Objective To construct the recombined DNA pcDNA3.1-hBMP-2 and transfect into human marrow stromal stem cells (MSCs) in vitro, and to explore theeffects of transfection on cellular proliferation and expression of vascular endothelial growth factor (VEGF). Methods The expression of human bone morphogenetic protein 2(hBMP-2) in these cells after transfection was determined by in situ hybridization and immunohistochemical analysis and Western blot analysis. The changes of cell proliferation were observed by flow cytometry. The effects of BMP-2 gene transfection on expression of VEGF in the cells were analyzed by in situ hybridization of VEGF cDNA probe. Results Stable expressionof hBMP-2 in pcDNA3.1-hBMP-2 transfected MSCs was confirmed in the levels of mRNA and protein.Cellular proportion in S period increased, which indicated that the synthesis of cell DNA increased. The expression of VEGF in the cells increased obviously. Conclusion With the help of lipofectamine, the pcDNA3.1-hBMP-2 were transfected into human MSCs successfully. hBMP-2 plays an important role in promoting cellular proliferation and vascular generation during bone repair.
Objective To explore a new method of treating early avascular necrosis of femoral head (AVNFH). Methods Sixty-nine New Zealand adult rabbitswith a mean weight of 2.8 kg after AVNFH presenting were randomly divided into three groups. In group A, deproteinized bone(DPB) combined with the recombinant plasmid pcDNA3.1/vascular endothelial growth factor 165(VEGF165) was implanted in the drilled channel of the necrotic femoral head. In group B, only DPB was implanted. In group C, channel was drilled without DPB or plasmid implanted. Femoral head specimens were obtained 3 days, 1, 2, 4, 8 and 16 weeks after operation. The expression of VEGF165 was examined by RT-PCR, Western blot and immunohistochemical techniques. X-ray testedbone formation generally. Angiogenesis and repair of the femoral head were observed by histological and histomorphometric analysis. Results In group A, the expressions of VEGF165 mRNA and protein were detected 3 days postoperatively, reached apex 1 week and lasted more than 3 weeks after implantation. The ratios of IOD of collagen type Ⅰ were 0.29±0.11, 0.55±0.13 and 0.67±0.10 IOD/μm2 respectively at 2, 4 and 8 weeks postoperatively and the ratios of IOD of new capillary vessels were 0.33±0.10and 0.57±0.16 IOD/μm2 respectively at 2, 4 weeks postoperatively in group A, showing statistically significant difference (Plt;0.01) when compared with groups B and D. X-ray test indicated much bone callus formed early. Conclusion Transfection of the VEGF165 gene can enhance local angiogenesis at early stage andDPBVEGF165 compound can improve bone formation. Deproteinized bone combined with VEGF165 gene provides a potential method for therapy of osteonecrosis.
Objective To evaluate the effect of vascular endothelial growth factor (VEGF) on tumor angiogenesis, and its usage in tumor therapy.Methods The recent literatures about VEGF and angiogenesis were reviewed and analyzed. The advances of VEGF study were summarized. The effects of anti-angiogenesis in tumor biological therapy were introduced.Results Angiogenesis had been identified as an important factor for promoting tumor growth. VEGF was a basic and pivotal factor in tumor angiogenesis. The anti-angiogenesis treatments aimed at VEGF, including the applications of VEGF inhibitor and gene therapy of adenovirus medium, had got great progress. Conclusion VEGF is a leading factor of tumor angiogenesis, the anti-angiogenesis therapy aimed at VEGF has probably provided a new chance to malignant tumor treatment.
For research the relationship between the expression of vascular endothelial growth factor (VEGF) and the cell proliferation. The expression of VEGF was evaluated while the cell cycle of hepatoma cell line Hep G2, which was synchronized at G0 phase with serum deprivation, and reinitiated with TPA and blocked with antisense oligonucleotides of c-jun. Results: Hep G2 cell did not express VEGF at G0 phase. TPA could induce the expression of VEGF as well as initiation of cell cycle. The antisense oligonucleotides of c-jun could prohibit the expression of VEGF and arrest the cell cycle at G0 phase. Conclusion: The fact that the expression of VEGF accompanies the initiation of cell cycle suggests that they be regulated by the same singnal pathway, the expression of VEGF may be a marker indicating the proliferation of hepatoma cells.
Objective To investigate the synergistic effect of a combination of grafted olfactory ensheathing cells (OECs) from the olfactory bulbs and intrathecal injection of vascular endothel ial growth factor (VEGF) on repairing spinal cord injury, and to explore the neuroprotection on both neurons and nerve fibers. Methods OECs from neonatal rats were cultured, purified, and collected with 0.25% trypsin after 9 days. A total of 75 adult female Wistar rats (weighing 200-250 g) were randomly divided into 5 groups: group A was sham-surgery group receiving laminectomy; the spinal cord injury model was establ ished with weight-dropped apparatus in the rats of groups B, C, D, and E. Then group B was injected with 10 μL DMEM-F12 medium without serum at injury site on the 1 day and was intrathecally administrated with 10 μL sal ine solutiontwice a day during the following 1 week; group C was injected with 10 μL DMEM-F12 medium and 25 ng recombined ratVEGF165 (rrVEGF165); group D was injected with 10 μL DMEM-F12 medium containing 1 × 105 OECs and 10 μL sal ine solution; group E was injected with 10 μL DMEM-F12 medium containing 1 × 105 OECs and 25 ng rrVEGF165. The functional recovery of hindl imb was evaluated by the Basso-Beattie-Bresnahan (BBB) score at 1 day and each week from 1 to 8 weeks. The histological changes and the changes of ultrastructure were observed at 8 weeks after operation by HE and electron microscope, and the immunohistochemistry staining was used for p75 nerve growth factor receptor (p75NGFR), Caspase-3, and von Willebrand factor (vWF). Results The function of hindl imb recovered rapidly in group E; the BBB score reached the peak at 8 weeks, and it was significantly higher than those in other groups (P lt; 0.05). The histology and ultrastructure observation showed that nerve fibers and neurons were damaged seriously in group B, oderately in groups C and D, and sl ightly in group E. Numerous spared tissue between nerve stumps, fibers with regular myel ination, and neurons with l ittle vacuolar mitochondria were observed in group E. The immunohistochemistry staining revealed that Caspase-3 positive cells in groups B, C, D, and E were significantly more than that in group A (P lt; 0.05); more Caspase-3 positive cells were found in groups B and D than in groups C and E (P lt; 0.05), while no significant difference was found between groups C and E (P gt; 0.05). And more vessels per high field were examined in groups C and E than in groups A, B, and D (P lt; 0.05), while no significant difference was found between groups C and E (P gt; 0.05). The p75NGFR positive results showed the survival of OECs in groups D and E at 8 weeks after OECstransplantation. Conclusion Grafted OECs combined with intrathecal injection of VEGF has significant promotive effects on restoration of spinal cord injury in rats, can improve part function of nerve fibers, and shows neuroprotection on damaged cells and fibers, which have a synergistic effect.
Objective To investigate the effect of small interfering RNA(siRNA) targeting hypoxia inducible factor1alpha; (HIF1alpha;) and vascular endothelial growth factor (VEGF) on expression of VEGF in human vascular endothelial cells. Methods HIF-1alpha; siRNA recombinant plasmid was constructed. Human vascular ndothelial cells were cultured in vitro and divided into normoxia group (20% O2) and hypoxia group (1% O2). Hypoxia group was then divided into control group, vector group, HIF-1alpha; group (HIF-1alpha; siRNA), VEGF group ( VEGF165 siRNA) and cotransfection group (HIF-1alpha; siRNA+VEGF165 siRNA). LipofectamineTM 2000 (LF2000) mediated vector plasmid was transfected to cells in each group except the control group. The expression of HIF-1alpha; siRNA and VEGF165 siRNA recombinant plasmid were identified by reverse transcriptasepolymerase chain reaction (RT-PCR). The expression of VEGF mRNA and protein were detected by RTPCR and immunocytochemical method. Results The expression of HIF-1alpha; siRNA and VEGF165 si RNA recombinant plasmid were detected 24 hours after transfected. The expression of VEGF mRNA and protein was faint in the normoxia group, but increased obviously in hypoxia group. The expression of VEGF mRNA and protein in the HIF1alpha;, VEGF and cotransfection groups were lower than which in the control group. Cotransfection group showed the highest inhibitory effect. Conclusion HIF-1alpha; and VEGF165 siRNA can effectively inhibit the expression of VEGF in human vascular endothelial cells.
Abstract: Objective To evaluate the significance of expression of vascular endothelial growth factor-C （VEGF-C） and cytokeratin 19 （CK19） in patients with stage I non-small cell lung cancer （NSCLC）. Methods A total of 269 patients with NSCLC who underwent standard lobectomy and lymph node dissection by the same surgical team in our hospital from January 2004 to June 2005 were included in this study. All the clinical data and follow-up results were complete, and all the pathological specimens were well kept. No preoperative or postoperative adjuvant therapy such as radiotherapy and chemotherapy was administered to those patients. Expressions of VEGF-C in cancer tissues was detected by immunohistochemical streptavidin-peroxidase （S-P） method, and CK19 was marked to examine micrometastasis in hilar and mediastinal lymph nodes. Clinical outcomes, pathological results and follow-up data were analyzed in combination with VEGF-C and CK19 expression. Results VEGF-C expression was not statistically different between different category in sex（Hc=1.722，P=0.084）, age （Hc=0.914，P=0.360）, smoking （Hc=2.440，P=0.295）, pathology type （Hc=5.668，P=0.058）or tumor size （Hc=0.165，P=0.920） . VEGF-C expression was statistically different between different groups of pathological differentiation （Hc=29.178，P=0.000). CK19 expression was not statistically different between different category in sex（χ2=0.000，P=0.999）, age （χ2=0.005，P=0.999）, smoking （χ2=2.294，P=0.317）, pathology type （χ2=0.573，P=0.289）, tumor size（χ2=0.006，P=0.999）, and pathological differentiation （χ2=2.927，P=0.231）. Five-year survival rate was statistically different between different grade of VEGF-C expression （χ2=37.318，P=0.000), and was also statistically different between positive group and negative group of CK19 （χ2=39.987，P=0.000). There was statistical difference between different grade of VEGF-C expression and positive rate of CK19 （χ2=25.954，P=0.000). Conclusion Expression of VEGF-C and CK19 is closely related to postoperative 5-year survival of patients with stage I NSCLC. Detection of VEGF-C and CK19 is of great clinical significance as it is helpful to predict patient prognosis and choose proper postoperative adjuvant therapy.
Esophageal cancer is one of the common malignant tumors with high incidence and poor prognosis. Angiogenesis-related pathways play an important role in the occurrence and development of esophageal cancer. Vascular endothelial growth factor (VEGF) is the main mediator of angiogenesis. In addition to promoting angiogenesis and maintaining the survival of neovascularization, VEGF can also directly act on esophageal cancer cells and promote the occurrence and development of tumors. This article reviews the biology of VEGF and its effect on blood vessels, the expression of VEGF in esophageal cancer cells and its influencing factors, the role of VEGF in esophageal cancer cells, the immunomodulatory activity of VEGF and the clinical study of VEGF inhibitors. The purpose of this study is to provide a basis for more rational use of VEGF inhibitors in the treatment of esophageal cancer.
ObjectiveTo observe the expression of vascular endothelial growth factor (VEGF) and aquaporin 4 (AQP4) in the inner limiting membrane (ILM) of diabetic retinopathy (DR) with macular edema, and analyze the correlation between VEGF and AQP4 expression. Methods A cross-sectional study. From September 2019 to September 2020, 38 eyes of 38 patients with DR and idiopathic macular hole (iMH) who underwent vitrectomy (PPV) combined with ILM stripping at the Hangzhou campus of The Affiliated Eye Hospital of Wenzhou Medical University at Hangzhou were included in the study. Among them, there were 25 males and 13 females who aged 37-76 years old, average age was 59±10 years old; All eye included 15 right eyes and 23 left eyes. iMH and DR included 9 eyes in 9 cases and 29 eyes in 29 cases, respectively, and they were divided into iMH group and DR group. The DR group was divided into DME group and no DME group according to whether it was accompanied by diabetic macular edema (DME), with 14 eyes and 15 eyes respectively. After the stripped ILM tissue was fixed, immunofluorescence analysis was performed to obtain a picture of the fluorescence mode of AQP4 and VEGF, and the fluorescence intensity value of VEGF and AQP4 was measured by Image J software. The differences of VEGF and AQP4 immunofluorescence values in the specimens between groups were compared by one-way analysis of variance. The correlation between the fluorescence intensity of AQP4 and the fluorescence intensity of VEGF was analyzed by Pearson correlation analysis. Results The average fluorescence intensity valuesof VEGF and AQP4 in ILM specimens of DME group, no DME group and iMH group were 38.96±7.53, 28.25±3.12, 30.07±4.84 and 49.07±8.73, 37.96±6.45, 38.08±5.04, respectively. The average fluorescence intensity of VEGF and AQP4 in the ILM specimens of the DME group was significantly higher than that of the no DME group and iMH group, and the difference was statistically significant (F=13.977, 9.454; P＜0.05). The average fluorescence intensity values of VEGF and AQP4 on IML specimens in the DR group were 33.80±7.91, 43.76±9.44, respectively. The results of Pearson correlation analysis showed that the fluorescence intensity of VEGF and AQP4 in the ILM specimens of the DR group was significantly positively correlated (r=0.597, P=0.003). ConclusionsThe expressions of VEGF and AQP4 in ILM of eyes with DR and DME are significantly increased compared with those without DME. The expression of VEGF and AQP4 in ILM of eyes with DR is positively correlated.
ObjectiveTo detect the expressions of microvessel density (MVD)-CD34 and vascular endothelial growth factor (VEGF) in hepatic alveolar hydatid tissue in gerbil model and explore their clinical significances. MethodsSixty health gerbils were randomly equally divided into two groups, an experimental group and a sham operation group, each gerbil was given liver vaccination by opening their abdominal. Each gerbil in the experimental group was injected with approximately 400 echinococcus protoscoleces (0.1 mL), and each gerbil in the sham operation group received a corresponding volume of physiological saline. Six gerbils were sacrificed on day 20, 40, 60, 80, and 100. The hepatic alveolar hydatid tissue (AE) and its surrounding liver tissue (HSAE) were collected from the experimental group and the normal liver tissue (NL) was collected from the sham operation group, and the expressions of MVD-CD34 and VEGF were detected by immunohistochemistry staining (EnVision method). ResultsEchioncoccus multilocularis hydatid tissues were observed over the liver and in the partly abdominal cavity in the experimental group each gerbil by general observation. The expressions of CD34 and VEGF were observed in the AE at each time point after infection and located in the cytoplasmic of endothelial cells. The number of MVD-CD34 of AE at each time point in the AE was (9.83±3.87)/HP, (25.33±6.71)/HP, (34.50±5.50)/HP, (37.67±5.71)/HP and (44.67±4.93)/HP, respectively, which were significantly higher than those in the HSAE〔0/HP, (1.17±0.98)/HP, (3.50±1.38)/HP, (5.83±2.71)/HP, and(8.83±2.48)/HP, respectively〕and NL (all were 0), P < 0.05. The point of VEGF at each time point in the AE was 2.95±0.46, 3.90±0.68, 4.27±1.05, 5.33±0.95, and 4.50±0.81 respectively, which were significantly higher than those in the HSAE(1.07±0.63, 1.38±0.75, 1.55±0.83, 1.67±0.47, 2.10±0.55, respectively) and NL (1.02±0.83, 1.12±0.63, 1.26±0.26, 1.20±0.74, 1.21±0.28), P < 0.05. ConclusionAngiogenesis might be involved in infiltrated growth of alveococcus, and VEGF might contribute to angiogenesis of alveolar hydatid tissue.