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。     

绒毛膜促性腺激素对双侧隐睾大鼠睾丸生殖细胞凋亡的影响

目的探讨绒毛膜促性腺激素(HCG)对双侧隐睾生殖细胞凋亡的影响。方法SD雄性幼鼠于日龄22d制备双侧隐睾模型(模型组)后开始隔日肌注HCG20U,共7次。假手术组作为对照。日龄35、60d时处死取睾丸,采用生物素-dUTP/酶标亲和素(TUNEL)法检测生殖细胞凋亡情况。结果模型组双侧隐睾睾丸凋亡指数(AI)高于假手术组。35d假手术HCG治疗组AI高于假手术HCG未治疗组(P<0.05)。60d各HCG治疗组睾丸的AI高于相应HCG未治疗组,组间比较无显著差异(P>0.05)。结论隐睾时睾丸生殖细胞凋亡增加;HCG可增加生殖细胞的凋亡。     

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。     

不同缝线行睾丸固定复位术对隐睾模型大鼠生精能力的影响

目的建立内分泌型双侧隐睾大鼠模型,通过手术复位睾丸,检测、对比应用不同缝线固定睾丸对隐睾模型大鼠生精能力的影响。方法取孕SD大鼠10只,自然分娩产仔鼠,随机选取新生雄性仔鼠40只为正常对照组,并选取80只为双侧隐睾模型组。双侧隐睾模型组大鼠从出生第3天起皮下注射17-β雌二醇,第26天观察双侧睾丸均未降入阴囊内为隐睾模型制作成功。将双侧隐睾大鼠随机分为1号丝线手术组(采用1号丝线固定睾丸)和6-0可吸收线手术组(采用6-0可吸收线固定睾丸),各40只。于日龄45 d、56 d时采集血清后处死。采用ELISA间接法测定其血清抗精子抗体(AsAb)水平。睾丸切片苏木精-伊红染色光镜下观察曲细精管生育力指数(TFI)、平均曲细精管直径(MTD)。结果 1号丝线手术组隐睾大鼠45 d、56 d时TFI、MTD水平均低于6-0可吸收线手术组(Pa<0.05);45 d、56 d时1号丝线手术组大鼠血清AsAb IgG、IgA、IgM水平均显著高于6-0可吸收线手术组(Pa<0.01)。结论采用6-0可吸收线固定睾丸可以减少AsAb的产生。     

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.     

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。     

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。     

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.     

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.     

生脉注射液对青春前期大鼠睾丸扭转/复位缺血再灌注的保护作用

目的观察青春前期大鼠睾丸单侧扭转复位后对双侧睾丸氧自由基的远期影响,探讨生脉注射液对其的保护作用。方法 5周龄健康SD雄性大鼠24只,随机分为实验组(生脉注射液组)、对照组(9 g.L-1盐水组)和假手术组,每组8只。实验组和对照组建立左侧睾丸扭转复位模型,假手术组游离睾丸,不予扭转。于术后7周取各组大鼠双侧睾丸,分别测定大鼠睾丸组织内超氧化物歧化酶(SOD)、一氧化氮合酶(NOS)活性和丙二醛(MDA)水平。结果与对照组比较,实验组和假手术组大鼠双侧睾丸中SOD活性升高,NOS活性和MDA水平下降(Pa<0.05)。与假手术组比较,实验组大鼠双侧睾丸组织中SOD活性下降,NOS活性和MDA水平升高(Pa<0.05)。结论生脉注射液可提高大鼠睾丸组织中的SOD活性,提高机体抗氧化能力,清除氧自由基,抑制脂质过氧化反应,减轻MDA对细胞膜的损伤,对睾丸缺血再灌注损伤具有一定的保护作用。     

Testicular histology related to fertility outcome and postpubertal hormone status in cryptorchidism

BACKGROUND: Early surgical correction of an undescended testis is performed to prevent the development of male infertility. However, in boys with cryptorchidism early successful surgery cannot prevent infertility if they lack Ad spermatogonia. In this study, sperm concentrations and postpubertal hormone levels were correlated to bilateral testicular histology. The aim was to define the risk of future infertility via a testis biopsy program for boys with cryptorchidism. METHODS: Eighty-nine boys who had an orchidopexy were subjected to bilateral testicular biopsy. Histological analysis of 178 biopsies indicated three groups of high, intermediate, and low risk of infertility according to the presence of Ad spermatogonia. After puberty, sperm concentrations were analysed and correlated with plasma gonadotropin and testosterone levels. FINDINGS: In patients with unilateral cryptorchidism 70% of scrotal testes had an impaired transformation of Ad spermatogonia, indicating that cryptorchidism is a bilateral disease. Sperm concentrations correlated to the number of Ad spermatogonia found at the time of orchidopexy (p<0.001). All males in the high risk of infertility group were oligospermic (mean: 8.9x10 (6) sperm/ejaculate) and 20% were azoospermic. These patients had 25 times less sperm compared to the group with presence of Ad spermatogonia in both testes (p<0.001). Correlations between testicular histology and postpubertal hormone levels confirmed a relative gonadotropin deficiency in the majority of males with cryptorchidism. INTERPRETATIONS: Ad spermatogonia proved to be a discriminating factor for the fertility outcome in cryptorchidism. Gonadotropin treatment following orchidopexy should be considered in cryptorchidism when no Ad spermatogonia are found in undescended gonads and scrotal testis have Ad germ cell counts <0.005 per tubule.     

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.     

隐睾儿童血清苗勒管抑制物质的研究

目的探讨苗勒管抑制物质(MIS)在隐睾的诊断以及睾丸发育评价中的作用。方法选择54例单纯性隐睾患儿为观察组,36例无生殖器病变的儿童为对照组,分别检测两组儿童血清MIS水平,并比较两者之间的差异。结果观察组与对照组按年龄分组,各组间年龄比较,差异无统计学意义(P>0.05)。观察组血清MIS为48.1±26.2 ng/ml,对照组为76.3±22.8 ng/ml,两组比较,差异有统计学意义(P<0.05)。双侧隐睾患儿血清MIS为39.2±28.3 ng/ml,单侧隐睾患儿为56.5±30.1 ng/ ml,两者比较,差异有统计学意义(P<0.05)。MIS水平与睾丸发育情况呈正相关(r=0.35,P<0.05),与治疗年龄相关(r=0.19,P<0.05)。结论MIS可作为反映隐睾患儿睾丸发育状况的一项重要指标。     

Inguinal approach for the management of unilateral non-palpable testis: Is diagnostic laparoscopy necessary?

ObjectiveDuring the last decade laparoscopy has increasingly been advocated as the primary investigative procedure for the management of the non-palpable testis. We reviewed the medical records in a consecutive series of boys with non-palpable testis to examine the contribution of the initial inguinal approach in the management of unilateral non-palpable testis.Materials and methodsAmong the 183 consecutive patients with cryptorchidism from 2003 to 2012, there were 21 patients with unilateral and three with bilateral non-palpable testes. All unilateral patients then underwent inguinal and scrotal exploration through an inguinal incision. For those patients with an intra-abdominal peeping testis, the gonad was placed into the scrotum after meticulous cranial mobilization of the spermatic cord.ResultsPatient age ranged from 11 months to 144 months (mean age: 23 months). Among the 21 unilateral cases, testicular absence or atrophy was confirmed in seven patients with a scrotal nubbin in six, and blind-ending vas and vessels at the external inguinal ring in one patient. Among the remaining 14 patients with sizeable testes, 12 testes were intra-abdominal peeping testes and two testicles were seen within the distal inguinal canal, which may be missed on physical examination owing to patient obesity. The intra-abdominal peeping testicle had the opened processus vaginalis entering the internal ring in which testicle was found. These were fixed into the scrotum successfully by cranial mobilization of spermatic vessel sometimes cutting the internal oblique muscle and by Prentiss and Fowler-Stephen's maneuver. Diagnostic laparoscopy was done on three patients with bilateral cases.ConclusionsGiven the result that most of nubbins are within the scrotum and testes with intra-abdominal peeping testes are fixed down safely into the scrotum, the inguinal approach may suffice for the management of unilateral non-palpable testis. Laparoscopy should be reserved for patients with bilateral non-palpable undescended testes.     

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.     

隐睾症中激素和雌雄激素受体的研究

目的　从激素和雌雄激素受体变化探讨隐睾症发病机制。方法　化学发光免疫技术检测30例隐睾症和16例斜疝(对照组)患儿血清促卵泡生成激素(FSH)、促黄体生成激素(LH)和睾丸酮(T)激素水平;免疫组化技术观察隐睾症患儿睾丸引带、阴囊肉膜、阴囊皮肤和斜疝患儿疝囊组织雄激素受体(AR)和雌激素受体(ER)表达情况。结果　①激素水平:隐睾症组血清LH和T水平与斜疝组二者之间无显著性差异(P<0.05);双侧隐睾组血清FSH水平高于斜疝组(0.01相似文献   

神经细胞粘附分子及OCT-4蛋白在隐睾中的病理表达及意义

目的 探讨神经细胞粘附分子(NCAM)及转录因子OCT-4在人隐睾中的病理表达及意义.方法 48例隐睾患儿,年龄6个月～13.1岁,平均3岁,单侧43例,双侧5例,共50个隐睾组织活检样本.免疫组化法检测生精小管和间质内NCAM、OCT-4的表达情况.结果 50个样本中,NCAM生精小管内阴性、睾丸间质细胞(Lc)阳性、OCT-4阴性的牛理规律表达率为38%(19/50).NCAM牛精小管、Lc中病理表达率为58%(29/50).NCAM生精小管内阳性病理表达率为30%(15/50),与年龄、睾丸位置无相关性(P>0.05).NCAM于Lc内阴性病理表达率为32%(16/50),与睾丸位置无相关性(P>0.05),但随年龄增加,阳性表达率增加(P<0.05).OCT-4阳性病理表达率为12%(6/50),与睾丸位置无相关性(P>0.05).结论 NCAM和OCT-4在隐睾中的病理性表达反映了生殖母细胞的迁移分化障碍或延迟,可以被作为预测隐睾生殖潜能和癌变趋势的可靠分子标志物.