BACKGROUND & AIMS: BACKGROUND:Curcumin has anti-inflammatory, antioxidant and anticancer properties. Despite the considerable evidence showing that curcumin is an efficacious and safe compound for multiple medicinal benefits, there are some demerits with respect to the therapeutic effectiveness of curcumin, namely, poor stability and solubility, and its role in angiogenesis in vivo is still not yet clear. More recently, the biodegradable polymer nanoparticles have been developed. This offers promise for the therapeutic effectiveness of curcumin by increasing its bioavailability, solubility and retention time. METHODS:Here, we compared the medicinal effectiveness of curcumin and nanocurcumin (NC), and found that nanocurcumin can inhibit angiogenesis more effectively than curcumin in zebrafish. Tests of proliferation and apoptosis showed no difference between nanocurcumin-treated and wildtype embryos. RESULTS:qPCR and in situ hybridization experiments indicated that the VEGF signaling pathway genes, vegfa, VEGF-C and flt4 were all down-regulated after nanocurcumin treatment, and vegfa over-expression rescued the vascular defective phenotype. Moreover, hif1a expression also decreased and hif1a over-expression also rescued the vascular defective phenotype but the Notch signaling pathway had no difference after nanocurcumin treatment. CONCLUSION:These results indicate that nano curcumin inhibits angiogenesis in zebrafish by downregulating hif1a/vegfa signaling pathway. Hence, our work reveals the key role of nanocurcumin in angiogenesis in vivo.

背景与目标：

BACKGROUND & AIMS: :Matrix vesicles (MVs) are well positioned in the growth plate to serve as a carrier of morphogenetic information to nearby chondrocytes and osteoblasts. Bone morphogenetic proteins (BMPs) carried in MVs could promote differentiation of these skeletal cells. Vascular endothelial growth factor (VEGF) in MVs could stimulate angiogenesis. Therefore, a study was undertaken to confirm the presence of bone morphogenetic protein (BMP)-1 through-7, VEGF, and the noncollagenous matrix proteins, bone sialoprotein (BSP), osteopontin (OPN), osteocalcin (OC), and osteonectin (ON) in isolated rat growth plate MVs. MVs were isolated from collagenase-digested rachitic rat tibial and femoral growth plates. The presence of BMP-1 through BMP-7, VEGF, BSP, ON, OPN, and OC was evaluated by Western blot, plus ELISA analyses for BMP-2 and-4 content. The alkaline phosphatase-raising ability of MV extracts on cultured rat growth plate chondrocytes was measured as a reflection of MV ability to promote chondroosseous differentiation. BMP-1 through-7, VEGF, BSP, ON, OPN, and OC were all detected by Western blot analyses. Chondrocytes treated with MV extracts showed a two-to threefold increase in alkaline phosphatase activity over control, indicating increased differentiation. Significant amounts of BMP-2 and BMP-4 were detected in MVs by ELISA. Combined, these data suggest that MVs could play an important morphogenetic role in growth plate and endochondral bone formation.

背景与目标： : 基质囊泡 (mv) 位于生长板中，可作为向附近的软骨细胞和成骨细胞提供形态发生信息的载体。MVs中携带的骨形态发生蛋白 (bmp) 可以促进这些骨骼细胞的分化。MVs中的血管内皮生长因子 (VEGF) 可以刺激血管生成。因此，进行了一项研究以确认存在骨形态发生蛋白 (BMP)-1至-7，VEGF和非胶原基质蛋白，骨唾液酸蛋白 (BSP)，骨桥蛋白 (OPN)，骨钙素 (OC) 和骨连接蛋白 (ON) 在分离的大鼠生长板mv中。从胶原酶消化的rachitic大鼠胫骨和股骨生长板中分离出mv。通过Western blot和ELISA分析BMP-2和-4含量来评估BMP-1通过BMP-7，VEGF，BSP，ON，OPN和OC的存在。测量了MV提取物在培养的大鼠生长板软骨细胞上的碱性磷酸酶升高能力，以反映MV促进软骨骨分化的能力。BMP-1-7、VEGF、BSP、ON、OPN和OC均通过蛋白质印迹分析检测。用MV提取物处理的软骨细胞显示碱性磷酸酶活性比对照增加了两到三倍，表明分化增加。通过ELISA在MVs中检测到大量的BMP-2和BMP-4。综合起来，这些数据表明MVs可能在生长板和软骨内骨形成中起重要的形态发生作用。

BACKGROUND & AIMS: :Bone marrow-derived mesenchymal stem cells (MSC) are a promising source for cell-based treatment of myocardial infarction (MI), but existing strategies are restricted by low cell survival and engraftment. We examined whether vascular endothelial growth factor (VEGF) improve MSC viability in infarcted hearts. We found long-term culture increased MSC-cellular stress: expressing more cell cycle inhibitors, p16(INK), p21 and p19(ARF). VEGF treatment reduced cellular stress, increased pro-survival factors, phosphorylated-Akt and Bcl-xL expression and cell proliferation. Co-injection of MSCs with VEGF to MI hearts increased cell engraftment and resulted in better improvement of cardiac function than that injected with MSCs or VEGF alone. In conclusion, VEGF protects MSCs from culture-induce cellular stress and improves their viability in ischemic myocardium, which results in improvements of their therapeutic effect for the treatment of MI.

背景与目标： : 骨髓间充质干细胞 (MSC) 是基于细胞的心肌梗塞 (MI) 治疗的有希望的来源，但是现有的策略受到低细胞存活率和植入的限制。我们检查了血管内皮生长因子 (VEGF) 是否可以改善梗塞心脏的MSC活力。我们发现长期培养增加了MSC细胞应激: 表达更多的细胞周期抑制剂p16(INK)，p21和p19(ARF)。VEGF治疗可减少细胞应激，增加促生存因子，磷酸化-Akt和Bcl-xL表达和细胞增殖。与单独注射MSCs或VEGF相比，将MSCs与VEGF共同注射到MI心脏可增加细胞植入并改善心脏功能。总之，VEGF保护MSCs免受培养诱导的细胞应激，并改善其在缺血性心肌中的生存能力，从而改善其治疗MI的治疗效果。

BACKGROUND & AIMS: AIMS OF THE STUDY:Studies on circulating VEGF have reported mixed results, possibly due to a lack of standardization of the pre-analytical phase. The aim of our investigation was to standardize the sampling procedure for the determination of VEGF in different blood fractions. BASIC PROCEDURES:We evaluated various clotting times for obtaining serum in 30 subjects, as well as different procedures for the preparation of plasma Edinburgh anticoagulant mixture (EDTA, PGE1, theophylline) and CTAD. VEGF was also assayed in lysed whole blood. In vitro platelet activation was monitored by measuring the levels of PF4. VEGF and PF4 were measured using commercially available enzyme-linked immunoassays. MAIN FINDINGS:Clotting time increased the release of VEGF, which reached a plateau between 2 and 4 hours. The percent increase of VEGF at 2 hours ranged from 118% to 4,515% (median 327%) compared to samples centrifuged within 10 min from withdrawal. VEGF was not different and PF4 was very low or undetectable in Edinburgh plasma and CTAD plasma, while it was significantly higher in sodium citrate plasma. VEGF in CTAD plasma was not correlated with platelet count or leukocytes. Serum VEGF did not correlate with the leukocyte number, but it correlated significantly with the platelet count. PRINCIPAL CONCLUSIONS:The procedures for sample collection described above are highly standardized and easy to perform in a routine setting. We therefore suggest systematic evaluation of VEGF in CTAD plasma, in serum (clotting for 2 hours at room temperature) and in whole blood, until prospective controlled clinical studies will have clarified in which blood compartment(s) VEGF provides clinically relevant information.

背景与目标：

BACKGROUND & AIMS: :Current models of brain development support the view that VEGF, a signaling protein secreted by neuronal cells, regulates angiogenesis and neuronal development. Here we demonstrate an autonomous and pivotal role for endothelial cell-derived VEGF that has far-reaching consequences for mouse brain development. Selective deletion of Vegf from endothelial cells resulted in impaired angiogenesis and marked perturbation of cortical cytoarchitecture. Abnormal cell clusters or heterotopias were detected in the marginal zone, and disorganization of cortical cells induced several malformations, including aberrant cortical lamination. Critical events during brain development-neuronal proliferation, differentiation, and migration were significantly affected. In addition, axonal tracts in the telencephalon were severely defective in the absence of endothelial VEGF. The unique roles of endothelial VEGF cannot be compensated by neuronal VEGF and underscores the high functional significance of endothelial VEGF for cerebral cortex development and from disease perspectives.

背景与目标： : 目前的大脑发育模型支持以下观点: VEGF是神经元细胞分泌的信号蛋白，可调节血管生成和神经元发育。在这里，我们证明了内皮细胞衍生的VEGF的自主和关键作用，对小鼠大脑发育具有深远的影响。内皮细胞中Vegf的选择性缺失导致血管生成受损和皮质细胞结构的明显扰动。在边缘区检测到异常的细胞簇或异型，皮质细胞的紊乱引起了几种畸形，包括异常的皮质层合。大脑发育过程中的关键事件-神经元增殖，分化和迁移受到显着影响。此外，在没有内皮VEGF的情况下，端脑中的轴突束严重缺陷。内皮VEGF的独特作用不能通过神经元VEGF来补偿，并从疾病的角度强调了内皮VEGF对大脑皮层发育的高度功能意义。

BACKGROUND & AIMS: BACKGROUND:Intracellular signaling responsible for gastrin-releasing peptide (GRP) receptor-mediated neovascularization is not clearly understood. We sought to determine the cellular mechanisms involved in the GRP receptor regulation of vascular endothelial growth factor (VEGF) release in neuroblastoma cells. MATERIALS AND METHODS:BE(2)-C cells were treated with bombesin (BBS), the amphibian equivalent of GRP, Phorbol myristate acetate (PMA) a PKC agonist, or GF109293X (GFX), and analyses were performed for VEGF secretion, phosphorylated protein kinase B (AKT), extracellular signal-regulated kinases (ERK) and protein kinase D (PKD) expression. RESULTS:BBS rapidly increased VEGF secretion at 30 min. Pre-treatment with PMA alone produced similar results; this effect was synergistic with the addition of GRP. Conversely, GFX blocked PMA-stimulated increase in VEGF secretion. Immunofluorescent staining for VEGF correlated to BBS, PMA and GFX. CONCLUSION:PKC is critically responsible for rapid VEGF secretion by GRP receptor signaling in neuroblastoma cells. Inhibition of VEGF significantly reduced GRP-mediated cell proliferation, suggesting its crucial role in neuroblastoma tumorigenesis.

背景与目标：

BACKGROUND & AIMS: :This study aimed to identify and describe anatomical and functional changes on short (1-3 months) and medium (6-12 months) term after intravitreal injections of bevacizumab (Avastin, Genentech) in patients with choroidal neovascularization (CNV) in the context of exudative form of age-related macular degeneration (AMD). We performed a retrospective, analytical, interventional study, based on a series of cases with exudative form of AMD, which also comprised a prospective component related to the inclusion and treatment of the patients with a very new interventional method for that time (2006) and the follow-up of the effects of intravitreal injection of bevacizumab (1.25 mg) therapy in three monthly doses for short (1-3 months) and medium (6-18 months) periods of time. The follow-up of these patients was made by determining visual acuity (VA) as best corrected visual acuity (BCVA) at baseline and at every visit, slit lamp examination with contact or noncontact lenses each time, and optical coherence tomography and/or angiofluorography, applied only for certain patients, at various times of the study. In total, 376 intravitreal injections were administered to 117 eyes of 96 patients. The VA improved in the assessment of 3 months in 77 eyes (66%), either subjective (by the patient) or objectively quantified (by the physician). In 40 eyes (34%), there was no change in VA. In patients for whom optical coherence tomography could be performed, a significant reduction of the macula's thickness was found. The use of bevacizumab in subretinal neovascular membrane treatment is effective and safe on short and medium term, with the improvement of BCVA and reduction of macular edema in a significant number of cases.

背景与目标： : 本研究旨在识别和描述玻璃体内注射贝伐单抗 (Avastin，genentech) 在渗出形式的年龄相关性黄斑变性 (AMD) 的情况下的脉络膜新生血管 (CNV) 患者中。我们进行了一项回顾性，分析，干预性研究，基于一系列渗出性AMD病例，其中还包括与当时非常新的介入方法 (2006) 的患者的纳入和治疗有关的前瞻性成分，以及玻璃体内注射贝伐单抗 (1.25 mg) 治疗的效果的随访 (3个月短 (1-3个月) 和中 (6-18个月) 的时间段。这些患者的随访是通过在基线和每次访视时确定视力 (VA) 为最佳矫正视力 (BCVA)，每次使用隐形眼镜或非接触镜片进行裂隙灯检查，以及光学相干断层扫描和/或血管荧光成像，仅适用于某些患者，在研究的不同时间。总共对96例患者的117只眼睛进行了376次玻璃体内注射。在77眼 (66%) 的3个月评估中，VA改善了主观 (由患者) 或客观量化 (由医生)。在40眼 (34%) 中，VA没有变化。在可以进行光学相干断层扫描的患者中，发现黄斑厚度显着减少。在视网膜下新生血管膜治疗中使用贝伐单抗是短期和中期安全有效的，在大量病例中可以改善BCVA并减少黄斑水肿。

BACKGROUND & AIMS: :Hepatocellular carcinoma (HCC) is the third-leading cause of cancer-related deaths with 750,000 newly diagnosed cases each year. Surgery, radiotherapy, and chemotherapy constitute the main treatment modalities for HCC, but liver cirrhosis and damage often occur. Molecular targeted drugs have been recently developed to treat HCC. Vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR) autocrine signaling is closely related to the growth, progression, and metastasis of HCC, making the VEGF/VEGFR axis an ideal target for the development of molecular targeted agents. Here, we report the effects of the novel anti-VEGF humanized monoclonal antibody BD0801 on the growth of HCC cells in vitro and in vivo as well as the underlying mechanisms. BD0801 significantly inhibited the proliferation of HepG2, SMMC-7721, and Bel7402 cells in vitro, accompanied with an induction of apoptosis and cell cycle arrest at the G1 phase. BD0801 potently suppressed AKT, Erk1/2, and retinoblastoma (Rb) phosphorylation, while increasing p21 and decreasing cyclin D1 protein levels. BD0801 significantly inhibited growth in mouse tumor xenografts and induced cell apoptosis of HepG2 and SMMC-7721 tumor xenografts. Furthermore, BD0801 effectively reduced the vascular density and tumor tissue microvessel density (MVD). Similarly, BD0801 decreased AKT, Erk1/2, and Rb phosphorylation and cyclin D1 expression whereas it increased p21 protein expression in mouse HCC tumor xenografts. Importantly, BD0801 showed a better effect than Bevacizumab (Bev) on the inhibition of cell growth and induction of apoptosis in HCC cells in vitro and in vivo. These findings suggest that BD0801 is a potent anti-VEGF monoclonal antibody for the treatment of HCC.

背景与目标： : 肝细胞癌 (HCC) 是癌症相关死亡的第三大原因，每年有750,000例新诊断病例。手术，放疗和化疗是肝癌的主要治疗方式，但经常发生肝硬化和损害。最近开发了分子靶向药物来治疗HCC。血管内皮生长因子 (VEGF)/VEGF受体 (VEGFR) 自分泌信号与HCC的生长、进展和转移密切相关，使得VEGF/VEGFR轴成为开发分子靶向药物的理想靶点。在这里，我们报告了新型抗VEGF人源化单克隆抗体BD0801对体外和体内HCC细胞生长的影响以及潜在的机制。BD0801在体外显着抑制HepG2，SMMC-7721和Bel7402细胞的增殖，并在G1期诱导凋亡和细胞周期停滞。BD0801有效抑制AKT，Erk1/2和视网膜母细胞瘤 (Rb) 的磷酸化，同时增加p21并降低细胞周期蛋白D1蛋白水平。BD0801显著抑制小鼠肿瘤异种移植物的生长，并诱导HepG2和SMMC-7721肿瘤异种移植物的细胞凋亡。此外，BD0801有效降低了血管密度和肿瘤组织微血管密度 (MVD)。同样，BD0801降低了小鼠HCC肿瘤异种移植物中AKT，Erk1/2和Rb磷酸化和cyclin D1的表达，而增加了p21蛋白的表达。重要的是，在体外和体内，BD0801在抑制HCC细胞生长和诱导凋亡方面显示出比贝伐单抗 (Bev) 更好的效果。这些发现表明BD0801是一种有效的抗VEGF单克隆抗体，可用于治疗HCC。

BACKGROUND & AIMS: :Interleukin-1 beta (IL-1 beta) is a multipotent cytokine participating in a variety of cardiovascular diseases. In this study, we examined the effects of IL-1 beta on the expression of vascular endothelial cell growth factor (VEGF) and pursued the molecular mechanisms underlying this effect. Treatment of cultured neonatal rat cardiac myocytes with IL-1 beta increased the levels of VEGF mRNA in a time- and a concentration-dependent manner. These effects were completely abolished by SB203580 and SB202190 (p38 MAPK inhibitors) but not by PD98059 (MEK1 inhibitor), calphostin C (protein kinase C inhibitor), or genistein (tyrosine kinase inhibitor). While IL-1 beta phosphorylated c-Jun N-terminus protein kinase (JNK) rapidly and transiently, the effect of IL-1 beta on p38 mitogen-activated protein kinase (MAPK) was gradual and persistent. Transient transfection assays showed that IL-1 beta increases the transcription from the VEGF promoter. A series of 5;-deletion and site-specific mutation analyses indicated that IL-1 beta as well as overexpression of p38 MAPK and JNK activate VEGF promoter activity through two G+C-rich sequences located at -73 and -62. Electrophoretic mobility shift and supershift assays showed Sp1 and Sp3 proteins specifically bind to the G+C-rich sequences. The half-life of VEGF mRNA was significantly increased in cells treated with IL-1 beta. Together, these results indicate that IL-1 beta induces VEGF gene expression at both transcriptional and post-transcriptional levels, and IL-1 beta evokes p38 MAPK and JNK signalings, which in turn stimulate the transcription of the VEGF gene through Sp1-binding sites. These findings suggest the role of IL-1 beta as a cytokine inducing VEGF in cardiac myocytes, and imply that activation of stress-activated MAP kinases regulate Sp1 sites-dependent transcription.

背景与目标： : Interleukin-1 β (IL-1 β) 是一种参与多种心血管疾病的多能细胞因子。在这项研究中，我们研究了IL-1 β 对血管内皮细胞生长因子 (VEGF) 表达的影响，并探讨了这种作用的分子机制。用IL-1 β 处理培养的新生大鼠心肌细胞以时间和浓度依赖性方式增加VEGF mRNA的水平。SB203580和SB202190 (p38 MAPK抑制剂) 完全消除了这些作用，而PD98059 (MEK1抑制剂)，钙磷蛋白C (蛋白激酶C抑制剂) 或染料木黄酮 (酪氨酸激酶抑制剂) 则没有。虽然快速且短暂地IL-1 β 磷酸化的c 6月N末端蛋白激酶 (JNK)，但IL-1 β 对p38丝裂原活化蛋白激酶 (MAPK) 的影响是渐进且持续的。瞬时转染测定表明，IL-1 β 增加了来自VEGF启动子的转录。一系列5; 缺失和位点特异性突变分析表明，IL-1 β 以及p38mapk和JNK的过表达通过位于-73和-62的两个富含G C的序列激活VEGF启动子活性。电泳迁移率偏移和超移测定显示Sp1和Sp3蛋白与富含G C的序列特异性结合。在用IL-1 β 处理的细胞中，VEGF mRNA的半衰期显着增加。总之，这些结果表明IL-1 β 在转录和转录后水平上诱导VEGF基因表达，IL-1 β 引起p38mapk和JNK信号，这反过来通过Sp1-binding位点刺激VEGF基因的转录。这些发现表明IL-1 β 作为诱导心肌细胞中VEGF的细胞因子的作用，并暗示应激激活的MAP激酶的激活调节Sp1位点依赖性转录。

BACKGROUND & AIMS: BACKGROUND:While randomized controlled trials (RCTs) are based on strict inclusion/exclusion criteria, non-interventional studies (NISs) might provide additional information to guide management in patients more representative to the real-world setting. The aim of this study was to compare baseline characteristics of patients receiving intravitreal treatment in the NIS OCEAN with those from published RCTs. METHODS:The ongoing OCEAN study enrolled patients treated with ranibizumab for neovascular age-related macular degeneration (nAMD), diabetic macular oedema (DME) or branch/central retinal vein occlusion (B/CRVO). Baseline patient characteristics were compared by indication within the OCEAN cohort. Furthermore, the characteristics were set in reference to those of published RCTs in the same indications. Confidence intervals (CIs) were calculated and assessed for statistically significant differences as indicated by non-overlapping CIs. RESULTS:Patient characteristics in the NIS OCEAN were evaluated for 3,614 patients with nAMD, 1,211 with DME, 204 with BRVO and 121 with CRVO. Between these groups, significant differences in mean age, gender distributions, and mean baseline VA were seen, reflecting known differences between the indications. Compared to the patient characteristics of published RCTs (trials selected by literature search: nAMD: 13 RCTs, DME: 9, RVO: 5), the OCEAN patients' mean age was significantly higher in every indication. The gender distributions across the trials were comparable, with only few differences between OCEAN and the RCTs. Regarding the mean baseline VA, notable differences were found in nAMD and in DME, with VA significantly higher in some RCTs and lower in others. CONCLUSIONS:The described differences underline the complementarity of NISs and RCTs. OCEAN covers a broader spectrum and more variability of patients than do RCTs. As baseline values may have impact on the treatment response (ceiling effect), there is an ongoing need for research in all patient subgroups. Country-specific assessments of patient populations can better reflect the real-world situation. NISs can deliver insights that RCTs may not, as NISs can include non-typical patients, patients with comorbidities, a broader age spectrum and patients of various disease stages. TRIAL REGISTRATION:The NIS OCEAN was registered on www.clinicaltrials.gov (identifier: NCT02194803 ).

背景与目标：

BACKGROUND & AIMS: AIMS:This study was designed to evaluate the antiangiogenic properties of emodin and its ability to inhibit tyrosine-kinase-mediated phosphorylation of vascular endothelial growth factor (VEGF) receptors in colon cancer cells. METHODS:The effects of emodin on VEGF-receptor (VEGFR) phosphorylation were determined by assaying the tyrosine kinase activity and by Western blot analysis. The antiproliferative and proapoptotic activities of emodin were evaluated by soft agar colony formation, flow cytometric analysis of cell cycle, and by apoptotic assay. RESULTS:Emodin causes a dose-dependent inhibition of VEGFR phosphorylation in colon cancer cells. Treatment with 40 muM of emodin decreased the relative activity of VEGFR-1 to 22.4%, when compared to the control group (assigned a value of 100%); VEGFR-2 and -3 showed a similar reduction in relative activity at 58.5% and 31.6%, respectively (p < 0.01, in each case). Treatment with emodin reduced VEGFR phosphorylation, as evidenced by Western blot analysis. Flow cytometric analysis showed that, upon treatment with emodin, the HCT116 cell cycle was blocked at the G2/M phase. Emodin also increased the apoptosis of HCT116 cells in a dose-dependent manner; treatment with 40 muM emodin increased the apoptotic rate from 8.1% +/- 2.7% in the control group to 27.8% +/- 10.9% in the treated group (p < 0.01). CONCLUSIONS:These studies demonstrate that emodin may inhibit cancer-cell growth by blocking VEGFR signaling and indicate that emodin can be used as a potential inhibitor for tumor angiogenesis.

背景与目标：

BACKGROUND & AIMS: :The angiogenic factor vascular endothelial growth factor (VEGF) predicts outcome in primary breast carcinoma. Alteration of the p53 gene causes down-regulation of the expression of thrombospondin-1, a natural inhibitor of angiogenesis. This study was conducted to investigate the association between mutant p53 protein and VEGF expression, and the prognostic value of these factors. VEGF165 and p53 protein were measured in tumour cytosols by enzyme immunoassays. Recurrence-free survival (RFS) and overall survival (OS) were estimated in 833 consecutive patients, 485 node-negative (NNBC) and 348 node-positive (NPBC) with primary invasive breast cancer. A significant association was found between mutant p53 protein and VEGF expression. Univariate analysis showed both p53 and VEGF to be significant predictors of survival. Similar correlation was seen when p53 was combined with VEGF. Univariate analysis of NNBC showed significant prognostic value of p53 for OS, also when combined with VEGF expression; for NPBC, significant reductions in RFS and OS were seen for p53-positive patients, and these findings were enhanced when combined with VEGF, also in the sub-group receiving adjuvant endocrine treatment. Multivariate analysis showed both p53 and VEGF as independent predictors of OS in all groups. When the 2 factors were combined, an increased relative risk of 2.7 was seen for OS in the group with both p53 positivity and high VEGF content, as compared with 1.7 in the group with one risk factor. The results suggest an association between loss of wt-p53 and increased VEGF expression, indicating that angiogenic activity may depend, at least partly, on altered p53-protein function. Combination of these 2 biological markers appears to give additional predictive information of survival. A high-risk group of patients was associated with p53 positivity and higher VEGF content.

背景与目标： 血管生成因子血管内皮生长因子 (VEGF) 可预测原发性乳腺癌的预后。p53基因的改变导致血管生成的天然抑制剂thrombospondin-1的表达下调。这项研究旨在研究突变型p53蛋白与VEGF表达之间的关系，以及这些因素的预后价值。通过酶免疫测定法在肿瘤细胞质中测量了VEGF165和p53蛋白。在833位连续患者中估计无复发生存率 (RFS) 和总生存率 (OS)，485淋巴结阴性 (NNBC) 和348淋巴结阳性 (NPBC) 原发性浸润性乳腺癌。发现突变型p53蛋白与VEGF表达之间存在显着关联。单因素分析显示p53和VEGF都是生存的重要预测指标。当p53与VEGF结合时，观察到类似的相关性。NNBC的单因素分析显示，p53对OS具有重要的预后价值，同时与VEGF表达相结合; 对于NPBC，p53-positive患者的RFS和OS显著降低，当与VEGF结合时，这些发现增强，同样在接受辅助内分泌治疗的亚组中。多变量分析显示，所有组中p53和VEGF都是OS的独立预测因子。当将这2个因素合并时，在p53阳性和高VEGF含量的组中，OS的相对2.7风险增加，而在具有一个危险因素的组中，与1.7相比。结果表明wt-p53损失与VEGF表达增加之间存在关联，表明血管生成活性可能至少部分取决于p53-protein功能的改变。这两种生物标记的组合似乎提供了生存的其他预测信息。高危组患者与p53阳性和较高的VEGF含量有关。

BACKGROUND & AIMS: :Vascular endothelial growth factor (VEGF) signaling is critical for both normal and disease-associated vascular development. Dysregulated VEGF signaling has been implicated in ischemic stroke, tumor angiogenesis, and many other vascular diseases. VEGF signals through several effectors, including the Rho family of small GTPases. As a member of this family, Rac1 promotes VEGF-induced endothelial cell migration by stimulating the formation of lamellipodia and membrane ruffles. To form these membrane protrusions, Rac1 is activated by guanine nucleotide exchange factors (GEFs) that catalyze the exchange of GDP for GTP. The goal of this study was to identify the GEF responsible for activating Rac1 in response to VEGF stimulation. We have found that VEGF stimulates biphasic activation of Rac1 and for these studies we focused on the peak of activation that occurs at 30 min. Inhibition of VEGFR-2 signaling blocks VEGF-induced Rac1 activation. Using a Rac1 nucleotide-free mutant (G15ARac1), which has a high affinity for binding activated GEFs, we show that the Rac GEF Vav2 associates with G15ARac1 after VEGF stimulation. Additionally, we show that depleting endothelial cells of endogenous Vav2 with siRNA prevents VEGF-induced Rac1 activation. Moreover, Vav2 is tyrosine phosphorylated upon VEGF treatment, which temporally correlates with Rac1 activation and requires VEGFR-2 signaling and Src kinase activity. Finally, we show that depressing Vav2 expression by siRNA impairs VEGF-induced endothelial cell migration. Taken together, our results provide evidence that Vav2 acts downstream of VEGF to activate Rac1.

背景与目标： 血管内皮生长因子 (VEGF) 信号对正常和疾病相关的血管发育都至关重要。VEGF信号异常与缺血性中风，肿瘤血管生成和许多其他血管疾病有关。VEGF通过几种效应器发出信号，包括Rho家族的小gtp酶。作为该家族的一员，Rac1通过刺激片状脂质和膜皱褶的形成来促进VEGF诱导的内皮细胞迁移。为了形成这些膜突起，Rac1被鸟嘌呤核苷酸交换因子 (gef) 激活，该因子催化GDP交换GTP。这项研究的目的是确定负责激活Rac1以响应VEGF刺激的GEF。我们已经发现VEGF刺激Rac1的双相激活，对于这些研究，我们专注于在30分钟时发生的激活峰。抑制VEGFR-2信号传导阻断VEGF诱导的Rac1激活。使用对结合活化的GEFs具有高亲和力的无Rac1核苷酸突变体 (G15ARac1)，我们表明Rac GEF Vav2在VEGF刺激后与G15ARac1缔合。此外，我们显示用siRNA耗尽内源性Vav2的内皮细胞可防止VEGF诱导的Rac1激活。此外，Vav2在VEGF处理后被酪氨酸磷酸化，这在时间上与Rac1激活相关，并且需要VEGFR-2信号传导和Src激酶活性。最后，我们显示siRNA抑制Vav2表达会损害VEGF诱导的内皮细胞迁移。综合而言，我们的结果提供了Vav2作用于VEGF下游激活rac1的证据。

BACKGROUND & AIMS: BACKGROUND AND OBJECTIVES:The application of microencapsulated stem cells has been shown to have many advantages in various fields of medical research. However, optimal modes for preparation of microencapsulate stem cells need to be improved, and expression and release of products of microencapsulated gene modified stem cells need to be studied in vitro. AIM OF THE STUDY:To explore the optimal parameters when preparing microencapsulated stem cells, and to investigate the effect of microencapsulation on growth, secretion, and metabolism of genetically modified human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs). MATERIALS AND METHODS:In this study, the parameters of preparation were regulated by observing the microcapsule shape and size. Live/dead cell viability kits and fluorescein isothiocyanate-labeled dextrans (FD) were used to detect the microencapsulated cell viability, and the permeability of microcapsules, respectively. Vascular endothelial growth factor (VEGF) production in the supernatant of microencapsulated and non-microencapsulated VEGF gene-modified hUCMSCs cultures was measured by ELISA. RESULTS:The optimal parameters of preparing microcapsules were regulated as followed: bolus velocity was 6 ml/h, and airflow velocity was 3 L/min. The morphology of microcapsules was a spherical structure with a diameter of 450 ± 30 µm. More than 90% of the cells were viable after 21 days of culture. Low and middle molecular weight FD was able to pass through the microcapsules; however, high molecular weight FD was not. Also, the VEGF concentration in microencapsulated and non-microencapsulated cell culture supernatants exhibited no significant difference at each time point. CONCLUSIONS:Microencapsulated stem cells can be ideally prepared via specifically regulated preparation. Lastly, microencapsulation does not alter growth, secretion, and metabolism of the genetically modified hUCMSCs.

背景与目标：

BACKGROUND & AIMS: :The objective of this study was to investigate the expressions of angiogenic factors and elucidate their angiogenic and prognostic roles in hepatocellular carcinoma (HCC) with background of hepatitis B virus (HBV). We evaluated microvessel density (MVD) of HCC, and investigated immunohistochemical expression of vascular endothelial growth factor (VEGF), angiopoietins (Ang-1 and Ang-2), and matrix metalloproteinases-9 (MMP-9) in 67 specimens of surgically resected HCC, which were all positive for hepatitis B surface antigen. We investigated the relationship between their expressions and clinicopathological factors or prognosis. The microvessel density (MVD) of tumor tissue and surrounding normal liver tissue was 93.1 +/- 43.8/mm2 and 30.4 +/- 14.8/mm2, respectively. The MVD of well-differentiated HCC was significantly less than that of poorly differentiated HCC. MVD was positively correlated with VEGF and Ang-2 expression (P = 0.0023 and 0.0265, respectively). There was less tumor recurrence in low Ang-2 and low MMP-9 group than high Ang-2 and/or high MMP-9 group (P = 0.002). In Cox regression model, portal vein thrombus and intrahepatic metastasis was the risk factors of tumor recurrence (P = 0.003 and 0.001, respectively). Our study showed that the expression of VEGF and Ang-2 were positively correlated with MVD. Ang-2 expression and/or MMP-9 expression might be a significant predictive factor for recurrence after resection in HCC patients with the background of HBV.

背景与目标： : 这项研究的目的是研究血管生成因子的表达，并阐明其在具有乙型肝炎病毒 (HBV) 背景的肝细胞癌 (HCC) 中的血管生成和预后作用。我们评估了HCC的微血管密度 (MVD)，并在67例手术切除的HCC标本中研究了血管内皮生长因子 (VEGF)，血管生成素 (Ang-1和Ang-2) 和基质metalloproteinases-9 (MMP-9) 的免疫组织化学表达。乙肝表面抗原。我们调查了它们的表达与临床病理因素或预后之间的关系。肿瘤组织和周围正常肝组织的微血管密度 (MVD) 分别为93.1 +/- 43.8/mm2和30.4 +/- 14.8/mm2。高分化HCC的MVD明显小于低分化HCC。MVD与VEGF和Ang-2表达呈正相关 (分别为P = 0.0023和0.0265)。低Ang-2和低MMP-9组的肿瘤复发率低于高Ang-2和/或高MMP-9组 (P = 0.002)。在Cox回归模型中，门静脉血栓和肝内转移是肿瘤复发的危险因素 (分别为P = 0.003和0.001)。我们的研究表明，VEGF和Ang-2的表达与MVD呈正相关。Ang-2表达和/或MMP-9表达可能是HBV背景的HCC患者切除后复发的重要预测因素。