睾丸肿瘤的病因、分类及诊断

睾丸既是男性的生殖腺，又具有内分泌功能。睾丸由生精小管及其周围的间质组成。生精小管内衬生精上皮，由不同发育阶段的生殖细胞和支持细胞组成。间质中有血管、巨噬细胞、肥大细胞及间质细胞。睾丸的生精功能和产生雄激素的功能受到垂体促性腺激素的调控。促滤泡激素（FSH）促使睾丸生成精于，促黄体激素（LH）促使睾丸合成雄激素。而FSH发挥效应的靶细胞即睾丸的支持细胞，LH的靶细胞是睾丸间质中的间质细胞。睾丸生殖细胞肿瘤是源于生精小管中的一些异形细胞，其属全能的未分化细胞，可以发展为复杂的多样化的肿瘤。因此，睾丸肿瘤…     

Leydig细胞睾酮合成的调节

睾丸由间质和生精小管两部分组成.间质中的内含物主要包括Leydig细胞、巨噬细胞、淋巴管和血管等;生精小管由支持细胞(Sertoli细胞)和不同发育期的生精细胞共同组成生精上皮,其外包有基膜和类肌细胞.哺乳动物体内的雄激素95%由睾丸Leydig细胞分泌,肾上腺皮质的网状带仅分泌少量雄激素.     

睾丸间质干细胞分化和移植

睾丸间质细胞是男性体内合成雄激素的主要细胞,胚胎发育期中肾胚的间质细胞及生精小管周成纤维样细胞可能是睾丸间质细胞的干细胞。在胚胎期间质干细胞分化为胎儿型间质细胞;出生后间质干细胞经间质祖细胞、未成熟间质细胞分化为成熟间质细胞。老年期间质细胞数量可能不变,但雄激素合成下降。间充质干细胞及脂肪干细胞等干细胞经诱导可分化为分泌雄激素的睾丸间质细胞,因此,间质干细胞移植可望成为治疗男性性腺功能不全和中老年雄激素缺乏的创新方法,本文对睾丸间质干细胞的分化及移植方面研究进行综述。     

睾丸间质细胞的发生与调控

睾丸间质细胞分布于生精小管的疏松间质组织中,是产生雄性激素的主要场所。睾丸间质细胞的功能障碍是导致男性原发性性腺功能低下、隐睾、尿道下裂等疾病的重要原因,因此睾丸间质细胞对男性生殖具有重要意义。本文对近年来关于睾丸间质细胞的发生、发育及调节的研究进展予以综述。     

高氟对C57BL/6J小鼠睾丸中AQP1、AQP4表达的影响

目的观察和分析氟化钠对C57BL/6J小鼠睾丸中水通道蛋白AQP1和AQP4表达的影响。方法选择健康C57BL/6J雄性小鼠24只,随机分为2组:对照组(n=12)和氟化钠干预组(n=12)。用放射免疫法测定C57BL/6J小鼠血清中的睾酮水平。免疫组织化学法检测AQP1和AQP4蛋白在睾丸中白膜、生精小管内的支持细胞以及睾丸间质中的表达情况。结果 AQP1在睾丸中白膜、生精小管内的支持细胞以及睾丸间质中均有表达,但在氟化钠干预组中表达相对较弱,睾丸间质细胞表达最强,依次是白膜和支持细胞,与对照组比较差异具有统计学意义(P0.05);AQP4在睾丸中白膜、生精小管以及睾丸间质中均没有观察到,氟化钠干预组与对照组比较差异无统计学意义(P0.05);对照组和氟化钠干预组的睾酮变化具有差异性(P0.05)。结论 AQP1在睾丸中的表达要比AQP4强,AQP1在氟化钠干预组睾丸中的变化比较明显,与睾酮的变化具有正相关性,说明在氟中毒致睾丸损伤中AQP1可能起主要的调节作用。     

睾丸体积与睾丸功能的相关性

本研究的目的旨在确定用一种新型睾丸计测量睾丸体积与生精小管功能和间质细胞功能的相关性,而且确定睾丸功能每一指标正常时的临界睾丸体积。一般认为,在男性学临床中,睾丸大小是初步评价生精功能的最好指标,因为生精小管等生发组织约占睾丸体积的98％,精细胞的减少意味着睾丸发育不良,因此人们认为只有睾丸大小正常或接近正常,才能产生足够量的精子,但认为睾丸体积小于正常时,其间质细胞的功能是正常的。然而具有正常生精功能和间质细胞功能的临界睾丸体积是多少,尚无人报     

Attractin蛋白和mRNA在不同日龄大鼠睾丸中的表达

目的:已知成熟大鼠睾丸组织中有Attractin蛋白的广泛表达,本文旨在进一步探讨Attractin蛋白和AttractinmRNA在各年龄段大鼠睾丸组织中的分布。方法:取出生第1d(新生鼠)、第5d(青春前期)、第20d(青春中期)、第50d(青春后期)及第70d(成年期)大鼠睾丸和附睾组织固定,采用酶免疫组织化学和原位杂交方法检测Attractin和AttractinmRNA在大鼠睾丸组织中的表达。结果:免疫组化显示各年龄段大鼠睾丸生精小管和间质细胞、管周肌样细胞上均有Attractin蛋白的表达,分布于胞膜和胞质,精子和附睾上未见表达。随着日龄的增加,间质细胞的表达逐渐增强。原位杂交显示各年龄段大鼠睾丸生精小管和间质细胞、管周肌样细胞、支持细胞上也均有AttractinmRNA阳性棕色颗粒杂交信号,主要表达于胞核及胞质。且随着日龄的增加,睾丸生精细胞表达略强于间质细胞。精子和附睾上未见表达。结论:AttractinmRNA和Attractin蛋白在各日龄大鼠睾丸组织中均有广泛的分布,且分布一致。提示各日龄大鼠睾丸组织都具有合成Attractin蛋白的能力。其生理功能和机制尚有待进一步探讨。     

小鼠睾丸消化细胞异体移植重构生精小管的研究

目的:采用免疫缺陷小鼠作为受体,通过对小鼠睾丸消化细胞异位移植后不同时期移植物的研究,观察生精小管重构、生精细胞归巢及精子发生情况。方法:取新生ICR小鼠的睾丸消化成单细胞悬液,将其与Matrigel基质胶混匀后移植于雄性裸鼠背部皮下,术后裸鼠行去势。移植后分别于4、6、8、10周处死5只裸鼠,计算移植成功率,取移植物测量直径,并进行HE染色和免疫组化检测,观察生精小管的重构、生精细胞归巢及精子发生情况。结果:20只受体鼠接受睾丸消化细胞移植后全部存活。睾丸消化细胞移植后10周内可见明显隆起的包块,包块直径由第4周的(3.91±0.71)mm增加到(6.69±0.50)mm,移植物表面有血管生成。对移植物石蜡切片进行HE染色可见生精小管样结构,部分生精小管管腔内可见由精原细胞发育至精子细胞的各级生殖细胞,未见明显精子产生。对8周移植物进行免疫组化观察,可见生殖细胞标志物Mvh、支持细胞标志物Gata4和间质细胞标志物P450Scc表达。结论:新生小鼠睾丸消化细胞移植于裸鼠背部皮下后可重构生精小管,为研究睾丸组织工程及睾丸发育和精子发生过程中睾丸各组成细胞之间的相互作用提供了理想的研究模型。     

长期饮酒对小鼠生长发育及其睾丸功能的影响

目的利用未成熟雄性小鼠为动物模型研究饮用酒精对哺乳动物生长发育及生殖的毒性作用。方法选用24只21d龄断奶未成熟小鼠随机分为3组：啤酒组、白酒组和对照组。啤酒组自由饮用啤酒，白酒组饮用稀释的白酒（5％），对照组饮用自来水。实验5周后对体重、脏器重等指标进行称量，对附睾中精子密度、精子畸形率等进行检测，对睾丸的组织学形态进行观察。结果整个实验期啤酒组体重显著增加，脏器重受到不同程度的影响；白酒组的精子密度显著降低，在这同时精子畸形率也明显升高，睾丸形态发生了不同程度的病理变化。啤酒组睾丸间质问隙增宽，间质细胞增生；白酒组睾丸问质问隙进一步增宽，生精小管内生精细胞排列紊乱疏松，生精细胞问出现空隙，有的甚至变性、坏死、脱落，部分生精小管生精细胞全部脱落或退化消失，生精小管管壁萎缩变薄。结论长期饮酒对小鼠的生长发育及睾丸功能具有毒性作用。     

神经生长因子mRNA水平与睾丸生精细胞成熟和功能的关系

以地高辛素标记人β－神经生长因子ＤＮＡ为探针，采用原位杂交的方法观察了小鼠睾丸曲精小管中ＮＧＦｍＲＮＡ的分布情况，发现阳性细胞分布于大部分曲精小管基底至管腔的各级生精细胞中，随着生精细胞成熟度的增加，ＮＧＦｍＲＮＡ表达水平逐步增加，细胞核与间质呈阴性。结果提示：ＮＧＦ在睾丸中具有自分泌与旁分泌作用的可能，不断促进精子自身的成熟与活力。     

Ultrastructure of the seminiferous tubules in oligoasthenoteratozoospermic men associated with varicocele

Varicocele is associated with venous reflux that may cause increased heat and interstitial pressure within the testes, with variable pathological effects on spermatogenesis. This study aimed to study the ultrastructural testicular changes in the seminiferous tubules of 20 infertile severe oligoasthenoteratozoospermia (OAT) men associated with varicocele and five patients with obstructive azoospermia without varicocele as controls. They were subjected to testicular biopsy which was evaluated by transmission electron microscopy. Ultrastructurally, the seminiferous epithelium in the testicular biopsies of infertile severe OAT men associated with varicocele was variably affected in the form of thickening of the peritubular connective tissue, vacuolation of Sertoli cell and germ cell cytoplasm, presence of degenerated and apoptotic cells among the germinal epithelium, altered spermatids and abnormal spermatozoa. It is concluded that varicocele in severe OAT men is associated with ultrastructural changes in the seminiferous tubule.     

大鼠睾丸减数分裂相关基因的初步筛选

目的:比较成年大鼠睾丸生精小管处于减数分裂期的发育片段和睾丸间质细胞所表达基因的差异,初步筛选出与减数分裂相关的基因,为进一步研究减数分裂相关基因对精子发生的调控奠定基础。方法:运用在透射光解剖显微镜下区分和显微分割的方法,将成年大鼠新鲜睾丸生精小管第X III~I期处于减数分裂阶段的片段分离出来,同时分离睾丸间质细胞,将二者进行mRNA差异显示逆转录聚合酶链反应(DDRT-PCR)分析,所得差异片段进行纯化回收,然后进行反向斑点杂交。结果:经mRNA差异显示,第X III~I期生精小管片段共回收到7个差异cDNA片段,而间质细胞共回收到9个差异cDNA片段。经反向斑点杂交,共获得11个初步鉴定的特异表达增加的差异cDNA片段,片段大小为200~500 bp,其中6个从第X III~I期生精小管片段获得,5个从间质细胞获得。结论:这些差异片段可作为睾丸减数分裂表达的序列标签进行更深入地研究。     

Seasonal changes in testicular structure and function in the blue fox (Alopex lagopus), as quantified by morphometric analysis and measurement of adenylate cyclase activity

The volume of the blue fox testis showed 5-fold changes during the year, associated with considerable changes in cellular composition. The seminiferous epithelium was maximally regressed in August, when 94% of tubules contained only spermatogonia. By late October, approximately 6 months before the mating season, 40% of tubules contained primary spermatocytes. From the middle of January until the end of April all tubules contained spermatids or more advanced haploid cells. Tubular diameter increased by 73% during testicular re-development, and epithelial height increased 3-fold. Regression to the basal state occurred during May to July. The volume densities of the seminiferous epithelium and of interstitial tissue remained approximately constant throughout the year. Soluble Mn2+-dependent adenylate cyclase activity showed seasonal variations that paralleled those of the haploid germ cell population and testicular volume, whereas somatic cell adenylate cyclase activity was relatively constant.     

Bee glue (propolis) improves reproductive organs,sperm quality and histological changes and antioxidant parameters of testis tissues in rats exposed to excess copper

This study was designed to determine the effects of propolis on the sperm quality, antioxidant and histological parameters in the testicular tissues of male Sprague Dawley rats exposed to excessive copper (Cu). In this aim, 24 rats were randomly divided into four groups as follows: the control, Cu, Propolis and Cu+Propolis. When compared to control group, Cu administration significantly decreased sperm motility and concentration, increased total abnormal sperm rate. It caused a significant induction the malondialdehyde (MDA), and reduction the superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) in testicular tissues. Also, it caused loss, disorganisation and vacuolation of the germinal epithelium, oedema of the interstitial tissues, proliferation of the interstitial cells, spilled immature spermatogenic cells in the lumen of some seminiferous tubules. A large number of active caspase-3-positive stained apoptotic cells and a significant decrease in Johnsen's testicular score were determined. However, significant ameliorations were observed in all sperm characteristics, MDA, SOD, CAT, GSH, seminiferous tubules, number of apoptotic cells and Johnsen's testicular score in Cu+Propolis group. Our results showed that oral supplementation of propolis had curative effect on the sperm quality, antioxidant and histological parameters in the testicular tissues of male Sprague Dawley rats exposed to Cu.     

Mechanism of alcoholic testicular damage]

To observe the mechanism of alcoholic testicular damage, in a previous experiment we used weanling male SD rats aged 45 days, weighing about 200 g, and fed a liquid diet (Lieber's) containing 5% ethanol. The latter accounted for 36% of total caloric intake for 7 weeks, but did not result in testicular atrophy. In a later experiment, we used a liquid diet in which ethanol accounted for 46% of the total calorie count. It provided a high-fat, low-protein content which simulated the nutritional background of patients with alcoholic liver diseases. This diet resulted in testicular atrophy. Histological and biochemical changes accompanying this experimental testicular atrophy included the following: 1) The testes of alcohol-fed animals contained smaller seminiferous tubules with reduced numbers of total cells, but no degeneration was seen in the spermatids. 2) In the peritubular wall of the seminiferous tubules, we observed curvature, irregularities, infolding of the basement membrane, and lamellation of the lamina densa, as well as hyperplasia of collagen fibers in the tunica propria. 3) In the cytoplasm of the Sertoli cells, deposits of gigantic fat droplets and stratification of the mitochondria were observed. The permeability of the Sertoli cell tight junction was confirmed using the Lanthanum method. 4) Testosterone levels in both the serum and testes declined. 5) Lactate dehydrogenase-X (LDH-X) activity in the testes declined. 6) Low Km alcohol dehydrogenase (ADH) activity localized in the testicular interstitial tissue was increased. These results indicate that the composition of three major nutritional elements as well as alcohol concentration are important in the mechanism of alcoholic testicular damage, and this damage affects both the testicular interstitial cell and the seminiferous tubules, particularly the Sertoli cells and peritubular wall of the latter. In addition, the findings suggest that ADH is involved in alcohol metabolism in the interstitial cells of the testes.     

Spermatogenesis in the aged--a borderland between normal and pathologic anatomy

Aging of the testis is a normal physiological process, which gradually proceeds with increasing age. Under normal conditions, an abrupt breaking off the spermatogenetic and hormonal functions of the testis does not occur. Already in the testis of every adult fertile man single degenerating germ cells are recognizable, the occurrence of which is understood as physiological germ cell loss. In men, older than 65 years, further degenerative changes of the germ cells and of the germinal epithelium regularly can be found, which have negative effects on the spermatogenetic activity of the germinal epithelium. These changes concern disturbances of the kinetics of spermatogenesis, disturbances of spermatogoniogensis and of meiosis, malformations of the spermatids, the release of immature germ cells from the germinal epithelium, the concentration of lipids within the Sertoli cells, the appearance of diverticles of the seminiferous tubules and possibly also atrophy of seminiferous tubules. In the course of regular testicular aging these degeneration phenomena can be observed to a relatively small extent diffusely distributed in the testis and exhibiting individual differences. A phatological alteration of the testicular tissue is under consideration as soon as a definitive pattern of damage predominates in the whole organ or when a focal degeneration of testicular tissue has appeared. The borderland between normal and pathological anatomy of testis tissue during aging is characterized.     

胎儿睾丸组织异种移植后生精细胞发育初探

目的 :以人胎儿睾丸组织为供体 ,免疫缺陷小鼠为受体 ,研究人类睾丸生精细胞异种移植后的继续发育情况。　方法 :将 2 6周胎儿的睾丸组织植入去势裸鼠背部 ,于移植后 1 35d取出移植物 ,进行组织形态学观察 ,分析原始生精细胞在异种异位的发育情况。　结果 :移植 1 35d后取出的移植物显示 ,其生长幅度已由移植前直径约 1mm和湿重约 5mg ,分别增加到移植后大于 3mm和 2 0mg。组织形态学观察发现 ,移植前的睾丸主要是由直径为(6 0± 1 5 ) μm的精曲小管索构成 ,其中包含的细胞主要是原始Sertoli细胞和少量原始生精细胞 ,细胞排列呈弥散无规则状态 ;而移植后 1 35d的精曲小管索已发育成具有管腔的精曲小管 ,出现由Sertoli细胞和生精细胞组成的完整生精上皮 ,直径增大到 (80± 2 5 ) μm。原本呈不规则分布的原始Sertoli细胞和生精细胞大部分已迁移到基膜处 ,其中有少数生精细胞发育成为精原细胞。　结论 :人胎儿睾丸组织移植到去势裸鼠背部后 ,可以继续存活并进一步生长发育。     

大鼠非特异性睾丸炎模型的建立及生精上皮的变化

目的:探讨大鼠生精上皮在非特异性炎症状态下的变化。方法:20只成年W istar雄性大鼠随机分为2组(对照组和实验组),对照组腹腔注射生理盐水(1 m l/kg),实验组注射细菌内毒素(LPS),剂量为1 mg/kg,两组均隔1 d注射1次,距第1次注射10 d后取材。HE染色观察睾丸组织的病理改变,免疫组化方法探讨生精上皮内增殖细胞核抗原(PCNA)、α连接素的变化。结果:实验组睾丸组织有明显炎性改变;生精上皮内PCNA表达量显著降低:每个生精小管(仅精原细胞着色的小管)阳性细胞个数为(59±5)个,比对照组明显减少(P<0.01),而这种小管占总管数的比例显著升高(P<0.01),为0.673±0.054;实验组大鼠睾丸内α连接素表达量亦减少(P<0.01),阳性反应颗粒平均光密度为0.150±0.014。结论:睾丸炎症时精原细胞增殖和生精上皮内细胞间粘附力度均减弱,这可能是睾丸炎导致男性不育的原因之一。     

A morphological study of the testes in patients with idiopathic male infertility--immunohistochemical analysis of collagens and laminin in human testes

In the human testis, the distribution of extracellular components, such as types I, III, IV and V collagens and laminin, was investigated immunohistochemically by light microscopy. Specimens were obtained by testicular biopsy from 40 patients with idiopathic male infertility and 14 normal adult males. In the normal testes, the basement membrane was positive for types I, III, IV and V collagens and laminin. However, the distribution patterns of these components were different. Furthermore, a reactivity for types I and III collagens was found in the interstitial connective tissue matrix. Immunoreactivity for types I and III collagens was markedly positive in the limiting membranes around the Leydig cells. In the pathological testes, all the layers of the basement membrane of both thickened and obstructed tubules were positive for types I and III collagens. On the other hand, reaction products of type IV collagen were localized in the inner layer of the basement membrane and the peritubular cell (myoid cell) layer, and those of laminin were only found in the inner layer. Type V collagen-reactivity was observed in the basement membrane of thickened tubules. Positive reactions for types IV and V collagens and laminin were seldom recognized in the obstructed tubules. In the interstitial space, the connective tissues were significantly increased as compared with normal testes, which included extracellular components that reacted for types I and III collagens. Histological findings in normal adult testes and pathological testes were compared. Quantitative analysis of mean thickness of the basement membrane (W), mean seminiferous tubular diameter (T), T/W ratio and Leydig cell index demonstrated significant differences between the two groups.(ABSTRACT TRUNCATED AT 250 WORDS)     

Attractin蛋白在不同病理分型无精子症患者睾丸组织中的表达

目的探讨Attractin蛋白在不同生精功能状态的人睾丸组织中的表达情况。方法对31例无精子症患者进行睾丸组织病理检查诊断,根据病理形态的不同分为:唯支持细胞综合征型(n=4),生精功能低下型(n=12),生精阻滞型(n=5),生精功能基本正常型(n=10)。选择上述各型结构完整的睾丸组织,分别应用免疫组化法(SP)观察Attractin蛋白在不同生精功能状态的睾丸组织中的表达。结果各组睾丸组织内均存在Attractin蛋白的表达,其分布特点为:睾丸生精小管和间质细胞、管周肌样细胞、支持细胞上均有Attractin蛋白的表达,主要表达于胞膜和胞质,胞膜表达强于胞质。Leydig细胞、Sertoli细胞、精原细胞、精母细胞及精子细胞均为阳性表达,呈棕黄色着染。Attractin的表达与生精功能有关,正常组表达明显高于其它组,唯支组表达明显低于其他组。结论 Attractin蛋白与男性生殖密切相关,但其具体作用环节尚有待进一步研究。