Egfl7 knockdown causes defects in the extension and junctional arrangements of endothelial cells during zebrafish vasculogenesis.

The endothelial cell (EC) -specific secreted protein EGFL7 is important for tubulogenesis in newly forming blood vessels. We studied its role in vascular tube formation by a quantitative ultrastructural analysis of Egfl7-knockdown zebrafish embryos. At 24 hours postfertilization, the endothelia of dorsal aorta (DA) and posterior cardinal vein (PCV) were correctly anchored to the hypochord and endoderm, respectively, but failed to expand into the vascular area. This resulted in vessels with reduced or split lumen and open sheets of ECs. Concomitantly, the organization of hematopoietic cells-identified by the presence of previously undescribed membrane tubules-between DA and PCV, and within the vessels, was severely disturbed. Strikingly, ectopic cell junctions occurred across the obstructed vessel lumen, on the luminal EC surfaces, which in control conditions never display junctions of any kind. These data suggest that Egfl7 provides ECs with a cue for their extension into the vascular area and in establishing EC cell polarity.     

Plasminogen activator inhibitor-1 as a measure of vascular remodelling in breast cancer

The generation of urokinase plasminogen activator (uPA) by tumours is an important pathway for neoplastic cell invasion and metastasis. Indeed in several tumour types, elevated levels of uPA, its receptor (uPAR) or its inhibitor plasminogen activator inhibitor-1 (PAI-1) is associated with a poorer prognosis. Since endothelial cells also use this proteolytic system to remodel the extracellular matrix during angiogenesis and since angiogenesis, as assessed by microvessel density, is also a predictor of patient survival, this study was designed to investigate the relationship between angiogenesis and the urokinase system in breast tumours. The aims were to assess whether the uPA, uPAR and/or PAI-1 correlates with angiogenic activity and could therefore be a useful objective clinical measure of tumour neovascularization; and to clarify whether the poor outcome associated with high levels of the urokinase system is due to its association with angiogenesis. The study also sought to examine the relationship between the uPA system and vessel remodelling using loss of a basement membrane epitope (LH39) normally associated with established capillaries. The cytosolic levels of uPA, PAI-1 and uPAR were therefore measured by enzyme linked immunoabsorbent assay, together with tumour vascularity, in 136 well-characterized invasive breast carcinomas. There were significant relationships between uPA and uPAR (Spearman r=0.37, p<0.0001), uPA and PAI-1 (Spearman r=0.19, p=0.03) and between uPAR and PAI-1 (Spearman r=0.23 p=0.01). A significant correlation was also observed between PAI-1 and vessel remodelling (Spearman r=0.34, p=0.04), patient age (p=0.01), nodal status (p=0.047) and tumour grade (p=0.04), but no association between tumour vascularity and PAI (p=0.96), uPA (p=0.69) or uPAR (p=0.81) was present. No significant association was seen between any of the urokinase variables and expression of the angiogenic factor thymidine phosphorylase. Furthermore, no significant associations were found between any of the studied parameters and overall survival in a univariate analysis of the cancer patients. A multivariate Cox proportional hazard model of overall survival showed that uPA (p=0.15), but not uPAR (p=0.52) or PAI-1 (p=0.61), gave no additional prognostic information. These findings show that uPA may work via an independent pathway to angiogenesis and therefore combined blockade of uPA and angiogenesis may have additional therapeutic benefits. It also shows, as recently demonstrated in animal models, that PAI-1 may be a key regulator of vascular remodelling in human cancer.     

基于“阴火”理论探讨内皮细胞代谢重塑与肿瘤血管化关系及中医药干预思考

血管内壁的内皮细胞借助调节血管稳态、血管张力、细胞黏附、细胞增殖、凝血抗性、炎症因子等来响应机体的物理和化学信号,进而维持血管处于衡动稳定的状态。血管生成是肿瘤发生发展所需的关键条件,肿瘤血管化的病理模式为瘤体生长提供所需的营养和氧气,以促进其增殖。近年来,内皮细胞参与肿瘤血管渗入,驱动血管生成被认为是促进肿瘤转移的关键环节。血管内皮细胞可通过调控代谢重塑以满足肿瘤血管化过程中对物质和能量的需要,其异常的代谢特征使得其能更好地适应肿瘤微环境的变化,这种代谢的改变常被看作是肿瘤血管化进程的重要基础。中医“阴火”理论源自《黄帝内经》,原指“阴虚生内热”,属内伤之火范畴,后经历代医家推演变化,逐步形成“脾胃气虚-阴火上乘-气火失调”的病机认识。内皮细胞代谢重塑的发病过程是“阴火”病理表征的客观体现,而阴火造化生息、燔灼妄动之性又与肿瘤新生血管增生无制、亢变为害的疾病状态相吻合。脾胃亏虚与气机失衡的改变可致水谷精微不得布散,发生代谢障碍,停久为痰为瘀,与血脉相搏结,致使局部环境异化（肿瘤微环境形成）、脉道不利（内皮细胞代谢失衡）,异变形成肿瘤新生血管。在“阴火”证候要素指导下,围绕气与火的相关性,运用“扶正调气”的治法遣方用药,可调节内皮细胞代谢功能,达到机体相对平衡状态,进而抑制肿瘤血管新生。基于以上认识,该文围绕内皮细胞代谢重塑与阴火相关理论,阐发学术微旨,以期为中医药干预肿瘤血管化进程提供一定的理论支持。     

微血管定量和血管内皮生长因子表达在非小细胞肺癌中的意义

目的 :研究非小细胞肺癌 (NSCL C)的预后与血管内皮生长因子 (VEGF)表达及肿瘤微血管定量间的关系。方法 :对81例行手术治疗的 NSCL C患者运用免疫组织化学的方法检测 VEGF的表达及微血管定量 ,检测 VEGF使用抗 VEGF单克隆抗体 ;检测微血管定量用抗 CD34 单克隆抗体。结果 :在术后半年出现转移的 32例 NSCL C中微血管定量为 (32± 15 .4)根/ mm3比术后未出现转移的 49例患者 (12± 8.6 )根 / mm3明显增高 ,经统计学处理 (P=0 .0 0 1)。在术后半年出现转移的 32例中有 2 4例 VEGF表达阳性 (75 % ) ,在术后未出现转移的 49例中有 2 5例 VEGF表达阳性 (5 1% ) ,经统计学处理 (P=0 .0 31)。结论 :NSCL C的转移形式为血管形成依赖性 ,VEGF是 NSCL C的血管形成的一个重要因素。VEGF的表达是检测NSCL C预后的一个有意义的指标     

血管内皮生长因子在乳腺癌中的表达及其与腋窝淋巴结转移的关系

目的：探讨血管内皮生长因子（ＶＥＧＦ）和微血管密度（ＭＶＤ）与乳腺浸润性导管癌腋窝淋巴结转移的关系。方法：采用ＬＳＡＢ法对６５例浸润性导管癌术后根治标本,进行ＶＥＧＦ的表达和微血管密度的测定。结果：６５例乳腺癌的 ＭＶＤ为（４７．６±２２．４）个,ＶＥＧＦ总阳性率为 ８０．００％;ＶＥＧＦ、ＭＶＤ与淋巴结转移有关,在腋窝淋巴结转移的乳腺癌组织中,ＶＥＧＦ的阳性率 ８８．８９％（３２／３６）,ＭＶＤ（５６．６±２０．５）,均明显高于无腋窝淋巴结转移者ＶＥＧＦ的阳性率 ６８．９６％（２０／２９）,ＭＶＤ（３１．３±１８．９）．差异有显著性（Ｐ＜０．０５）;ＶＥＧＦ和 ＭＶＤ在乳腺浸润性导管癌中的表达具有显著的正相关（ｒ＝８.２１３,Ｐ＜０．０５）。结论：ＶＥＧＦ与乳腺癌血管生成密切相关;ＶＥＧＦ表达的增高及 ＭＶＤ的增加对乳腺癌腋窝淋巴结转移可能有促进作用。     

重组血管生成抑制因子r—K4K5的分离纯化与功能鉴定

目的 分离纯化r-K4K5,探讨r-K4K5对牛毛血管内皮（BCE）细胞、鸡胚绒毛尿囊膜（CAM）新生血管生成及实验性人肺腺癌SPC-A1生长的抑制作用。方法 通过盐析、凝胶过滤提纯r-K4K5,BCE细胞在含r-K4K5的DMEM中培养24、48、72h后分别计数;孵化7d的鸡胚加r-K4K5后继续孵育72h,观察新生血管生成;已经接种入SPC-A1肺腺癌组织的荷瘤裸小鼠（Balb/c,nu/nu）,瘤旁注射r-K4K5继续饲养,观察肿瘤生长变化。结果 r-K4K5抑制BCE细胞增殖,48～72h作用明显;r-K4K5处理的CAM组中直径小于50um的小血管明显减少;高剂量r-K4K5治疗的荷瘤裸小鼠组,平均瘤重与对照组比较有统计学意义。结论 r-K4K5能够抑制BCE细胞增殖,抑制鸡胚CAM新生血管生成,抑制实验性人SPC-A1肺腺瘤生长。     

血管内皮细胞生长因子及其受体在大肠癌组织中的表达

目的观察血管内皮生长因子（ＶＥＧＦ）及其受体（ＫＤＲ）在大肠癌组织中的表达情况。方法利用免疫组化ＳＡＢＣ 法检测８０例大肠癌组织及１０例正常大肠组织中的ＶＥＧＦ及ＫＤＲ的表达。结果ＶＥＧＦ染色性物质主要位于肿瘤细胞 膜及胞浆,ＫＤＲ染色性物质主要位于癌组织及癌组织旁的血管内皮细胞上,大肠癌组织中ＶＥＧＦ和ＫＤＲ的表达阳性 率显著高于正常对照组织（Ｐ＜０．０１）,ＶＥＧＦ和ＫＤＲ的表达与肿瘤的分化程度及Ｄｕｋｅｓ分期密切相关（Ｐ＜０．０５）,与病理 分型无关（Ｐ＞０．０５）。结论ＶＥＧＦ和ＫＤＲ在大肠癌的发生和发展中起着重要作用,可反映大肠癌的恶性程度和进展情 况,并作为预后的指标。     

激素性股骨头坏死血管新生相关基因的鉴定及其靶向中药成分筛选验证

目的 基于生物信息学分析筛选激素性股骨头坏死（steroid induced osteonecrosis of femoral head,SONFH）中涉及血管生成相关的基因,进一步探讨其作用机制并筛选靶向中药活性成分。方法 通过Perl语言和R软件处理GEO数据,获得SONFH的关键差异基因,使用David工具进行基因本体（gene ontology,GO）和京都基因与基因组百科全书（Kyoto encyclopedia of genes and genomes,KEGG）分析。然后使用STRING平台和Cytoscape软件分析蛋白互作关系,用MCODE和CytoHubba插件识别重要基因簇模块和核心基因,并进一步验证。使用CTD数据库搜索靶向核心基因的中药活性成分并用autodock vina软件进行分子对接,以及探索核心基因与SONFH的相互作用。利用激素诱导人股骨头骨微血管内皮细胞（bone microvascular endothelial cells,BMECs）,通过CCK-8、细胞划痕实验和Transwell实验观察白藜芦醇对细胞增殖及迁移的影响,通过Western ...     

直流电场对鸡胚绒毛尿囊膜血管生成作用的研究

目的：探讨直流电场（electric fields．EF）对血管生成的影响及作用机制。方法：于发育第8天的鸡胚绒毛尿囊膜（chick embryo chorioallantoic membrane，CAM）上植入明胶海绵载体，其中加载不同强度EF，计数长入载体的血管和载体内的微血管密度（microvascular densily．MVD）。结果：150mV／mm、200mV／mm组在CAM上长入载体边缘的血管数和MVD与对照组间的有显著差别（p〈0．01）。结论：强度为150mV／mm、200mV／mm的直流电场能促进CAM的血管生成。     

Aberrant gene expression associated with recurrent pregnancy loss

Recent studies indicate that a number of factors including chromosomal abnormalities, immunological feto-maternal rejection, hormonal irregulation and anatomical factors are involved in provoking recurrent pregnancy loss (RPL). This indicates that normal cellular regulation of these factors is required for maintaining normal pregnancy. In addition, it is expected that biological processes for maintaining normal pregnancy require a series of differential gene expression. As expected, our previous investigations revealed that there are >/=30 genes showing different levels of expression between normal and RPL patients. In addition, other research groups have also identified a number of genes that are expressed aberrantly in pregnancy failure. In this review, recent study on aberrant expression levels of genes, which are grouped as immunity-related, angiogenesis-related, apoptosis-related and other groups of genes, will be discussed.