目的 通过复制人肝癌细胞株HepG2裸鼠皮下移植瘤模型，观察绿茶提取物表没食子儿茶素没食子酸酯（EGCG）干预对HepG2移植瘤新生血管生成的影响。 方法 瘤体接种复制HepG2移植瘤模型，荷瘤裸鼠20只随机分组，实验组给予EGCG溶液每日20 mg/（kg·只），腹腔注射3周，对照组给予等量灭菌注射用水3周，末次用药24 h，后处死裸鼠，剥离移植瘤。常规病理切片观察移植瘤组织结构；逆转录-聚合酶链式反应和免疫组织化学法检测移植瘤缺氧诱导因子-1α（HIF-1α）、血管内皮生长因子（VEGF）mRNA及蛋白表达，并通过检测CD34表达计数瘤组织微血管密度（MVD）。 结果 组织病理学观察实验组移植瘤见大量坏死区，瘤体内血管数量明显少于对照组；实验组HIF-1α、VEGF mRNA及蛋白表达水平比对照组均明显下调（P＜0.05），实验组MVD比对照组明显下降（P＜0.05）。 结论 EGCG可抑制荷瘤裸鼠HepG2移植瘤新生血管生成。
Objective To investigate the effect of the vascular endothelial growth factor (VEGF) gene therapy, the surgical delay, and the combination of the two therapeutic approaches on the survival of the rat over-area abdominal axial skin flap. Methods In 48 male Wistar rats (weight, 400-450 g), a model of the abdominal axial skin flap supplied by the superficial epigastric blood vessel was created. The rats were randomly divided into 6 groups: Group A (the blank group), Group B (the gene-therapy-during-operation group), Group C (the gene-therapy-before-operation group), Group D (themerely-surgical-delay group), Group E (the gene-therapy-during-surgical-delay group), and Group F (the gene-therapy-aftersurgical-delay group). Seven days after operation, the survival rate of the skin flap was measured; the specimens were harvested from the skin flap for a histological investigation of themicrovessels and for an immunohistochemical staining to observe the expression of VEGF165. Results The average survival rate of the skinflap was significantly greater in each of the treated groups than in Group A (Plt;0.05); the rate was the greater in Group E (Plt;0.05), but with no statistically significant difference between the other treated groups (Pgt;0.05). The average number of the microvessels was significantly greater in Groups B, C, E andF than in Groups A and D (Plt;0.05), but with no statistically significant difference between Groups B, C, E and F and between Groups A and D (Pgt;0.05). The lumen diameter of the microvessels was significantly greater in Group D than in Groups E and F (Plt;0.05), and the diameter was significantly greater in Groups D, E andF than in the other groups (Plt;0.05). More deposition of VEGF DNA detected by the immunohistochemical staining was in Groups B, C, E and F than in Groups A and D. There was no newly-formed blood vessel in the rat cornea in the treated groups.Conclusion Both the administration of pcDNA4-VEGF165 and the surgical delay can improve the survival of the rat abdominal axial skin flap, but the mechanism of the effect is different in explanation. The combination of the two therapeutic approaches can achieve a better effect.
【Abstract】Objective To investigate the change of vascular endothelial growth factor (VEGF) expression in HepG2 cells under hypoxia. Methods HepG2 cells were cultured under hypoxia（hypoxia group） and normal condition (control group). VEGF expression of HepG2 cells was examined by immunohistochemical staining. The growth of HepG2 cells was examined by MTT colorimetry and cell count. VEGF level in the culture medium was measured by ELISA.Results After 48 h and 72 h of culture, the growth rate of HepG2 cells in hypoxia group was lower than that in control group (P＜0.05). The cell count in hypoxia group (2.51×104/μl and 2.69×104/μl, respectively) was much lower than that in control group(3.01×104/μl and 3.52×104/μl) after 48h and 72h of culture (P＜0.05). In hypoxia group, VEGF level in the culture medium after 24 h and 48 h was higher than that in control group (P＜0.05, P＜0.01). Conclusion Hypoxia may enhance the VEGF expression in HepG2 cells and this could be the reason of high expression of VEGF after transcatheterized hepatic arterial chemoembolization.
Objective To observe the degradation of the polyactic glycolate acid (PLGA) microparticles with releasing-slowly vascular endothelial growth factor(VEGF) synthesized by the method of emulsification-diffusion. Methods The method of emulsification-diffusion is to incorporate VEGF into microparticles composed of biodegradable PLGA. The controlled release of microparticles are acquired. The content of the VEGF released slowly from PLGA microparticles in vitro was detected with ELISA at different time. Results We synthesized 100 releasing-slowly VEGF PLGA microparticles with the size of 0.20-0.33 μm by 5 times. The contents were 62±11 ng/L, 89±14 ng/L, and 127±19 ng/L in the 1st, the 2nd and the 3rd months after degradation, respectively. Conclusion The PLGAmicroparticles with releasing-slowly VEGF can be synthesized by the method of emulsification-diffusion.
ObjectiveTo observe the serum vascular endothelial growth factor (VEGF), apelin and heme oxygenase-1 (HO-1) levels in patients with type 2 diabetes mellitus (T2DM) and to explore their their relationship with diabetic retinopathy (DR).MethodsA total of 208 patients with T2DM and 50 healthy subjects (control group) from the Central Hospital of Western Hainan during January 2014 and December 2017 were selected in this study. Vision, slit lamp microscope, indirect ophthalmoscope and FFA examinations were performed on all the subjects. According to the results of the examinations combined with the DR clinical staging criteria, the patients were divided into non-DR (NDR) group, non-proliferative DR (NPDR) group, and proliferative DR (PDR) group, with 72, 76 and 60 patients in each, respectively. The clinical data of each group were recorded, and the levels of fasting blood glucose (FPG), HbA1c, total cholesterol (TC), three acylglycerol (TG), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), VEGF, apelin and HO-1 were detected in each group. The receiver operating characteristic curve (ROC) were used to analyze the value of VEGF, apelin and HO-1 in predicting the occurrence of PDR. Correlation analysis of serum VEGF, Apelin and HO-1 with clinical parameters in PDR patients by Pearson correlation analysis.ResultsThe level of VEGF (56.82±10.16 vs 91.74±22.83, 140.15±36.40, 195.28±42.26 pg/ml) and apelin (2.95±0.53 vs 4.68±0.74, 7.25±1.13, 10.16±1.35 ng/ml) in PDR group were significantly higher than those in NPDR, NDR and control groups (F=17.306, 21.814; P＜0.05). The level of HO-1 (50.37±10.14 vs 43.58±8.16, 30.25±6.28, 22.60±4.72 mmol/L) in PDR group was significantly lower than those in NPDR, NDR and control groups (F=15.827, P＜0.05). The ROC curve analysis showed that the best cut-off values of serum VEGF, apelin and HO-1 were 162.50 pg/ml, 8.30 ng/ml, 27.13 mmol/L, and the three combined to predict PDR of AUC (95%CI) was 0.906 (0.849−0.962), and their sensitivity (90.3%) and specificity (83%) were better. The correlation analysis showed that the VEGF, apelin and HO-1 of PDR patients were correlated with the course of diabetes (r=0.382, 0.416, −0.36; P＜0.05), FPG (r=0.438, 0.460, −0.397; P＜0.05) and HbAlc (r=0.375, 0.478, −0.405; P＜0.05), and the serum VEGF were correlated with apelin and HO-1 (r=0.793, −0.594; P＜0.01).ConclusionElevated serum VEGF and apelin levels and reduced HO-1 levels are associated with the progression of DR, and the three combination helps predict the occurrence of PDR.
Objective To observe the effect of RNA interference (RNAi) on HepG2 hepatic cancer cell by small interfering RNA (siRNA). Methods siRNA targeting vascular endothelial growth factor (VEGF) gene was transfected into HepG2 cells by LipofectimineTM 2000. The VEGF mRNA and protein were respectively detected by real-time quantitive PCR and Western blot, and the concentration of VEGF protein in the cell culture supertant was determined by ELISA at 48 h after culture. Results The average efficiency of siRNA transfection was (90.4±2.9)% after 6 h cell culture. The expressions of VEGF mRNA and protein in HepG2 cells could be effectively suppressed by siRNA, and the concentration of VEGF protein in the cell culture supertant was also decreased. Conclusion siRNA can knock down the expression of VEGF gene and decrease the concentration of VEGF protein in HepG2 cells.
【Abstract】Objective To understand the features of lymphatic vessel, and to summarize the foundation and mechanism of the promotion and inhibition of tumor lymphangiogenesis recorded on the current studies of animal experiments and clinical researches. Methods The related literatures of the structural features of lymphatic vessel, lymphatic endothelial molecular markers, the origin of lymphatic tumors, the molecular mechanisms and regulatory factors were reviewed, and the relationship between tumor lymphangiogenesis and lymphatic metastasis, the treatment targeting at the formation of the anti-tumor lymphatic vessel and its existing problems were also analyzed. Results Hyperplasia of lymphatic vessels occurred during the process of tumor formation and progression. The structural features of the lymphatic vessels in the tumor were conducive to tumor lymphatic metastasis. In recent years, methods of anti-lymphangiogenesis and inhibition of tumor lymphatic metastasis had achieved considerable success in animal experiments. However, there were still a lot of problems need to be solved. Conclusion Tumor lymphangiogenesis has a significantly positive correlation between tumor lymphatic metastasis and patients’ prognosis, which may indicate that treatment against the formation of tumor lymphatic vessel maybe effective.
【Abstract】Objective To evaluate the status of vascular endothelial growth factor (VEGF) expression in breast carcinoma and benign disease and define the relationship with age,menopause, tumor size，clinical stage，distant metastasis and lymph node metastasis. Methods Seventy cases of invasive ductal breast carcinomas,30 benign breast diseases and 7 adjacent nonneoplastic specimens were assessed for VEGF protein expression by immunohistochemistry LSAB method. Results VEGF were expressed more frequently in breast cancer than in benign diseases.VEGF was significantly correlated with axillary lymph node metastasis and distant metastasis,whereas no statistical correlation with other factors. Conclusion VEGF status has certain value to make differential diagnosis between malignant and benign breast diseases and predict the possibilities of distant and lymph node metastasis.
Objective To establish the overall diagnostic accuracy of the measurements of vascular endothelial growth factor (VEGF) for malignant ascites. Methods After a systematic review of current studies, sensitivity, specificity, and other measures of the value of ascites concentrations of VEGF in the diagnosis of malignant ascites were pooled by using random effects models. Qualified studies on evaluation of VEGF in diagnosis of malignant ascites in English and Chinese published from January 1990 to December 2009 were retrieved from The Cochrane Library, Cochrane Central Register of Controlled Trials, MEDLINE, EMbase, China National Knowledge Infrastructure (CNKI) databases, WanFang Data, and VIP Information. Two reviewers independently assessed the methodological quality of each study with the tool of QUADAS. Statistical analyses were performed by employing Meta-Disc 1.4 software. Meta-analyses of the reported sensitivity and specificity of each study and Summary Receiver Operating Characteristic (SROC) curve were performed. Results Seven studies met the inclusion criteria for the analysis. After testing the heterogeneity of the included studies, a random effect model was selected to calculate the pool weighted sensitivity and specificity with 95% confidence interval: the sensitivity was 0.81 (95%CI 0.75 to 0.85), the specificity was 0.90 (95%CI 0.86 to 0.94), the DOR was 50.45 (95%CI 28.37 to 89.73), and the AUC of SROC was 0.9507 (SE=0.013 0). The subgroups were analyzed to identify the sources of heterogeneity according to race and agent sources. There was homogeneity among the three studies with agents from Ramp;D company (χ2=0.05, P=0.9750; I2=0.0%), and the AUC of SROC were 0.9675 (SE=0.016 7). Conclusion VEGF has a highly accurate sensitivity and specificity with a b ROC curve, which makes it a new marker to differentiate malignant ascites from the benign.