Objective To study the effect of vascular endothelial cell growth factor (VEGF) on repair of bone defect with cortical bone allograft. Methods Forty five New Zealand white rabbits, weighted 2.5-3.0 kg, were made bone defect model of 1.5 cm in length in the bilateral radii and then were randomly divided into 3groups. The defect was repaired with only cortical bone allograft in the control group, with the cortical bone allograft and local injection of human recombinantVEGF in the experimental group, and with the cortical bone allograft and abdominal injection of VEGF PAb3 in the antagonist group. Roentgenography, immunohistochemical staining and tetracycline labelling were carried out to evaluate the reparative results 1, 3, 5, 8 and 16 weeks after operation. Results Immunohistochemical staining results showed that a great deal of blood vessels formed in the experimental group, and the number of blood vessels increased gradually with the time and reached the highest value at the 8th week. Tetracyclinelabelling showed the same result.The best results in callus formation, ossification rate and count of microvascular density were shown in the experimental group, while those in the control group were significantly better than those in the antagonist group (Plt;0.05),but there was no significant difference between the experimental group and the control group at the 8th week and the 16th week (Pgt;0.05). Conclusion VEGF can accelerates the bone formation and angiogenesis in the bone allografts, thus it can promote the repair of bone defects.
In order to study the effect of vascular endothelial cell growth factor (VEGF) on the survival of skin flap 30 SD rats were used. A randomized flap measuring 7.5 cm x 3.0 cm was created on the back of each SD rat. The treatment group (n = 10) received VEGF 40 ng/flap by subcutaneous injection with microinjector during and 24 hours after operation. The control groups received heparin 16 U/flap (n = 10) or normal saline 800 microliters/flap (n = 10). After operation, on the 3rd and 11th day, the survival rate of the skin flaps and the dermovascular density of each flap were investigated by histological and histo-morphometrical examination. The results showed that there was no significant difference in the survival rate between the treatment group and the controls on the 3rd day after operation, while on the 11th day, there was a significant difference between them, and the survival rate was much higher in the treatment group. Besides, dermovascular density was much more increased in the treatment group than that in the controls, especially in the distal 1/3 of the flap (P lt; 0.02). The conclusion was that VEGF could .
ObjectiveTo investigate the expression of vascular endothelial growth factor （VEGF） and proliferating cell nuclear antigen (PCNA) in colorectal cancer and its relationship with metastasis and recurrence. MethodsParaffinembedded specimens from 59 patients with colorectal cancer, 16 patients with adenomas and 12 normal colonic tissues were examined and compared by SP immunohistochemical method. ResultsThe positive rate of VEGF in colorectal cancer were significantly higher than that in adenomas (P＜0.05). The positive rate of VEGF in Dukes A and B stage of colorectal cancer were significantly higher than those in Dukes C and D (P＜0.05). Expression of VEGF in postoperative recurrence group was markedly higher than that in the group with no recurrence (P＜0.05). Proliferative activity expression suggested that the poorer the differentiation, the more PCNA increased in case of lymphnode or hepatic metastasis. The PCNA showed marked difference between postoperative and nonpostoperative recurrences (P＜0.05). Conclusion The expression of VEGF and PCNA is closely related to the invasion and metastasis of tumor during the operation. The increased VEGF and high PCNA implies that there may be some potential metastasis present.
Abstract: Objective To study the pathophysiological mechanism of the morphological change of immature pulmonary vessels in the piglet model of congenital heart defect with decreased pulmonary blood flow established with balloon atrial septostomy and pulmonary artery banding. Methods Twenty piglets at an age of one to two months were divided into three groups with random number table. For the control group (group C,n=6), small incisions were carried out on the right chest to produce a transient reduction in the pulmonary blood; for the lowmedium pulmonary artery stenosis group (group T1, n=7), the balloon dilator was delivered through the surface of the right atrium and septostomy and pulmonary artery banding were performed, and the systolic transpulmonary artery banding pressure (Trans-PABP) was controlled to be 20.30 mm Hg; For the severe pulmonary artery stenosis group (group T2, n=7), the same surgical procedures with group T1 were performed while TransPABP was controlled to be more [CM(159mm]than 3050 mm Hg.At 2 months after surgery respectively,a lung tissue of 1.0 cm×0.8 cm×0.8 cm from the lateral segment of the right middle lobe was taken out to be observed under optic microscope. The morphological change of the distal arterioles was detected. Furthermore, the content of vascular endothelial growth factor (VEGF) and matrix metalloproteinase2( MMP2) were also examined by the method of enzymelinked immunosorbent assay (ELISA). Results The model was successfully established in all the survival piglets of the group T1 and group T2. Two months after operation, the inner diameter of the pulmonary arterioles in group T1 was significantly higher than that in group C (82.89±10.72 μm vs.74.12±9.28 μm;t=－5.892, Plt;0.05), so as group T2 (85.47±5.25 μm vs.74.12±9.28 μm;t=-6.325, Plt;0.05); the number of arterioles per square centimeter (NAPSC) of group T1 was significantly lower than that of the group C (229.70±88.00 entries/cm 2 vs. 431.50±40.60 entries/cm2; t=39.526, Plt;0.05), so as group T2 (210.00±40.30 entries/cm2 vs. 431.50±40.60 entries/cm2; t=67.858, Plt;0.05). Two months after operation, the lung expression of MMP -2 and VEGF in group T1 was significantly lower than that in group C (58.30±19.60 ng/ml vs. 81.20±16.70 ng/ml, t=14.261, Plt;0.05; 17.80±3.00 pg/ml vs. 21.40±3.80 pg/ml, t=8.482, P＜0.05), so does group T2 (42.10±15.20 ng/ml vs. 81.20±16.70 ng/ml, t=27.318, P＜0.05; 12.30±3.20 pg/ml vs. 21.40±3.80 pg/ml, t=15.139, P＜0.05). Conclusion Structural remodeling of pulmonary extracellular matrix is an important feature of the piglet model of congenital heart defect with decreased pulmonary blood flow. The arterioles show significant hypoplasia or degradation. Change in the structural proteins and cytokines during the reduction of blood in the lung is the key to structural remodeling.
Objective To observe the effects of vascular endothelial growth factor antisense oligonucleotide (VEGF-ASODN) on expression of vascular endothelial growth factor (VEGF) and growth in gastric cancer cells. Methods The VEGF-ASODN was synthesized artificially with phosphorothioic acid. After transfecting with VEGF-ASODN in gastric cancer cells SGC-7901, the initial copy number of mRNA was detected by real-time RT-PCR, and the quantity of VEGF protein in both cell and supernatant were detected by ELISA. The levels of expression of survivin protein in cells were measured by Western blot. FCM and MTT method were used to detect cellular apoptosis and the activity of cells, respectively. The effect of transfection on the growth of cells was evaluated by growth curve. Results The copy number of VEGR mRNA, protein levels of VEGF in the cells and in culture fluid all decreased when the concentration of transfected VEGF-ASODN increased, as well as the levels of survivin protein （P＜0.05）. The ratio of apoptosis increased, the activity of cells also decreased as the concentration of transfected VEGF-ASODN increased （P＜0.05）. Conclusion Transfection with VEGF-ASODN in gastric cancer cells SGC-7901 can inhibit the expressions of VEGF and survivin remarkably. It can enhance cellular apoptosis and suppress growth of cells.
OBJECTIVE To review the fundamental research and the experimental study of vascular endothelial growth factor (VEGF). METHODS The laboratory information and experimental study of VEGF were extensively reviewed. RESULTS VEGF, as a mitogen specifically for endothelial cells, could bly stimulate angiogenesis as well as vascular permeability, especially in hypoxia state. Experimentally, survival area and viability of flaps could be increased significantly when exogenous VEGF was applied in the form of protein or cDNA. CONCLUSION VEGF provides a new way for therapeutic angiogenesis in reparative and reconstructive surgery.
Objective To observe the impact of collagen patches using 1-ethyl-3- （3-dimethylaminopropyl) carbod-iimide hydrochloride chemistry （EDC） to conjugate vascular endothelial growth factor （VEGF） ＋ basic fibroblast growth factor （bFGF） or VEGF alone on the survival rate of transplanted human bone morrow mesenchymal stem cells （hBM-MSCs）in vitro and in vivo. Methods Collagen patches which were activated by EDC were used as the control group，and EDC activated collagen patches that were conjugated with VEGF or VEGF ＋ bFGF were used as the experiment groups（VEGF group and VEGF ＋ bFGF group）. hBM-MSCs （0.5×106/patch） were used as seeding cells to construct engineered heart tissue （EHT）. MTT assay was performed to assess in vitro proliferation of hBM-MSCs on 3 different collagen patches. Ventricular aneurysm model after myocardial infarction was created by left anterior descending artery （LAD） ligation in male SD rats，and EHT which were constructed with 3 different patches were used for ventricular plasty. Four weeks later，immunofluorescence staining was used to examine arteriole density （anti-α-SMA staining） and transplanted cell survival （anti-h-mitochondria staining）. Results （1） hMSCs proliferation in VEGF group and VEGF ＋ bFGF group was significantly better than that in the control group on the 2nd and 4th day after cell transplantation （P＜0.05）； （2） Four weeks afterEHT implantation，immunofluorescence staining for α-SMA revealed that arteriole density of VEGF group and VEGF ＋ bFGF group was significantly higher than that of the control group （P＜0.05）； （3） Immunofluorescence staining forh-mitochondria showed that survival rates of transplanted hBM-MSCs of VEGF group and VEGF ＋ bFGF group were significantly higher than that of the control group （P＜0.05）； （4） There was a significantly positive correlation between survival rate of hBM-MSCs and arteriole density （r 2=0.99，P=0.02）. Conclusion VEGF or VEGF ＋ bFGF conjugated collagen patch can significantly improve hBM-MSCs proliferation in vitro and enhance survival rate of transplanted hBM-MSCs by accelerating revascularization of EHT in vivo.
ObjectiveTo investigate the role of protease-activated receptor-2 (PAR-2) activation on the expression of vascular endothelial growth factor (VEGF) in MKN28 gastric cancer cells. Methods①MKN28 cells were treated with increased concentrations of trypsin (0, 0.1, 1.0, 10.0, and 100.0 nmol/L respectively) for 6 hours, or treated with 10.0 nmol/L trypsin for 3, 6, 12, and 24 hours (blank control group was treated with PBS) respectively, then the expression levels of VEGF mRNA and its protein in MKN28 cells were detected by real-time reverse transcription polymerase chain reaction (qRT-PCR) and Western bolt method, with the concentration of VEGF protein in broth was detected by enzyme linked immunosorbent assay (ELISA) method.②MKN28 cells were divided into blank control group (treated with PBS), trypsin group, trypsin+PD98059 group, trypsin+SB203580 group, PD98059 group, and SB203580 group, then the expression levels of VEGF mRNA and its protein in MKN28 cells were detected by qRT-PCR method and Western bolt method respectively. Results①The effect of different concentration of trypsin. Compared with blank control group, the expression levels of VEGF mRNA and its protein in 0.1, 1.0, 10.0, and 100.0 nmol/L group were higher (P < 0.05); compared with 0.1 nmol/L group, the expression levels of VEGF mRNA and its protein in 1.0, 10.0, and 100.0 nmol/L group were higher (P < 0.05); compared with 1.0 nmol/L group, the expression levels of VEGF mRNA and its protein in 10.0 and 100.0 nmol/L group were higher (P < 0.05); but there was no significant difference between 10.0 nmol/L and 100.0 nmol/L group (P > 0.05). The broth concentration of VEGF protein in blank control group, 0.1, 1.0, 10.0, and 100.0 nmol/L group crept upward (P < 0.05).②The effect of different treated time of 10.0 nmol/L trypsin. The expression levels of VEGF mRNA in blank control group, 3, 6, 12, and 24 hours group crept upward, and there was significant difference between any 2 groups (P < 0.05). But the expression of VEGF protein was not similar with VEGF mRNA. Compared with blank control group, the expression levels of VEGF protein in 3, 6, 12, and 24 hours group were higher (P < 0.05); compared with 3 hours group, the expression levels of VEGF protein in 6, 12, and 24 hours group were higher (P < 0.05); but there was no significant difference among 6, 12, and 24 hours group (P > 0.05). The broth concentration of VEGF protein in blank control group, 3, 6, 12, and 24 hours group crept upward, and there was significant difference between any 2 groups (P < 0.05).③The effect of extracellular regulated protein kinase (ERK) inhibitor (PD98059) and p38 inhibitor (SB203580). The expression levels of VEGF mRNA and its protein in trypsin group were all higher than corresponding indexes of blank control group, trypsin+PD98059 group, trypsin+SB203580 group, PD98059 group, and SB203580 group (P < 0.01), but there was no significant difference among blank control group, trypsin+PD98059 group, trypsin+SB203580 group, PD98059 group, and SB203580 group (P > 0.05). ConclusionActivation of PAR-2 can induce the expressions of VEGF mRNA and its protein in MKN28 gastric cancer cells, that is mediated by ERK1/2-and p38-dependent pathway.