Gene Expression ; Graft Occlusion, Vascular ; prevention & control ; Humans ; Liver ; blood supply ; Stents ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in hippocampus of rats with aging.

METHODSParaffin sections of brain tissue of rats at the age of 3, 18, 24, 30 months were stained by immunohistochemistry, the expression of VEGF and MVD was quantitatively analyzed.

RESULTSInnunohistochemical staining showed that the VEGF-positive cells were mainly pyramidal neuron in hippocampus; the intensity of VEGF-positivity in neuron cells was decreased with the aging (P<0.05). The MVD in hippocampus was also decreased with the aging of rats (P<0.05).

CONCLUSIONIncreasing VEGF contents and improving blood circulation in brain tissue may prevent or treat vascular dementia and cerebrovascular diseases.

Aging ; Animals ; Capillaries ; pathology ; Hippocampus ; blood supply ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo investigate the differences in the expression of angiogenesis-related molecules between benign and malignant breast neoplasms.

METHODSThirty breast cancer patients (33 foci) and 30 with benign breast neoplasms (34 foci) were examined for CD34, vascular endothelial growth factor (VEGF) and Flk-1/KDR expressions using immunohistochemistry.

RESULTSIn patients with breast cancer, the microvessels densely distributed around the cancer nest. The microvessel density (MVD) in the cancer patients was significantly higher than that in patients with benign tumors (34.48+/-8.34 vs 18.65+/-4.69, P<0.05). In the breast cancer patients, strong VEGF expression was found in the epithelial cells and vascular endothelial cells around the breast carcinoma, and Flk-1/KDR was also strongly expressed in the vascular endothelial cells. The expressions of VEGF and Flk-1/KDR were hardly detectable in the benign tumors.

CONCLUSIONVEGF is an important regulatory factor in promoting breast tumor angiogenesis.

Breast Neoplasms ; blood supply ; metabolism ; Carcinoma, Ductal, Breast ; blood supply ; metabolism ; Female ; Humans ; Neovascularization, Pathologic ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVE: To analyze the expression of vascular endothelial growth factor (VEGF), miR-205, Ezrinand Lamin A/C in ovarian cancer tissues. METHODS: The expression of VEGF in the serum of epithelial ovarian cancer and that of healthy volunteers were detected by enzyme-linked immunosorbent assay; the expressions of vascular endothelial growth factor receptor 1 (VEGFR-1), VEGFR-2, Ezrin and Lamin A/C were detected by immunohistochemistry and the micro-vessel density (MVD) of CD31 was detected by immunohistochemistry in epithelial ovarian cancer, benign ovarian and normal ovarian specimens; and the expression of miR-205, Ezrin and Lamin A/C were detected by real-time PCR in epithelial ovarian cancer, benign ovarian and normal ovarian specimens. RESULTS: The expression of VEGF in the serum of epithelial ovarian cancer patients (116.10± 11.94) was significantly higher than that of healthy volunteers (40.04±4.97, P<0.05). The positive expression rates of VEGFR-1 and VEGFR-2 in the epithelial ovarian cancer specimens were 75.9% and 91.4% respectively, which were significantly higher than that in the benign ovarian and the normal ovarian specimens (P<0.05). No differences were observed in the positive expression rates of VEGFR-1 and VEGFR-2 between the benign ovarian and the normal ovarian specimens (P>0.05). The average length of MVD in the epithelial ovarian cancer specimens (7.56±0.51), was significantly higher than that in the normal ovarian specimens (1.22±0.56, P<0.05) and in the benign ovarian specimens (0.7±0.39, P<0.05). No differences were observed in the average length of MVD between the benign ovarian and the normal ovarian specimens (P>0.05). The relative expression level of miR-205 was 0.106±0.035 in the epithelial ovarian cancer specimens, which was significantly higher than that in the normal ovarian specimens (0.0007±0.0005, P<0.05); the relative expression level of miR-205 in the benign ovarian specimens was (0.0002±0.0002), higher than that in the normal ovarian specimens, but with no significance (P>0.05). The positive expression rates of Ezrin and Lamin A/C in the epithelial ovarian cancer specimens were 51.7% and 60.3%, respectively, which were significantly lower than those in the benign ovarian and the normal ovarian specimens (P<0.05). No differences were observed in the positive expression rates of Ezrin and Lamin A/C between the benign ovarian and the normal ovarian specimens (P>0.05). The relative expression levels of Ezrin and Lamin A/C mRNA in the epithelial ovarian cancer specimens were (0.026±0.003) and (0.060±0.007), respectively, which were significantly lower than those in the normal ovarian specimens (P<0.05). There was no statistical significance between the relative expression level of Ezrin and Lamin A/C mRNA in the epithelial ovarian cancer specimens and that in the benign ovarian specimens (0.029± 0.011, 0.089 ± 0.019; P>0.05) . CONCLUSION VEGF is significantly expressed in the serum of epithelial ovarian cancer patients; and miR-205 is up-regulated in the epithelial ovarian cancer specimens. Ezrin and Lamin A/C are down-regulated in the epithelial ovarian cancer samples. VEGF, miR-205 and target protein may be associated with the invasion and metastasis of epithelial ovarian cancer.
Carcinoma, Ovarian Epithelial ; Cytoskeletal Proteins ; genetics ; metabolism ; Down-Regulation ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunohistochemistry ; Lamin Type A ; genetics ; metabolism ; MicroRNAs ; genetics ; metabolism ; Neoplasms, Glandular and Epithelial ; genetics ; metabolism ; Ovarian Neoplasms ; genetics ; metabolism ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; blood ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

OBJECTIVETo conduct a preliminary study on the effect of curcumol in promoting angiogenesis activity and its mechanism in zebrafishes, in order to provide basis for clinical prescription.

METHODZebrafishes biological model was established to, observe curcumol's effect on embryo blood vessel growth, blood vessel regeneration of adult fishes after tail-cutting and tissue regeneration of fish fries after tail-cutting. The relative fluorescence quantitative PCR method was adopted to determine the gene expression of vascular endothelial growth factor (VEGFA) and receptor VEGFR2 of fish fries after tail-cutting.

RESULTCurcumol contributed to angiogenesis of intersegmental blood vessels in zebrafishes embryos and speed up regeneration of blood vessels in adult fishes after tail-cutting. Furthermore, curcumol can increase the gene expression of VEGFA and VEGFR2 in fish fries.

CONCLUSIONCurcumol can promote angiogenesis in zebrafishes, and enhance the gene expression of VEGFA and VEGFR2 in fish fries after tail-cutting and speed up the regeneration of their tails.

Animals ; Embryo, Nonmammalian ; blood supply ; drug effects ; metabolism ; Gene Expression Regulation ; drug effects ; Neovascularization, Physiologic ; drug effects ; Sesquiterpenes ; pharmacology ; Vascular Endothelial Growth Factor A ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Zebrafish ; embryology ; genetics ; physiology

OBJECTIVETo reseach the correlations between cyclooxygenase-2 (COX-2) and vascular endothelial growth factor (VEGF) expressions and angiogenesis in pharyngeal tissue of patients with obstructive sleep apnea hypopnea syndrome (OSAHS).

METHODSBiopsies were obtained by uvulopalatopharyngoplasty from 40 patients with mild to severe OSAHS. Control specimens of palatopharyngeal and palatoglossal arch mucosa were retreved from 6 patients with chronic tonsillitis and proved have no related disorders. HE was used to observe the changes of pharyngeal tissue, immunohistochemical staining with antibodies against COX-2, VEGF, microvessel density (MVD) (marked with CD34).

RESULTSCOX-2 and VEGF mainly expressed at pavement-epithelium and glandular epithelium of pharyngeal tissue, and stronger COX-2 and VEGF expression was found in midrange and severe OSAHS than mild and control group (P < 0.01), so as MVD. COX-2 expression was correlated positively with VEGF expression, and had significant correlation with MVD. VEGF expression had the same correlation with MVD. These three targets had considerable relation with apnea hypopnea index (AHI) and lowest O2 saturation at night.

CONCLUSIONThere was angiogenesis which had important relationship with hypoxia degree in patients of OSAHS, and COX-2 and VEGF play a crucial role in its development.

Adult ; Cyclooxygenase 2 ; genetics ; metabolism ; Female ; Humans ; Male ; Middle Aged ; Neovascularization, Pathologic ; Pharynx ; blood supply ; metabolism ; Sleep Apnea, Obstructive ; metabolism ; physiopathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo study the effects of prolonged 85% oxygen exposure on lung vascular development and the expression of angiopoietin-1 (Ang-1) in the neonatal rat lungs.

METHODSNinety-six Sprague-Dawley rat pups were randomly exposed to air (control group) and 85% oxygen (experimental group) 6 hrs after birth. The rats were sacrificed 3, 7 and 14 days after exposure and their lungs were sampled. The lung sections were stained with hematoxylin and eosin for histological evaluation and analysis of vessel volume density. Expressions of angiopoietin-1 (Ang-1) in lung tissue were measured by immunohistochemistry. Expression of Ang-1 protein and mRNA was detected by Western Blot and Real time-PCR.

RESULTSAfter being exposed to 85% oxygen for 14 days, lung tissues had pathological changes as "new" bronchopulmonary dysplasia (BPD). The RAC on day 7 and day 14 in experimental group decreased significantly as compared with the control group [(10.55 ± 0.13) vs. (11.74 ± 0.19), (12.47 ± 0.05) vs. (15.03 ± 0.16), P < 0.05]. The X-ray showed that the diameter of lung vessel was much smaller and the vessels had less branches in experimental group compared with the control group on day 14. The vessel volume density on day 14 in experimental group decreased significantly as compared with the control group [(3.55 ± 0.09) vs. (6.03 ± 0.16), P < 0.05]. Immunohistochemistry and Western blotting showed that the expressions of Ang-1 protein on day 7 and day 14 in the experimental group decreased significantly as compared with the control group [(4.27 ± 0.34) vs. (3.10 ± 0.29), P < 0.05, (5.65 ± 0.49) vs. (3.21 ± 0.28), P < 0.01], [(0.88 ± 0.31) vs. (0.41 ± 0.12), P < 0.05, (0.90 ± 0.29) vs. (0.21 ± 0.06), P < 0.01]. The expressions of Ang-1 mRNA on day 7 and day 14 in the experimental group also decreased significantly as compared with the control group [(0.85 ± 0.14) vs. (0.44 ± 0.21), P < 0.05, (0.87 ± 0.24) vs. (0.24 ± 0.05), P < 0.01].

CONCLUSIONSProlonged exposure of high concentration of oxygen may cause impairment of lung vascular development by inhibiting expression of Ang-1 in neonatal rats, which is likely to contribute to pathogenesis of BPD.

Angiopoietin-1 ; metabolism ; Animals ; Animals, Newborn ; Hyperoxia ; Lung ; blood supply ; metabolism ; Pulmonary Artery ; growth & development ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo verify whether the extracellular domain of kinase domain region (KDR) has anti-angiogenesis activity in vivo.

METHODScDNA was cloned into adeno-associated virus (AAV) vector pSNAV and transfected to baby hamster kidney (BHK) cells. Recombinant AAV was obtained from the cell culture supernatant after adding helper virus. Recombinant AAV-infected human bladder cancer EJ cell line (EJ cells) were injected subcutaneously into Balb-c nude mice. Tumor specimens were removed from the mice, paraffin-embedded and sliced, then stained by immunohistochemistry. Microvessel density (MVD) was determined under a microscope.

RESULTSThe tumor volume developed by EJ cells transfected with the extracellular domain of KDR was significantly smaller (1.70 +/- 0.18 cm(3)) compared with that in the control (5.62 +/- 0.67 cm(3)) (P < 0.05), although tumor developed to be detectable on almost the same time (14.7 +/- 2.4 days vs 14.1 +/- 3.2 days). Further, MVD in the experimental group was lower than that in the control (41.3 +/- 4.8 vs 6.2 +/- 2.1, P < 0.05).

CONCLUSIONThe extracellular domain of KDR could be expressed in nude mouse bladder cancer tissue and inhibit tumor angiogenesis.

Animals ; Cloning, Molecular ; Cricetinae ; Dependovirus ; genetics ; Endothelial Growth Factors ; metabolism ; Female ; Genetic Therapy ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neovascularization, Pathologic ; prevention & control ; Urinary Bladder Neoplasms ; blood supply ; therapy ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the effect of vascular endothelial growth factor 165 (VEGF 165) gene on vascularization of dermal substitute in vivo.

METHODSHuman umbilical vein endothelial cells (HUVECs) were cultured in M199 medium containing FBS in the volume fraction of 10% (briefly called complete medium). (1) HUVECs were divided into non-transfection group (without transfection), empty vector group [transfected with pIRES2-enhanced green fluorescent protein (EGFP) plasmid], and VEGF plasmid group (transfected with pIRES2-EGFP-VEGF plasmid) according to the random number table, with 6 wells in each group. At post transfection hour (PTH) 24, the expression of green fluorescent protein (GFP) in each group was observed under inverted phase contrast fluorescence microscope, and the expression rate of GFP was detected with flow cytometer. Cells in non-transfection group were tested with the same methods as listed above. The cells in stable transfection in empty vector group and VEGF plasmid group were sifted by neomycin. The mRNA and protein expression levels of VEGF 165 in cells and the protein amount of VEGF 165 in the supernatant of cell culture medium in 3 groups were respectively determined by real-time fluorescent quantitation PCR, Western blotting, and enzyme-linked immunosorbent assay. (2) Forty-eight male nude mice were divided into 4 groups according to the random number table, with 12 mice in each group. Mice in saline group were subcutaneously implanted with dermal substitutes which had been cultured in saline for 2 days on both sides of back (the same site below); mice in medium group were subcutaneously implanted with dermal substitutes which had been cultured in complete medium for 2 days; mice in non-transfected cells group were subcutaneously implanted with dermal substitutes that had been cultured in complete medium with non-transfected HUVECs for 2 days; mice in transfected cells group were subcutaneously implanted with dermal substitutes that had been cultured in complete medium with HUVECs stably transfected with VEGF plasmid for 2 days. The dermal substitutes in every group were taken out on post operation day (POD) 3, 7, 14, and 21. Distributions of microvessels and HUVECs in dermal substitutes were observed by immunohistochemical staining, and the microvessel number was counted on POD 14; the expression level of VEGF 165 protein in dermal substitutes was determined by Western blotting. The experiments were all done in triplicate. Data were processed with one-way analysis of variance and LSD method.

RESULTS(1) Obvious green fluorescence was only observed in the two groups with transfected cells at PTH 24. Expression rates of GFP in the cells of non-transfection group, empty vector group, and VEGF plasmid group were respectively 0, (85.2 ± 3.2) %, and (93.1 ± 2.4) %. In the non-transfection group, empty vector group, and VEGF plasmid group, the relative expression amounts of VEGF 165 mRNA were respectively 1, 1.05 ± 0.09, and 3.02 ± 0.13 (F = 5.28, P < 0.05); the relative expression amounts of VEGF 165 protein were respectively 0.78 ± 0.16, 0.76 ± 0.13, and 1.92 ± 0.18 (F = 7.62, P < 0.05); the protein quantity of VEGF 165 in cell supernatant was respectively (62.4 ± 2.7), (73.1 ± 3.8), (117.5 ± 3.1) pg/mL (F = 15.08, P < 0.05). The mRNA and protein levels of VEGF 165 and VEGF 165 protein amount in supernatant were significantly higher in VEGF plasmid group than in the other two groups, with P values all below 0.05. (2) The number of HUVECs in dermal substitutes of transfected cells group was significantly higher than that of the other three groups on POD 14. The numbers of microvessels of dermal substitutes on POD 14 in saline group, medium group, non-transfected cells group, transfected cells group were respectively 4.2 ± 1.1, 5.2 ± 1.1, 6.6 ± 0.9, 13.8 ± 0.8 per 200 times visual field (F = 17.96, P < 0.01). The microvessel number in transfected cells group was significantly higher than that of the other three groups, with P values all below 0.05. The relative expression ratio of VEGF 165 protein of dermal substitutes in transfected cells group was significantly higher than that in saline group as of POD 7. On POD 14 and 21, the relative expression ratios of VEGF 165 proteins in non-transfected cells group (1.652 ± 0.086, 2.152 ± 0.062) and transfected cells group (2.403 ± 0.091, 2.879 ± 0.047) were significantly higher than those of saline group (1.299 ± 0.027, 1.362 ± 0.103), with P values all below 0.05. And the index level of transfected cells group was significantly higher than that in non-transfected cells group (with P values below 0.05). The VEGF 165 protein content in dermal substitutes increased with time extension in all groups.

CONCLUSIONSTransfection of VEGF 165 gene in HUVEC could effectively facilitate vascularization of dermal substitutes in vivo by high expression of VEGF 165 protein.

Animals ; Cells, Cultured ; Dermis ; blood supply ; Human Umbilical Vein Endothelial Cells ; Humans ; Male ; Mice ; Mice, Nude ; Plasmids ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo explore the protective effect of electroacupuncture (EA) pretreatment on ovarian function in rats with ovulation induction.

METHODSThirty SD female rats were numbered according to random number table. According to vaginal smear method, rats of estrus were divided into a normal group (10 rats) and cohabitated with male SD rats with the proportion of 1:1. With computer-generated random number, the remaining rats were divided into a model group and an EA group, 10 rats in each one. The model of superovulation was established with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (HCG) in the model group and EA group. Before model establishment and cohabitation, rats in the EA group were treated with EA at "Guanyuan (CV 4)" and "Sanyinjiao (SP 6)", once for 15 min, for consecutive 7 days. Rats in the normal group and model group received no further treatment. The third day 23:00 pm after cohabitation, blood samples in three groups were collected to test the level of estradiol (E₂) and progesterone (P). After the rats were sacrificed, the HE staining method was applied to observe the morphological changes of ovarian tissue; the immunohistochemical method was applied to measure the expression of vascular endothelial growth factor (VEGF) and its receptor VEGFR-2; the real-time quantitative PCR technique was applied to measure the gene expression of VEGF and VEGFR-2.

RESULTSThe number of ovarian follicle in the EA group was higher than that in the model group and normal group (all P < 0.05); the ratio of corpus luteum size to ovarian size in the EA group was lower than that in the model group (P < 0.01). The ratio of plasma estradiol to progesterone in the EA group tended to be normal group (P < 0.05) and lower than that in the model group (P < 0.01). The protein expression of VEGF and VEGFR-2 in lutein granulosa cell and follicular fluid in the EA group was lower than that in the model group (P < 0.05); gene level of VEGF and VEGFR-2 in ovarian tissue in the EA group was lower than that in the model group (P < 0.05, P < 0.01).

CONCLUSIONEA pretreatment has certain protective effect on ovarian function in rats with ovulation induction, which is likely to be related to regulation of VEGF and its receptor.

Acupuncture Points ; Animals ; Chorionic Gonadotropin ; blood ; Electroacupuncture ; Estradiol ; blood ; Female ; Male ; Ovary ; physiology ; Ovulation Induction ; Pregnancy ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Vascular Endothelial Growth Factor A ; genetics ; metabolism