短暂脑缺血发作患者红细胞与损伤内皮细胞粘附研究

采用流室系统定量地研究了短暂脑缺血发作患者红细胞与损伤内皮细胞的粘附特性．结果表明短暂脑缺血发作患者红细胞与损伤内皮细胞的粘附较红细胞与正常内皮细胞的粘附数目增多，强度增加，提示内皮细胞损伤促进了红细胞与内皮细胞粘附，而红细胞与内皮细胞粘附作用增强又加重了内皮细胞损伤，最终造成血流低灌注，导致神经功能缺损．同时也提示内皮细胞因素是红细胞与内皮细胞粘附的重要影响因素之一．     

短暂脑缺血发作患者经细胞与损伤内皮细胞粘附研究

采用流室系统定量地研究了短暂脑缺血发作患者红细胞与损伤内皮细胞的粘附特性。结果表明短暂脑缺血发作患者红细胞与损伤内皮细胞的粘附较红细胞与正常内皮细胞的粘附数目增多，强度增加，揭示内皮细胞损伤促进了红细胞和内皮细胞粘附，而红细胞与内皮细胞粘附作用增强了又加重内皮细胞损伤，最终造成血流低灌注，导致神经功能缺损。     

正常红细胞与血管内皮细胞粘附特性

用流室法研究正常红细胞（ＲＢＣ）与人工培养的脐静脉内皮细胞之间的粘附特性。结果表明在低切应力下，正常红细胞与内皮细胞之间存在一定数量的粘附，随着切应力的逐步增大，ＲＢＣ—内皮细胞的粘附越来越减少。自体血浆的参与能增加ＲＢＣ—内皮细胞之间的粘附数量约３～５倍，并且也增强了粘附的强度。本文给出了反映切应力与ＲＢＣ—内皮细胞粘附数关系的经验公式，得到粘附特征参数ａ，ｂ值。此ａ，ｂ值能反映不同条件下的ＲＢＣ—内皮细胞粘附特性，本文结果还提出，在体内微静脉部位，健康人红细胞与内皮细胞之间可能会出现粘附     

脑血栓形成病人自体血浆对红细胞与内皮细胞粘附的影响

本文采用流室系统，定量地研究了脑血栓形成病人和健康人自体血浆对红细胞与内皮细胞粘附特性影响的差异，并深入探讨了脑血栓形成病人自体血浆纤维蛋白原和自体血小板对红细胞与内皮细胞粘附的影响。通过显微观察和计数，结果表明：１）血浆能增强红细胞与内皮细胞的粘附，这在脑血栓形成病人表现得尤为明显；２）脑血栓形成病人自体血浆纤维蛋白原促进了红细胞与内皮细胞的粘附，自体血小板的作用有待进一步研究。     

脑血栓形成病人自休血浆对红细胞与内皮细胞粘附的影响

本文采用流室系统，定量地研究了脑血栓形成病人和健康人自体血浆对红细胞与内皮细胞粘附特性影响的差异，并深入探讨了脑血栓形成病人自体血浆纤维蛋白原和自体血小板对红细胞与内皮细胞粘附的影响，通过显微观察和计数，结果表明：（１）血浆能增强红细胞与内皮细胞的粘附，在脑务栓形成病人表现得尤为明显（２）脑血栓形成病人自体血浆纤维蛋白原促进了红细胞与内皮细胞的粘附，自体血小板的作用有待进一步研究。     

病理情况下红细胞与血管内皮细胞粘附特性的研究进展

正常生理情况下红细胞几乎不与血管内皮细胞粘附，但在某些病理条件下，红细胞与内皮细胞间的粘附增强。粘附有其固有的分子基础，并受红细胞，内皮细胞，血浆以及管壁管因素的影响。粘附引起了血管阻塞，内皮损伤等病现结果，并表现出相应的临床症状。     

藻酸双酯钠对健康人红细胞与培养内皮细胞粘附的影响

本研究采用流室系统，定量地研究了体外藻酸双酯钠注射液对健康人红细胞与培养的人脐静脉内皮细胞粘附的影响。结果发现：藻酸双酯钠通过同时作用于红细胞与内皮细胞，从而使红细胞与内皮细胞粘附的数目减少，强度减弱，这可能是藻酸双酯钠能改善血液循环状况的又一重要机制；本研究还发现藻酸双酯钠在对抗红细胞与内皮细胞粘附方面，预防用药较治疗用药效果更佳，这具有重要的临床学意义。     

右旋糖酐40、70对红细胞与内皮细胞粘附的影响

采用流室显微观察系统，定量研究右旋糖酐４０（ＤＸ４０）和右旋糖酐７０（ＤＸ７０）对红细胞与内皮细胞动态粘附特性的影响。统计分析处理数据，得到反映切应力与细胞间动态粘附数关系的经验公式及粘附特征常数ａ，ｂ值。ａ值反映红细胞的初始粘附数，ｂ值反映随切应力增大，粘附数减小的速率。ＤＸ７０实验组的ａ值大于ＤＸ４０组的，而ｂ值的情况相反。故结果显示切应力越大，粘附的红细胞越少；ＤＸ４０使红细胞的粘附明显减少；ＤＸ７０使红细胞粘附显著增加。由此提示不同长度的细胞桥接分子对红细胞的粘附影响不同，且临床上选用ＤＸ４０作血浆扩容剂较ＤＸ７０优越得多     

内皮细胞脂质过氧化损伤对单核细胞粘附的影响

以巯基氧化剂联胺作用于培养人血管内皮细胞，引发细胞过氧化脂质蓄积与质过氧化损伤，研究内皮细胞脂质过氧化损伤对人血单核细胞粘附的影响，结果发现，内皮细胞脂质过氧化损伤使单核细胞对内皮细胞的粘附增加，单核细胞主要粘附于受损伤的内皮细胞表面和间隙，提示单核细胞对内皮细胞粘附增加，是内皮细胞脂质过氧化损伤促进和加重动脉粥样硬化病变形成的一个重要机制。     

血管内皮细胞的活化有利于造血干细胞的归巢

目的:探讨影响造血干细胞归巢的因素,为提高造血干细胞移植的效率奠定基础。方法:传代培养的血管内皮细胞长成单层后,用血管内皮细胞生长因子(VEGF)、粒细胞集落刺激因子(G-CSF)和脂多糖(LPS)激活内皮细胞,再将经过去除红细胞、粒细胞、单核细胞和T淋巴细胞而富集的脐血造血干细胞与内皮细胞共同培养,使之发生粘附。用双抗体夹心ELISA法测定C-Kit⁺的造血干细胞与血管内皮细胞的粘附情况。并比较VCAM-1单克隆抗体封闭血管内皮细胞对造血干细胞粘附的影响。结果:静息状态的血管内皮细胞能够与C-Kit⁺的造血干细胞发生粘附,但粘附率较低。用VEGF、G-CSF和LPS激活血管内皮细胞后,造血干细胞的粘附显著增加。用粘附分子VCAM-1单克隆抗体预处理激活血管内皮细胞后,造血干细胞的粘附明显下降。结论:激活的血管内皮细胞明显增加对造血干细胞的粘附,这种粘附与粘附分子的作用有关。     

补肾宁心方对人单核-血管内皮细胞粘附的影响

目的:探讨补肾宁心方对人单核-血管内皮细胞粘附的影响及机理。方法:以培养人脐静脉内皮细胞(HUVECs)作为靶细胞,在内皮细胞培养基中加入氧化的低密度脂蛋白(ox-LDL)或在试验体系中加入灌服补肾宁心方的兔血清,以孟加拉玫瑰红活细胞染色法测定人单核细胞系U937与HUVECs的粘附,并用流式细胞仪检测内皮细胞表面粘附分子细胞间粘附分子-1(ICAM-1)、血管细胞粘附分子-1(VCAM-1)以及E-选择素的表达。结果:ox-LDL显著增强单核U937细胞与内皮细胞之间的相互粘附,如在试验体系中加入灌服补肾宁心方的动物血清,则粘附率明显降低(P＜0.01)。流式细胞仪分析结果显示ox-LDL能明显促进内皮细胞表面ICAM-1、VCAM-1以及E-选择素的表达,补肾宁心方中药灌服血清可显著下调内皮细胞表面ICAM-1、VCAM-1以及E-选择素的表达(P＜0.01)。结论:补肾宁心方含药血清可能通过下调内皮细胞表面粘附分子的表达抑制单核-血管内皮细胞粘附,从而发挥对血管内皮细胞的保护作用。     

E-selectin mediates Porphyromonas gingivalis adherence to human endothelial cells

Porphyromonas gingivalis, a major periodontal pathogen, may contribute to atherogenesis and other inflammatory cardiovascular diseases. However, little is known about interactions between P. gingivalis and endothelial cells. E-selectin is a membrane protein on endothelial cells that initiates recruitment of leukocytes to inflamed tissue, and it may also play a role in pathogen attachment. In the present study, we examined the role of E-selectin in P. gingivalis adherence to endothelial cells. Human umbilical vein endothelial cells (HUVECs) were stimulated with tumor necrosis factor alpha (TNF-α) to induce E-selectin expression. Adherence of P. gingivalis to HUVECs was measured by fluorescence microscopy. TNF-α increased adherence of wild-type P. gingivalis to HUVECs. Antibodies to E-selectin and sialyl Lewis X suppressed P. gingivalis adherence to stimulated HUVECs. P. gingivalis mutants lacking OmpA-like proteins Pgm6 and -7 had reduced adherence to stimulated HUVECs, but fimbria-deficient mutants were not affected. E-selectin-mediated P. gingivalis adherence activated endothelial exocytosis. These results suggest that the interaction between host E-selectin and pathogen Pgm6/7 mediates P. gingivalis adherence to endothelial cells and may trigger vascular inflammation.     

HUVECs from newborns with a strong family history of myocardial infarction overexpress adhesion molecules and react abnormally to stimulating agents

Atherosclerosis is a complex disease involved in major fatal events such as myocardial infarction and stroke. It is the result of interactions between metabolic, dietetic and environmental risk factors acting on a genetic background that could result in endothelial susceptibility. Our aim was to determine the patterns of expression of adhesion molecules and whether phosphatidylserine is translocated to the cell surface of human umbilical vein endothelial cells (HUVECs) isolated from healthy newborns born to parents with a strong family history of myocardial infarction under TNF-alpha or oxLDL stimulated conditions. Compared to control HUVECs, experimental cords showed: (a) a four-fold increase in VCAM-1 expression under basal conditions, which showed no change after stimulation with the pro-atherogenic factors; (b) a two-fold increase in basal P-selectin expression that reached a 10-fold increase with any of the pro-atherogenic factors; (c) a basal ICAM-1 expression similar to P-selectin that was not modified by the pro-atherogenic molecules; (d) a similar PECAM-1 expression. Unexpectedly, phospathidylserine expression in experimental cord HUVECs was significantly increased (211 817 versus 3354 TFU) but was not associated to apoptotic death as the percentage of dead cells induced by TNF-alpha treatment was very low (0.55 versus 9.87% in control HUVECs). The latter result was corroborated by TUNEL staining. T cell adherence to HUVECs was highly up-regulated in the genetically predisposed samples. The analysis of nonpooled HUVECs, from newborns to family predisposed myocardial-infarction individuals, might represent a useful strategy to identify phenotypical and functional alterations, and hopefully, to take early preventive actions.     

蛋白激酶C在脂多糖诱导内皮细胞β-1,4-半乳糖基转移酶-Ⅰ表达中的作用

目的 研究蛋白激酶c(PKC)对脂多糖(LPS)诱导内皮细胞β-1,4-半乳糖基转移酶-Ⅰ(β-1,4-galactosyltransferase-Ⅰ,β-1,4-GalT-Ⅰ)表达的调节作用,以及对内皮细胞骨架结构改变及其黏附能力的影响.方法 分别用PKC激动剂或几种不同类型的PKC抑制剂预处理人脐静脉内皮细胞(HUVECs)30 min,LPS刺激HUVECs 4 h后,RT-PCR、Western blot方法 检测β-1,4一GalT-Ⅰ表达变化,细胞荧光染色观察β-1,4-GalT-Ⅰ催化的糖链表达变化及细胞骨架结构的改变,通过内皮-单核细胞黏附试验观察HUVECs黏附能力的改变.结果 几种不同类型的PKC抑制剂均能不同程度地抑制LPS刺激HUVECs引起的β-1,4-GalT-Ⅰ表达的上调,PKC激动剂能够使上调的β-1,4-GalT-Ⅰ的表达进一步增加;在HUVECs中β-1,4-GalT-Ⅰ与细胞骨架有共同定位,PKC抑制剂显著抑制LPS诱导的内皮细胞骨架蛋白的重构和β-1,4-GalT-Ⅰ细胞内的再分布;PKC抑制剂显著抑制LPS诱导的内皮-单核细胞黏附能力的上调.结论 PKC可能参与调节LPS诱导的HUVECs β-1,4-GalT-Ⅰ的表达,可能多种类型的PKC参与了这一调节过程;PKC可能通过对β-1,4-GalT-Ⅰ的调节进而影响炎症过程中内皮细胞骨架蛋白的重构及内皮细胞与单核细胞的黏附能力.     

Cell-surface thioredoxin-1: possible involvement in thiol-mediated leukocyte-endothelial cell interaction through lipid rafts

Human thioredoxin-1 (hTrx) exhibits a disulfide reducing activity and was originally identified as a soluble cytokine-like factor secreted from cells of a human T-cell leukemia virus type I (HTLV-I)-transformed cell line. Recent studies have revealed that endogenous Trx plays an important role in cytoprotection against various oxidative stress-associated disorders. However, the function of exogenous Trx is still not fully understood. We report here that a cysteine-modified mutant of recombinant human Trx (rhTrx-C35S) binds to human umbilical vein endothelial cells (HUVECs) as well as stimulated T cells and rapidly enters these cells via lipid rafts. In addition, we found that endogenous Trx is expressed on the surface of HUVECs, including lipid rafts. These events suggest cell-surface Trx as a possible target of rhTrx-C35S. Furthermore, we found that anti-human Trx mouse monoclonal antibody inhibits adherence of LPS-stimulated human peripheral blood polymorphonuclear cells (PMNs) to HUVECs. This adherence was also suppressed by a recombinant human Trx (rhTrx), but not by a mutant rhTrx (rhTrx-C32S/C35S) with no reducing activity. Cell-surface Trx may be involved in the process of interaction between PMNs and HUVECs and a possible target of cysteine-modified exogenous Trx as well as wild-type exogenous Trx through redox regulation.     

胎儿有核红细胞的可识别标记

[摘要] 从母血循环中分离、富集和提取胎儿有核红细胞,进行无创性产前基因诊断，是近年来医学界具有重大意义的研究课题。胎儿有核红细胞由于自身特点较其它胎儿细胞更适用于产前诊断。有多种方法可用于分离胎儿有核红细胞，本文主要就胎儿有核红细胞特异性标记进行综述。目前使用最多的阳性标记是CD71、CD36、GPA、血红蛋白等，同时使用CD45 等阴性标记可以提高分选效率及纯化率。随着分子生物学、分子免疫学等学科的发展，会有更多的胎儿有核红细胞（Nucleated Red Blood Cells,NRBCs）特异性标记被发现，从而促进产前诊断的不断发展。     

Phosphorylation and disorganization of vascular-endothelial cadherin in interaction between breast cancer and vascular endothelial cells.

Adherens junctions of the endothelium play a key role in the maintenance of endothelial permeability and are composed of the vascular endothelial (VE)-cadherin/catenin adhesion complex. We report that following tumour cell (MDA MB231 cells) adherence to the HUVECs, there was a rapid (within 5 min) redistribution of VE-cadherin, resulting in its transient loss from regions of endothelial cell-cell contact. The molecule gradually reorganised within the endothelial cell contacts after this time. It was further shown that the overall expression of VE-cadherin did not change, however, the amount of alpha- and beta-catenins coprecipitated with VE-cadherin markedly decreased after 5 min of tumour cell adhesion to the HUVECs. Immunoprobing of these samples with anti-phosphotyrosine antibodies demonstrated that the tyrosine phosphorylation of VE-cadherin was significantly increased following 5 min of tumour cell adhesion. Together, these results suggest that the adhesion of tumour cells to HUVEC promotes the redistribution of VE-cadherin from interendothelial adherens junctions, an effect that may be attributed to the increase in tyrosine phosphorylation of members of the VE-cadherin/catenin adhesion complex. This, in turn, may increase vascular endothelial permeability and facilitate the transendothelial migration of tumour cells during extravasation.     

Alkali-Treated LPS of Yersinia enterocolitica does not Induce Expression of E-Selectin, ICAM-1 or VCAM-1 on Endothelial Cells but may Mediate Antibody- and Complement-Dependent Cell Injury

Lipopolysaccharide (LPS) prepared from a rough mutant of Salmonella typhimurium and deacylated enzymatically (dLPS) does not promote neutrophil adherence to human umbilical vein endothelial cells (HUVECs). This paper reports that similarly, a smooth form of LPS prepared from Yersinia enlerocolitica O:3, a serotype known to trigger reactive arthritis in humans, and treated with alkali (yersinia LPS-OH) failed to augment neutrophil adherence to HUVECs. Studies of the mechanism underlying the poor augmentation revealed that neither enzymatically deacylated LPS from Escherichia coli J5 (J5 dLPS) nor yersinia LPS-OH stimulated expression of endothelial cell adhesion molecules E-selectin, VCAM-1 and ICAM-1, whereas both Intact J5 LPS and yersinia LPS were stimulatory. Impaired up-regulation could not be explained by decreased binding of yersinia LPS-OH to HUVECs. Furthermore, ⁵¹Cr-labelled HUVECs treated with different concentrations of yersinia LPS-OH released ⁵¹Cr in the presence of anti-yersinia anti-0 antibody and complement. J5 dLPS and yersinia LPS-OH inhibited up-regulation of the adhesion molecules induced by J5 LPS and yersinia LPS but not that induced by tumour necrosis factor alpha.
Taken together, the results suggest that although yersinia LPS-OH can depress development of acute inflammation by inhibiting up-regulation of etidothelial-cell adhesion molecules, sufiicient LPS-OH is bound to induce cell injury and thereby inflammation in the prescence of specific antibody and complement. The findings may have pathogenetic implications in yersinia-triggered reactive arthritis characterized by dissemination of yersinia LPS throughout the body.     

人脐静脉内皮细胞与表面改性的PHBHHx膜的体外相容性

目的 评价人脐静脉内皮细胞（HUVECs）在含有不同组分中药涂层、碱处理前后的3-羟基丁酸和3-羟基已酸共聚酯（PHBHHx）膜上的生长状况,为研究PHBHHx膜与内皮细胞的生物相容性提供实验依据。 方法 用Baudine改良法分离培养人脐静脉内皮细胞,并用免疫组织化学方法鉴定;将已传至第3代人脐静脉内皮细胞接种于材料表面,培养8、12、24h,用扫描电镜观察人脐静脉内皮细胞在不同表面上黏附形态的变化,同时用细胞免疫荧光标记法比较其在不同膜上的分布情况,最后用四甲基偶氮唑盐(MTT)法检测各组细胞的活力。 结果 成功分离出人脐静脉内皮细胞,Ⅷ 因子及血管内皮细胞受体Ⅱ(FLK-1)染色阳性;人脐静脉内皮细胞培养结果表明,细胞在各组材料表面均呈良好生长,并大量增殖。MTT分析结果显示,碱处理的PHBHHx膜及添加5%、10%中药涂层的材料对人脐静脉内皮细胞的体外生长、增殖有促进作用;扫描电镜和荧光显微镜下可见,碱处理的PHBHHx膜及添加5%、10%中药涂层的材料表面的人脐静脉内皮细胞活力旺盛、形态饱满、分布均匀。 结论 经表面改性处理的、且加入5%、10%中药涂层的PHBHHx膜,具有良好的人脐静脉内皮细胞相容性,在体外细胞培养的环境下,有利于细胞的生长、贴附和增殖。