卵圆细胞转化过程中生物学标志物的表达变化及其意义

卵圆细胞为肝内的干细胞,大量研究认为,卵圆细胞参与了肝脏肿瘤的发生,可能是肝癌发生的始动细胞.鉴于卵圆细胞在原发性肝癌的发生中具有特殊作用,将其体外培养,使之成为细胞癌变多步骤发生发展的体外模型,动态观察其在转化过程中的生物学特征和蛋白表达变化,对于探讨卵圆细胞分化调控的机制以及进一步研究肝癌的发生机制都具有重要意义.     

肝细胞肝癌中卵圆细胞的组织学与超微结构研究

目的：探讨人肝癌肝组织存在卵圆细胞的可能性。方法：对２０例人肝细胞肝癌的手术标本行常规组织学和超微结构观察，并用胆管上皮分化标记ＣＫ７和肝细胞分化标记白蛋白对以上组织作免疫组化染色，同时对其中５例作免疫电镜标记。结果：光镜下，１４／２０例癌肿边缘常可见到增生的小胆管样结构。电镜下，１４／２０例可找到三型卵圆细胞。其中Ⅰ型细胞体积较小、核大、胞质少，此为较为原始的卵圆细胞。Ⅱ型细胞体积稍大，胞质稍多     

人输卵管粘膜上皮细胞的培养、纯化及生长特性

早期胚胎的发育过程在输卵管中进行,与输卵管微环境相互作用,并按其特定的程序逐渐发生[1,2]。因此,输卵管在人类生殖过程中占有重要的地位。目前,输卵管上皮细胞的形态及功能研究已成为生殖医学领域的一个研究热点[3~5],但对妊娠早期输卵管的功能的研究尚不完全[6],在体的研究     

大鼠羊膜上皮细胞移植对损伤脊髓生长相关蛋白43表达的影响

目的:探讨羊膜上皮细胞(AECs)移植对大鼠损伤脊髓生长相关蛋白43(GAP-43)表达和神经功能恢复的影响.方法: 取E12d～14d的SD大鼠AECs体外培养;改良-Allen撞击法制备大鼠脊髓损伤(SCI)模型,并随机分成假手术组、SCI+生理盐水对照组和SCI+AECs移植组;分别于术后第1、3、7、14和28天通过BBB 神经行为学评分法对神经功能进行评估,观察其恢复状况,并采用免疫组织化学和免疫印迹法检测脊髓组织GAP-43表达的变化.结果: GAP-43的表达于脊髓损伤后第7天显著增加,AECs移植后损伤脊髓中的GAP-43持续高表达至第28天,而盐水对照组的表达于术后14d左右逐渐恢复至正常水平.与盐水对照组相比较,AECs移植后2～4周脊髓损伤大鼠的后肢运动功能得到一定的改善.结论: AECs移植后可使损伤大鼠脊髓GAP-43的表达增高,并在一定程度上促进了大鼠后肢运动功能的恢复.     

卵丘细胞凋亡对体外培养卵母细胞发育潜能的影响

目的 研究体外培养中卵丘细胞凋亡对卵母细胞结构的影响,探讨体外受精中可用卵丘细胞凋亡率预测卵母细胞发育潜能的原因.方法 对GV期人卵丘-卵母细胞复合体进行体外成熟培养,用HE染色、DAPI染色和原位末端标记(TUNEL)法标记3种方法对单卵卵丘细胞凋亡率进行检测.分为凋亡率高和低的2组,用光镜和透射电镜观察卵母细胞的结构.对MⅡ期卵母细胞进行体外受精,培养2d后在光镜下评价胚胎质量.结果 卵丘细胞凋亡率低的卵母细胞结构和发育潜能良好;卵丘细胞凋亡率高的卵母细胞发育潜能差,存在各种结构异常,包括围卵周隙不均,对应不同部位的细胞质发育不同步;细胞质中出现堆积不均的细胞器团、次级溶酶体、及可能由溶酶体降解造成的大量空隙;线粒体外膜和嵴模糊、膨大,呈现凋亡迹象;第一极体碎裂;透明带异常增厚或变薄.相应的卵丘细胞微绒毛和细胞连接减少.结论 揭示了卵丘细胞凋亡对卵母细胞结构的影响,揭示了卵丘细胞凋亡率影响卵母细胞发育潜能的原因.     

小白鼠甲状腺滤泡上皮细胞发育的组织化学与立体计量学研究

对ＢＡＬＢ／ｃ小白鼠甲状腺滤泡上皮细胞从胚胎１４ｄ（Ｅ１４ｄ）至成年期的发育过程，进行了组织化学与立体计量学研究。结果表明：（１）Ｅ１４ｄ～１５ｄ，细胞排列成不规则的团和索状。Ｅ１６ｄ～Ｅ１７ｄ，细胞围成原始的甲状腺滤泡。Ｅ１８ｄ，滤泡腔内出现ＰＡＳ反应阳性胶体，成熟的甲状腺滤泡结构基本形成。（２）胚胎期滤泡上皮细胞核较大，核／质比大于１．０，分裂像易见。出生后２１ｄ，核／质比稳定在０．３５，滤泡上皮细胞形态结构接近成年期状态。（３）Ｅ１６ｄ，ＡｃＰ、ＡＴＰａｓｅ、ＭＡＯ和ＳＤＨ均呈阳性反应，随后酶活性逐步增强，至生后２１ｄ均达强阳性反应。     

乳腺癌中肌上皮细胞与预后的关系

通常,在几乎所有的乳腺癌中均存在肌上皮细胞.因此,我们通过对乳腺癌病理标本采用免疫组织化学方法显示肌上皮细胞,观察乳腺癌内的肌上皮细胞在各种病理类型中的分布情况,并分析其与乳腺癌预后的关系.     

肝硬化组织中卵圆细胞的光镜、电镜与免疫电镜观察

目的：探讨人肝硬化肝组织是否存在卵圆细胞。方法：对30例人肝细胞肝癌手术标本的癌旁肝硬化组织行常规组织学观察,并用胆管上皮分化标记CK7和肝细胞分化标记白蛋白对组织作免疫组化染色,同时对其中10例作了超微结构,5例作免疫电镜标记和观察。另外对5例正常肝脏和5例肝炎后肝硬化组织同时作了光镜、免疫组化和电镜观察,对2例肝炎后肝硬化作了免疫电镜标记和观察。结果：癌旁肝硬化组织的30例光间和10例电镜观察以及5例肝炎肝硬化光、电镜观察,均可以再生的肝细胞结节边缘见到散在的小细胞和增生的小胆管样结构。这些小细胞和增生的小胆管内少数小细胞,为卵圆形,体积较小,核大,胸质少,胞质内含较多的游恼核糖体,仅含少量粗面内质网和线粒体,质内可见张力微丝结构,这类细胞与邻近细胞间均有细胞间连接。免疫电镜示,卵圆细胞均表达CK7和白蛋白,但有些细胞内表达CK7多些,有些细胞则表达白蛋白多些。在正常肝组织内未见到类似细胞。结论：与动物实验性肝癌模型肝组织一样,人类肝硬化肝组织中出存在同样细胞形态和免疫表型特点的卵圆细胞。结果支持卵圆细胞可能为肝前体细胞或干细胞的假设。     

NK细胞的发育分化与功能极化

近十年来,NK细胞的研究有两大重要进展,一是发现NK细胞的受体多样性,二是发现NK细胞的功能多样性[1,2]。NK细胞受体主要由两个基因复合体集中编码,分别是人第19号染色体上的白细胞受体复合体(Leukocyte receptor complex,LRC)和第12号染色体上的NK细胞受体复合体(NK re-     

卵丘细胞分泌的趋化因子5与辅助生育结局的相关性

目的探讨卵丘细胞分泌的趋化因子5能否预测卵母细胞质量和辅助生育妊娠结局。方法收集75个体外受精周期中获得的562枚卵母细胞的卵泡液、受精液和其后的胚胎培养液,根据卵母细胞的发育命运进行分组：A组为未受精卵母细胞;B组受精后发育为非优质胚胎;C组受精后发育为优质胚胎;D组受精后发育为妊娠胚胎。ELISA方法测定4组卵泡液、受精液和胚胎培养液中CCL5的含量,并将结果进行比较。并分析卵母细胞成熟与CCL5水平的关系。结果A组卵泡液和受精液中CCL5的含量最低。C组与B组卵泡液和受精液中CCL5的含量比较有显著差异,但与D组比较无明显差别。B、C、D3组胚胎培养液中CCL5含量比较无明显差异。MII期卵子卵泡液中CCL5含量明显高于MI期和GV期卵子。结论卵丘细胞分泌的CCL5水平与卵母细胞的成熟和妊娠结局相关,可为选择移植胚胎和预测妊娠结局提供新的参考标准。     

老龄小鼠胸腺内皮细胞的变化

目的研究老化引起胸腺萎缩和功能减低的相关机制。方法用组织化学和免疫荧光染色方法分析胸腺皮质和髓质结构;用BrdU标记流式细胞仪方法研究胸腺内皮细胞增殖;细胞凋亡测定用TUNNEL流式细胞仪方法。结果老龄小鼠胸腺内皮细胞增殖阳性率6.7%,对照年轻小鼠为9.2%。老龄小鼠胸腺内皮细胞凋亡率为11.3%,对照年轻小鼠为2.6%。老龄小鼠胸腺结构出现退行性改变。结论老化引起胸腺内皮细胞增殖能力下降、凋亡率增加和胸腺结构出现退行性改变,可能是老年胸腺萎缩免疫力降低的原因之一。     

In Vitro Studies of Human Prostatic Epithelial Cells: Attempts to Identify Distinguishing Features of Malignant Cells

Abstract

Recent advances in culture techniques have enabled routine establishment and propagation of epithelial cells derived from normal and malignant tissues of the human prostate. Comparative studies of the responses of normal and cancer-derived cell populations to various growth and differentiation factors in vitro were undertaken to examine the possibility that cancer cells might respond differentially. Clonal growth assays in serum-free medium demonstrated that optimal proliferation of normal as well as cancer cell strains was generally dependent on the presence of cholera toxin, epidermal growth factor, pituitary extract, hydrocortisone, insulin, and high levels of calcium in the culture medium, and on the use of collagen-coated dishes. Only one cancer strain responded aberrantly to epidermal growth factor and hydrocortisone. Putative differentiation factors (transforming growth factor-β and vitamin A) inhibited the growth of all normal and cancer strains. The origin of a cancer-derived cell strain that responded similarly to normal strains was verified by positive labeling with a prostate cancer-specific antibody, validating the conclusion from these studies that normal and cancer prostatic epithelial cells are not distinguishable on the basis of responses to the tested factors.     

表皮生长因子单抗对人牙龈上皮细胞增殖的影响

本文采用体外人牙龈上皮细胞 (HGEC)培养技术和四唑盐 (MTT)比色法 ,探讨表皮生长因子 (EGF)对HGEC的作用 ,并评价抗EGF受体 (EGFR)单克隆抗体对它的影响。结果发现 ,从第 2天开始 ,1μg/L的EGF对HGEC有明显的促增殖作用(P <0 0 5 ) ,第 4天起 ,促增殖非常显著 (P <0 0 1) ,并持续到第 8天 ;EGFR抗体浓度在 1∶10 0 0～ 1∶10时 ,能显著地抑制EGF对HGEC的促增殖作用 (P <0 0 1) ,1∶10 0浓度抑制作用最大 ,并持续作用至第 8天 (P <0 0 1)。从而表明 ,EGF对HGEC有显著的促增殖作用 ,这种作用能被EGFR抗体所阻断。     

第3～5周人胚肝的细胞特征和生长因子及受体表达的研究

用第 3～ 5周人胚 ,石蜡切片 ,免疫组化染色 ,光镜下观察人胚肝的细胞特征和HGF、IGF -I、TGFβ1等生长因子及其受体、PCNA、AFP、CK19等的表达。结果发现第 3周末肝芽形成 ,第 4周肝索开始形成 ,第 3～ 4周人胚肝由同一类具有幼稚细胞形态学特征的细胞构成。这些细胞为AFP、c Met阳性反应。第 5周时肝索细胞的数量增加 ,开始出现PCNA的表达 ,仍仅为同一类细胞。第 5周肝索细胞呈IGF -I、TGFβ1及其受体免疫反应阳性 ,HGF阴性 ,其周围的心肌细胞及间充质细胞为HGF阳性反应。结果提示第 3～ 5周 ,组成肝芽和肝索的细胞属于肝干细胞 ,其形态和因子表达的差异说明肝干细胞可能处于不同的发育阶段 ,AFP、c Met可以作为此阶段肝干细胞的标记物 ,HGF、IGF -I、TGFβ1及其受体可能参与对早期人胚肝发育的调节。     

胎儿侧脑室脉络丛上皮细胞发育特点

目的观察脉络丛上皮细胞发育特点.方法在光镜下对43例胎儿侧脑室脉络丛上皮细胞进行观察和形态定量分析.结果第一阶段和第二阶段脉络丛柱状上皮细胞占脉络丛上皮细胞总数71.2%和52%;柱状上皮细胞高度分别为17.4±0.7μm和16±0.8μm,宽度分别为8.6±0.9μm和9.5±0.5μm.第一、第二和第三阶段脉络丛立方上皮细胞占脉络丛上皮细胞总数分别为28.8%、47.8%和100%;立方上皮细胞高度分别为15.6±0.9μm、10.4±0.6μm和8.6±0.7μm,宽度分别为14.2±1.7μm、10.5±0.5μm和8.4±0.9μm.结论侧脑室脉络丛上皮细胞(1)从柱状上皮细胞分化成立方上皮细胞;(2)随着胎龄增大,细胞逐渐变小.     

Diesel Exhaust Particle-Exposed Human Bronchial Epithelial Cells Induce Dendritic Cell Maturation and Polarization via Thymic Stromal Lymphopoietin

Human exposure to air pollutants, including ambient particulate matter, has been proposed as a mechanism for the rise in allergic disorders. Diesel exhaust particles, a major component of ambient particulate matter, induce sensitization to neoallergens, but the mechanisms by which sensitization occur remain unclear. We show that diesel exhaust particles upregulate thymic stromal lymphopoietin in human bronchial epithelial cells in an oxidant-dependent manner. Thymic stromal lymphopoietin induced by diesel exhaust particles was associated with maturation of myeloid dendritic cells, which was blocked by anti-thymic stromal lymphopoietin antibodies or silencing epithelial cell-derived thymic stromal lymphopoietin. Dendritic cells exposed to diesel exhaust particle-treated human bronchial epithelial cells induced Th2 polarization in a thymic stromal lymphopoietin-dependent manner. These findings provide new insight into the mechanisms by which diesel exhaust particles modify human lung mucosal immunity.     

Increased IgG Antibody-Induced Cytotoxicity Against Airway Epithelial Cells in Patients with Nonallergic Asthma

Background  IgG autoantibodies to airway epithelial cell proteins have been detected in patients with nonallergic asthma. Objective and Methods  To evaluate the functional significance of these autoantibodies, we examined the presence of IgG antibody-induced cytotoxicity against airway epithelial cells (A549) by the microcytotoxicity assay using IgG antibodies purified from patients with nonallergic asthma. Results  IgG antibody-induced cytotoxicity (expressed as percent cell lysis) was significantly increased in nine patients with nonallergic asthma (mean ± standard deviation; 30.6 ± 7.3%) as compared with eight healthy controls (13.9 ± 5.1%) and nine patients with allergic asthma (20.3 ± 10.4%; p < 0.05). In addition, IgG antibody-induced cytotoxicity was significantly inhibited when IgG antibodies from patients with nonallergic asthma were pre-incubated with recombinant human airway epithelial cell autoantigens (cytokeratin 18 or alpha-enolase proteins; p < 0.05). Conclusion  These results suggest a possible involvement of IgG autoantibody-induced cytotoxicity against airway epithelial cells in the pathogenesis of nonallergic asthma.     

The Limbal Epithelial Progenitors in the Limbal Niche Environment

Limbal epithelial progenitors are stem cells located in limbal palisades of vogt. In this review, we present the audience with recent evidence that limbal epithelial progenitors may be a powerful stem cell resource for the cure of human corneal stem cell deficiency. Further understanding of their mechanism may shed lights to the future successful application of stem cell therapy not only to the eye tissue, but also to the other tissues in the human body.     

Effect of Cholesterol Depletion on Interleukin-8 Production in Human Respiratory Epithelial Cells

Purpose

The lipid entities of cell membranes are components of the immune system and important mediators of inflammation. Despite increasing interest in the function of epithelial cells in inflammation, the role of cholesterol in this process has not been described. Here, we investigated the effect of cholesterol depletion on the inflammatory process in airway epithelial cells via the expression of interleukin (IL)-8 as a marker of inflammation.

Methods

A 549 cells were treated with 0.5% methyl-β-cyclodextrin as a selective cholesterol extractor. The IL-8 level was assessed by enzyme-linked immunosorbent assay and reassessed after cholesterol repletion. Mitogen-activated protein kinase (MAPK) inhibitors were used to determine the upstream signaling pathway for IL-8 production in cholesterol-depleted cells.

Results

We found a relationship between the amount of cholesterol in A 549 cells and inflammation of the airway. IL-8 production was increased in cholesterol-depleted A 549 cells and restored by cholesterol repletion. IL-8 production was decreased by pretreatment with the extracellular signal-regulated kinase (ERK) inhibitor U0126 but not with JNK inhibitor II or the p38 MAPK inhibitor SB202190.

Conclusions

Our findings suggest that inflammatory responses are increased in cholesterol-depleted epithelial cells via the MAPK signaling system, predominantly by the ERK pathway. We conclude that the lipid components of airwayepithelial cells may play a role in the inflammatory process.     

Matrigel‐Induced Tubular Morphogenesis of Human Eccrine Sweat Gland Epithelial Cells

Human eccrine sweat glands are tubule‐structured glands of the skin that are vital in thermoregulation, secretion, and excretion of water and electrolytes. A study of tubular morphogenesis in vitro would facilitate the development of a tissue engineering model for eccrine sweat glands and other tubule‐structured glands. Matrigel, a basement membrane matrix, has been shown to promote differentiation and morphogenesis of many different cell types, including tubular cells. This study investigated the growth, differentiation, and tubular morphogenesis of human eccrine sweat gland epithelial cells cultured in Matrigel. Human eccrine gland epithelial cells were isolated and cultured in vitro. The cell growth in Matrigel was evidenced by the formation of cell clusters, which were observed under an inverted microscope. The internal structure of the cell clusters was further investigated by hematoxylin–eosin (HE) staining and confocal laser scanning microscopy (CLSM) of propidium iodide‐stained nuclei. The results demonstrated that although on a plastic surface or in a collagen gel the cells could not form tubular structures, they formed tubular structures when cultured in Matrigel. Consequently, we conclude that Matrigel can promote tubular morphogenesis of human eccrine sweat gland epithelial cells. Anat Rec, 2011. © 2011 Wiley‐Liss, Inc.