循环内皮细胞在狼疮性肾炎血管病变诊断中的应用

目的：检测狼疮性肾炎（LN）患者循环血内皮细胞（CECs），探讨其在肾脏血管病变的诊断和判断病情中的作用。方法：60例经肾活检诊断为LN患者，其中30例伴血管病变患者，30例不伴血管病变患者。正常对照为30名健康志愿者。抽取外周血，采用免疫磁珠分离方法计数CECs的数量，同时进行血清肌酐、尿蛋白和尿红细胞等临床项目检查。结果：不伴血管病变组LN患者CECs数目与正常对照组相比差异无统计学意义，伴血管病变组患者CECs数目显著高于不伴血管病变组和正常对照组（P〈0．01）。伴血管病变组患者CECs与血清肌酐水平呈正相关（r=0．513，P〈0．01）。在伴血管病变LN患者中，合并血栓性微血管病变（TMA）患者CECs数量明显高于不伴有TMA患者（P〈0．01）。结论：CECs检测不仅能反映LN患者血管病变的存在，而且还能作为判断血管病变严重程度的指标。     

Circulating tumour cells: insights into tumour heterogeneity

Tumour heterogeneity is a major barrier to cure breast cancer. It can exist between patients with different intrinsic subtypes of breast cancer or within an individual patient with breast cancer. In the latter case, heterogeneity has been observed between different metastatic sites, between metastatic sites and the original primary tumour, and even within a single tumour at either a metastatic or a primary site. Tumour heterogeneity is a function of two separate, although linked, processes. First, genetic instability is a hallmark of malignancy, and results in ‘fixed’ genetic changes that are almost certainly carried forward through progression of the cancer over time, with increasingly complex additional genetic changes in new metastases as they arise. The second type of heterogeneity is due to differential but ‘plastic’ expression of various genes important in the biology and response to various therapies. Together, these processes result in highly variable cancers with differential response, and resistance, to both targeted (e.g. endocrine or anti‐human epithelial growth receptor type 2 (HER2) agents) and nontargeted therapies (e.g. chemotherapy). Ideally, tumour heterogeneity would be monitored over time, especially in relation to therapeutic strategies. However, biopsies of metastases require invasive and costly procedures, and biopsies of multiple metastases, or serially over time, are impractical. Circulating tumour cells (CTCs) represent a potential surrogate for tissue‐based cancer and therefore might provide the opportunity to monitor serial changes in tumour biology. Recent advances have enabled accurate and reliable quantification and molecular characterization of CTCs with regard to a number of important biomarkers including oestrogen receptor alpha and HER2. Preliminary data have demonstrated that expression of these markers between CTCs in individual patients with metastatic breast cancer reflects the heterogeneity of the underlying tumours. Future studies are designed to determine the clinical utility of these novel technologies in either research or routine clinical settings.     

Circulating endothelial cells in patients with acute myeloid leukemia

OBJECTIVES: The circulating endothelial cells (CEC) are proposed to be a non-invasive marker of angiogenesis. The level of CEC in peripheral blood (PB) of acute myeloid leukemia (AML) patients has not been investigated prior to this study. We evaluated the count of resting (rCEC), activated (aCEC) and endothelial progenitor cells (CEPC) in the PB of AML and healthy subjects. In addition we correlated the levels of CEC with disease status, known prognostic factors and response to treatment. METHODS: CEC were quantified by utilizing four-color flow cytometry procedures in 48 AML patients at the time of diagnosis and 29 healthy controls. Additionally, measurements were again taken after the first course of induction treatment in 12 of the patients. RESULTS: The numbers of aCEC, rCEC and CEPC were significantly higher in the AML patients than in the controls (P < 0.0001, P < 0.0001 and P < 0.001, respectively). The CEC count was significantly higher in the AML patients with white blood cell count (WBC) >15 G/L, elevated lactic dehydrogenase (LDH) levels and a higher (over median) absolute blasts count (ABC) in PB than in the group with WBC <15 G/L (P < 0.03), a normal LDH level (P < 0.03) and a lower (相似文献   

Increased circulating activated endothelial cells,vascular endothelial growth factor,and tumor necrosis factor in thalassemia

An increased number of circulating endothelial cells (CECs) was demonstrated in alpha- and beta-thalassemic patients, beta-thalassemia/hemoglobin E (BE), both splenectomized (BE[S]) and non-splenectomized (BE[NS]), had higher numbers of CECs than alpha-thalassemia, both HbH (alpha-thal l/alpha-thal 2; H) and HbH with hemoglobin Constant Spring (alpha-thal 1/CS; H/CS). CECs were also increased in heterozygous HbE (EA) and homozygous HbE (EE). The highest level of tumor necrosis factor-alpha (TNF-alpha) was found in HbH/CS patients, whereas the highest levels of vascular endothelial growth factor (VEGF) was observed in BE[S] patients. Significant decreases, in protein C and protein S levels were found in both alpha- and beta-thalassemia compared with normal. Good correlations between the numbers of CEC and TNF-alpha, VEGF, protein C, and protein S levels were demonstrated in this study. In addition, markers for endothelial cell activation and injury (intercellular adhesion molecule-1, ICAM-1/CD54; vascular cell adhesion molecule-1, VCAM-1/CD106; and E-selectin, ELAM-1/CD62E) were detected on the surface of isolated CECs using immunofluorescence technique. Appearance of CECs with markers for endothelial cell activation, together with increased levels of TNF-alpha and VEGF and decreased levels of protein C and protein S in the circulation, may account for the propensity of vascular perturbation in thalassemic subjects.     

Surface markers: An identity card of endothelial cells

All endothelial cells have the common characteristic that they line the vessels of the blood circulatory system. However, endothelial cells display a large degree of heterogeneity in the function of their location in the vascular tree. In this article, we have summarized the expression patterns of a number of well‐accepted endothelial surface markers present in normal microvascular endothelial cells, arterial and venous endothelial cells, lymphatic endothelial cells, tumor endothelial cells, and endothelial precursor cells.     

黑龙江立克次体感染血管内皮细胞的初步研究

目的 通过黑龙江立克次体感染体外培养人脐静脉内皮细胞（human umbilical vein endothelial cells, HUVEC）探讨其在内皮细胞内的生长规律。方法 将泛影葡胺密度梯度超速离心纯化的黑龙江立克次体（HLJ-054株）感染体外培养的HUVEC,通过间接免疫荧光和扫描电镜检测与观察不同时相黑龙江立克次体在内皮细胞内的生长状况。热灭活黑龙江立克次体做平行对照。 结果 立克次体粘附并侵入内皮细胞,在感染后第6h及第24h分别为感染高峰;感染5 d后细胞内立克次体逐渐增殖,第8～9d细胞内立克次体急剧增殖,并见细胞核内有少量立克次体,细胞出现病变,第12d细胞内充满立克次体,大部分细胞皱缩脱落。结论 黑龙江立克次体能够感染血管内皮细胞,在血管内皮细胞内不断增殖而使细胞死亡。     

小干扰RNA对大鼠血管内皮细胞ICAM-1表达的抑制作用

目的:探讨小干扰RNA对大鼠血管内皮细胞细胞间黏附分子-1(ICAM-1)表达的抑制作用.方法:使用 ICAM-1siRNA及错配siRNA转染肿瘤坏死因子-α刺激过的大鼠血管内皮细胞,聚合酶链反应和蛋白印迹检测转染后48 h ICAM-1 mRNA和蛋白表达水平.结果:与空白对照组比较,大鼠血管内皮细胞在转染48 h后ICAM-1 mRNA表达减少90.0％,蛋白表达减少83.0％.结论:使用小干扰RNA能有效抑制大鼠血管内皮细胞ICAM-1表达.     

The mechanisms of hepatic sinusoidal endothelial cell regeneration: A possible communication system associated with vascular endothelial growth factor in liver cells

Vascular endothelial growth factor (VEGF) has been shown to induce proliferation of sinusoidal endothelial cells in primary culture. To elucidate the mechanisms of sinusoidal endothelial cell regeneration in vivo, mRNA expression of VEGF and its receptors, flt-1 and KDR/flk-1, were studied in rat livers. Northern blot analysis revealed that VEGF-mRNA was expressed in hepatocytes immediately after isolation from normal rats. In contrast, non-parenchymal cells, including sinusoidal endothelial cells, expressed VEGF receptor-mRNA. Vascular endothelial growth factor-mRNA expression in hepatocytes was decreased during primary culture, but increased following a peak of DNA synthesis, induced by addition of epidermal growth factor or hepatocyte growth factor to the culture medium at 24 h of plating. In a 70% resected rat liver, VEGF-mRNA expression increased with a peak at 72 h after the operation, and mRNA expression of VEGF receptors between 72 and 168 h. In such a liver, mitosis was maximal in hepatocytes at 36 h and in sinusoidal endothelial cells at 96 h. Also, mRNA expression of both VEGF and its receptors was significantly increased in carbon tetrachloride-intoxicated rat liver compared with normal rat liver. Vascular endothelial growth factor expression was minimal in Kupffer cells isolated from normal rats, but marked in activated Kupffer cells and hepatic macrophages from the intoxicated rats. Vascular endothelial growth factor-mRNA expression was also increased in activated stellate cells from these rats and in the cells activated during primary culture compared with quiescent cells. We conclude that increased levels of VEGF expression in regenerating hepatocytes may contribute to the proliferation of sinusoidal endothelial cells in partially resected rat liver, probably through VEGF receptors up-regulated on the cells. Also, VEGF derived from activated Kupffer cells, hepatic macrophages and stellate cells may be involved in this proliferation in injured rat liver.     

A prospective pilot study of circulating endothelial cells as a potential new biomarker in portal hypertension

Background/Aims: Peripheral circulating endothelial cells (CEC) have been proposed as a prognostic marker in cardiovascular diseases. Cirrhosis and portal hypertension are associated with vascular injury yet little is known about CEC count in these conditions. Therefore, we evaluated CEC count in patients with cirrhosis, and correlated it with markers of portal hypertension/disease severity. Patients/Methods: Fifteen patients with cirrhosis/portal hypertension and 15 matched controls were prospectively recruited for study participation. An automated rare cell analysis system was used to enumerate CEC from peripheral blood and correlated with clinical features. Results: Median CEC levels were significantly higher in patients with cirrhosis as compared with controls (median [interquartile range (IQR)]; cirrhosis: 73.7 cells/4 ml [53.7–140.3]; controls: 28.7 cells/4 ml [21–58.7]; P=0.021). Ratio of CEC to platelet count (CEC/PC) also distinguished patients with cirrhosis from controls (IQR; cirrhosis: 0.723 [0.396–1.672]; controls: 0.126 [0.103–0.333]; P<0.001). Receiver operator characteristic analysis revealed that CEC cut‐off of 42 cells/4 ml showed sensitivity of 87% and specificity of 74% for differentiating cirrhosis from controls (AUC: 0.74), while CEC/PC ratio at 0.21 showed sensitivity of 100% and specificity of 73% (AUC: 0.89). Furthermore, CEC/PC index was significantly elevated in patients with hepatic decompensation as defined by Child B/C (P<0.05). The intra‐ and interobserver variability correlation coefficients for CEC measurement were 0.9989 and 0.9986 respectively. Conclusion: Median CEC count and CEC/PC ratio are significantly elevated in patients with cirrhosis, with CEC/PC also increased in patients with decompensated cirrhosis. These data provide rationale for larger validation studies to assess if CEC may have prognostic utility in patients with cirrhosis and portal hypertension.     

Increased circulating endothelial cells in sickle cell crisis

To determine whether increased numbers of circulating endothelial cells, a possible indicator of endothelial injury, are present in subjects with sickle cell disease, we measured circulating endothelial cells in 30 normal subjects and in 23 subjects with sickle cell anemia. Mean circulating endothelial cells were significantly higher (P less than 0.025) in the sickle cell subjects than in the normal subjects. Circulating endothelial cells were significantly higher than normal in 10 sickle cell subjects studied during painful crisis (P less than 0.01) but not in 13 sickle cell subjects studied while in the steady state. To control for the known stimulatory effect of cigarette smoking on circulating endothelial cells, we analyzed the results for smokers and nonsmokers separately. Mean circulating endothelial cells were not significantly higher in sickle cell subjects who smoked (n = 10) than in normal subjects who smoked (n = 8), but were significantly higher (P less than 0.05) in sickle cell nonsmokers (n = 13) than in normal nonsmokers (n = 22). Among nonsmoking sickle cell subjects, mean circulating endothelial cells were significantly higher than normal (P less than 0.01) during painful crisis (n = 7), but not in the steady state (n = 6). We conclude that circulating endothelial cells are significantly increased in sickle cell crisis, and may indicate the occurrence of acute endothelial injury during episodes of microvascular occlusion.     

Circulating endothelial (progenitor) cells reflect the state of the endothelium: vascular injury, repair and neovascularization

An increase in the number of circulating endothelial cells (CEC) and of bone marrow-derived endothelial progenitor cells (EPC) in the peripheral blood is associated with vascular injury, repair and neovascularization. The phenotype and number of CEC may serve as diagnostic or prognostic parameters of vascular injury and tumour growth. An increase in the number of EPC may reflect repair of ischaemic vascular injury, a finding which has resulted in the initiation of clinical cardiovascular pilot trials using cell therapy. However, there is no consensus on the exact phenotype of the EPC and haematopoietic stem cells (HSC) and therefore the best candidate cell for transplant has not been established. Although the use of peripheral blood stem cells following mobilization, or of ex vivo-expanded cells, may improve EPC-mediated vascular graft endothelialization or tissue vascularization, sustained EPC-induced neovascularization still needs to be proven. Flow cytometric characterization, in combination with functional assays, will further elucidate the phenotype of the CEC and EPC, thereby providing reliable detection to appreciate their role in vascular diseases and cancer and to evaluate and, if possible, improve their therapeutic potential.     

Increased circulating endothelial cells in acute heart failure: comparison with von Willebrand factor and soluble E-selectin

BACKGROUND: Circulating endothelial cells (CECs) in the peripheral blood, probably representing the most direct evidence of endothelial cell damage, are increased in myocardial infarction, unstable angina and critical limb ischaemia. As chronic heart failure is also associated with endothelial abnormalities, we hypothesised that CECs are raised in acute heart failure and that they would correlate with plasma indices of endothelial perturbation, that is, von Willebrand factor (vWf) and soluble E-selectin. METHODS: We studied 30 patients with acute heart failure (venesected within 24 h of emergency hospital admission), 30 patients with chronic stable heart failure (venesected as out-patients, all patients in sinus rhythm with ejection fraction < or = 40%) and 20 healthy controls. CECs were quantified using epifluorescence microscopy after CD146-immunomagnetic separation and phenotyped by streptavidin/biotin immunocytochemistry. Citrated plasma was analysed for soluble E-selectin and vWf by ELISA. RESULTS: Levels of CECs, vWf and soluble E-selectin were significantly higher (all p<0.01) in patients with heart failure compared to controls, with no significant differences between acute and chronic heart failure. CECs correlated with plasma vWf (p<0.0001) and soluble E-selectin (p = 0.022) but not ejection fraction or NYHA class. In multiple regression analysis, heart failure was the only independent predictor of raised CECs (p<0.0001). Immunoperoxidase-defined surface expression of CD34, CD45 and CD36 by CECs was <2%, 0% and 8%, respectively. CONCLUSION: CECs, a possibly heterologous population, may be used as a novel measure of endothelial damage in acute heart failure and may have implications for the thrombotic risk associated with acute and chronic heart failure and prognosis in this condition.     

Plasma microparticles and vascular disorders

Microparticles are circulating, phospholipid rich, submicron particles released from the membranes of endothelial cells, platelets, leucocytes and erythrocytes. Investigation into their biological activity has revealed diverse actions in coagulation, cell signalling and cellular interactions. These actions are mediated through their phospholipid rich surfaces and the expression of cell surface molecules which reflect their cell of origin and its state of activation. Microparticle numbers are reported to be elevated in a number of conditions where vascular dysfunction and inflammation are important pathophysiological mechanisms, for example coronary artery disease or thrombotic microangiopathies. Currently, there are a variety of different methods used for the quantitation of circulating microparticles; however with standardisation their assessment may prove to be of clinical value, reflecting the state of the vasculature. Knowledge of the functional properties of microparticles will contribute to our understanding of the mechanisms underlying vascular dysfunction and prothrombotic states.     

缺氧对血管内皮细胞内Ca2+的影响以及合贝爽的保护作用

目的观察缺氧对人大血管内皮细胞内Ca2 水平的影响以及合贝爽的保护作用。方法将培养的血管内皮细胞分为对照组、缺氧组和治疗组(加入合贝爽再缺氧),应用激光共聚焦显微镜测量各组内皮细胞内Ca2 水平。结果与对照组及治疗组比较,缺氧组细胞内Ca2 水平显著升高(P<0.01),治疗组与对照组比较无统计学差异(P>0.05)。结论缺氧可引起血管内皮细胞内钙超载,合贝爽对其具有保护作用。     

睾酮对血管内皮细胞表达血栓调节蛋白的影响

目的　观察睾酮对内皮细胞表达血栓调节蛋白的作用。方法　用不同浓度的睾酮 (2、6、10、10 0、30 0、4 0 0μg L)处理传代培养内皮细胞株ECV 30 4细胞 2 4h后 ,用四唑盐比色试验检测细胞活力 ;用酶联免疫吸附法检测上层培养液中的可溶性血栓调节蛋白及细胞总血栓调节蛋白的含量。结果　 2、6、10 μg L的睾酮均促进细胞表达血栓调节蛋白 (2 .89± 0 .96vs1.77± 0 .13,3.0 8± 0 .77vs1.77± 0 .13,2 .72± 0 .4 5vs1.77± 0 .13) ,其中以 6 μg L的睾酮作用最大 ,高浓度的睾酮对血栓调节蛋白的表达没有作用 ,不同浓度的睾酮对可溶性血栓调节蛋白的分泌均没有作用。结论　睾酮促进细胞血栓调节蛋白表达增加 ,可增强细胞的抗凝能力。     

血管内皮细胞衰老与心血管疾病的相关性

血管内皮细胞(VEC)是覆盖于血管内膜表面的单层扁平鳞状上皮细胞,其构成血管壁的生物屏障,不仅属于一种保护性屏障,还能够产生一些自体分泌物用于调节体内平衡和血管紧张度。VEC衰老可导致血管功能受损,是心血管系统(CVS)主要的危险因素,并与心血管疾病(CVD)有着密切的关系。然而,VEC衰老的机制以及VEC衰老对血管功能的影响尚不完全清楚。本综述总结了VEC衰老的特征及其相关分子机制,并对年龄相关CVD进行了阐述。     

低氧对培养心肌细胞表达和分泌血管内皮生长因子的影响

目的　探讨低氧对心肌细胞血管内皮生长因子 (VEGF)表达的影响。方法　采用Kuwabara等并加以改进的方法对心肌细胞进行低氧培养 ,并于低氧培养的即刻、2 4、4 8h分别测定培养液中的氧分压。使用逆转录聚合酶链反应 (RT PCR)、免疫组织化学和酶联免疫吸附测定 (ELISA)法分别检测低氧培养的不同时间点VEGFmRNA及VEGF蛋白的表达和分泌。结果　与常规培养相比 ,简易低氧培养培养液氧分压降至 5 8mmHg ,并于 3个时间点保持这一水平 (P >0 .0 5 )。从mRNA水平和蛋白水平我们检测到VEGF表达的增加 ,于低氧培养 2 4h达峰值 ,培养液中的峰值浓度为 (72 4 .6 7± 5 6 .87)pg ml。结论　低氧促进心肌细胞表达VEGFmRNA、VEGF蛋白和分泌该蛋白     

老年高血压病并下肢动脉硬化症的血管内皮功能

目的了解老年高血压病及高血压病合并下肢动脉硬化症(LEASD)患者内皮依赖性舒张功能(FMD)、非内皮依赖性舒张功能(NMD)以及血浆一氧化氮(NO)、内皮素(ET)的水平。方法采用高分辨率超声诊断系统分别检测36例老年高血压病(老年高血压组)患者以及49例老年高血压病合并LEASD(老年高血压合并LEASD组)患者肱动脉的FMD及NMD;同时采用硝酸盐镉还原、比色法测定NO水平(以硝酸盐浓度表示);采用放射免疫法检测ET水平,并分别与40例健康老人(健康老年组)进行对照研究。结果老年高血压合并LEASD组患者肱动脉的FMD及NMD均显著低于老年高血压组和健康老年组(P<0.05),而老年高血压组患者的FMD和NMD亦显著低于健康老年组(P<0.05);老年高血压合并LEASD组患者的NO水平显著低于老年高血压组和健康老年组,而ET却显著高于老年高血压组和健康老年组,而老年高血压组患者NO水平同样显著低于健康老年组,ET水平显著高于健康老年组(P<0.05)。结论老年高血压病及高血压病合并LEASD患者肱动脉FMD及NMD均受损;老年高血压病及高血压病合并LEASD患者均存在血管内皮功能失调,且高血压病合并LEASD患者内皮功能紊乱更严重。