血管内皮生长因子及其受体在结直肠癌中的表达及其意义

目的：探讨血管内皮生长因子（ＶＥＧＦ）及其受体（ＦＬＴ、ＦＬＫ－１）在结直肠癌组织中的表达及其对肿瘤新生血管生成及转移的影响。方法应用免疫组化技术,检测５０例人结直肠癌组织ＶＥＧＦ、ＦＬＴ、ＦＬＫ－１的表达和微血管密度（ｍｉｃｒｏｖｅｓｓｅｌｓ ｄｅｎｓｉｔｙ,ＭＶＤ）,并分析其与病理分型、分级、浸润深度、淋巴结、淋巴管和血管转移的相关。结果ＶＥＧＦ、ＦＬＴ、ＦＬＫ－１主要表达于癌浸润边缘和坏死组     

基于脑微血管内皮细胞保护的注射用丹参多酚酸质量生物标志物研究

目的 基于血管内皮保护的关键药效环节,发现能够反映注射用丹参多酚酸（SAFI）的质量生物标志物（Qbiomarkers）,为完善其质量控制评价提供参考。方法 90℃加热处理SAFI 4、18、60 h,得到不同化学质量的SAFI（SAFI4、SAFI18、SAFI60）,高效液相色谱（HPLC）法分析紫草酸、丹酚酸B、丹酚酸Y、丹参素钠、原儿茶醛、丹酚酸D、迷迭香酸变化趋势。体外培养小鼠脑微血管内皮细胞（bEnd.3）,建立氧糖剥夺/复氧（OGD/R）模型,给予SAFI、SAFI4、SAFI18、SAFI60处理18 h,CCK-8法检测细胞相对存活率,试剂盒法检测上清液乳酸脱氢酶（LDH）、NO水平,划痕实验检测细胞迁移率;通过DIA蛋白质组学分析不同化学质量的SAFI给药后产生的差异蛋白,结合药效学及偏最小二乘回归（PLSR）分析筛选SAFI的Q-biomarkers。结果 与SAFI相比,SAFI4、SAFI18、SAFI60样品中的紫草酸、丹酚酸B和丹酚酸Y含量下降,丹参素钠含量上升,原儿茶醛、丹酚酸D、迷迭香酸变化不明显。药效学结果表明,与模型组相比,SAFI、SAFI4及SAFI18组细胞相对存活率显著上升、LDH泄漏量显著下降、NO水平显著下降、细胞迁移率显著升高（P＜0.05、0.01、0.001） ,对细胞具有显著保护作用;而SAFI60组无明显保护作用。蛋白质组学结果表明,18个差异蛋白的含量随药效学和成分的变化而发生一致性上升或下降,与药效进行相关性分析发现Kdr、Cxcl12的变量重要性投影值（VIP）皆大于1,尤其Kdr ［血管内皮生长因子受体2 （VEGFR2）］与血管内皮功能高度相关,可能是潜在的SAFI的Q-biomarkers。结论 通过不同化学质量的SAFI的内皮保护药效及差异蛋白关联分析发现,VEGFR2可以作为SAFI的Q-biomarker。     

VEGF mRNA、KDR mRNA及蛋白在非小细胞肺癌中表达的意义探讨

目的探讨非小细胞肺癌中血管内皮生长因子(VEGF)、受体KDRmRNA和蛋白表达的意义。方法应用原位杂交技术及免疫组织化学PV-9000法分别检测30例新鲜非小细胞肺癌标本中VEGFmRNA、KDRmRNA及蛋白的表达。结果VEGF、KDRmRNA和蛋白均主要分布在肺癌细胞、血管内皮细胞及纤维母细胞,肺癌细胞VEGFmRNA和蛋白的阳性率分别为66.67%(20/30)和60.00%(18/30),两者的表达有极显著正相关性(r=0.86,P<0.01);KDRmRNA和蛋白的阳性率分别为73.33%(22/30)和76.67%(23/30),两者的表达亦有极显著正相关性(r=0.7366,P<0.01);VEGFmRNA、KDRmRNA及相应蛋白在肺癌细胞中的表达均呈极显著性正相关(原位杂交r=0.853,免疫组化r=0.5148,均P<0.01);肺癌细胞VEGFmRNA、KDRmRNA及相应蛋白的阳性表达率在不同大小的3组肺癌标本间,差异均具有显著性(确切P=0.014,P=0.002;P=0.0087,P=0.0016);淋巴结有转移组的VEGFmRNA、KDRmRNA及相应蛋白在肿瘤细胞中的阳性表达率显著高于淋巴结无转移组者(确切P=0.007,P=0.003;P=0.0008,P=0.0073)。结论肺癌细胞产生的VEGF通过自分泌环作用于癌细胞上的KDR受体促进癌细胞生长;VEGF及KDR的检测可作为肺癌淋巴结转移的有效评估指标;VEGF及KDR有望成为肺癌治疗的新靶点。     

Tumor angiogenesis--a potential target in cancer chemoprevention.

Tumor angiogenesis is critically important for the growth of solid tumors as tumors remain in dormant phase for a long time in the absence of the initiation of blood vessel formation. Tumors can grow up to approximately 2mm size without requirement of blood supply as diffusion is sufficient at this level to support the removal of wastes from and supply of nutrients to tumor cells. Therefore, angiogenesis process could be an important target to suppress tumor growth and metastasis. Angiogenesis is required at almost every step of tumor progression and metastasis, and tumor vasculature has been identified as strong prognostic marker for tumor grading. Endothelial cells are the main players of angiogenesis process and could be peculiar target for antiangiogenic therapy because they are non-transformed and easily accessible to achievable concentrations of antiangiogenic agents, and also are unlikely to acquire drug resistance. Several antiangiogenic strategies have been developed to inhibit tumor growth by targeting different components of tumor angiogenesis. Chemopreventive agents have been shown to target and inhibit different aspects and components of angiogenesis process and can be used conveniently as they are mostly non-toxic natural compounds and could be part of our daily diet. However, a risk assessment for the use of antiangiogenic phytochemicals is needed as they can also disrupt normal physiologic angiogenesis such as wound healing and endometrium development processes. This review focuses on how different chemopreventive phytochemicals target various components of angiogenesis, including angiogenic signaling, which usually starts from tumor cells producing angiogenic factors and affecting endothelial cells growth, migration and capillary vessel organization for tumor angiogenesis.     

VEGF及其受体抗肿瘤血管治疗应用新进展

肿瘤的抗血管治疗是目前的研究热点之一.本文主要综述VEGF及VEGFR抗肿瘤血管治疗的临床应用及临床前实验阶段的研究进展,并对当前抗血管药物进行了比较.     

短肽库中血管内皮细胞生长因子受体Flt-1拮抗剂筛选条件的改进

目的:改进噬菌体肽库的筛选条件,提高筛选阳性率及阳性克隆重复性.使血管内皮细胞生长因子受体Flt-1拮抗剂的筛选更为有效.方法:通过对血管内皮细胞生长因子受体Flt-1的原核表达产物的变性复性处理,获得相对纯化的可溶性Flt-1受体.将该可溶性受体固相化,对噬菌体表达12肽库进行4 ～ 5轮Bio-panning后,挑取单个噬菌斑制备高滴度噬菌体上清.用ELISA法初步鉴定各噬菌体克隆与sFlt-1的特异性结合并测定阳性克隆的DN A 顺序.在Bio-panning过程中增加非特异性蛋白的预吸附,并减少输入噬菌体数量, 再次筛选短肽库.结果:经过第一次筛选,获得了一系列具有一定重复性与同源性的噬菌体克隆. 改进筛选条件后,筛选阳性率由2%提高到8.3%,阳性克隆的重复性也得到显著提高.该高重复阳性克隆的氨基酸顺序与前一次筛选获得的阳性克隆有较高同源性.结论:Bio-panning 条件的改进提高了筛选效率,为不纯蛋白筛选短肽库积累了经验,并为血管内皮细胞生长因子受体拮抗剂的研制奠定了基础.     

Simultaneous blockade of programmed death 1 and vascular endothelial growth factor receptor 2 (VEGFR2) induces synergistic anti‐tumour effect in vivo

Recent basic and clinical studies have shown that the programmed death ligand (PD‐L)/PD‐1 pathway has a significant role in tumour immunity, and its blockade has a therapeutic potential against several human cancers. We hypothesized that anti‐angiogeneic treatment might augment the efficacy of PD‐1 blockade. To this end, we evaluated combining the blockade of PD‐1 and vascular endothelial growth factor receptor 2 ( VEGFR2) in a murine cancer model using Colon‐26 adenocarcinoma. Interestingly, simultaneous treatment with anti‐PD‐1 and anti‐VEGFR2 monoclonal antibodies (mAbs) inhibited tumour growth synergistically in vivo without overt toxicity. Blocking VEGFR2 inhibited tumour neovascularization significantly, as demonstrated by the reduced number of microvessels, while PD‐1 blockade had no impact on tumour angiogenesis. PD‐1 blockade might promote T cell infiltration into tumours and significantly enhanced local immune activation, as shown by the up‐regulation of several proinflammatory cytokine expressions. Importantly, VEGFR2 blockade did not interfere with T cell infiltration and immunological activation induced by PD‐1 blockade. In conclusion, simultaneous blockade of PD‐1 and VEGFR2 induced a synergistic in‐vivo anti‐tumour effect, possibly through different mechanisms that might not be mutually exclusive. This unique therapeutic strategy may hold significant promise for future clinical application.     

Sirolimus inhibits lymphangiogenesis in rat renal allografts,a novel mechanism to prevent chronic kidney allograft injury

Lymphangiogenesis occurs in renal allografts and it may be involved in the maintenance of the alloreactive immune response and thus participate in the development of chronic kidney allograft injury. Sirolimus (SRL) has been shown to inhibit lymphangiogenesis. The aim of this study was to describe lymphangiogenesis and its regulation during the development of chronic kidney allograft injury and to investigate the effect of SRL on allograft lymphangiogenesis and chronic kidney allograft injury. A rat renal transplantation model was used. Allografts treated with cyclosporine A or with SRL were analyzed in various time points. Syngenic transplantations were used as controls. Kidney function was followed with serum creatinine. Histology was analyzed by Chronic Allograft Damage Index (CADI). Immunohistochemistry was used to detect lymphatic vessels, VEGF‐C and VEGFR‐3. In cyclosporine‐treated allografts VEGF‐C/VEGFR‐3 pathway was strongly upregulated leading to extensive lymphangiogenesis 60 days after transplantation. Lymphangiogenesis correlated positively with the CADI score. Sirolimus efficiently inhibited lymphangiogenesis, improved graft function and attenuated the development of chronic kidney allograft injury when compared with cyclosporine. In conclusion, lymphangiogenesis is associated with chronic kidney allograft injury and SRL is a potent inhibitor of lymphangiogenesis in renal allografts. Inhibition of lymphatic proliferation could mediate the nephroprotective properties of SRL.     

The classification and diagnostic algorithm for primary lymphatic dysplasia: an update from 2010 to include molecular findings

Historically, primary lymphoedema was classified into just three categories depending on the age of onset of swelling; congenital, praecox and tarda. Developments in clinical phenotyping and identification of the genetic cause of some of these conditions have demonstrated that primary lymphoedema is highly heterogenous. In 2010, we introduced a new classification and diagnostic pathway as a clinical and research tool. This algorithm has been used to delineate specific primary lymphoedema phenotypes, facilitating the discovery of new causative genes. This article reviews the latest molecular findings and provides an updated version of the classification and diagnostic pathway based on this new knowledge.