隐丹参酮对鸡胚尿囊膜血管生成的抑制作用

目的评价隐丹参酮对鸡胚尿囊膜（CAM）新生血管生成的抑制作用。方法在六日龄CAM上植入含隐丹参酮的甲基纤维素膜片并孵化48h。解剖显微镜下计数加药组和对照组局部血管数，用抑制率反映药物对CAM新生血管形成的影响。结果隐丹参酮0．1μg／egg和0．2μg／egg剂量相关性减少CAM局部的血管数，抑制率分别达到65％和72％；与对照组相比用药组局部血管明显变细受损。结论隐丹参酮对CAM血管生成有显著的抑制作用。     

微创内固定技术治疗跟骨骨折的基础研究

目的：测量跟骨轴长、宽度和高度，探讨螺钉入路及固定点，为微创内固定技术治疗跟骨骨折治疗提供解剖学依据。方法：260份无跟骨损伤的轴位及侧位X线片和120份无跟骨损伤的CT片，在X线片及CT上做其纵轴长度为跟骨轴长L，取其中点做跟骨纵轴的垂线为宽度W。在跟骨侧位片上，从跟骨丘部顶点向下做垂线，于跟骨下缘相交的距离为跟骨丘部高度H。成人跟骨标本60个，测量跟骨轴长、宽度、高度；跟骨结节入钉点区域及该部位入钉固定跟骨前部、后关节面、载距突。结果：X线片跟骨测量跟骨轴长平均为（68．02±8．24）mm，宽度平均为（30．86±6．26）mm，高度平均为（40．66±7．48）mm；CT片跟骨测量跟骨轴长平均为（67．82±6．38）mm，宽度平均为（30．16±4．62）mm，高度平均为（40．02±3．34）mm；跟骨标本测量跟骨轴长平均为（69．12±7．32）mm，宽度平均为（32．18±5．34）mm，高度平均为（42．02±6．62）mm；各数值均为非正态分布，因此未进行均数的显著性检验。跟骨各数值之比：宽度／轴长为0．45，高度／轴长为0．60，宽度／高度为0．76。跟骨结节外侧区域入钉可以牢固地与跟骨前部、后关节面及载距突固定为一体。结论：跟骨宽度、高度与轴长的测量可为跟骨骨折治疗及手术器械研制提供解剖学参数，通过跟骨结节外侧区域入钉可以固定跟骨前部、后关节面、载距突为一体，可有效维持跟骨的外形。     

VEGF、ANG-1、ANG-2、TSP-1的表达与胆管细胞性肝癌血管生成和侵润转移的关系

目的：研究血管内皮生长因子（VEGF）、血管生成素-1（ANG-1）、血管生成素-2（ANG-2）、血小板反应蛋白-1（TSP-1）的表达与胆管细胞性肝癌(CCC)血管生成和侵润转移的关系。方法： 对33例手术切除的CCC标本进行CD34、VEGF、 ANG-1、 ANG-2 和TSP-1的免疫组化染色，研究VEGF、ANG-1、ANG-2、TSP-1的表达与胆管细胞性肝癌血管生成和肿瘤门静脉侵犯、肝内转移、淋巴结转移以及肿瘤分化水平之间的关系。 结果： 本组CCC的微血管密度（MVD）为(87.2±52.6)/mm2，VEGF、ANG-1、ANG-2 和TSP-1的阳性率分别为75.6%、36.0%、57.6%和45.5%。VEGF和ANG-2的阳性表达与高MVD相关，TSP-1则与MVD负相关（P<0.01,P<0.05,P<0.01）。阳性TSP-1与肝内转移正相关（46.7% vs 5.6%，P<0.05）。结论： CCC瘤内的血管新生活跃，VEGF和ANG-2的阳性表达与CCC血管生成正相关，TSP-1则与其负相关，TSP-1的阳性表达还与肝内转移相关，VEGF、ANG-1、ANG-2的表达与肿瘤的侵润转移未见显著相关。     

克拉霉素抑制肺癌细胞诱导血管内皮细胞迁移的实验研究

目的 观察克拉霉素（CAM）对人小细胞癌细胞表达VEGF其诱导血管内皮细胞迁移的影响，探讨CAM抗血管生成的机制。方法 采用免疫组化和图象分析技术，观察不同浓度的CAM作用下人小细胞肺癌细胞（NCI－H446）中VEGF蛋白表达的变化。采用共培养法（Marigel invasion chamber)，观察CAM为NCI－H446细胞诱导人血管内皮细胞（ECV－304）迁移的抑制作用。结果 CAM达到30mmol/L对其诱导的ECV－304细胞迁移表现出明显的抑制作用，达到40mmol/L可以抑制CI－H446细胞表达VEGH，CAM浓芳在30，40和50mmol/L时，抑制率分别为19．7％，24．3％和25．0％，呈现出明显剂量－反应关系（r=-0.764,P=0.001)。结论 CAM能够抑制肺癌细胞诱导的血管内皮细胞迁移，对其表达VEGF也具有抑制作用，以上作用可能是CAM抗血管生成的机制之一。     

灵芝多糖抑制鸡胚尿囊膜模型中的血管生成及细胞黏附

目的 进一步了解灵芝多糖抗肿瘤的作用机制。方法 应用鸡胚尿囊膜（Chicken chorioallantoic membrane,CAM）血管生成模型观察灵芝多糖对血管生成的作用,及对高转移潜能的人前列腺癌PC-3M-1E8细胞黏附于基质成分层黏连蛋白（Laminin,LN）的影响。结果 灵芝多糖在一定剂量范围内（0．2～5μg/鸡胚）可明显抑制CAM的血管生成及抑制细胞黏附,在0．33～33g／L剂量范围内,呈现出量一效依赖关系。结论 灵芝多糖的抗肿瘤作用与其抑制血管生成及细胞黏附有关。     

抗血管生成在胃癌治疗中的研究进展

胃癌的生长、转移依赖于肿瘤血管生成。当血管生成开关打开,肿瘤内有足够的新生血管长入,提供营养并带走代谢产物,肿瘤便获得了进一步生长、转移的能力。已知包括血管内皮细胞生长因子、整合素、基质金属蛋白酶和表皮生长因子受体在内的20多种细胞因子与胃癌微血管密度（microvessel density, MVD）相关,是胃癌血管靶向治疗的有效靶点。     

人脑星形细胞肿瘤IL-8的表达及其与血管生成的关系

目的：探讨IL-8,VEGF在脑胶质瘤中的表达及与血管生成的关系。方法：应用由45例人脑星形细胞肿瘤和6例正常脑组织组成的组织芯片，采用免疫组织化学技术分别进行IL-8，VEGF，CD34标记并进行半定量，观测在不同病理分级胶质瘤中的表达及与微血管密度（MVD）之间的关系。结果：6例正常脑组织中不表达IL-8，VEGF。45例脑胶质瘤中，27例IL-8呈阳性表达，32例VEGF呈阳性表达，Ⅲ，Ⅳ级脑胶质瘤中IL-8，VEGF的表达比Ⅰ级、Ⅱ级明显增强，IL-8，VEGF评分为强阳性的胶质瘤内MVD明显高于评分为阴性和阳性的MVD（P<0.01）。IL-8表达与MVD呈正相关（rs=0.64，P<0.01）。VEGF表达与MVD之间呈正相关（rs=0.44，P<0.01）。IL-8表达与VEGF表达之间亦呈正相关（rs=0.56，P<0.01）。结论：IL-8，VEGF的表达与胶质瘤病理分级、MVD密切相关，二者在胶质瘤的血管生成中可能相互关联、共同调节肿瘤血管生成。     

X线填充剂新载体羧甲基纤维素最佳配比的实验研究

目的：研究一种适宜于标本管道灌注后X线摄影和CT扫描三维成像的新型X线填充剂载体。方法：（1）6％、8％、10％、和12％羧甲基纤维素水溶液和4种不同比例的氧化铅／水：50g／L，100g／L，150g／L，200g／L，按正交设计调配成16种氧化铅／羧甲基纤维素水溶液配比悬浮液，做成20ml棒状铸件封装行CT扫描，以获得最佳配比。（2）使用最佳氧化铅／羧甲基纤维素配比悬浮液，进行SD大鼠全身血管灌注后，摄X线片和CT扫描三维成像。结果：SD大鼠X线摄影血管清晰，填充良好，光滑连续；小血管显示良好。血管三维图像主干及大分支饱满清晰，立体感强；边缘连续平滑，无齿状伪影；小血管显示三级以上。结论：最佳氧化铅／羧甲基纤维素配比：羧甲基纤维素的水溶液浓度为12％、氧化铅／羧甲基纤维素的配比为200g／L，羧甲基纤维素／氧化铅水凝胶可作为一种适宜于标本管道灌注后X线摄影和CT扫描三维成像的新型X线填充剂。     

婴幼儿桡动脉穿刺的应用解剖学研究

目的：积累婴幼儿桡血管的解剖学资料，为临床婴幼儿桡动脉穿刺提供形态学基础。方法：在20例40侧婴幼儿尸体上肢防腐标本上和32例64侧婴幼儿活体上肢上，分别解剖和超声观测桡动脉、桡静脉和桡神经浅支的位置关系及血管的内径。结果：（1）桡动脉两侧有伴行桡静脉，桡神经浅支偏向桡动脉的外侧；超声诊断仪显示桡动脉、桡静脉无重叠现象。（2）桡动脉在前臂前区下1／3部变浅，可供穿刺的桡动脉暴露长度为（50．6±5．1）mm。（3）桡动脉、桡静脉的内径18～24月龄及25～36月龄分别为（1．43±0．08）mm、（1．52±0．07）mm及（0．78±0．07）mm、（0．87±0．12）mm。（4）皮肤表面距桡动脉前壁的深度18～24月龄及25～36月龄分别为（5．21±0．19）mm及（5．54±0．19）mm。结论：穿刺部位宜选在桡动脉暴露段上1／3处。     

TSP-1和VEGF在胃癌微血管生成中的调控作用及其临床意义

目的：探讨TSP-1和VEGF在胃癌微血管生成中的调控作用及临床意义.方法：采用免疫组织化学SP法检测85例胃癌组织（实验组）以及癌缘外≥5 cm远隔胃黏膜组织（对照组）中TSP-1、VEGF和CD31表达水平,研究TSP-1和VEGF的表达与微血管密度（MVD）的相关关系.结果：（1）实验组TSP-1、VEGF阳性率分别为40%（34/85）、81.2%（69/85）,较对照组明显增高（p＜0.01）;（2）TSP-1的表达与MVD呈负相关（p＜0.01）,VEGF的表达与MVD的表达呈正相关（p＜0.01）.结论：胃癌中TSP-1和VEGF的表达与血管新生关系密切,是调控血管生成的重要因素,两者表达平衡决定胃癌新生血管生成.     

鸡胚尿囊绒毛膜法研究细胞间黏附分子-1在血管生成中的作用

目的　探讨细胞间黏附分子 1(intercellularadhesionmolecule 1,ICAM 1)在血管生成中的作用。　方法采用鸡胚尿囊绒毛膜 (chorioallantoicmembrane ,CAM)法进行在体血管生成实验。　结果　 1 10d鸡胚的尿囊绒毛膜经ICAM 1作用 3d后 ,明胶海绵周围放射状走行的微血管非常明显 ,似车辐 ,显微镜下明胶海绵内有垂直长入的微血管 ,明胶海绵周边CAM间充质内微血管数目显著多于对照组 (P <0 0 1)。 2 6d鸡胚的尿囊绒毛膜经Anti ICAM 1作用 3d后 ,明胶海绵周围放射状走行的微血管极不明显 ,显微镜下明胶海绵内几乎没有新生的微血管 ,明胶海绵周边CAM间充质内微血管数目显著少于对照组 (P <0 0 1)。　结论　结果提示 1 ICAM 1有诱导微血管生成的作用 ;2 ICAM 1参与胚胎的血管生成。     

In vivo evidence of angiogenesis inhibition by β2-glycoprotein I subfractions in the chorioallantoic membrane of chicken embryos

The vascular network expansion and functioning are important factors affecting normal intra-uterine fetal development. This study addressed the previously reported antiangiogenic potential of beta-2-glycoprotein I (β₂GPI) in vivo in the chick embryo model of angiogenesis. The effects of two naturally occurring β₂GPI forms on the development of the chorioallantoic membrane (CAM) vessels and the chicken embryo were investigated. β₂GPI monomers and dimers were obtained by fractioned purification and characterized using SDS-PAGE, immunoblot, and ELISA. The egg exposure was performed by injection of small volumes of 2.5 µg/mL solutions of the β₂GPI subfractions. Angiogenesis was evaluated through quantitative measurements of vascular architecture parameters in the captured CAM images, using computational analysis of texture contrasts and computer vision techniques. Quantitative information was assigned to the CAM vasculature modifications. In vivo, the β₂GPI dimer completely halted the formation of CAM vessels and led to embryo death after 48 h of exposure. The β₂GPI monomer allowed the embryo to develop up to the 10th day, despite early changes of CAM vessels. The impaired normal vessel growth proceeded as a self-limited effect. The β₂GPI monomer-exposed eggs showed reduced vascularization on the 6th day of incubation, but embryos were viable on the 10th day of incubation, with ingurgitated CAM vessels implying sequelae of the angiogenesis inhibition. Both subfractions impaired CAM vasculature development. The β₂GPI dimer proved to be largely more harmful than the β₂GPI monomer. β₂GPI modification by cleavage or dimerization may play a role in angiogenesis control in vivo.     

An experimental study in the chick embryo chorioallantoic membrane of the anti-angiogenic activity of cyclosporine in rheumatoid arthritis versus osteoarthritis

OBJECTIVE AND DESIGN: Angiogenesis plays an important role in the pathogenesis of rheumatoid arthritis (RA) and correlates with clinical score, synovial hyperplasia and infiltration of inflammatory cells. Many of the available treatments for RA have been shown to possess some degree of anti-angiogenic activity. Here, we studied the effect of cyclosporine, which exerts anti-angiogenic activity in vitro and in vivo [1] on angiogenesis induced in vivo in the chick embryo chorioallantoic membrane (CAM) by synovial RA and osteoarthritis (OA) tissues. MATERIAL AND METHODS: Wet synovial biopsies from 10 RA and 6 OA patients were treated with vehicle alone or with cyclosporine and implanted on the CAM at day 8 of incubation. On day 12, CAM tissues were assessed for the extent of angiogenesis and mononuclear cell infiltration. RESULTS: Cyclosporine inhibited angiogenesis and reduced the number of mononuclear cells in the CAM extracellular matrix only in RA implants. CONCLUSIONS:These data provide further evidence for a central role of new-formed blood vessels in RA. Moreover, cyclosporine on account of both its immunosuppressive and its anti-angiogenic activity can be proposed for the treatment of RA.     

Evaluation of metastatic and angiogenic potentials of human colon carcinoma cells in chick embryo model systems

Increased metastatic and angiogenic potentials of aggressive human colon carcinoma cells were verified in independent chick embryo models by comparing in vivo highly metastatic SW620 colon carcinoma cell line with its isogenic, non-metastatic SW480 cell variant. In the experimental metastasis model, both cell types rapidly arrested in the chorioallantoic membrane (CAM) vasculature as demonstrated by quantitative PCR and immunohistochemistry. Live cell imaging also indicated that both SW620 and SW480 cells efficiently extravasated from the CAM capillary system. However, only few SW480 cells were present in the CAM tissue after 24–48 h. In contrast, the numbers of SW620 cells increased exponentially, indicating proliferative and survival advantages of metastatic colon carcinoma cells in vivo. Multicellular SW620 foci were identified in close proximity to CAM blood vessels. A positive correlation between increased metastatic ability and VEGF-expression of colon carcinoma SW620 cells was demonstrated by the substantial inhibitory effects of anti-VEGF treatment on the levels of metastatic colonization and density of blood vessels adjacent to tumor cell foci. Furthermore, the chick embryo angiogenesis model confirmed high levels of VEGF-dependent angiogenesis induced by SW620 cells, but not SW480 cells. Thus, chick embryo experimental metastasis and CAM angiogenesis models appear to coordinately reflect critical features of advanced colon carcinomas, i.e., the acquisition of enhanced survival and increased angiogenic potentials, both constituting critical determinants of colon cancer progression. The use of rapid and quantitative chick embryo models might provide alternative approaches to conventional mammalian model systems for screening anti-cancer agents.     

Promotion of angiogenesis by low energy laser irradiation

The effect of low energy laser (He-Ne) irradiation (LELI) on the process of angiogenesis in the infarcted rat heart and in the chick chorioallantoic membrane (CAM), as well as the proliferation of endothelial cells in tissue culture, was investigated. Formation of new blood vessels in the infarcted rat heart was monitored by counting proliferating endothelial cells in blood vessels. In the CAM model, defined areas were laser-irradiated or nonirradiated and blood vessel density was recorded in each site in the CAM at various time intervals. Laser irradiation caused a 3.1-fold significant increase in newly formed blood vessels 6 days post infarction, as compared with nonirradiated rats. In the CAM model, a slight inhibition of angiogenesis up to 2 days post irradiation and a significant enhancement of angiogenesis in the laser-irradiated foci as compared with control nonirradiated spots were evident. The LELI caused a 1.8-fold significant increase in the rate of proliferation in endothelial cells in culture over nonirradiated cells. It is concluded that LELI can promote the proliferation of endothelial cells in culture, which may partially explain the augmentation of angiogenesis in the CAM model and in the infarcted heart. These results may have clinical significance by offering therapeutic options to ameliorate angiogenesis in ischemic conditions.     

Angiogenic activity in injured rat corneas as assayed on the chick chorioallantoic membrane

The temporal appearance of an angiogenic effect in chemically cauterized rat corneas was determined by studying the responses that they induced in the vessels of the chick chorioallantoic membrane (CAM). Injured rat corneas were grafted to the CAM from 90 minutes to 7 days after cautery. As controls, uninjured rat corneas and corneas of healthy rats cauterized immediately after death were also grafted. The vascular responses to the grafts were graded in a masked fashion by stereoscopic biomicroscopy on a five-tiered scale, by evaluations of projected colored photographs on the same scale, and by histologic examination of the grafts. Separate coefficients of angiogenesis were determined for the stereoscopic and photographic evaluations. We detected significant differences between corneas of healthy rats that were uninjured or cauterized chemically immediately after death and those that were cauterized in the living rat. Uninjured corneas and corneas cauterized postmortem elicited a mild vascular response in the CAM, as reflected by low coefficients of angiogenesis. Whereas blood vessels were not detected in corneas injured postmortem, some normal corneas vascularized but only after being on the CAM for at least 7 days. The coefficients of angiogenesis of corneas that were cauterized during life were significantly higher than those of both control groups prior to grafting after comparable times on the CAM. Corneas vascularized on the CAM included those that were cauterized as soon as 90 minutes prior to grafting. The strongest vascular responses, as reflected by coefficient of angiogenesis and the frequency of histologically confirmed nucleated avian erythrocytes within intracorneal blood vessels, were found with corneas that were grafted to the CAM 3 days after chemical cauterization. Corneas that vascularized on the CAM were associated with a prominent leukocytic infiltrate suggestively derived from the chick embryo. The results suggest that chemically cauterized rat corneas contain a chemoattractant for polymorphonuclear leukocytes within 90 minutes of injury and that such polymorphonuclear leukocytes or other components of the injured corneas possess the ability to stimulate angiogenesis on the CAM.     

Alterations of blood vessel development by endothelial cells overexpressing fibroblast growth factor-2

A close relationship exists between angiogenesis and the formation of vascular lesions. The development of the vascular system in the chick embryo chorioallantoic membrane (CAM) may thus represent a model to study the effects of the deregulation of endothelial cell behaviour. Alterations of the developing vascular tree of the CAM were observed after exposure to murine aortic endothelial (MAE) cells overexpressing human fibroblast growth factor-2 (FGF2) cDNA (pZipFGF2 MAE cells), or to their conditioned medium (CM). pZipFGF2 MAE cells injected into the allantoic sac or applied on to the CAM of day 8-9 chick embryos induce neovascularization and the appearance of haemangioma-like lesions. This activity was not prevented by anti-FGF2 antibodies. The CM from pZipFGF2 MAE cells was also active when adsorbed into a gelatin sponge and applied on to the CAM, both in the absence and in the presence of anti-FGF2 antibodies. No effects on vessel development were exerted by parental MAE cells, FGF2-transfected NIH 3T3 fibroblasts, or their conditioned media. In vitro, pZipFGF2 MAE cell CM caused parental MAE cells to invade fibrin gels and to undergo morphogenesis on Matrigel. This activity was not mimicked by recombinant FGF2 nor affected by anti-FGF2 antibodies, and depended on a M (r) approximately 45 000 heat-labile heparin-binding factor. Size exclusion chromatography of pZipFGF2 MAE cell CM demonstrated that the in vitro activity co-purified with an in vivo angiogenic capacity. Thus, FGF2 overexpression in mouse endothelial cells induces the production of an angiogenic activity distinct from FGF2, which may contribute to the genesis of angioproliferative lesions.     

Aquaporin‐1 expression in the chick embryo chorioallantoic membrane

The chick embryo chorioallantoic membrane (CAM) is commonly used in vivo to study both angiogenesis and anti‐angiogenesis. Rapid membrane water transport is mediated by a family of molecular water channels, called aquaporins (AQPs), which have been identified in the epithelial and endothelial cells of higher vertebrates. AQP1, expressed in adsorptive and secretory epithelia, is also expressed in endothelial cells of capillaries and arteries. Its mRNA has been found in vascular smooth muscle cells (VSMCs) of arteries and capillaries, as well as in a subset of VSMCs of human atherosclerotic plaques. This study investigated the developmental expression of AQP1 in the chick CAM by Western blot and immunohistochemistry. Western blot results show that a major nonglycosylated band was observed with electrophoretic mobility of approximately 28 kDa in the three developmental stages examined. Immunohistochemistry data demonstrate that AQP1 was clearly expressed in the ectodermal and endodermal epithelia, the vascular endothelium, and the VSMCs. Because little information is available on the behavior of microvessel AQP1 during angiogenesis in normal and pathological conditions, our data relative to the pattern of expression of AQP1 in CAM blood vessels in normal conditions may be considered a useful tool to further investigate its modifications in several experimental conditions implying a stimulation or an inhibition of angiogenesis in the CAM assay. Anat Rec 268:85–89, 2002. © 2002 Wiley‐Liss, Inc.     

Ultrastructural changes in blood vessels in epidermal growth factor treated experimental cutaneous wound model

This study investigates the impact of epidermal growth factor (EGF) on blood vessels, specifically on the development of intussusceptive angiogenesis in cutaneous wound healing. Excisional wounds were formed on both sides of the medulla spinalis in dorsal location of the rats. The control and EGF-treated groups were divided into two groups with respect to sacrifice day: 5 d and 7 d. EGF was topically applied to the EGF-treated group once a day. The wound tissue was removed from rats, embedded in araldite and paraffin, and then examined under transmission electron and light microscopes. The ultrastructural signs of intussusceptive angiogenesis, such as intraluminal protrusion of endothelial cells and formation of the contact zone of opposite endothelial cells, were observed in the wound. Our statistical analyses, based on light microscopy observations, also confirm that EGF treatment induces intussusceptive angiogenesis. Moreover, we found that induction of EGF impact on intussusceptive angiogenesis is higher on the 7th day of treatment than on the 5th day. This implies that the duration of EGF treatment is important. This research clarifies the effects of EGF on the vessels and proves that EGF induces intussusceptive angiogenesis, being a newer model with respect to sprouting type.     

An evaluation of methods to quantitate the chick chorioallantoic membrane assay in angiogenesis

The vascular responses by chick chorioallantoic membranes (CAM) to more than 150 normal and chemically injured rat corneas grafted to shell-less chicken CAMs were evaluated independently by three observers in a masked fashion by in vivo stereomicroscopy, projections of colored transparencies, and by light microscopy of tissue sections of the grafts. The experience gained from this study is reviewed as a point of focus for the strengths and weaknesses of the CAM technique in the assay of potential angiogenic substances. Despite certain shortcomings, the CAM technique can provide useful information relevant to studies on angiogenesis, particularly when the subjective CAM method is supplemented by histological evaluation of grafted tissues.