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Background:Diabetic retinopathy (DR) is a sight-threatening microvascular complication of diabetes in which the vascular endothelium is damaged due to oxidative stress and inflammation, and vitreous VEGF concentration becomes elevated. The aim of the present study was to assess the association of DR with genetic variations of the MnSOD, a major antioxidant enzyme, and VEGF, an important mediator of neovascularisation, in northern Iran. Methods: 70 patients with DR and 70 healthy control subjects matched for age and sex was recruited for this study. PCR-based RFLP assay was used to determine the genotypes of MnSODA16V and VEGF+405 C/G polymorphisms. Results and Conclusions:A higher frequency of the AV genotype (71.43%) of the MnSODA16V polymorphism was found in the patients compared with controls which had a 8.33-fold increase in risk of DR (OR= 8.33, 95% CI= 2.56-27.13, P= 0.0004). The frequency of GG, GC, and CC genotypes of VEGF +405 C/G polymorphism in controls were 42.86%, 45.71% and 11.43%, respectively, while in DR patients were 18.57%, 48.57%, and 32.86%, respectively.The +405C allele was considered as a high risk factor of DR (OR= 2.55, 95% CI= 1.57-4.14, P= 0.0001). In conclusion, It is suggested that the MnSODA16V and the VEGF+405 C/G polymorphisms may be associated with the risk of DR in northern Iran.

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Background: Henoch-Schönlein purpura (HSP) is an lgA mediated small vessel systemic vasculitis disease in children. The etiology and pathogenesis of HSP disease remain unknown. However, environmental and genetic risk factors could play important roles in susceptibility to HSP disease. In this study we investigated the association of 5՛-untranslated region polymorphism (-634G/C) of VEGF gene with HSP among Iranian Azeri Turkish population. Methods: Thirty unrelated Iranian Azeri Turkish children with HSP and fifty healthy unrelated subjects without HSP and other inflammatory diseases were enrolled in this population. -634G/C polymorphism of VEGF gene was genotyped by polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) technique. Results: The distribution of CC genotype in VEGF -634G/C polymorphism statistically showed a significant difference in HSP patients in compare to that of control group (P= 0.009). Conclusions: The CC genotype of VEGF -634G/C polymorphism could be associated with susceptibility to HSP disease in Iranian Azeri Turkish ethnic group.

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Regarding the importance of inhibiting VEGF and unique features of VHHs as a new generation of antibody-based therapeutics, the present study aimed to generate VHHs against the receptor binding domain of VEGF, thereby blocking of VEGF binding to its receptor. After preparing the gene repertoire of VHH fragments from an immunized camel, a VHH phage display library was constructed. We adopted a stringent successive biopanning to isolate the phages displaying VHH with high affinity to VEGF-RBD.A significant enrichment of phages that specifically bound to the target protein was obtained after six rounds of panning. Of the specific clones with high binding affinity screened by monoclonal phage ELISA, 52% shared the same VHH sequence, showing its high enrichment. Using molecular simulation of antigen-antibody interaction based on the crystallographic information of VEGF/VEGFR2, molecular dynamics simulations and MM/PBSA free energy calculations, we provide a reliable picture of the binding site of antibody on antigen. The key residues in the VEvhh1-VEGF interface were dissected and the energetics was analyzed by MM/PBSA. The results of studies revealed that VEvhh1 binds to the receptor binding site of VEGF with high binding energy and showed the highest affinity to the residues of VEGF which are responsible for VEGF binding to VEGFR2. Also the antibody potently covers these key functional residues of VEGF, thereby inhibiting VEGF binding to its receptor and probably abrogating its biological activity. This study may represent VEvhh1 as an anti-VEGF and anti-angiogenic candidate.

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Abnormal angiogenesis is associated with various diseases such as solid tumors and metastasis, retinopathies, and rheumatoid arthritis. VEGF-A is the most important mediator of angiogenesis among all growth factors. The bioactivity of VEGF is mediated by two tyrosine kinase receptors VEGFR-1 and VEGFR-2 present on endothelial cells. VEGF signaling through VEGFR-2 is the major angiogenic pathway that leads to stimulation of endothelial cell ingrowths into the tumor. It comprised of an extracellular portion, a cytoplasmic portion, and a short transmembrane domain. The extracellular portion consists of seven Ig-like domains (D1–D7), of which the 1st to 3rd domains function as ligand-binding sites. In the present work, a soluble recombinant extracellular domain 1-3 of VEGFR-2 was expressed in Pichia pastoris to inhibit the VEGF-induced angiogenesis. The 975 bp DNA fragment containing extracellular domain 1-3 kdr, was designed according to the nucleotide sequence at GenBank and protein sequence at Swiss-Prot. The recombinant secretory expression vector (pPinkαHC/KDR1-3) was constructed and transferred into yeast by electroporation. The high expression transformants were identified through complementation of adenine auxotrophy and cultured. KDR1-3 was expressed under the induction of %1 methanol and confirmed by using SDS-PAGE and western blot techniques. After being purified by affinity chromatography using Ni-NTA resins, binding of expressed product to hVEGF165 was proved by two direct ELISA and ELISA receptor binding assays. The data showed that human VEGFR-2 extracellular domain 1-3 with eukaryotic protein structure, that there is no reported paper about, was successfully expressed. 

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Abstract
Introduction: Cervical cancer is the fourth most common cancer among women. In recent years, attention has increased to natural products such as curcumin with anti-cancer potential as a therapeutic supplement. However, due to its poor solubility, its clinical use is limited. In this regard, in this study, to improve clinical parameters, the effects of nanocurcumin on the angiogenesis inhibition of cervical cancer were investigated and compared with free curcumin.
Materials and Methods: MTT method was used to evaluate the proliferation of the HeLa cell line with free curcumin and nanocurcumin at different doses and time intervals and the rate of apoptosis was assessed by flow cytometry. Then, the expression of the vascular endothelial growth factor (VEGF-A) gene in HeLa cells was measured by Real-Time PCR and Western blotting, respectively.
Results: According to IC50 for 48 hours in the HeLa cell line, which was 15 μM/ml and 50 μM/ml for nanocurcumin and free curcumin, respectively, the nanocurcumin showed a greater lethal effect. VEGF-A gene expression (p <0.0001) and protein level (p <0.01) were significantly lower following nano-curcumin treatment than free curcumin.
Conclusion: Nanocarrier increased the solubility and further inhibited the proliferation of cervical cancer HeLa cells and was three times more effective than curcumin in inhibiting angiogenesis at the same concentration. Therefore, nanocurcumin can be a good option for drug supplementation along with routine cervical cancer treatment.
Keywords: Cervical cancer, Nanocurcumin, HeLa cell, VEGF-A.

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Objective: The use of stem cells, particularly mesenchymal stem cells (MSCs), with genes and various growth factors as treatments for myocardial infarction and various other diseases is highly regarded. However these cells meet with inflammation and a hypoxic environment in the target tissue. Hence, treatment with factors that increase the resistance of these stem cells is of importance. Stem cells also can be used as carriers for gene therapy. The aim of the present research is to produce VEGF expressing MSCs. We investigate the effect of stromal derived factor 1 on MSC survival in order to use these cells in a future rat myocardial infarction model.  Methods: MSCs were purified from young male rats by aspirating the cavity of femurs and tibias. After characterization, MSCs were transduced with VEGF using lipofectamine. Expression and function of VEGF was confirmed. Next, we treated MSCs with SDF1α at various time points. The effect of this chemokine was investigated using the LDH assay and by viable cell counts. Results: The experiments confirmed the production and function of VEGF by MSCs. The LDH levels decreased significantly in SDF1α treated MSCs. Cell viability increased significantly in the presence of this chemokine. Conclusion: Treatment of MSCs with the SDF1α chemokine has increased the survival of these cells. These MSCs are proper candidates for increasing angiogenesis and for further analysis in a rat model of myocardial infarction.  

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Objective: Elderly age is accompanied with reduced muscle strength and mass. Vascular endothelial growth factor (VEGF) is the most important stimulant of angiogenesis in muscles which, by increasing blood flow to the muscles, causes muscle strength and endurance. Exercise training leads to increased nitric oxide (NO) and release of VEGF by raising the shear stress during proliferation of endothelial cells and the process of angiogenesis. Garlic also modulates blood flow. This study has assessed the effects of regular swimming training combined with the consumption of garlic extract on the levels of NO plasma and VEGF tissue of soleus slow-twitch and gastrocnemius fast-twitch muscles in aged rats.
Methods: Male aged rats (40-50 weeks) with an average weight of 250-300 g were randomly divided into 5 groups (n=7 rats per group) - control, saline, aerobic training, garlic, and aerobic training + garlic. The swimming training program was scheduled for 50 m daily, 3 times per week for 8 weeks. The groups that received supplements and supplementary training were given a daily dose by gavage of 1 ml/kg body weight of the garlic extract for 8 weeks. We collected tissue and blood samples 48 h after the last training session and after a 10-12 h fasting period. The amount of NO and VEGF were detected by colorimetry and ELISA, respectively.
Results: One-way ANOVA revealed that regular aerobic training increased plasma NO (P=0.001), VEGF of the soleus muscle tissue (P<0.001), and VEGF of the gastrocnemius muscle (P=0.004) in the aged rats. In the group that received garlic extract, there was a significant increase in NO levels (P=0.001), and VEGF in the soleus muscle tissue (P=0.007) and gastrocnemius muscle tissue (P=0.015). Combined training and garlic supplement significantly increased the plasma level of NO (P<0.001), and VEGF in the soleus muscle tissue (P<0.001) and gastrocnemius muscle tissue (P=0.001) compared to the control and saline groups.
Conclusion: Consumption of garlic extract alone and combined with aerobic training significantly increased plasma NO levels, and VEGF of the soleus and gastrocnemius muscle tissues. However, while the extents of increase in the training combined with garlic extract group was higher than that of the garlic group, there was no significant difference observed between the two groups. The lack of significant difference between these two groups might be due to the intensity and type of the training and would need additional research.

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Objective: Diabetes is known to accelerate endothelial cell dysfunction. Meanwhile, an adequate physical activity is recommended as a nonpharmacological treatment. Hence, the aim of the present study is to evaluate the effect of two different time periods of regular swimming exercise on serum levels of NO, VEGF, and TGF-β1 in diabetic male rats.
Methods: We randomly divided 28 male Wistar rats into 4 groups of 7 rats per group - control, diabet, diabet-train 30, and diabet-train 60. Diabetes was induced using alloxan (90 mg/kg, intraperitoneal injection) in rats. The animals performed swimming training for 30 and 60 min for 5 weekly sessions, for a total of 8 weeks. The rats were killed 72 h after the last training session and was measured the levels of glucose, insulin, NO, VEGF, and TGF-β1. One-way ANOVA was used for statistical analysis (p≤0.05).
Results: Diabetes increased the levels of glucose, insulin and TGF-β1, whereas it significantly reduced NO and VEGF levels. After 8 weeks, 30 min- swimming training, causing reverse of health changes compared to diabetes (p<0.05). 60 min- training did not make significant change (p>0.05). But there were significant differences in the levels of glucose, NO, and TGF-β1 between the two groups that trained for 30 and 60 min.
Conclusion: It seems swimming training with enough time, which is sought for diabetic patients, can lead to a protective effect against cardiovascular disease by impacting the endothelial function factors in these patients.