Intrastriatal Gene Transfer of Vascular Endothelial Growth Factor Rescues Dopaminergic Neurons in a Rat Parkinson's Disease Model

Parkinson's disease (PD) is a chronic progressivegerontic disease, which is characterized by the degeneration of dopaminergic neurons in substantia nigra. Efforhas been made to find effective treatment to arrest oslow down the degeneration of dopaminergic neuronsOnce regarded as an endothelial cell-specific factorvascular endothelial growth factor (VEGF), in recenyears, was unexpectedly found to be a critical player inneurodegeneration, and have direct neurotrophic andneuroprotective effects…     

Serum Vascular Endothelial Growth Factor-C and Vascular Endothelial Growth Factor Level in Patients with Colorectal Carcinoma and Clinical Significance

The opti mal staging approachin patients withcolorectal carcinoma(CRC)who have been in-volved in lymph node metastasis on computedtomography(CT)scans or magnetic resonance i m-aging scans(MRI)is far less reliable and contro-versial,because the i maging criteria for tumor in-volvement are relied on the size and shape of thelymph nodes.But as we known,accurate tumorstaging is essential for choosing the appropriatetreat ment strategy for patients with CRC.The in-volvement of lymph nodesis ani…     

Regulation of Hypoxic Response Elements on the Expression of Vascular Endothelial Growth Factor Gene Transfected to Rat Skeletal Myoblasts under Hypoxic Environment

The regulation of hypoxic response elements on the expression of vascular endothelial growth factor （VEGF） gene transfected to primary cultured rat skeletal myoblasts under hypoxic environment was investigated. pEGFP-C3-9HRE-CMV-VEGF vector was constructed with molecular biology technique and transfected to primary cultured rat skeletal myoblasts by lipofectamine in vitro. Gene expression of transfected myoblasts was detected by RT-PCR, Western blot and fluorescence microscope under different oxygen concentrations and different hypoxia time. The results showed that in hypoxia group, the VEGF gene bands were seen and with the decrease of oxygen concentrations and prolongation of hypoxia time, the expression of VEGF mRNA was obviously increased. Under hypoxic environment, the expression of VEGF protein in the transfected myoblasts was significantly increased. EGFP was expressed only under hypoxic environment but not under normoxic environment. It was concluded that hypoxia promoter could be constructed with HRE and regulate the expression of VEGF gene under hypoxic and normoxic environment, which could enhance the re- liability of gene therapy.     

Effect of Antisense Oligodeoxynucleotide of Vascular Endothelial Growth Factor C on Lymphangiogenesis and Angiogenesis of Pancreatic Cancer

In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular en-dothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografted human pancreatic cancer cells (Panc-1) was established. Thirty nude mice were randomly divided into 3 groups: PBS control group (group A), scramble-sense con-trol group (group B) and antisense group (group C). All nude mice were treated once every 2 days as 3 times per week, for 3 weeks (oligonucleotide 10 mg/kg every time). After treatments were com-pleted, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohis-tochemical method to examine microvessel density (MVD), lymphtic vessel density (LVD) of pan-creatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5±41.5, 221.5±52.3 and 108.6±14.9 pg/mL in groups A, B and C re-spectively (P<0.01). LVD in groups A, B and C was 13.8±2.1, 12.4±1.9 and 4.2±1.6 respectively (P<0.01). MVD in groups A, B and C was 27.5±8.7, 25.9±4.2 and 19.4±5.6 respectively with no sig-nificant difference among the groups (P>0.05). It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis.     

Resveratrol Inhibits the Secretion of Vascular Endothelial Growth Factor and Subsequent Proliferation in Human Leukemia U937 Cells

This study examined the effect of resveratrol on the secretion of vascular endothelial growth factor(VEGF) and subsequent proliferation of human leukemia U937 cells,and explored the mechanisms involved.Human leukemia U937 cells were treated with resveratrol of different concentrations(12.5-200 μmol/L) for different time lengths(12-48 h).The proliferation of the U937 leukemic cells was determined by MTT assay.Apoptosis was observed by Annexin-Ⅴ-FIFC/PI double staining and flow cytometry(FCM).Cells cycle was analyzed by PI staining and FCM.The content of VEGF was determined by ELISA.Human umbibical vein endothelial cells were examined for vasoformation in vitro after exposures to resveratrol of various concetrations.The results showed that resveratrol inhibited the proliferation of U937 leukemia cells in a dose-and time-dependent manner.Resveratrol induced apoptosis and S-phase cell cycle arrest in human leukemic U937 cells.Resveratrol inhibited the secretion of VEGF in U937 cells.Resveratrol inhibited the vasoformation of human vein endothelial cells in a dose-dependent manner.It was concluded that resveratrol could down-regulate the secretion of VEGF,induce apoptosis and suppress the proliferation of U937 cells.     

Serum Level of Vascular Endothelial Growth Factor in Patients with Obstructive Sleep Apnea Hypopnea Syndrome

To explore the relationship between the serum vascular endothelial growth factor (VEGF) level and the severity of obstructive sleep apnea hypopnea syndrome (OSAHS), the concentrations of serum VEGF in 40 OSAHS patients and 9 healthy controls were measured by using ELISA method. Meanwhile the correlation between the concentration of VEGF and parameters of polysomnography (PSG) was examined. Our results showed that the concentrations of VEGF were significantly higher in OSAHS patients with severe hypoxia (536.8±334.7 pg/mL) than in those with mild hypoxia (329.2±174.7 pg/mL) and healthy controls (272. 8±211.0 pg/mL) (P<0.05 for both). The concentra- tions of VEGF were also significantly higher in OSAHS patients with hypertension (484.5±261.4 pg/mL) than in those without hypertension (311.0±158.4 pg/mL) and healthy controls (272. 8± 211.0 pg/mL) (P<0.05 for both). There was a positive correlation between the concentration of VEGF and the apnea hypopnea index (AHI) (r=0.34, P<0.05). It is concluded that the concentration of the serum VEGF is positively related to the severity of OSAHS. The elevated serum VEGF level may be involved in the pathogenesis of the complications of obstructive sleep apnea hypopnea syndrome.     

Expression of Human Vascular Endothelial Growth Factor （VEGF165）in pichia pastoris and Its Biological Activity

Objective :To express human vascular endothelial growth factor(hVEGF165) cDNA in Pichia pastroris,purify the expressed product and detect the biological activity of it.Methods:By inserting hVEGF165 cDNA coding 165 amino acid esidues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides,a recombinant expression plasmid pPIC9K/hVEGF165 was constructed and transformed to yeast host strain KM71,then multiple-copy insert transformants were screened out and cultured in flasks,and hVEGF165 was expressed under the induction of 1% methnol.Results:SDS-PAGE showed that after being induced with 1% methanol for 4d,the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expressed.Western blot showed good antigenicity and specificity of expressed product.After being purified by Heparin-Sepharose CL6B affinity chromatography,the purity of expressed product reached above 90%,Biological assays proved that the expressed product could stimulate the proliferation of HUVEC.Conclusion:hVEGF165 was successfully expressed.The study opened up a wide prospect for the application of VEGF165 in the prevention and treatment of ischemic heart disease and other tissue ischemic diseases such as secondary arterial occlusion in limbs.     

Expression of Vascular Endothelial Growth Factor and Cyclooxygenase-2 in Laryngeal Squamous Cell Carcinoma and Its significance

n order to study the expressions of vascular endothelial growth factor （VEGF） and cyclooxygenase-2 （COX-2） in human laryngeal squamous cell carcinoma （LSCC） and its significance, the expression of VEGF mRNA and COX-2 mRNA in 62 cases of LSCC and 54 adjacent noncancerous laryngeal tissues and 9 normal human laryngeal mucous tissues was detected by using techniques of semi-quantitative RT-PCR. It was found that the expression level of VEGF and COX-2 mRNA was significantly increased in LSCC as compared with that in the normal human laryngeal mucous tissues （both P〈0. 01）, and the expression level of VEGF and COX-2 mRNA were significantly increased in stage Ⅲ＋ Ⅳtissues of LSCC as compared with the stage Ⅰ ＋ Ⅱ tissues of LSCC （P 〈0.01）. There was a high positive correlation between VEGF and COX-2 expression in LSCC （r= 0. 756,P〈0.01）. These data raise the possibility that VEGF and COX-2 may play key roles in the growth, invasion and metastasis of LSCC.     

Expression of Vascular Endothelial Growth Factor C and Its Correlation with Lymph Node Metastasis in Colorectal Carcinoma

Lymphnodemetastasiswastheunderlyingcauseforincurablenessandthehighmortalityratesofmalignantsolidtumor.Asaconsequence,detectionoftumorcellsinlymphaticvesselsandregionallymphnodeswasakeyfactorinthestagingandtreatmentofhumantumors[1],buttheexactmechanismands…     

Expression of Vascular Endothelial Growth Factor （VEGF） in Human Osteosarcoma Cells Transfected with Adeno-associated Virus-antisense VEGF

Ithasbecomeacommonmethodforcancertreatmentthroughantivascularizationinthepresent.Somestudieshaverevealedthatlivercancer ,lungcancerandcervicalcancercanbetreatedsuccessfullythroughholdingbackvascularendothelialgrowthfac tor (VEGF) proteinbecauseofitsimport…     

Gene polymorphism of alpha-2 macroglobulin in patients with Parkinson＇s disease

Objective： To explore the relationship between polymorphism of α2-macroglobulin (A2M) gene and Parkinson‘s disease(PD)in Han Nationality in Shanghai. Methods：The distributions of A2M gene polymorphism (a Val1000l1e in exon24, V/I)were detected in 66 PD patients and 120 healthy controls using polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) method. Results：The I allelic frequency in A2M exon24 gene of PD patients (90.9 % ) was significantly lower than that of the healthy controls(96.3%)(OR=0.39,P=0. 033),so was the I/I genotype (OR=0.32,P=0. 015),especially in the patients more than 60 years old(OR=0.31 ,P=0.04). Conclusion：The I allele in exon24 of A2M gene might inhibit the onset of PD in Han Nationality in Shanghai.     

Effects of Vascular Endothelial Cell Growth Factor on Fibrovascular Ingrowth into Rabbit＇s Hydroxyapatite Orbital Implant

The effects of different concentrations of vascular endothelial cell growth factor (VEGF) on the fibrovascular ingrowth into rabbits hydroxyapatite orbital implant were investigated. Twelve New Zealand white i‘abbits were divided into 3 groups and received hydroxyapatite orbital implant surgery in their right eyes. Before and after the operation, the implants were treated with I0 ng/ml VEGF, 100 ng/ml VEGF, or normal saline as control group. The animals received technetium bones scan at 2, 4, and 6 weeks postoperatively. The mean radioactivity counts within region of interest (ROI) of the surgery eye (R) and the non-surgery eye (L) in the same animal were tested,and the R/L ratios were calculated. The implants were harvested at 6th weeks and examined histopathologically. The results showed that at second week, there was no significant difference in mean R/L ratios between VEGF group and control group (F=2.83, P=0. 111); At 4th week (F=7. 728, P=0. 011) and 6th week (F=7. 831, P=0. 011) postoperatively, the mean ratios in VEGF groups were significantly higher than that in control group. At 6th week postoperatively,the fibrovascularization rates in VEGF groups were higher than in control group significantly (F=8. 711, P=0. 008). It was suggested that VEGF could promote the fibrovascular ingrowth into hydroxyapatite orbital implant, thus might shorten the time required for complete vascularization of the HA orbital implant.     

Clinical Value of Vascular Endothelial Growth Factor Combined with Interferon-γ in Diagnosing Malignant Pleural Effusion and Tuberculous Pleural Effusion

In order to investigate the clinical value of vascular endothelial growth factor （VEGF） combined with interferon-γ （IFN-γ） in diagnosing malignant pleural effusion and tuberculous pleural effusion, 42 cases of malignant pleural effusion and 45 cases of tuberculous pleural effusion in Tongji Hospital, from March 2004 to May 2005, were included, The carcinoembryonic antigen （CEA）, VEGF and IFN-γ levels of pleural effusion were detected by using ELISA, and adenosine deaminase （ADA） activity was determined by using enzyme kinetic analytical method. The sensitivity, specificity, accuracy and area under the curve （AUCR^ROC） of CEA and VEGF, VEGF/IFN-γ ratio, ADA and IFN-γ were measured by receiver operating characteristic curve （ROC）, The results showed that CEA, VEGF levels and VEGF/IFN-γ ratio were significantly higher and the ADA and IFN-γ levels were significantly lower in malignant group than those in tuberculous group （P〈0,01）, The sensitivity, specificity, accuracy and AUCR^ROC of VEGF/IFN-γ ratio （88,7%, 99,8%, 94,4%, 0.96 respectively） were higher than those of CEA （67.8%, 96.1%, 82,4%, 0.78 respectively） and VEGF （81,5%, 84,3%, 82.9%, 0.79 respectively）. The sensitivity, specificity, accuracy and AUCR^ROC of IFN-γ （85.7%, 96,4%, 90.9%, 0.94 respectively） were higher than those of ADA （80,2%, 87,6%, 83.8%, 0,81 respectively）. It was concluded that VEGF/IFN-γ ratio and IFN-γ could be used as valuable parameters for the differential diagnosis of malignant pleural effusion and tuberculous pleural effusion.     

Dynamic Expression of Tumor Necrosis Factor-α and Vascular Endothelial Growth Factor in Rat Model of Pulmonary Emphysema Induced by Smoke Exposure

In order to explore the roles of tumor necrosis factor-α(TNF-α) and vascular endothelial growth factor(VEGF) in the pathogenesis of pulmonary emphysema,male Wistar rats were randomized into group A1,group A2.5 and group A4,each with smoke exposure for 1 month,2.5 months or 4 months,respectively.Group B1,group B2.5 and group B4 were used as non smoking controls at corresponding time points.TNF-α in bronchoalveolar lavage fluid(BALF) and expression of VEGF in lung tissue was determined by ELISA or by SABC immunohistochemistry assay either.Lung slices were stained with hematoxylin and eosin(HE).Results showed that in animal with smoke exposure the mean linear interceptor(Lm),an index of pulmonary emphysema and the content of TNF-α in BALF increased gradually,on contrary,the expression of VEGF in lung tissue decreased(P<0.05).This phenomenon was not obvious in animals without smoke exposure.Lm was negatively correlated to the VEGF expression(γ=-0.81,P<0.01) and positively correlated to TNF-α concentration(γ = 0.52,P<0.004),which implies that smoke exposure decreased the expression of VEGF and increased the expression of TNF-α.It is plausible to speculate that the imbalance of TNF-α and VEGF may play an important role in the pathogenesis of smoke-induced pulmonary emphysema.     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Expression of Proliferating Cell Nuclear Antigen and Vascular Endothelial Growth Factor in Primary Culture Cells Derived from Human Ovarian Mucinous Cystadenocarcinoma

The effects of antisense oligodeoxynucleotide （antisense ODN） to follicle-stimulating hormone receptor （FSHR） and follicle-stimulating hormone （FSH） on the expression of proliferating cell nuclear antigen （PCNA） and vascular endothelial growth factor （VEGF） were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma （OMC）. The primary （）MC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co-cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups （P〈0.05 or P〈 0.01）, while decreased significantly in antisense ODN groups （P〈0.05 or P〈 0.01） and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly （P〈0.01）. It was suggested that FSH might promote the development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH.     

Expression of Human Vascular Endothelial Growth Factor Receptor Flt—1 Extracellular Domain 1—3 Loop cDNA in Pichia pastoris,Purification of the Expressed Product and Detection of Its Biological Acti

Objective:to express human vascular endothelial growth factor receptor Flt-1 extracellular domain 1-3 loop cDNA in Pichia.pastroris,and to purify the expressed product and detect its biological activity.Methods:By inserting human Flt-1(1-3 loop)cDNA coding 316 amino acid residues into Pichia pastoris expression vector pPIC9K containing AOX1 promoter and the sequences of α secreting signal peptides,a recombinant expression plasmid pPIC9K/Flt-1(1-3) was constructed and transformed to yeast host strain GS115,then His^ Muts phenotype transformant was screened out and cultured in flasks,and Flt-1(1-3) was expressed under the induction of 1% methanol.Results SDS-PAGE showed that after being induced with 1% methanol for 4d ,the expressed product existed in supernatant in the form of soluble molecule and contained 60% of total protein expresses.Western blot showed good antigenicity and specificity of expressed product.After being purifed by CM-Sepharose FF and Sephacry1 S-100 chromatography,the purity of the expressed product reached obove 90%,Biological assay proved that the expressed product could bind to hVEGF165 and inhibit the proliferation of HUVEC stimulated by hVEGF165.Conclusion:Human vascular endothelial growth factor receptor Flt-1 exracellular domian 1-3 loop was successfully expressed.The study lays f foundation for further application of the expressed product in the troduct in the treatment of vasoformation related diseases,such as tumor and diabetic retinopathy.