犬腺垂体远侧部P物质样免疫反应神经纤维的超微结构研究

本室曾在光镜水平上显示,犬腺垂体远侧部P物质样免疫反应多膨体神经纤维与腺细胞关系极为密切。本实验应用包埋前免疫组织化学电镜法,进一步研究了这种神经纤维的超微结构,及其与腺细胞的关系。本文发现P物质样免疫反应多膨体神经纤维与各种腺细胞以及滤泡星形细胞发生直接接触,与促生长激素细胞和促肾上腺皮质激素细胞有典型的突触结构,其中与后者构成的突触较多。大多数突触为不对称型,含有圆形或椭圆形清亮小泡和大致密芯泡。本实验结果表明,犬腺垂体远侧部P物质样免疫反应神经纤维对腺细胞活动有直接调节作用。     

狗腺垂体远侧部降钙素基因相关肽免疫反应神经纤维的超微结构研究

本实验应用包埋前免疫电镜法,研究了狗腺垂体远侧部降钙素基因相关肽(CGRP)免疫反应神经纤维的超微结构,及其与腺细胞的关系。发现CGRP免疫反应神经纤维与促肾上腺皮质激素细胞和促生长激素细胞构成典型的突触结构,其中与前者之间的突触较多。大多数突触为非对称型,神经终末内含有清亮小泡和大致密芯泡。还观察到许多未被免疫染色的神经终末,它们与促肾上腺皮质激素细胞、促生长激素细胞、催乳激素细胞和促甲状腺激素细胞形成突触。我们还应用包埋后免疫双标记法,发现在狗腺垂体远侧部神经纤维中,P物质样免疫反应物质与CGRP免疫反应物质共存,但它们分布于不同大致密芯泡内。     

培养鼠腺垂体细胞的形态和分泌功能实验研究

目的利用细胞培养技术,对大鼠腺垂体细胞进行体外培养,进而确定腺垂体细胞在体外生长的最佳时机,为垂体移植细胞库的建立以及垂体移植的应用提供实验依据.方法取大鼠腺垂体细胞做体外培养,动态观察培养细胞的生长状态,通过RIA和免疫化学染色技术,评价体外培养的垂体细胞的功能状态.结果培养液中GH、LH的含量在培养的第7天最高,为9.49±2.21(ng/ml)和7.83±1.51(ng/ml),以后逐渐下降.免疫化学染色证实分散培养后的垂体细胞中有多种激素细胞的生长.结论培养至第7天的腺垂体细胞分泌GH、LH的功能状态为最佳,提示适于进行移植或保存.     

培养鼠腺垂体细胞的形态和分泌功能实验研究

目的　利用细胞培养技术 ,对大鼠腺垂体细胞进行体外培养 ,进而确定腺垂体细胞在体外生长的最佳时机 ,为垂体移植细胞库的建立以及垂体移植的应用提供实验依据 .方法　取大鼠腺垂体细胞做体外培养 ,动态观察培养细胞的生长状态 ,通过RIA和免疫化学染色技术 ,评价体外培养的垂体细胞的功能状态 .结果　培养液中GH、LH的含量在培养的第 7天最高 ,为 9.4 9± 2 .2 1(ng/ml)和 7.83± 1.5 1(ng/ml) ,以后逐渐下降 .免疫化学染色证实分散培养后的垂体细胞中有多种激素细胞的生长 .结论　培养至第 7天的腺垂体细胞分泌GH、LH的功能状态为最佳 ,提示适于进行移植或保存     

一种新的精液脱落细胞染色方法的探讨

随着社会的发展 ,科技的进步 ,现在男性学的研究工作不断开展新的检查项目 ,评价男性生育力的各项检测手段日益更新 ,沿用九十年代的精液常规检查已经不适用今天的医疗。尤其是镜检把精液内脱落细胞一律视为白细胞很显然是不科学的。很多病人经抗感染治疗几个月无效 ,延误了诊断和治疗。所以 ,对精液脱落细胞正确检查就象分析血细胞那样对精子发生感染 ,抗精子免疫等生殖道状况作出比较详细的了解 ,对发生在睾丸、附睾和附性腺等不同性质疾病能反应出来。另一方面 ,对睾丸病变所进行的睾丸活检是一次手术 ,会给病人带来创伤和感染的机会 ,不…     

两种染色方法对脑垂体三种嗜色细胞显示效果的比较

脑垂体由腺垂体和神经垂体两部分组成,其中腺垂体内有3种类型细胞：嗜酸性细胞、嗜碱性细胞和嫌色细胞。在常规的苏木精-伊红染色切片中,脑垂体3种嗜色细胞显示的效果一直不甚理想,学生在显微镜下进行观察辨认比较困难,为了更清晰地显示脑垂体3种嗜色细胞,我们采用不同的染色方法对脑垂体进行染色,经过多次实验显示,采用Mallory氏法染色,获得了较满意的染色效果,现介绍如下：     

棉酚对大鼠垂体远部细胞超微结构的影响

本文对服棉酚30mg/kg体重/d共5周的成年雄性大鼠垂体远部细胞的超微结构进行了观察。结果表明,促性腺激素细胞的分泌活性显著增强,表现为功能不太活跃的Ⅰ、Ⅱ型细胞比对照组减少;而高尔基复合体发达,粗面内质网扩张为小泡的Ⅲ、Ⅳ型细胞增多,并出现类阉割细胞(Ⅴ型)和脱颗粒细胞(Ⅵ型)。垂体远部其他类型促激素细胞未见明显变化。本文还讨论了垂体促性腺激素细胞的分型、棉酚引起促性腺激素细胞变化的可能机制。     

吖啶橙／溴乙锭双荧光染色检测细胞凋亡的形态学方法

吖啶橙／溴乙锭双荧光染色检测细胞凋亡的形态学方法南京医科大学第一附属医院临床实验研究室（南京２１００２９）汪承亚盛瑞兰汪凡丁小健邱宁雷细胞凋亡（ａｐｏｐｔｏｓｉｓ）是不同于细胞坏死的一种细胞主动性死亡，近年已引起人们广泛重视。Ｋｅｒ等［１］首先用电子...     

吉姆萨染色法对大脑组织凋亡细胞的染色

根据资料记载凋亡细胞的染色可以用吉姆萨染色法,但具体的操作方法却未见报道。我们根据吉姆萨染料染血涂片的原理,将其用于大鼠大脑海马区细胞的染色,探索出其染色程序,发现染色效果优于常规HE染色,核仁和凋亡细胞着色清晰,细胞与背景对比明显。现介绍如下：     

大鼠实验性腺胃癌过程中脑垂体远侧部FSH细胞的免疫组织化学观察

目的：观察腺垂体ＦＳＨ细胞在大鼠实验性腺胃癌的免疫组织化学变化。材料和方法：ＭＮＮＧ诱发断乳雄性Ｗｉｓｔａｒ大鼠的实验性脾胃癌模型,免疫组织化学、形态计量及图像分析技术。结果：实验性腺胃癌组大鼠脑垂体ＦＳＨ细胞与正常组相比较,免疫反应减弱,面数密度减少（Ｐ〈０．０５）,平均光密度（ＭＯＤ）减弱（Ｐ〈０．０５）。结论：在大鼠实验性胃癌过程中,腺垂体ＦＳＨ阳性细胞数量减少的同时,可能伴有合成和分泌功能     

吗啡依赖及乌拉地尔戒断大鼠垂体远侧部TSH细胞免疫组织化学研究

用皮下注射吗啡 ,建立雄性大鼠吗啡依赖模型。戒断组给吗啡依赖大鼠侧脑室注射乌拉地尔( IVC)。阳性对照组给正常大鼠 IVC,阴性对照组给正常大鼠 IVC相同体积生理盐水。用免疫组织化学方法观察吗啡依赖大鼠 ,IVC戒断大鼠和 IVC正常大鼠垂体远侧部 TSH细胞的变化。结果发现 ,吗啡依赖大鼠和IVC戒断大鼠 TSH免疫反应细胞的面数密度减少 ,免疫反应减弱 ( P <0 .0 1)。阳性对照组大鼠 TSH免疫反应细胞的面数密度增大 ,免疫反应增强 ( P <0 .0 5 )。本文对吗啡及乌拉地尔所引起大鼠垂体远侧部 TSH免疫反应细胞变化机理进行了探讨     

Pituitary Folliculo-Stellate-Like Cells Stimulate Somatotroic Pituitary Tumor Growth in Nude Mice

A new cell line (TtT/GF) established from a murine pituitary thyrotropic tumor having characteristics similar to those of pituitary folliculo-stellate cell (FS cell) was implanted into nude mice together with cells from a rat pituitary somatotrophic tumor cell line (MtT/S) to determine whether the former enhances pituitary tumor growth. For as long as 2-3 mo after implantation, MtT/S cells implanted either alone or together with fibroblasts formed either no tumors or only very small tumors in the nude mice. In contrast all of the nude mice that had received MtT/S cells implanted together with TtT/GF cells developed large tumors. Furthermore, the mice bearing the MtT/S and TtT/GF implants showed a significantly higher body weight and serum growth hormone level than those bearing only MtT/S cells or a combination of MtT/S cells and fibroblasts. The TtT/GF cell line itself had no tumorigenicity during the experimental period. Therefore, the TtT/GF cell line as a model of FS cells enhanced pituitary endocrine cell tumor formation. Additionally, immunocytochemistry showed that TtT/GF cells positive for glial fibrillary acidic protein (GFAP) or S-100 protein were present in the parenchymatous tissue elements or connective tissue surrounding the tumor nests. In the parenchymatous tissue, the TtT/GF cells exhibited a stellate appearance and surrounded neighboring tumor cells with their long cell processes. These results suggest that TtT/GF cells can serve as a model for pituitary FS cells, and are capable of stimulating pituitary tumor growth either by modifying the microenvironment or producing growth factors.     

Intercellular Communication Within the Rat Anterior Pituitary: XIV Electron Microscopic and Immunohistochemical Study on the Relationship Between the Agranular Cells and GnRH Neurons in the Dorsal Pars Tuberalis of the Pituitary Gland

Although numerous investigators in 1970s to 1980s have reported the distribution of LH‐RH nerve fibers in the median eminence, a few LH‐RH fibers have been shown to be present in the pars tuberalis. The significance of the finding remains to be elucidated, and there are few studies on the distribution of LH‐RH neurons in the pars tuberalis, especially in the dorsal pars tuberalis (DPT). Adult male Wistar‐Imamichi rats were separated into two groups: one for electron microscopy and the other for immunohistochemistry to observe LH‐RH and neurofilaments. Pituitary glands attached to the brain were fixed by perfusion, and the sections were prepared parallel to the sagittal plane. The typical glandular structure of the pars tuberalis was evident beneath the bottom floor of the third ventricle, and the thick glandular structure was present in the foremost region. Closer to the anterior lobe, the glandular structure changed to be a thin layer, and it was again observed at the posterior portion. Then the pituitary stalk was surrounded with the dorsal, lateral, and ventral pars tuberalis. LH‐RH and neurofilaments fibers were noted in the bottom floor, and some of them vertically descended to the gland. Adjacent to the glandular folliculostellate cells in the pars tuberalis, Herring bodies with numerous dense granules invading into the gland were present between the pituitary stalk and DPT. It was postulated that the “message” carried by LH‐RH might have been transmitted to the cells in the DPT to aid in the modulation of LH release. Anat Rec, 290:1388–1398, 2007. © 2007 Wiley‐Liss, Inc.     

Allotransplantation of Pituitary Cells to Rat Testicle

Fifty-three transplantations of pituitary tissue (intact and minced with scissors) under the tunica albuginea testis were carried out. In both series pituitary cells retained viability for up to 6 months. Transplantation of cell suspension was not accompanied by the formation of necrotic zone in the transplant; the adjacent testicular tubules and the entire testicular structure remained unchanged, while transplantation of large fragments of the pituitary tissue impaired of spermatogenesis in the adjacent testicular tubules, although most of them remained unchanged. Hence, cell suspension is preferable for transplantation of pituitary tissue into the testicle.     

细胞内抗原免疫金银染色技术的改进

为解决细胞内抗原应用免疫金银法染色时背景过重的问题,建立了甘氨酸二次阻断的处理方法,效果较好。     

In Search of a Prognostic Classification of Endocrine Pituitary Tumors

Pituitary tumors, the most frequent intracranial tumors, are historically considered benign. However, various pieces of clinical evidence and recent advances in pathological and molecular data suggest the need to consider that these tumors are more than an endocrinological disease despite the low incidence of metastasis. We present here a historical and critical review of the classifications of pituitary tumors, including a new prognostic clinicopathological classification based on tumor size, immunohistological subtype (prolactin (PRL), growth hormone (GH), follicle-stimulating hormone and luteinizing hormone (FSH-LH), adrenocorticotropic hormone (ACTH), and thyroid-stimulating hormone (TSH)), and five grades, which take into account invasion and proliferation of the tumors. We also present a brief review of the main markers of tumor behavior. We believe that a better classification of these tumors and the identification of prognostic markers will help the clinician to choose the appropriate therapeutic management.     

Clinical Impact of the Current WHO Classification of Pituitary Adenomas

WHO classifications should be used for comparing the results from different groups of pathologist and clinicians by standardized histopathological methods. Our present report describes the important parameters of pituitary adenoma pathology as demand of the WHO classification for correlation to endocrine data and prognosis. The combination of HE stain based structures with immunostainings for pituitary hormones allows subclassification of adenomas as the best method not only for correlations to clinical hyperfunctions but also for statements to the sensitivity of drug therapies (somatostatin analogs, dopamine agonists). GH-, PRL- and ACTH-secreting pituitary adenomas are further classified based on the size and number of their secretory granules by electron microscopy, or as is mostly the case nowadays by cytokeratin staining pattern, into densely and sparsely granulated. Granulation pattern may be considered for the prediction of treatment response in patients with GH-secreting adenomas, since the sparsely granulated subtype was shown to be less responsive to somatostatin analog treatment. For prognosis, it is important to identify aggressive adenomas by measurements of the Ki-67 index, of the number of mitoses, and of nuclear expression of p53. Among the criteria for atypical adenomas, high Ki-67 labeling index and invasive character are the most important adverse prognostic factors. Promising molecular markers have been identified that might supplement the currently used proliferation parameters. For defining atypical adenomas in a future histopathological classification system, we propose to provide the proliferative potential and the invasive character separately.     

Laminin Isoforms and Laminin-Producing Cells in Rat Anterior Pituitary

Laminin is a key component of the basement membrane and is involved in the structural scaffold and in cell proliferation and differentiation. Research has identified 19 laminin isoforms, which are assemblies of α, β, and γ chains (eg, the α1, β1, and γ1 chains form the laminin 111 isoform). Although laminin is known to be present in the anterior pituitary, the specific laminin isoforms have not been identified. This study used molecular biological and histochemical techniques—namely, RT-PCR, immunohistochemistry, and in situ hybridization—to identify the laminin isoforms and laminin-producing cells in rat anterior pituitary. RT-PCR showed that laminin α1, α3, and α4 genes were expressed in anterior pituitary. Immunohistochemistry revealed laminin α1 in gonadotrophs and laminin α4 in almost all vascular endothelial cells. Laminin α3 was seen in a subset of vascular endothelial cells. We then performed in situ hybridization to localize β and γ chains in these cells and found that laminin β1, β2, and γ1 were expressed in gonadotrophs and that laminin β1 and γ1 were expressed in endothelial cells. In conclusion, we identified gonadotroph-type (laminin 111 and 121) and vascular-type (laminin 411 and 311) laminin isoforms in rat anterior pituitary.     

Ultrastructural Morphology of Folliculo-Stellate Cells in Human Pituitary Adenomas

Fifty-six pituitary adenomas were studied by electron microscopy in a search for the presence of folliculo-stellate cells (FSCs) with the aim of evaluating their prevalence and ultrastructural morphology. FSCs were scattered in two adenomas (one oncocytoma and one densely granulated GH cell adenoma) and were numerous in a sparsely granulated GH cell adenoma; their overall prevalence was 5.4%. Ultrastructural examination of the three neoplasms revealed that FSCs were hypertrophic element with abundant cytoplasm and organelles (in contrast to FSCs of the normal pituitary) and no obvious signs of neoplastic transformation. Junctional complexes between FSCs were similar to those described in the normal gland. Numerous follicular structures were lined by FSCs. FSCs in pituitary adenomas are probably nonneoplastic, reactive cells showing signs of hyperactivity, similar to FSCs found during pituitary hypersecretion and in estrogen-induced tumor.