环磷酰胺代谢产物丙烯醛对未成熟睾丸损伤的实验研究

目的探讨环磷酰胺（cyclophosphamide，CP）代谢产物丙烯醛（acrolein，ACR）对未成熟睾丸Sertoli细胞的氧化应激损伤及其机制。方法建立新生SD大鼠Sertoli细胞原代培养模型，实验组分别给予不同浓度的ACR溶液，对照组给予PBS溶液，3h、12h后分别用MTT法检测各组细胞活力，依据CD50分别采用50、100μmol／L浓度的ACR进行干预。免疫细胞化学法检测丙烯醛-蛋白质加合物（ACR-Pro），硫代巴比妥法和Fenton法检测细胞丙二醛（MDA）、羟自由基（OH·）含量，三价铁还原法、黄嘌呤氧化酶法和NADPH还原法测定细胞总抗氧化能力（T-AOC）、超氧化物歧化酶（SOD）和谷胱甘肽还原酶（GR）活性。结果①ACR可明显降低Sertoli细胞的活力，呈剂量时间依赖性（P〈0．05），3h、12h的CD50分别为105、200μmol／L；②ACR干预后可见细胞内ACR—Pro显著增多（P〈0．05）；③各实验组中氧化应激损伤的标志物MDA和OH·含量分别较对照组显著升高（P〈0．05）；④各实验组反映抗氧化能力的指标T-AOC、SOD和GR均较对照组明显降低（P〈0．05）。结论体外实验证实ACR对未成熟睾丸Sertoli细胞具有明显的氧化应激损伤，提示CP对未成熟睾丸的损伤很可能即是其代谢产物ACR的氧化应激损伤所致。     

环磷酰胺代谢产物丙烯醛对未成熟睾丸支持细胞骨架的影响及机制研究

目的 探讨环磷酰胺(cyclophosphamide,CP)代谢产物丙烯醛(acrolein,ACR)对未成熟睾丸Sertoli细胞骨架的损伤及其机制.方法 建立新生SD大鼠Sertoli细胞原代培养模型,实验组给予100 μmol/L浓度的ACR溶液,对照组给予PBS溶液,12 h后分别用超氧化物阴离子荧光探针染色观察Sertoli细胞内超氧化阴离子的变化,透射电子显微镜观察ACR对细胞超微结构的影响,免疫荧光染色观察细胞骨架中F-actin的分布变化,Western blot方法测定ERK和p38的表达水平.结果 ①ACR能增强sertoli细胞内活性氧的水平;②ACR处理后,实验组细胞线粒体发生了肿胀,染色质发生了凝集,出现胞质浓缩、核溢裂及并且空泡化现象,而微丝发生聚集、边缘化和区域化;③ACR干预后实验组ERK和p38MAPK表达活性明显上升.结论 CP的主要毒性代谢产物ACR通过引起未成熟睾丸Sertoli细胞的氧化应激反应损伤支持细胞骨架,从而导致儿童生精功能障碍,而其可能通过活化ERK和p38的表达使MAPK信号途径激活来实现这一损伤过程.

Abstract：

Objective To investigate the mechanism and the effect of Acrolein on cytoskeleton of Sertoli cells. Methods We isolated and identified Sertoli cells from 8-day-old SD rats' testes. The experimental groups were treated with Acrolein, The control group were treated with phosphate buffered solution. After 12 hours, the treated cells were dyed with DHE to assay the distribution of superoxide anion, and the ultrastructure of the Sertoli cells was examined with transmission electron microscope, F-actin was stained with fluorescent antibody, and the expression of ERK and p38 in Sertoli cells were assayed by Western blot. Results The photodensity of superoxide anion, the expression of ERK and p38 were increased in the experimental groups. After treatment with Acrolein, the ultrastructure of the Sertoli cells was changed, manisfested as chondriosome swelling; chromatin clumping, condensed endochylema; split nuclei and vacuolization. All of these changes were followed by a F-actin accumulation, marginalization and regionalization. Conclusions Acrolein, the main toxic metabolite of cyclophosphamide, damages the cytoskeleton of immature Sertoli cells through oxidization stress, which might activate the MAPK signaling pathway mainly by increasing the expressions of ERK and p38.     

端粒、端粒酶活性与血液系统肿瘤

本文着重阐述端粒假说及其血液系统肿瘤性疾病中的端粒长度及酶活性变化，并展望了抗端粒酶治疗肿瘤的可能性     

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。     

儿童霍奇金病组织中MDM2和p53蛋白表达与端粒酶活性的相关性研究

本文通过检测ＭＤＭ2蛋白和 ｐ5 3蛋白的表达及端粒酶 (ＴＬＭＡ)活性 ,研究其与儿童霍奇金病 (ＨＤ)发生、发展的关系。材料与方法一、临床资料收集 1997年 1月～ 2 0 0 0年 5月本院及外院手术切除ＨＤ标本 2 8例 ,均为淋巴结 ,其中男 18例 ,女 10例 ,平均年龄 8.8岁。 2 8例标本经病理诊断为ＨＤ ,按Ｒｙｅ标准分型 :淋巴细胞为主型 (ＬＰ)16例 ,混合细胞型 (ＭＣ) 8例 ,结节硬化型 (ＮＳ) 4例。临床分期 :Ｉ期 13例 ,Ⅱ期 6例 ,Ⅲ期 7例 ,Ⅳ期 2例 ,正常淋巴结 2 0例作为对照。二、检测方法ＭＤＭ 2蛋白和 ｐ5 3蛋白通过免疫组化方法检测…     

男童一侧睾丸缺失对另侧睾丸影响的研究

目的探讨男童一侧睾丸缺失后对另侧睾丸的生长、性激素水平及第二性征发育的影响。方法对61例一侧睾丸缺失的患儿进行随访，随访内容包括所剩睾丸体积、性激素水平及第二性征发育情况。同期检测150例各年龄段正常男孩相应指标。按≤10岁、10～12岁、〉12岁，分年龄段分别进行比较。结果各年龄段内，睾丸缺失组睾丸体积与对照组比较，差异均有统计学意义俨〈0．05），睾丸缺失的不同疾病组之间比较，无统计学意义（尸〉0．05）；睾丸缺失早期另侧睾丸肥大明显，并随缺失时间延长而减轻；10～12岁及〉12岁组肥大尤为明显，与≤10岁组比较，差异有统计学意义（P〈0．05）；性激素水平随年龄增长而增加，但相同年龄段内，睾丸缺失组与对照组比较，差异无统计学意义（P〉0．05）；睾丸缺失患儿第二性征的出现及发育与正常儿童无明显差异。结论男童一侧睾丸缺失可致另侧睾丸代偿性肥大（CTA）。术后早期睾丸代偿增大更为明显；睾丸发育启动及增长发育期睾丸代偿肥大较处于相对静止期更为明显。但其增大程度与引起缺失的原发疾病无明显关系，且CTA不影响其后的性激素水平及青春期第二性征的出现。     

环磷酰胺对不同发育时期睾丸生精细胞毒性损伤的动物实验研究

目的探讨抗肿瘤药物所致牛精细胞损害与年龄的关系，为性腺保护剂的运用寻找理论依据。方法将环磷酰胺分别作用于处于不同发育时期的1周龄、3周龄、5周龄、9周龄雄性大鼠，应用HE染色法、TUNEL法和免疫组化法检测急性期生精细胞凋亡，bcl-2蛋白表达，细胞增殖能力变化及远期组织学损害。结果用药后24h，除1周龄组外，各实验组生精细胞显著凋亡（P〈0．01），bcl-2蛋白表达显著下降（P〈0．01），生精细胞S期所占的比例显著下降（P〈0．01）。用药后9周，除1周龄组外，各实验组曲细精管面积、直径、生精上皮细胞计数、Johnsen’s评分均显著低于相应对照组（P〈0．01），并随年龄增大损害有增加趋势。结论环磷酰胺可诱导生精细胞增殖启动阶段以后的生精细胞显著凋亡，且伴有bcl-2明显下调。并可湿著抑制精原细胞和细线前期精母细胞增殖。远期组织学改变年龄越小所受的损害越小，提示为使睾丸免受抗肿瘤药物的伤害而采用性激素使生精细胞增殖分化处于不分化状态是有效的防护方法之一。     

单侧睾丸扭转后对另侧睾丸影响的研究进展

一侧睾丸扭转导致睾丸血供障碍而出现睾丸急性梗死时，另侧睾丸是否受到损害，其后是否影响生育能力均是医生和患者非常关注的问题。近来，单侧睾丸扭转后对另侧睾丸的影响已引起广泛的研究兴趣，本文就其研究进展作一综述。     

大鼠睾丸扭转复位及减压治疗对睾丸的影响

目的 研究睾丸扭转复位及减压治疗对睾丸的影响,为睾丸扭转的预后判断、治疗方法的选择等提供新的理论依据.方法 将30只SD雄性大鼠随机分成5组,制成睾丸扭转复位及复位+减压治疗的模型.分别设立空白对照组及实验A～D组,睾丸扭转/复位组(A组)、睾丸扭转/复位+减压治疗组(B组)喂养至术后1 d处死;睾丸扭转/复化组(C组)、睾丸扭转/复位+减压治疗组(D组)喂养至术后1个月处死.应用化学检测和组织学分析方法,观察睾丸大体标本的变化、睾丸组织内丙二醛(MDA)含量的变化及睾丸组织Johnsen's评分.结果 五组睾丸MDA分别为3.18±0.22(空白对照组)、9.54±2.05(A组)、7.92±1.38(B组)、6.67±0.61(C组)、4.30±1.81(D组),实验组术侧睾丸的MDA含量显著高于自身对侧(P<0.05).D组MDA含量比C组明显下降(P<0.05).单纯复位组的术后睾丸标本比自身对侧和减压治疗组有明显萎缩;五组Johnsen's评分分别为10±0、7.2±0.18、8.2±0.19、2.2±0.19、9.2±0.18.D组比C组明显提高(P<0.05).结论 睾丸扭转复位+减压治疗能明显减少扭转侧睾丸生殖细胞凋亡,减轻脂质过氧化程度,可能有利于睾丸组织结构与功能的恢复.     

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.     

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     

The effects of orchidopexy and orchidectomy on testes of rats subjected to ipsilateral abdominal testis with vas deferens obstruction

An experimental study was planned to evaluate and compare the effects of orchidopexy and orchidectomy on the testes of rats subjected to unilateral abdominal testis with vas deferens obstruction. Four groups were established. Rats in the control group underwent a sham procedure. While the testis was maintained in the abdomen with the vas deferens ligated for 8 weeks in group 2, rats in groups 3 and 4 underwent orchidopexy or orchidectomy after 4 weeks. Remaining testes were harvested at the end of the 8-week period. Testis and body weights were obtained during harvest. Samples were evaluated through DNA flow cytometry, and percentages of haploid cells were determined. Groups were compared through unpaired t-test, and p-values less than 0.05 were considered significant. All three treatments had decreased testis weight over body weight values of ipsilateral testes. Ipsilateral orchidectomy increased the value among contralateral testes. However, none of the groups had a contralateral testicular value less than the sham-operated group. All three treatments decreased the percentage of haploid cells among ipsilateral testes, but only an abdominal testis was associated with a decrease in the percentage of haploid cells among contralateral testes. Maintaining a testis with an obstructed vas deferens in the abdomen for 8 weeks damages both ipsilateral and contralateral testes. Orchidopexy, while showing amelioration of the ipsilateral testis, spares the contralateral testis as well as orchidectomy. Orchidopexy for an undescended testis with vas deferens obstruction is a rational approach.     

Hormonal therapy in undescended testes

A clinical study trial of HCG therapy in 78 selected prepubertal cryptorchids (40 unilateral and 38 bilateral) haveing a total of 116 undescended gonads revealed that complete descent occurred in 12.5% gonads (13% in unilateral and 12% in bilateral group). Good response was observed in only those gonads which were located in the inguinal canal and at the external ring, irrespective of it being right or left sided gonad. In patients with impalpable testes, though there was no complete descent, yet partial descent was noted in 18% (21/116 gonads). The response was better in the older children.     

Antenatal torsion of the testes,a cause of vanishing testis syndrome

During the years 1976–1988, 412 patients were operated upon for cryptorchid testes. In 28 unilateral and 2 bilateral cryptorchid patients, no testes could be found. Pathologic examination of the blind-ended vas deferns (usually a nodule) in 6 cases showed calcified or hemosiderin-containing tissue. In 9 patients an abundance of blood vessels was noted. In 27 patients signs of fibrosis and/or hyalinization were found. It is postulated that in most instances the vanishing testis syndrome is due to in-utero vascular occlusion of the testicular vessels, which is most likely caused by torsion of the testis. Correspondence to: J. A. Bar-Maor     

绒毛膜促性腺激素对睾丸促性腺激素受体影响的实验研究

目的 促性腺激素（HCG／LH）的应用是小儿泌尿外科较常用的诊断和治疗手段，但其临床效果往往差异很大，其确切机制不明，而且长期以来促性腺激素治疗的方案和剂量也都很不统一。本文研究了HCG对HCG／LH受体的影响，希望能从受体水平探讨此方面的问题。方法 应用放射性配体受体结合分析法对不同给药（HCG）时间的大鼠睾丸匀浆制剂进行HCG／LH受体分析。结果 单剂（20U）HCG注射后大鼠睾丸HCG／LH受体数目在第2d的观察中发现明显下降，降至对照组的5.0%以下。第6d测定时回升到正常的60.0%，第10d时基本回升至对照组水平（P＞0.05)。持续使用HCG者，睾丸HCG／LH受体数目持续处于低水平。结论 HCG的应用将下降调节大鼠睾丸HCG／LH受体的数目，其恢复正常的时间约需10d。HCG的持续应用将持续使睾丸HCG／LH受体数目维持在低水平。     

外源性雌激素对小鼠睾丸引带收缩和增殖活性的影响

目的研究外源性雌激素(EEs)对胚胎小鼠睾丸引带组织肌动蛋白(Actin)和增殖细胞核抗原(PCNA)的表达以探讨EEs对睾丸引带收缩和增殖活性的影响。方法昆明雌性小鼠180只,随机分成18组,于孕9~17d每天分别给予DES、17α-E2、17β-E2和E3,每种药物又分别给予不同剂量25、50、100、200μg·kg-1·d-1和等体积的DMSO、生理盐水作空白和正常对照。孕19d处死母鼠,取出活胎,取其下腹部,常规固定、制片。应用免疫组织化学方法检测睾丸引带组织中肌动蛋白(Actin)和增殖细胞核抗原(PCNA)的表达。结果实验组小鼠睾丸引带组织发育较差,其中Actin含量降低,药物剂量越大,Actin含量越低(P<0.05);较大剂量(如200和100μg·kg-1·d-1药物组)的雌激素处理组胎鼠睾丸引带中PCNA含量降低明显(P<0.05)。两指标均有显著的剂量效应关系。结论EEs可随药物剂量的不同而不同程度地降低睾丸引带中Actin和PCNA的含量。表明EEs影响胚胎期睾丸引带的收缩和增殖活性,并可能是EEs影响睾丸引带甚或睾丸正常发育(下降)的机制。     

Growth patterns in young men treated for undescended testes in childhood

Testicular maldescent may be caused by endocrinological deficiency in some cases, and, on the other hand, cryptorchidism itself may have an injurious effect on testicular development and function. The purpose was to examine whether testicular maldescent is associated with abnormal growth in various body dimensions in otherwise healthy males treated for the malformation during childhood. A total of 76 young men, 16–30 years of age, who were treated for undescended testes at an age ranging from 10 months to 13 years, were examined by measuring various body dimensions. Previously cryptorchid men were slightly taller than age-matched controls, except for those with bilateral cryptorchidism treated preoperatively with hCG. The most prominent finding was a high bihumeral to bicristal ratio. In addition, testicular volume was positively correlated to both the bicristal and bihumeral width, as well as to the sitting and total height.     

Use of laparoscopy as the initial surgical approach of impalpable testes:10-year experience

AIM: To review the experience in the management of impalpable testes using laparoscopy as the initial approach and the need for inguinal exploration.METHODS: From January 2004 to June 2014, 339 patients with undescended testes underwent operation in our institute. Fifty patients(15%) had impalpable testes. All children with impalpable testes underwent initial laparoscopy. A retrospective review was conducted on this group of patients and the outcome was analyzed.RESULTS: Forty children had unilateral impalpable testis. Ten children had bilateral impalpable testes. Thirty-one children(78%) in the unilateral group underwent subsequent inguinal exploration while 4 children(40%) in the bilateral group underwent inguinal exploration(P 0.05). Orchidopexy was performed in 16 children(40%) in the unilateral group and 9 children(90%) in the bilateral group(P 0.05). Regarding the 24 children with unilateral impalpable testis and underwent orchidectomy for testicular nubbin(n = 19) or atrophic testes(n = 2) or has vanishing testes(n = 3); contralateral testicular hypertrophy was noticed in 10(41%). No intra-operative complication was encountered. Two children after staged Fowler-Stephensprocedure and 1 child after inguinal orchidopexy had atrophic testes.CONCLUSION: The use of laparoscopy in children with impalpable testes is a safe procedure and can guide the need for subsequent inguinal exploration. Children with unilateral impalpable testis were associated with an increased need for inguinal exploration after laparoscopy. Orchidopexies could be performed successfully in 90% of children with bilateral impalpable testes.     

Gross morphologic variations and histologic changes in cryptorchid testes

Cryptorchid testes are frequently accompanied by gross morphologic changes with regard to the size and shape of the testis, various degrees of detachment between the epididymis and testis, elongation of the caudal epididymis and vas deferens, as well as histologic changes. This study examined the gross and microscopic testicular, epididymal, and vas changes in relation to various parameters in 824 patients operated upon in our clinic during the last decade. The major findings were the following: (1) most patients (78.8%) were older than 4 years, and many older than 8 years (39.8%); (2) the frequency of abdominal undescended testes decreased with increasing age (P <0.1), while the frequency of suprascrotal testes increased with increasing age (P <0.001); (3) most of the undescended testes in a suprascrotal position were normal or had mild atrophy (78.7%), while those in intracanalicular, internal ring, and abdominal positions frequently presented moderate or severe atrophy (71,38%; (P <0.0001); and (4) most suprascrotal testes were either normal or had mild extension and looping of the epididymis (63.8%), while most of the intracanalicular, internal ring, and abdominal testes showed moderate or profound epididymal anomalies (63.4%;P <0.0001). We conclude that even though the necessity for early surgical correction of cryptorchidism is well established, these patients at least in our region, still tend to come very late to surgery, with consequent increased risks of infertility and malignant transformation.