A total of 60 inbred male BALB/C mice 4-5 weeks of age were divided at random into 3 groups as follows: AM group ( n = 20), carvedilol group ( n = 20 ) and control group (n = 20).The mice were sacrificed after gathering blood specimens by taking out the eyeballs and hearts tissue.The histological and ultrastructural changes were observed under light microscope and electron microscope.The concentrations of cardiac troponin Ⅰ (cTn Ⅰ ) were detected by chemiluminescence immunoassay (CLIA).Immunohistochemistry (IHC) was performed to analyze the contents of Bcl-2, Bax and Fas, TUNEL to detect the apoptotic index in myocardial cells.Results There were large number of lymphocyte and monocyte infiltrates under light microscope and karyopyknosis and chromatin gathered along the nuclear membrane under electron microscope in AM group.There were no inflammations and chromatin gathering in group C.Compared with control group, the Bcl-2, Bax and Fas protein expression significantly elevated in AM group ( 23.48 ± 2.24 vs.6.64 ± 1.60,26.15 ± 2.02 vs.5.09 ± 0.85,21.22 ± 3.62 vs.5.86 ± 1.37, P ＜ 0.0l ) . The histopathologic scores ( 2.60 ± 0.31 vs.2.02 ± 0.26, P ＜ 0.05 ) and karyopyknosis of carvedilol group decreased as compared with AM group.The Bcl-2, Bax and Fas protein expression (17.13 ±1.94 vs.23.48 ±2.24,17.66 ±2.62 vs.26.15 ±2.02,16.79 ±2.83 vs.21.22 ±3.62, P ＜ 0.05 ), AI [( 16.61 ± 4.67 ) % vs.( 24.51 ± 4.70) %, P ＜ 0.05]and contents of cTnI [( 1.878± 0.48 ) ng/ml vs. ( 1.102 ± 0.23 ) ng/ml, P ＜ 0.05]also decreased in carvedilol group compared with AM group.Conclusion Carvedilol could protect against AM by alleviating cardiomyocyte apoptosis.     

雷公藤多苷对青春期大鼠睾丸组织及c-kit表达的影响

目的：探讨雷公藤多苷对青春期大鼠睾丸组织的近、远期影响及机制。方法：5周龄雄性Sprague-Dawley（SD）大鼠48只,随机分为对照组、小剂量和大剂量雷公藤多苷组（每组16只）,分别给予1%羧甲基纤维素钠、雷公藤多苷小剂量（每日6 mg/kg）和大剂量（每日12 mg/kg）灌胃4周,观察停药24 h和4周后大鼠的睾丸组织学改变及c-kit表达。结果：停药24 h,两个雷公藤多苷组睾丸生精细胞均减少、c-kit表达下调（P0.05）,大剂量组仍表现为生精上皮萎缩、间质水肿,睾丸重量、生精细胞 c-kit 表达下降（P<0.01）。结论：雷公藤多苷可致青春期大鼠睾丸组织损伤,小剂量引起睾丸组织近期可逆性损伤;大剂量可致睾丸组织远期损伤,提示睾丸间质病变可能是雷公藤多苷致青春期大鼠睾丸组织远期损伤的原因之一。     

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.     

高氧致慢性肺疾病新生鼠肺组织Smad4、Smad7表达及其可能作用

目的 探讨在高氧致新生鼠慢性肺疾病（chronic lung diseases,CLD）发生、发展过程中,肺组织Smad4、Smad7蛋白的变化及其可能的作用.方法 将出生12h内的64只新生Wistar大鼠,采用随机数字表法随机分为高氧组和空气组（对照组）,每组均为32只.采用高体积分数氧诱导CLD模型,于实验1、3、7、14d分离大鼠肺组织.应用Masson染色观察大鼠肺组织形态学改变,采用肺组织纤维化评分以确定肺纤维化程度.免疫组织化学、Western blot测定大鼠肺组织中Smad4、Smad7的表 达水平.结果 与空气组比较,高氧组大鼠肺组织纤维化程度明显增强（肺组织纤维化评分7 d：2.67±0.21 vs 0.58±0.17;14 d：4.48±0.24 vs 0.63±0.13,P＜0.05）;肺组织Smad4、Smad7蛋白主要定位于肺上皮细胞及间质成纤维细胞;Smad4蛋白表达强度明显增强（7 d：122.35±10.30 vs 140.08±7.77;14 d：129.70±7.33 vs 144.99 ±6.49,P＜0.05）;Smad7蛋白表达第14天明显减弱（132.16±4.38 vs126.22±6.49,P＜0.05）.结论 暴露高氧中的新生鼠,肺组织信号传导蛋白Smad4表达上调及Smad7表达下调可能与CLD肺纤维化的发生、发展密切相关.