骨髓间质干细胞移植治疗大鼠脑出血模型的实验研究

目的:研究大鼠骨髓间质干细胞(MSCs)移植治疗脑出血的可行性。方法:分离MSCs后,连续传代培养、扩增,Brdu标记的MSCs通过颈动脉、侧脑室2种途径移植入脑出血大鼠模型体内,采用爬行计分法评估神经功能的恢复程度,并观察脑部Brdu阳性细胞的分布。结果:通过侧脑室、颈动脉移植后大鼠神经功能改善,明显优于对照组(P<0.05);经颈动脉注射组较经侧脑室注射组爬杆实验评分低(P<0.05)。移植的MSCs主要迁移到出血灶、大脑皮层、海马区等处。结论:MSCs移植对脑出血具有保护作用,而经颈动脉给药疗效优于经侧脑室给药。     

巨噬细胞、内皮细胞对高密度脂蛋白的氧化修饰

为了探讨高密度脂蛋白 (HDL)在体内发生氧化修饰的部位及机制 ,分别观察了培养人动脉平滑肌细胞 (SMC)、动脉内皮细胞 (EC)及巨噬细胞 (MΦ)与HDL共同温育过程中 ,HDL的琼脂糖电泳相对迁移率 (REM)、硫代巴比妥酸反应物质 (TBARS)以及溶血卵磷脂 卵磷脂 (LPC PC)值等氧化指标的变化 .结果发现 ,HDL与 3种细胞温育 12h时 ,HDL几乎不发生氧化修饰 ,而HDL与MΦ和EC温育 2 4h后 ,其REM、TBARS、LPC PC值均显著上升 (p <0 0 1) ;而HDL与SMC温育后 ,其REM、TBARS值无显著改变 ,LPC PC值增加 (p <0 0 1) .结果提示 ,活体内HDL可能主要在动脉壁的内皮细胞及巨噬细胞的作用下发生氧化修饰     

猫嗅球外丛层和僧帽细胞层年龄相关形态学变化

分别用Nissl法及免疫组织化学ABC法标记青、老年猫嗅球中嗅觉二级神经元和外丛层胶质细胞,显微镜下观察其分布并计数,对嗅觉二级神经元胞体直径和外丛层厚度进行测量,比较其年龄相关性变化,研究神经元与胶质细胞之间的关系,探讨老年性嗅觉功能衰退的相关神经机理。结果显示,老年猫嗅觉二级神经元胞体直径和分布密度均有不同程度的显著性下降(P<0.05);外丛层厚度变化不明显(P>0.05);外丛层胶质细胞特别是星形胶质细胞显著性增生(P<0.05)。表明在衰老过程中嗅觉二级神经元有丢失,并呈现功能下降,可能是老年性嗅觉功能衰退的原因之一。同时外丛层胶质细胞增生以进一步保护神经元,延缓其衰老。     

Morphological variety of intracellular microcolonies of Legionella species in Vero cells

Intracellular microcolonies of six Legionella species growing in Vero cells showed distinctly varied morphologies. The varieties were observed by light microscopy of Gimenez-stained, Legionella-infected Vero cells and by electron microscopy (EM). Legionella pneumophila Philadelphia-1 formed needle-shaped crystal-like microcolonies. Legionella bozemanii WIGA formed microcolonies like wool balls containing filamentous cells. In EM, these organisms proliferated in endosomes, which were adjacent to swollen rough endoplasmic reticula. Legionella oakridgensis OR-10 showed serpentine chains. Many mitochondria were observed around the microcolonies. Legionella jordanis BL-540 formed spherical moss-like microcolonies which were or were not surrounded by endoplasmic membranes. Legionella feeleii WO-44C spread throughout the cytoplasm without making clusters. Legionella dumoffii Tex-KL made big clusters that spread in the cytoplasm, a portion of which was outside the endosome membranes. These different morphologies imply diversity in modes of intracellular multiplication of Legionella spp.     

大流行流感疫苗血清学检测中马血球和鸡血球血凝抑制试验方法的比较

血凝抑制试验(HI)是评价季节性流感疫苗免疫效果的经典方法。由于对人、禽流感病毒的受体不同,不同来源的红血球检测敏感性可能有差异。实验中比较了经典的鸡血球HI方法和国外报道的马血球HI方法,发现两种方法在检测大流行流感疫苗接种者血清时,于不同毒株抗原检测时表现各不相同,检测结果差异在2倍之内,说明经典的鸡血球HI方法仍适用于评价大流行流感疫苗的免疫效果。     

Human breast microvascular endothelial cells retain phenotypic traits in long-term finite life span culture

Summary Attempts to study endothelial-epithelial interactions in the human breast have been hampered by lack of protocols for long-term cultivation of breast endothelial cells (BRENCs). The aim of this study was to establish long-term cultures of BRENCs and to compare their phenotypic traits with the tissue of origin. Microvasculature was localized in situ by immunohistochemitry in breast samples. From this tissue, collagen-rich stroma and adipose tissue were dissected mechanically and further disaggregated to release microvessel organoids BRENCs were cultured from these organoids in endothelial specific medium and characterized by staining for endothelial markers. Microvessels were a prominent feature of intralobular tissue as evidenced by immunostaining against endothelial specific markers such as CD31, VE-cadherin, and von Willebrand factor (VWF). Double staining against VE-cadherin and lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1) showed that blood and lymphatic vessels could be distinguished. An antibody against CD31 was used to refine protocols for isolation of microvasculature from reduction mammoplasties. BRENCs retained critical traits even at high passage, including uptake of low-density lipoprotein, and had E-selectin induced upon treatment with tumor necrosis factor-α. The first signs of senescence in passage 14 were accompained by gain of trisomy 11. At passage 18 cells showed chromosomal aberrations and growth arrest as revealed by β-galactosidase staining. We demonstrate here that breast microvasculature may serve as a large-scale source for expansion of BRENCs with molecular and functional traits preserved. These cells will form the basis for studies on the role of endothelial cells in breast morphogenesis.     

人肝癌细胞株中EGFR的表达和EGF对肝癌细胞生长的促进

本文分析了人肝癌细胞株7404,7721细胞中表皮生长因子受体(EGFR)基因表达和EGF对肝癌细胞生长的促进作用。~(125)Ⅰ-EGF对7404细胞的结合试验表明结合是可饱和的和专一的,从~(125)Ⅰ-EGF对7404、7721细胞结合浓度曲线作Scatchard作图和计算,提示每个7404和7721细胞表面分别有1.1×10~5和0.7×10~5的EGFR分子。Northern杂交分析证明EGFR基因在7404,7721细胞中的转录产物主要是5.6 kb EGFR mRNA,免疫印迹分析证明7404细胞和7721细胞的EGFR为170kd的蛋白。EGF对培养于含10%或0.5%小牛血清的RPMI-1640培液中的7404、7721细胞的贴壁依赖性生长有促进作用,促进作用的程度与培液中CS含量有一定关系,提示EGF的促生长作用可能是EGF与血清中其他成分协同作用的结果。EGF对培养于软琼脂中的7404,7721细胞的贴壁不依赖性生长也有明显促进作用。综合上述实验结果说明EGFR基因在人肝癌细胞中是活跃表达的,EGF可能是肝癌细胞生长依赖的一个重要有丝分裂原。     

大鼠卵巢绒毛膜促性腺激素受体在CHO中的表达和扩增

本文报道了利用二氢叶酸还原酶放大系统将大鼠ＬＨ／ｈＣＧ受体（记为Ｆ－ｈＣＧＲ）及其胞外肽段（记为Ｔ－ｈＣＧＲ）在中国苍鼠卵巢细胞（ＣＨＯ）中的表达。ＳＤＳ－ＰＡＧＥ分析表明,Ｆ－ｈＣＧＲ为一条蛋白质带,其表观分子量为９２ｋｄ,而Ｔ－ｈＣＧＲ为３５ｋｄ和３７ｋｄ两条带。表达受体对其配基ｈＣＧ表现出高的亲合力,Ｆ－ｈＣＧＲ的解离常数为７×１０－９ｍｏｌ／Ｌ,Ｔ－ｈＣＧＲ为６．４×１０－９ｍｏｌ／Ｌ。表达Ｆ－ｈＣＧＲ的转染ＣＨＯ细胞可结合１２５Ｉ－ｈＣＧ,而表达Ｔ－ｈＣＧＢ者不结合１２５Ｉ－ｈＣＧ。这提示Ｆ－ｈＣＧＲ主要存在于细胞质膜表面上。表达Ｆ－ｈＣＧＲ的转染ＣＨＯ细胞能刺激。ｃＡＭＰ的形成,而表达Ｔ－ｈＣＧＲ者不能刺激ｃＡＭＰ形成。免疫荧光定位结果表明,Ｔ－ｈＣＧＲ主要分布于质膜的细胞质侧以及胞内其他一些细胞器膜上。用免疫亲和层析可以得到纯化的Ｔ－ｈＣＧＲ。     

Alcohol exposure alters the expression pattern of neural cell adhesion molecules during brain development

Neural cell adhesion molecules (NCAMs) play critical roles during development of the nervous system. The aim of this study is to investigate the possible effect of ethanol exposure on the pattern of expression and sialylation of NCAM isoforms during postnatal rat brain development because alterations in NCAM content and distribution have been associated with defects in cell migration, synapse formation, and memory consolidation, and deficits in these processes have been observed after in utero alcohol exposure. The expression of NCAM isoforms in the developing cerebral cortex of pups from control and alcohol-fed mothers was assessed by western blotting, ribonuclease protection assay, and immunocytochemistry. The highly sialylated form of NCAM [polysialic acid (PSA)-NCAM] is mainly expressed during the neonatal period and then is down-regulated in parallel with the appearance of NCAM 180 and NCAM 140. Ethanol exposure increases PSA-NCAM levels during the neonatal period, delays the loss of PSA-NCAM, decreases the amount of NCAM 180 and NCAM 140 isoforms, and reduces sialyltransferase activity during postnatal brain development. Neuraminidase treatment of ethanol-exposed neonatal brains leads to more intense band degradation products, suggesting a higher content of NCAM polypeptides carrying PSA in these samples. However, NCAM mRNA levels are not changed by ethanol. Immunocytochemical analysis demonstrates that ethanol triggers an increase in PSA-NCAM immunolabeling in the cytoplasm of astroglial cells, accompanied by a decrease in immunogold particles over the plasma membrane. These findings indicate that ethanol exposure during brain development alters the pattern of NCAM expression and suggest that modification of NCAM could affect neuronal-glial interactions that might contribute to the brain defects observed after in utero alcohol exposure.