PURPOSE: Anti-vascular endothelial growth factor (VEGF) agents have been used for the last 10 years, but their safety profile, including cytotoxicity against various ocular cells such as retinal pigment epithelial (RPE) cells, remains a serious concern. Safety studies of VEGF agents conducted to date have primarily relied on healthy RPE cells. In this study, we assessed the safety of three anti-VEGF agents, namely, ranibizumab, bevacizumab, and aflibercept, on senescent RPE cells. METHODS: Senescent human induced pluripotent stem cell-derived RPE cells were generated by continuous replication and confirmed with senescence biomarkers. The viability, proliferation, protein expression, and phagocytosis of the senescent RPE cells were characterized 3 days after anti-VEGF treatment with clinical doses of ranibizumab, bevacizumab, or aflibercept. RESULTS: Clinical doses of ranibizumab, bevacizumab, or aflibercept did not decrease the viability or alter proliferation of senescent RPE cells. In addition, the anti-VEGF agents did not induce additional senescence, impair the protein expression of zonula occludens-1 and RPE65, or reduce the phagocytosis capacity of senescent RPE cells. CONCLUSIONS: Clinical dosages of ranibizumab, bevacizumab, or aflibercept do not induce significant cytotoxicity in senescent RPE cells.
Aging ; Bevacizumab* ; Biomarkers ; Endothelial Growth Factors ; Epithelial Cells* ; Humans ; Phagocytosis ; Ranibizumab* ; Retinaldehyde* ; Vascular Endothelial Growth Factor A

PURPOSE: CD105 (endoglin) has been shown to be a more useful marker to identify the proliferating endothelium involved in tumor angiogenesis than are the panendothelial markers. The monoclonal antibody D2-40 is a specific lymphatic endothelial marker. METHODS: We investigated CD105, lymphatic vessel marker (D2-40), vascular endothelial growth factor (VEGD)-A and the VEGF-D expressions as possible prognostic markers in the endoscopic biopsy tissue of stomach cancer patients. The pre-operative endoscopic biopsies and surgical biopsies from 73 patients were immunostained for CD105, D2-40, VEGF-A and VEGF-D. Positively stained microvessels were counted in densely vascular foci (hot spots) at a x200 field in each specimen. RESULTS: The microvessel density (MVD) and lymphatic vessel density (LVD), according to the CD105 and D2-40 expressions of the endoscopic biopsies, showed a statistically significant correlation with the surgical biopsies. The MVD via CD105 a showed statistically significant correlation with the histologic differentiation, T-stage, nodal metastasis and stage in the endoscopic biopsies and surgical biopsies, respectively. The lympathic vessel density (LVD) via D2-40 showed a statistically significant correlation with T-stage, nodal metastasis and stage in the endoscopic biopsies. The expressions of VEGF-A and VEGF-D showed a statistically significant correlation with the MVD and LVD. CONCLUSION: The MVD, as determined by the CD105 expression and the LVD as determined by the D2-40 expression may be useful markers for predicting the invasiveness with using a pre-operative endoscopic biopsy of stomach cancer.
Biopsy* ; Endothelium ; Humans ; Lymphatic Vessels ; Microvessels ; Neoplasm Metastasis ; Stomach Neoplasms* ; Stomach* ; Vascular Endothelial Growth Factor A* ; Vascular Endothelial Growth Factor D ; Vascular Endothelial Growth Factors*

Here, we investigated the role of zerumbone, a natural cyclic sesquiterpene of Zingiber zerumbet Smith, on angiogenesis using human umbilical vein endothelial cells (HUVECs). Zerumbone inhibited HUVECs proliferation, migration and tubule formation, as well as angiogenic activity by rat aorta explants. In particular, zerumbone inhibited phosphorylation of vascular endothelial growth factor receptor-2 and fibroblast growth factor receptor-1, which are key regulators of endothelial cell function and angiogenesis. In vivo matrigel plug assay in mice demonstrated significant decrease in vascularization and hemoglobin content in the plugs from zerumbone-treated mice, compared with control mice. Overall, these results suggest that zerumbone inhibits various attributes of angiogenesis, which might contribute to its reported antitumor effects.
Animals ; Aorta ; Endothelial Cells ; Fibroblast Growth Factors ; Ginger* ; Human Umbilical Vein Endothelial Cells ; Mice ; Phosphorylation ; Rats ; Receptors, Vascular Endothelial Growth Factor ; Vascular Endothelial Growth Factor Receptor-2

OBJECTIVELooking for an objective biomedical index to distinguish types and phases of hemangioma in order to provide an objective basis for selecting clinical treatment to hemangioma.

METHODSELISA (enzyme-linked immunosorbent assay) was used to determine serum VEGF concentration of 15 patients with proliferative hemangioma, 6 with involuted hemangioma, 6 with vascular malformation and 8 infants of the control group.

RESULTSThe serum VEGF concentrations of 15 proliferative hemangioma patients were significantly higher than those of involuted hemangioma patients, vascular malformation patients and control group infants. The serum VEGF concentrations of involuted hemangioma patients were a little bit higher than those of vascular malformation patients and control group infants, but without statistic significance.

CONCLUSIONSELISA could easily and accurately determine the serum VEGF concentration of different types and different phases of hemangioma. The determination of serum VEGF concentration could provide guidance for selecting a protocol of systemic corticosteroid treatment for proliferative hemangioma. Combined with gene expression and distribution of VEGF and its receptors and some other cytokines, the determination of serum VEGF concentration could help elucidate the mechanism of proliferative hemangioma.

Endothelial Growth Factors ; blood ; Enzyme-Linked Immunosorbent Assay ; Hemangioma ; blood ; Humans ; Infant ; Lymphokines ; blood ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the value of application of recombinant human VEGF to accelerate flap viability in a rat model of non-ischemic prefabricated flap.

METHODSPrefabricated Flaps were created in 48 SD rats. An autologous tail artery loop was anastomosed to the femoral artery and vein, and implanted subcutaneously in the lower abdomen. Flaps were divided into four groups of 12 each. At the time of loop implantation, the control groups received 0.9% NaCl (Control 1) and 16% (V/W) polyvinyl alcohol (PVA) solution (Control 2). The treatment groups received VEGF in 0.9% NaCl (treatment 1) and VEGF in PVA (treatment 2). In each group, a 3 cm x 4 cm flap nurtured by the tail artery pedicle was elevated and resutured into place after 3, 4 and 5 weeks. The percentage of surviving skin of each flap was determined by planimetry 7 days after flap elevation.

RESULTSMean skin survival areas at 3, 4, and 5 weeks were 1%, 0%, 10% in control; 0%, 16%, 25% in control 2; 3.57%, 39.13%, 75.00% in treatment 1; 8.13%, 41.98%, 58.41% in treatment 2. VEGF significantly improved flap survival by 5 weeks (P < 0.05).

CONCLUSIONThese results suggest VEGF can accelerate maturation of prefabricated flaps.

Animals ; Endothelial Growth Factors ; pharmacology ; Female ; Lymphokines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Recombinant Proteins ; pharmacology ; Surgical Flaps ; physiology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVEGene therapy has been becoming one of the most attractive medical areas. But the using of gene therapy in plastic surgery is relatively scarce. Our purpose was to investigate the effect of naked plasmid encoding Vascular Endothelial Growth Factor on the viability of the random skin flap by directly injected subcutaneously.

METHODS30 female Sprague-Dawley rats randomly divided into three groups. A random dorsal skin flap of 3 cm x 9 cm was elevated in each of the rats. And 1 ml double-distilled water solution was injected subcutaneously, which was only water in group 1 during the operation, 200 micrograms VEGF cDNA plasmid in group 2 during the operation, 200 micrograms pcDNA3.1/zeo(+)--VEGF in group 3, 24 hours before the operation, respectively. 7 days after the operation, all the animals were sacrificed by overdose anesthetic. The survival tissue was measured with planimetry. Two samples were harvested from each group for pathological check and immunohistochemical test.

RESULTSImmunohistochemical staining demonstrated that there was human VEGF deposited around the capillary in the flaps treated with VEGF gene. The flaps treated with VEGF gene had a larger percentage of survival skin (group 1 = 47% +/- 5.4%, group 2 = 65.4% +/- 6.3%, group 3 = 72.3% +/- 8.5%, P < 0.05).

CONCLUSIONVEGF gene directly injected into subcutaneous can express VEGF. It makes the gene therapy simple and practical and will be promising future in the tissue transplantation.

Animals ; Endothelial Growth Factors ; genetics ; Female ; Genetic Therapy ; Injections, Subcutaneous ; Lymphokines ; genetics ; Rats ; Rats, Sprague-Dawley ; Surgical Flaps ; physiology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Vascular Endothelial Growth Factor (VEGF) is a potent angiogenic factor with a key role in several pathological processes, including wound repair as well as a effective vascular permeability factor. This article review the present study of VEGF in molecular biology, the connection with repair and expression regulation and so on.
Animals ; Endothelial Growth Factors/physiology* ; Forensic Medicine ; Humans ; Intercellular Signaling Peptides and Proteins/physiology* ; Lymphokines/physiology* ; Rats ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors ; Wound Healing/physiology*

Vascular endothelial growth factor (VEGF165) is a highly specific vascular endothelial growth factor that can be used to treat many cardiovascular diseases. The development of anti-tumor drugs and disease detection reagents requires highly pure VEGF165 (at least 95% purity). To date, the methods for heterologous expression and purification of VEGF165 require multiple purification steps, but the product purity remains to be low. In this study, we optimized the codons of the human VEGF165 gene (vegf165) according to the yeast codon preference. Based on the Pichia pastoris BBPB vector, we used the Biobrick method to construct a five-copy rhVEGF165 recombinant expression vector using Pgap as the promoter. In addition, a histidine tag was added to the vector. Facilitated by the His tag and the heparin-binding domain of VEGF165, we were able to obtain highly pure rhVEGF165 (purity > 98%) protein using two-step affinity chromatography. The purified rhVEGF165 was biologically active, and reached a concentration of 0.45 mg/mL. The new design of the expression vector enables production of active and highly pure rhVEGF165 ) in a simplified purification process, the purity of the biologically active natural VEGF165 reached the highest reported to date.
Codon/genetics* ; Humans ; Pichia/genetics* ; Recombinant Proteins/genetics* ; Saccharomycetales ; Vascular Endothelial Growth Factor A/genetics* ; Vascular Endothelial Growth Factors

OBJECTIVE: To investigate the expressions of stromal cell-derived factor (CXCL12), stromal cell-derived factor receptor (CXCR4), vascular endothelial growth factor (VEGF) and microvessel density (MVD) in bone marrow microsputum of patients with multiple myeloma (MM) and their correlation with the prognosis. METHODS: The expressions of CXCL12, CXCR4, VEGF and MVD in bone marrow microtubules of 57 newly diagnosed MM patients and 26 normal bone marrow samples were detected by immunohistochemistry. The rank sum test was used to compare the differences between the two groups. The clinical data of the patients were collected to analyze the correlation between the indicators of the MM group and the prognosis. RESULTS: The expressions of CXCL12, CXCR4, VEGF and MVD in the bone marrow biopsy of the patients in MM group were significantly higher than those in the normal control group (P＜0.05). The expressions levels of CXCL12, CXCR4, VEGF and MVD were in the bone marrow of the patients in MM group were correlated with the ISS stage, risk stratification and the proportion of plasma cells in the bone marrow (P＜0.05). Univariate analysis showed that age, ISS stage, risk stratification, plasma cell ratio, expressions of CXCL12, CXCR4, VEGF, and MVD associated with the prognosis of patients with MM (P＜0.05). Multivariate analysis found that expressions of CXCR4, VEGF, MVD, age, and plasma cell ratio were independent prognostic factors. CONCLUSION The expressions of CXCL12, CXCR4, VEGF and MVD are increase in the bone marrow of patients with multiple myeloma, and their expressions levels are associate with the occurrence and development of multiple myeloma, and their high expression may indicate a poor prognosis.
Chemokine CXCL12 ; Humans ; Multiple Myeloma ; Neovascularization, Pathologic ; Patients ; Prognosis ; Receptors, CXCR4 ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

BACKGROUND: Many studies have shown that angiogenesis plays an important role in the growth and progression of solid tumors, including gastric carcinoma. Vascular endothelial growth factor(VEGF) is the most potent known inducer of microvascular hyperpermeability; in addition, it is a selective mitogen for endothelial cells. In this study, we studied that the relationship between angiogenesis and the expression of VEGF in gastric carcinoma. METHODS: 23 early gastric carcinomas and 28 advanced gastric carcinomas obtained by surgical resection were studied. Expression of the VEGF was semiquantitatively analyzed in paraffin sections by immunohistochemical method. Histologic sections immunostained for CD31 antigen were evaluated for microvessel density. RESULTS: The VEGF was mainly localized to the cytoplasm of carcinoma cells. Expression of the VEGF was significantly higher in advanced gastric carcinoma than in early gastric carcinoma (p< 0.05). Expression of the VEGF was correlated with the tumor depth and the stage(p< 0.05). The VEGF positivity was significantly higher in moderately and poorly differentiated gastric carcinoma than in well differentiated gastric carcinoma(p< 0.05). But the expression of the VEGF by Lauren,s classification was not significant between intestinal type and diffuse type(p> 0.05). The expression of CD31 was higher in advanced gastric carcinoma than in early gastric carcinoma. In gastric carcinoma, a correlation was observed between CD31 expression and the stage of the disease, the degree of histological differentiation. Also VEGF and CD 31 expression was observed significant correlation. CONCLUSION: VEGF is an important angiogenic factor associated with progression of the gastric carcinoma. Neovascularization assessment by CD 31 immunostaining was another efficient method for defining groups of tumors with aggressive clinical course.
Angiogenesis Inducing Agents ; Antigens, CD31 ; Classification ; Cytoplasm ; Endothelial Cells ; Endothelial Growth Factors ; Microvessels ; Paraffin ; Stomach Neoplasms ; Vascular Endothelial Growth Factor A*