牦牛与迁饲黄牛及低海拔黄牛肺动脉干及肺组织形态的比较

目的:对牦牛和迁饲黄牛及低海拔黄牛肺动脉干与肺组织形态进行比较研究,获得高原土生或迁饲动物慢性低氧时适应进化的模式。方法:采集青海地区海拔3000~4000m的牦牛、2500 m的高山迁饲黄牛及甘肃平凉1300 m的低海拔黄牛的肺动脉干及肺组织,固定、切片、染色后行光镜和电镜比较观察。结果:55%的迁饲黄牛和98%牦牛肺动脉干中膜以弹性纤维及胶原纤维为主,而45%的迁饲黄牛和100%低海拔黄牛以弹性纤维及平滑肌增多为主,且平滑肌增多成团分布。牦牛和迁饲黄牛肺浆膜结构致密,间皮下以弹性纤维为主,而低海拔黄牛肺浆膜结构疏松,间皮下以胶原纤维为主。相比于迁饲黄牛,牦牛和低海拔黄牛肺终末细支气管黏膜纵行皱襞更明显,牦牛肺呼吸性细支气管黏膜上皮为单层或假复层柱状上皮,多数细胞可见到纤毛,并且黏膜仍有皱襞形成;而黄牛主要为单层柱状上皮,部分细胞可见纤毛。迁饲黄牛肺内小动脉血管壁和微动脉管壁厚度增加,与牦牛及低海拔黄牛比较差异有统计学意义。结论:牦牛肺动脉干和肺组织形态与结构有适应高原的独特形式,迁饲黄牛在适应高原时发生了与牦牛一样的趋同性改变,同时又兼具低海拔黄牛的特点。     

牦牛和柴达木黄牛颈动脉体的组织微细结构比较

目的比较牦牛和柴达木黄牛颈动脉体(CB)的组织微细结构特征。方法选择健康成年牦牛和柴达木黄牛各5头,采用组织学染色、免疫组织化学SP法和透射电子显微镜技术,观察CB的显微及超微结构特征,并运用显微体视学方法比较两种动物CB中Ⅰ型细胞(明细胞、暗细胞)、Ⅱ型细胞和血管的体积密度(Vv)、面密度(Sv)、比表面(δ)、面数密度(NA)。结果牦牛和柴达木黄牛CB均由实质和间质构成,实质由大量的圆形或椭圆形的小球密集组合而成,小球内有丰富的实质细胞。牦牛Ⅰ型细胞数量多于柴达木黄牛,但其胞质内含的线粒体和特有的电子致密核心囊泡(EDCV)相对小而多。柴达木黄牛小球之间的结缔组织及内含的间质细胞明显多于牦牛,而牦牛CB中血管数量多于柴达木黄牛,而且管腔较大。牦牛CB明细胞的Vv、Sv、NA显著大于柴达木黄牛(P0.01);牦牛δ小于柴达木黄牛(P0.05)。牦牛暗细胞的Vv、Sv显著小于柴达木黄牛(P0.05),两组中NA和δ差异不显著(P0.05)。两种动物CB中Ⅱ型细胞的Vv、Sv、NA、δ差异均不显著(P0.05)。牦牛微血管的Vv、Sv、NA显著大于柴达木黄牛(P0.01),而δ显著小于柴达木黄牛(P0.01)。结论牦牛CB形成了固有的组织细胞成分和血管网形态特征以增强对高原低氧环境的适应能力。     

一日龄高原牦牛肺泡组织结构特征

目的 探讨1日(d)龄高原牦牛肺泡组织结构特点及其与高原低氧适应性的关系. 方法 1d龄高原牦牛和1d龄平原黄牛各5头,HE染色、光镜、透射电镜观察和形态测量学分析. 结果 1d龄高原牦牛肺泡较同日龄平原黄牛扩张充分,肺组织中有大量类似肺泡囊的囊状结构,囊内可见大量 "芽"状结构,同日龄平原黄牛肺组织中少见此种结构;高原牦牛肺泡中Ⅱ型肺泡细胞数量较多,肺泡腔内可见大量排出的板层小体.高原牦牛单位面积内的肺泡数(MAN)较1d龄平原黄牛少(P<0.05),但单个肺泡的面积(MSAA)、肺泡隔厚度(MAST)和单位面积内的肺泡面积(SA)均大于平原黄牛的相应值(P<0.05);高原牦牛气-血屏障的算术平均厚度较平原黄牛大(P<0.05),但气-血屏障调和平均厚度两者差异不显著(P>0.05). 结论 1d龄高原牦牛的肺泡发育较平原黄牛完善.高原牦牛肺泡数将以"芽生"的形式快速增加.高原牦牛出生时肺泡相对完善的发育和肺泡数的快速增加以及肺泡隔相对较厚且厚薄不均等组织学特点是高原牦牛能很好适应高原低氧环境的组织学基础.     

低氧训练大鼠骨骼肌组织低氧诱导因子1对血管新生的影响*

背景：低氧诱导因子1是一种在氧平衡调节中起关键作用的转录因子,与机体的耐缺氧能力密切相关。 目的：观察低氧训练大鼠骨骼肌组织中低氧诱导因子1和血管内皮CD34的蛋白表达,探讨低氧诱导因子1在促进骨骼肌组织血管形成中的作用。 方法：将健康雄性SD大鼠60只,随机分为6组：常氧对照组、低氧不运动组、常氧训练组、低住高练组、高住低练组和高住高练低练组。运动组采用10 周递增负荷跑台运动训练,每周训练6 d,运动量由第1 周的速度为15 m/min、持续时间为25 min 递增至第10 周速度为28 m/min、持续时间为50 min,低练组每周二、四、六在相当于海拔1 500 m的低氧环境中训练,并且在低氧环境中居住,低氧程度由第1 周相当于海拔1 800 m 递增至第10 周相当于海拔3 600 m。 结果与结论：低氧状态下,低氧诱导因子1有大量的蛋白表达,低氧复合运动表达更多,而CD34 蛋白表达只发生在常氧运动组和低氧训练组。提示低氧诱导因子1是促进骨骼肌组织血管新生的一种重要因子,但须结合运动才能产生积极的作用。      

高原低氧大鼠肺组织超微结构及其低氧诱导因子1α的表达

背景：低氧诱导因子1α可介导哺乳动物细胞适应低氧环境。 目的：观察高原低氧对大鼠肺组织超微结构的影响及其低氧诱导因子1α表达变化。 方法：将SD大鼠分别为进行高原低氧干预1,2,3和30 d,并设置对照组。4个高原低氧组由海拔5 m的西安地区途中耗时1 d带到海拔2 700 m的青海格尔木地区、途中耗时2 d带到海拔5 000 m的唐古拉地区,途中耗时3,30 d分别带到海拔4 500 m的西藏那曲地区。 结果与结论：光镜及电镜观察显示,急性高原低氧2 d组肺组织出现明显的高原肺水肿,急性高原低氧30 d组低氧诱导因子1α mRNA的表达明显增高(P < 0.01),高原肺水肿现象则明显减轻。结果证实,低氧习服后肺组织低氧诱导因子1α mRNA表达的提高有利于减轻高原肺水肿。     

低氧训练时心肌组织细胞低氧诱导因子1α与凋亡相关蛋白的表达

背景：运动对心肌细胞凋亡的影响有了比较多的研究,但低氧训练对心肌细胞凋亡的研究不多见。低氧诱导因子1α是否控制了Bcl-2家族的表达而影响凋亡发生,尚未明确。 目的：分析低氧训练时心肌组织低氧诱导因子1α蛋白表达情况与凋亡蛋白的相互关系。 方法：将健康雄性SD大鼠60只,随机分为6组：①对照组常温常氧下喂养,不运动。②低氧组在低氧环境下喂养,不运动。③运动组常温常氧下运动。④低住高练组常氧下居住,低氧、常氧训练交替进行。⑤高住低练组低氧下居住,常氧下训练。⑥高住高练低练组低氧下居住,低氧、常氧训练交替进行。运动组大鼠负荷跑台训练8周,训练每周6 d,运动速度和运动时间连续递增,运动速度从开始的10 m/min、持续时间为15 min递增至第8周的25 m/min、持续时间为50 min。低氧程度由开始从海拔1 500 m增加到第8周达到海拔3 600 m。训练结束后,运用苏木精-伊红染色、TUNEL法和蛋白免疫组织化学方法检测大鼠心肌组织细胞低氧诱导因子1α、Bcl-2、Bax的表达变化。 结果与结论：①常氧时低氧诱导因子1α几乎不表达,低氧可增加低氧诱导因子1α的蛋白表达,低氧复合运动,表达更多。②低氧组和低氧运动组大鼠心肌组织Bax表达变化不是很明显,但Bcl-2表达就远远低于常氧不运动组。③低氧组、运动组与低氧运动组心肌组织细胞凋亡较明显,但三者相互之间比较差别较小,说明低氧结合运动不会使细胞凋亡更加严重。结果表明各实验组Bcl-2表达显著低于对照组,Bax表达变化不明显。但低氧可增加低氧诱导因子1α的蛋白表达,低氧复合运动,表达更多。说明低氧诱导因子1α有可能调节Bcl-2、Bax的表达,从而控制细胞凋亡的发生与否。 中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程      

MDA-MB-231乳腺癌细胞两种低氧模型的建立与比较分析

目的:由于实体瘤内的肿瘤细胞常处于低氧环境,本实验拟探究两种常用低氧模型对乳腺癌细胞影响的差异。方法:以不同浓度(0～300μmol/L)CoCl₂处理人乳腺癌MDA-MB-231细胞不同时间(24 h、48 h和72 h)建立化学低氧模型,以不同程度低氧(1%、2%和5%O₂)处理MDA-MB-231细胞不同时间(4 h、16 h和24 h)建立物理低氧模型。采用CCK-8法检测细胞活力,Western blot检测低氧诱导因子1α(HIF-1α)、Bcl-2及基质金属蛋白酶2(MMP-2)的蛋白表达;流式细胞术检测细胞凋亡;Transwell和划痕实验检测细胞的侵袭及迁移能力。结果:综合不同时间和浓度的HIF-1α蛋白表达结果,最终以100μmol/L CoCl₂处理24 h建立化学低氧模型,以2%O₂处理24 h建立物理低氧模型;物理低氧细胞的HIF-1α蛋白表达明显高于化学低氧,且物理低氧细胞处于复氧环境后HIF-1α蛋白快速降解;物理低氧细胞凋亡增加,抗凋亡蛋白Bcl-2表达降低,而化学低...     

低氧对细胞的影响及其适应机制

低氧(hypoxia)损伤是临床许多疾病中常见的病理过程,也是导致机体死亡的重要原因之一.随着分子生物学和细胞检测技术的快速发展,人们对细胞低氧损伤及其适应机制的研究日益深入.低氧作为细胞生长潜在的环境致死因素,影响着细胞周期、形态结构、代谢、信号通路、增殖、分化及凋亡等多个方面.另一方面,低氧又还是癌细胞转移及产生耐药的原因之一.机体细胞为适应低氧环境以减少损伤,诱导产生的低氧诱导因子(hypoxia-inducible factor,HIF)是低氧应答时基因表达和恢复细胞内环境稳定的中心调控因子;此外,nonsense mediated RNA的衰变、microRNA的诱导、染色质的核型改变和翻译调节等也是细胞对低氧的适应途径.阐明细胞低氧损伤及其适应机制具有重要意义,也可以为相关疾病的治疗提供新思路.     

一种改良的间歇性低氧细胞模型方法

 目的: 研制细胞氧舱,建立间歇性低氧(intermittent hypoxia,IH)细胞模型并进行验证。方法: 定制细胞实验舱和空气模拟对照舱,根据氧分压-时间曲线设计间歇低氧模式。将人肺腺癌细胞A549随机分为正常对照(Con)组、间歇低氧6 h(6IH)组、间歇低氧9 h(9IH)组、空气模拟对照6 h(6AC)组、空气模拟对照9 h(9AC)组、持续低氧4 h(4SH)组、持续低氧6 h(6SH)组。暴露结束后光镜下观察细胞形态改变,real-time PCR、免疫组化法检测缺氧诱导因子1α(HIF-1α)的mRNA和蛋白表达变化。结果: 该模型间歇低氧模式为5% O₂ 60 min-20% O₂ 30 min,6个循环。与Con组比较,6AC、9AC组为原本细胞形态,6IH、9IH和6SH组部分细胞出现突起、变圆,胞质中出现较多黑色颗粒,细胞边界模糊,而4SH组未见明显异常。与6IH组比较,9IH组HIF-1α的mRNA和蛋白表达量明显增加(P<0.05);6IH和9IH组HIF-1α的mRNA 和蛋白分别高于4SH和6SH组(P<0.05);6AC、9AC组与Con组比较差异不显著。结论: 5% O₂ 60 min-20% O₂ 30 min的间歇性低氧-复氧细胞模式能模拟阻塞性睡眠呼吸暂停低通气综合征病理生理过程,是研究该疾病较理想的细胞模型。     

低氧培养促进大鼠髓核间质干细胞增殖

目的研究低氧培养对大鼠髓核间质干细胞(NPMSCs)增殖的影响。方法分离大鼠尾椎髓核获得NPMSCs并进行体外扩增培养,观察细胞形态,流式细胞仪鉴定细胞免疫表型。取第3代NPMSCs分为常氧组和2%低氧组,分别培养7 d及14 d,观察细胞形态;MTT法检测细胞增殖;流式细胞仪检测细胞活力和RT-q PCR检测低氧诱导因子1α(HIF-1α)、葡萄糖转运蛋白1(GLUT-1)、血管内皮生长因子(VEGF)、沉默信息调节蛋白1(SIRT1)及SIRT6的mRNA表达情况。结果原代NPMSCs细胞早期可形成葵花样细胞集落,传代后细胞增殖明显加快,细胞形态以纺锤形为主。第3代细胞高表达干细胞相关阳性表面抗原分子,低表达干细胞相关阴性表面抗原分子。低氧组细胞形态以多角形为主,细胞更容易聚集成团块,且细胞增殖速度明显加快(P0.05),细胞凋亡率明显下降(P0.05)。低氧组HIF-1α、VEGF、GLUT-1、SIRT1及SIRT6表达量明显高于常氧组(P0.05)。结论 2%O2的低氧环境促进髓核间质干细胞增殖,其机制可能与SIRT1及SIRT6介导的HIF-1α信号通路有关。     

Comparative pathology of the enlarged carotid body

A histological study was made of the carotid bodies of man and various animal species from low and high altitudes. The animals studied were the alpaca, llama, cattle, guinea-pig, rabbit, dog, rat and man. As well as a qualitative microscopic study, a differential cell count was carried out to determine the percentage of the light and dark variants of chief cells and of sustentacular cells present. The investigation showed that the carotid bodies enlarge in cattle, guinea-pigs and rabbits living in the hypobaric hypoxia of high altitude to which they have to acclimatize. The carotid bodies are not enlarged in llamas and alpacas which show Darwinian adaptation to high altitude. There is no single histopathological appearance to be found with enlargement of the carotid body; on the contrary, there appears to be a characteristic histological reaction for different species. Thus, man shows hyperplasia of sustentacular cells, cattle show focal dark cell proliferation and rabbits, guinea-pigs, and dogs show striking hyperplasia and vacuolation of chief cells. In the rat, the enlargement of the carotid body is not characterized by the differential proliferation of any specific element and, as a result, it does not appear to be a good model for the human organ. In man and rat, carotid body enlargement occurs in response to systemic hypertension as well as to chronic hypoxaemia and the histological response to the 2 stimuli is the same, depending on the species. The normal rabbit carotid body is more reminiscent of that of man but, in this species, the reaction of glomic tissue differs from that of human glomic tissue.     

Paragangliomas of the head and neck: immunohistochemical neuroendocrine and intermediate filament typing

Twenty-nine paragangliomas of the head and neck region including 20 glomus jugulare (GJ) and nine carotid body (CB) tumors were evaluated for the presence of neuroendocrine and intermediate filament antigens. Immunohistochemistry on formalin-fixed, paraffin-embedded tissue was used to identify: S-100 protein (S-100); neuron-specific enolase (NSE); chromogranin A (CHA); serotonin (SER); synaptophysin (SYN); cytokeratin (CK); neurofilament (NF); desmin (DES); vimentin (VIM); and glial fibrillary acidic protein (GFAP). S-100 protein staining of sustentacular cell nuclei and cytoplasm was found in all tumors and was present in chief cells in 4 of 20 GJ and 3 of 9 CB tumors. All tumors stained with at least three neuroendocrine markers (29 of 29 NSE, 28 of 29 SYN, 26 of 29 CHA, 25 of 29 SER). CK was detected in 2 GJ and 1 CB tumor using anticytokeratins AE 1/3 and CAM 5.2. Neurofilament protein could not be demonstrated in fixed material, and all tumors were negative for GFAP and desmin. Vimentin was inconsistently detected in chief and sustentacular cells. We conclude that, in formalin-fixed material, paragangliomas have S-100 protein staining of sustentacular cells with chief cells containing antigens associated with neuroendocrine differentiation. The presence of CK in some paragangliomas is consistent with recent tissue culture studies demonstrating immunoblot confirmation of CK in pheochromocytomas and represents a potential source of immunohistologic misinterpretation in diagnosis, unless a panel of markers is utilized.     

Comparison of cell block preparation methods for nongynecologic ThinPrep specimens

The purpose of this study was to compare four cell block (CB) methods in the setting of nongynecologic ThinPrep (TP) specimens. 48 CBs were prepared from 12 nongynecologic TP specimens using the following CB methods: (1) Inverted filter sedimentation (IFS); (2) Thrombin method; (3) Albumin method; (4) Simple sedimentation. Each CB was assigned a cellularity score: 0 no cells, 1+ hypocellular, 2+ hypocellular with tissue fragments, 3+ cellular. A score of 2+ or 3+ was given for 11/12 of thrombin, 7/12 IFS, 5/12 albumin, and 2/12 simple sedimentation CBs. Thrombin CBs demonstrated a pale background clot with evenly distributed cells. Albumin CBs had a cracked uneven background. IFS CBs had a clear background, but were technically difficult and cells appeared artifactually crowded. In the setting of nongynecologic TP specimens, the thrombin CB was easily prepared and produced the best CB in regards to cellularity, cell distribution, and background quality.     

Bilateral carotid body tumor: the role of fine-needle aspiration biopsy in the preoperative diagnosis

Carotid body (CB) is a round to ovoid or flattened structure situated within the adventitia of the common carotid artery bifurcation on both sides of the neck. CB contains two basic types of cells: chief cells (or glomus type 1) and sustentacular cells (glomus type 2). Carotid body tumor (CBT) or paraganglioma arises from the chief cells of the carotid body. The diagnosis of CBT is typically made with radiological studies. Fine needle aspiration biopsy (FNAB) is seldom requested for this purpose due to rare but dreadful reported complications such as hemorrhage and damage to the carotid artery. In this report we discuss the cytological findings of a malignant CBT diagnosed by FNAB in a 22 year-old female.     

Hyperplasia of the carotid body

The histopathology of hyperplasia of the carotid bodies was studied in 6 cases of hypoxaemia and right ventricular hypertrophy secondary to pan-acinar emphysema, and in five cases of systemic hypertension with left ventricular hypertrophy. The features of the hyperplasia were the same in the two groups. There was proliferation of sustentacular (type II) cells and compression of central cores of chief (type I) cells. It is speculated that the hyperplasia of sustentacular cells is associated in some way with the prevention of retention of sodium ions and water which characterises hypoxic cor pulmonale in "blue bloaters", systemic hypertension, and ascent to high altitude with the complications of acute mountain sickness, and pulmonary and cerebral oedema.     

Multiparameter DNA flow-sorting demonstrates diploidy and SDHD wild-type gene retention in the sustentacular cell compartment of head and neck paragangliomas: chief cells are the only neoplastic component

Head and neck paragangliomas are considered to be biphasic tumours, composed of two distinct cell types: chief cells and sustentacular cells. A substantial number of these tumours show mutations in the SDHD gene located at chromosome 11q23. Although there is general agreement that paragangliomas are a neoplastic proliferation of chief cells, the nature of sustentacular cells is still a matter of debate. To clarify the nature of sustentacular cells further, multiparameter DNA flow cytometry was performed utilizing S-100 labelling as a selective marker of the sustentacular fraction simultaneously with DNA content measurement in six head and neck paragangliomas. S-100-positive fractions and other tumour-cell populations were flow-sorted and restriction digestion analysis of SDHD mutations was performed on each fraction. Flow cytometry demonstrated that the S-100 labelled cells were diploid. Restriction digestion analysis in informative cases revealed retention of the wild-type SDHD allele in S-100-positive fractions and loss of the wild-type allele in S-100-negative fractions. These data strongly suggest that sustentacular cells should be regarded as a non-neoplastic cell population that may be induced as a tumour-specific stromal component by chief cells.     

The distribution of enkephalins in human carotid bodies showing cellular proliferation and chronic glomitis

Human carotid bodies obtained at necropsy that showed prominence of either the sustentacular cell or the dark variant of chief cell or chronic carotid glomitis were studied by an immunogold labeling technique. The peptides methionine and leucine enkephalin had a similar distribution to that found in the normal human carotid body. They were localized prominently and predominantly in the dark and progenitor variants of chief (type I) cells. The sustentacular (type II) cells showed no immunoreactivity for the enkephalins. Cell counts on immunolabeled chief cells in cases of sustentacular cell hyperplasia and chronic carotid glomitis were found to be at the lower end of the normal range, whereas those in dark cell prominence occurred nearer the upper limit.     

Pulmonary endocrine cells in various species in the Himalaya

The numbers, morphology and distribution of pulmonary endocrine cells in goats, sheep and the yak and its interbreeds with cattle, dzos and stols, were studied after their demonstration by means of the peroxidase-antiperoxidase technique with a polyclonal antiserum raised in the rabbit to human neuron-specific enolase, a marker for neuroendocrine cells. The numbers, morphology and distribution were related to species and not to residence at high altitude. Pulmonary endocrine cells were common and mainly distributed as solitary cells in the epithelium of the bronchial tree in sheep. They were much less common and found mainly as clusters in the alveolar capillary walls in goats and in the yak and its interbreeds with cattle.