INTRODUCTIONVascular endothelial growth factor (VEGF) which is also known as vascular permeability factor (VPF) is a heparin-binding, dimeric polypeptide growth factor and a potent mitogen for endothelial cells.VEGF can stimulate the endothelial cell growth and enhance the motility through its two known receptors flt-1 and KDR[1]. Acting through these receptors, VEGF may stimulate angiogenesis and promote tumor progression. VEGF12l, as one of the four VEGF protein isoforms containing the least number of amino acids, has all the biological function of VEGF and is the ideal isoforms for further studying VEGF at molecular levels[2]. In this study, we cloned

Percutaneous microballoon compression of the trigeminal ganglion is a brand new operative technique for the treatment of trigeminal neuralgia. However, it is unclear how the procedure mediates pain relief, and there are no standardized criteria, such as compression pressure, com-pression time or balloon shape, for the procedure. In this study, percutaneous microballoon compression was performed on the rabbit trigeminal ganglion at a mean inlfation pressure of 1,005 ± 150 mmHg for 2 or 5 minutes. At 1, 7 and 14 days after percutaneous microballoon compression, the large-diameter myelinated nerves displayed axonal swelling, rupture and demy-elination under the electron microscope. Fragmentation of myelin and formation of digestion chambers were more evident after 5 minutes of compression. Image analyzer results showed that the diameter of trigeminal ganglion cells remained unaltered after compression. These experi-mental ifndings indicate that a 2-minute period of compression can suppress pain transduction. Immunohistochemical staining revealed that vascular endothelial growth factor expression in the ganglion cells and axons was signiifcantly increased 7 days after trigeminal ganglion compression, however, the changes were similar after 2-minute compression and 5-minute compression. The upregulated expression of vascular endothelial growth factor in the ganglion cells after percu-taneous microballoon compression can promote the repair of the injured nerve. These ifndings suggest that long-term compression is ideal for patients with recurrent trigeminal neuralgia.

Neural stem cells are characterized by the ability to differentiate and stably express exogenous ge-nes. Vascular endothelial growth factor plays a role in protecting local blood vessels and neurons of newborn rats with hypoxic-ischemic encephalopathy. Transplantation of vascular endothelial growth factor-transfected neural stem cells may be neuroprotective in rats with cerebral palsy. In this study, 7-day-old Sprague-Dawley rats were divided into ifve groups: (1) sham operation (control), (2) cerebral palsy model alone or with (3) phosphate-buffered saline, (4) vascular en-dothelial growth factor 165 + neural stem cells, or (5) neural stem cells alone. hTe cerebral palsy model was established by ligating the letf common carotid artery followed by exposure to hypox-ia. Phosphate-buffered saline, vascular endothelial growth factor + neural stem cells, and neural stem cells alone were administered into the sensorimotor cortex using the stereotaxic instrument and microsyringe. Atfer transplantation, the radial-arm water maze test and holding test were performed. Immunohistochemistry for vascular endothelial growth factor and histology using hematoxylin-eosin were performed on cerebral cortex. Results revealed that the number of vas-cular endothelial growth factor-positive cells in cerebral palsy rats transplanted with vascular endothelial growth factor-transfected neural stem cells was increased, the time for ifnding water and the ifnding repetitions were reduced, the holding time was prolonged, and the degree of cell degeneration or necrosis was reduced. hTese ifndings indicate that the transplantation of vascu-lar endothelial growth factor-transfected neural stem cells alleviates brain damage and cognitive deifcits, and is neuroprotective in neonatal rats with hypoxia ischemic-mediated cerebral palsy.

Previous studies have demonstrated the protective effect of hypoxic preconditioning on acute ce-rebral infarction, but the mechanisms underlying this protection remain unclear. To investigate the protective mechanisms of hypoxic preconditioning in relation to its effects on angiogenesis, we in-duced a photochemical model of cerebral infarction in an inbred line of mice (BALB/c). Mice were then exposed to hypoxic preconditioning 30 minutes prior to model establishment. Results showed significantly increased vascular endothelial growth factor and CD31 expression in the ischemic penumbra at 24 and 72 hours post infarction, mainly in neurons and vascular endothelial cells. Hy-Hypoxic preconditioning increased vascular endothelial growth factor and CD31 expression in the ischemic penumbra and the expression of vascular endothelial growth factor was positively related to that of CD31. Moreover, hypoxic preconditioning reduced the infarct volume and improved rological function in mice. These findings indicate that the protective role of hypoxic preconditioning in acute cerebral infarction may possibly be due to an increase in expression of vascular endothelial growth factor and CD31 in the ischemic penumbra, which promoted angiogenesis.

We previously showed that the repair of bone defects is regulated by neural and vascular signals. In the present study, we examined the effect of topically appliedβ-nerve growth factor (β-NGF) on neurogenesis and angiogenesis in critical-sized bone defects iflled with collagen bone substi-tute. We created two symmetrical defects, 2.5 mm in diameter, on either side of the parietal bone of the skull, and filled them with bone substitute. Subcutaneously implanted osmotic pumps were used to infuse 10 μgβ-NGF in PBS (β-NGF + PBS) into the right-hand side defect, and PBS into the left (control) defect, over the 7 days following surgery. Immunohistochemical staining and hematoxylin-eosin staining were carried out at 3, 7, 14, 21 and 28 days postoperatively. On day 7, expression of β III-tubulin was lower on theβ-NGF + PBS side than on the control side, and that of neuroiflament 160 was greater. On day 14,β III-tubulin and protein gene product 9.5 were greater on theβ-NGF + PBS side than on the control side. Vascular endothelial growth factor expression was greater on the experimental side than the control side at 7 days, and vascular endothelial growth factor receptor 2 expression was elevated on days 14 and 21, but lower than control levels on day 28. However, no difference in the number of blood vessels was observed between sides. Our results indicate that topical application ofβ-NGF promoted neu-rogenesis, and may modulate angiogenesis by promoting nerve regeneration in collagen bone substitute-iflled defects.

Oenanthe javanica is an aquatic perennial herb that belongs to theOenanthe genus in Apiaceae family, and it displays well-known medicinal properties such as protective effects against glu-tamate-induced neurotoxicity. However, few studies regarding effects ofOenanthe javanica on neurogenesis in the brain have been reported. In this study, we examined the effects of a normal diet and a diet containing ethanol extract ofOenanthe javanica on cell proliferation and neu-roblast differentiation in the subgranular zone of the hippocampal dentate gyrus of adolescent rats using Ki-67 (an endogenous marker for cell proliferation) and doublecortin (a marker for neuroblast). Our results showed thatOenanthe javanica extract signiifcantly increased the number of Ki-67-immunoreactive cells and doublecortin-immunoreactive neuroblasts in the subgranular zone of the dentate gyrus in the adolescent rats. In addition, the immunoreactivity of brain-derived neurotrophic factor was signiifcantly increased in the dentate gyrus of theOe-nanthe javanica extract-treated group compared with the control group. However, we did not ifnd that vascular endothelial growth factor expression was increased in theOenanthe javanica extract-treated group compared with the control group. These results indicate thatOenanthe javanica extract improves cell proliferation and neuroblast differentiation by increasing brain-de-rived neurotrophic factor immunoreactivity in the rat dentate gyrus.

Objective. To investigate the dynamic changes of serum vascular endothelial growth factor(VEGF) levels in a rat model of acute myocardial infarction. Materials and methods. Eighty-eight adult male Sprague-Dawley rats weighing approximately 270 g were used in this study. Eighty rats were subjected to left coronary artery ligation, with 8 rats for each different duration of infarct.Eight sham-operated animals in which the left coronary artery was surgically exposed without ligation were used as con-trols. Blood samples were drawn from the right atrium before ( sham animals ) and 1,3,6,12,24 h and 2,3,5,7,14 dafter myocardial infarction. The concentrations of serum VEGF were measured by a sensitive enzyme-linked im munosorbent assay with a rabbit polyclonal antibody specific for VEGF. Results. In the 8 control animals, the mean concentration of serum VEGF was 66.99 ± 17.83 pg/ml. Six hours after myocardial infarction, the level of serum VEGF significantly increased to 125.68 ± 28.07 pg/ml ( P ＜0.01 vs.sham controls), and reached a peak (240.61 ± 70.63 pg/ml. P ＜0.01 vs. sham animals) at 24 h after ligation andthen decreased gradually over the remaining 2 weeks. However, the level remained significantly elevated for 14 d(107.64±30.13pg/ml, P＜0.01 vs. sham controls). Condusion. The present study shows that the levels of serum VEGF are markedly increased until 14 d in the ratmodel of acute myocantial infarction. The increased serum VEGF level may play an important role in the angiogencsisassociated with myocardial infarction.

Objective. To determine whether serum vascular endothelial growth factor(VEGF)concentrations are altered in several kinds of coronary heart disease patients.Materials and methods. Using a VEGF enzyme-linked immunosorbent assay(ELISA), serum VEGF concentrations were determined in antecubital venous blood of 16 patients with stable angina pectoris(SAP), 16 with unstable angina pectoris(UAP) and 16 with acute myocardial infarction(AMI) before and after thrombolytic therapy, and of 16 age and sex-matched healthy volunteers who used as controls.Results. The concentrations of serum VEGF in patients with SAP(98.60 ± 26.99pg/ml) and UAP (103.61 ±24.89pg/ml) tended to be higher than those in control subjects(80.44 ± 24.57pg/ml), but the differences did not reach statistical significance (P ＞ 0.05 for each). Before throm bolytic therapy, the concentrations of serum VEGF in patients with AMI (285.92 ± 125.15pg/ml) were significantly higher than those in patients with SAP, UAP or control subjects ( P ＜ 0.01 ,respectively), and correlated with synchronous serum creatine kinase (CK) and its MB isoenzyme (CK-MB) contents(r=0.866, P ＜ 0.001 and r =0.948,P ＜ 0.001;respectively). Three hours after thrombolysis, the concentrations of VEGF had fallen to 111.57 ± 31.29pg/ml ( P ＜0.01 vs. before thrombolytic therapy and P＜0.05 vs .control subjects).Conclusion. The present study shows that serum concentrations of VEGF in patients with AMI are markedly elevated and that increased serum VEGF levels may be one of the most sensitive indexes in diagnosing AMI and judging reperfusion.

转染VEGF165的内皮祖细胞移植恢复糖尿病ED大鼠的勃起功能

海绵体神经损伤修复中的三要素:脑源性神经营养因子、血管内皮生长因子与JAK/STAT信号通路回顾

勃起功能障碍基因疗法的现状

Current available treatment options for erectile dysfunction (ED) are effective but not without failure and/or side effects. Although the development of phosphodiesterase type 5 (PDE5) inhibitors (I.e. Sildenafil, tadalafil and vardenafil) has revolutionized the treatment of ED, these oral medications require on-demand access and are not as effective in treating ED related to diabetic, post-prostatectomy and severe veno-occlusive disease states. Improvement in the treatment of ED is dependent on understanding the regulation of human corporal smooth muscle tone and on the identification of relevant molecular targets. Future ED therapies might consider the application of molecular technologies such as gene therapy. As a potential therapeutic tool, gene therapy might provide an effective and specific means for altering intracavernous pressure "on demand" without affecting resting penile function. However, the safety of gene therapy remains a major hurdle to overcome before being accepted as a mainstream treatment for ED. Gene therapy aims to cure the underlying conditions in ED, including fibrosis. Furthermore, gene therapy might help prolong the efficacy of the PDE5 inhibitors by improving penile nitric oxide bioactivity. It is feasible to apply gene therapy to the penis because of its location and accessibility, low penile circulatory flow in the flaccid state and the presence of endothelial lined (lacunar) spaces. This review provides a brief insight of the current role of gene therapy in the management of ED.

Objective:To investigate the relationships between the expressions of p15,p16 and vascular endothelial growth factor (VEGF) and gastric carcinoma(GC).Methods: Using immunohistochemical staining to examine the expressions of p15, p16and VEGF in archival wax-embedded specimens of 80 GC and 20 gastric benign disease(GBD). Results: The positive expression rate (PER) of p15 was significantly lower in GC than in GBD (43.75% VS. 69.23%, P<0.05). No relationship was found between PER of p15 and clinicopathologic factors. PER of p16 was 20% in GC, 55% in GBD (P<0.01).PER of p16 wasn't significantly different in gross types, histological types, with or without distant metastasis and pTNM stages. PER of p16 was 71.43% in invasive mucosa or suomucosa group, 17. 24% in invasive muscle group and 13. 64% in invoive serosa group (P<0.01); 12.96% in GC with lymph nodes metastasis, 34.62% in GC without lymph node metastasis (P<0.05). PER of VEGF in GC was 75. 00%, in GBD 7.69% (P<0. 001), in ulcerative type of GC and infiltrating type of GC were 81.97% and 40. 00%, respectively (P <0.05), in GC of invasive serosa was 95.45%, in GC of invasive muscle 51.72%(P<0.001), in GC of invasive mucosa or sulomucosa 42.86% (P<0.001). PER of VEGF in GC with lymph node metastasis was 82. 8%, without lymph node metastasis 54. 6%(P<0.05), in GC accompanied with distant metastasis was 100%, in GC without distant metastasis71.1% (P<0.05). PER of VEGF in pTNM Ⅰ and Ⅱ was 53.13%, in Ⅲ and IV 89.56% (P<0. 001). The expression of p15 correlated significantly With that of VEGF (P<0.001) and with that of p16 (P<0.01) in GC. Conclusion: p15 expression down-regulation has relationship with GC, but on relationship with the progress. p16 expression downregulation and VEGF expression up-regulation show significant relationships with clinicopathologic factors. There are significant relations between p15 and p16 negative expressionsand between p15 expression down-regulation and VEGF expression up-regulation.

Study of the Influence of Angiostatin Intravitreal Injection on Vascular Leakage in Retina and Iris of the Experimental Diabetic Rats

Purpose: To examine the effect of an intravitreal injection of angiostatin on vascular leakage in retina and iris of the diabetes and study its possible mechanism.Methods: Experimental diabetes was induced in 24 rats by an intravenous injection of streptozotocin (STZ) during 48 adult rats. Three groups were randomization distributed of them. There were 8 of both normal and diabetic rats in each group. STZ-diabetic rats and age-matched normal rats received an intravitreal injection of 5 μl of sterile PBS (Phosphate Buffered Saline) into the right eye, and the left eye was non-injected in the group A; Angiostatin was injected into the vitreous of the right eye (7.5 μg/5μl/eye), and the left eye received the same volume of sterile PBS as the control in the group B and C. The vascular permeability of retina and iris was measured using the Evans blue method at 2 days following the injection in the group A and B. Expres sion of VEGF in retina was evaluated using western blot analysis 24 hours following the injection in the group C.Results: Diabetic rats showed significant increases of vascular permeability in the retina (P＜0.01) and iris (P＜0.05). Angiostatin-injected eyes showed significant decreases in vascular permeability in the retina (P＜0.01) and iris (P＜0.05) comparing with the PBS-injected eyes in STZ-diabetic rats. In contrast, intravitreal injection of the same dose of angiostatin into the age-matched normal rats did not result in any significant reduction in vascular permeability in the retina and iris, when compared with the contralateral eye with PBS injection (P＞0.05). Angiostatin injection significantly reduced VEGF level in the retinas of STZ-diabetic rats but did not affect retinal VEGF level in normal rats.Conclusions: Angiostatin significantly reduce pathological vascular permeability in the retina and iris of STZ-diabetic rats but not in normal rats. Angiostatin down-regulates VEGF expression and thus, blocks the major cause of vascular leakage in the diabetic retina. Therefore, angiostatin may have a therapeutic potential in the treatment of diabetic macular edema, cystoid macular edema, uvietis and other diseases with vascular leakage.

Effects and mechanism of miRNA-24on the proliferation,migration and tube formation of vascular endothelial cells

Objective:To investigate the effect and mechanism of miRNA-24 on the proliferation,migration and tube forma tion of vascular endothelial cells.Methods:Human umbilical vein endothelial cells (HUVECs) were assigned into control,microRNA (miR) 24 overexpression and anti-miR-24 groups.The proliferation and migration abilities of HUVECs were detected by methyl thiazolyl tetrazolium (MTT) assay and scratch wound healing assay,respectively.The ability of HUVECs to form tubular structures was evaluated by a tube formation assay.The mRNA and protein expressions of vascular endothelial growth factor (VEGF) and transcription factor Sp1 were determined by RT-PCR,immunocytochemis try and western blotting,respectively.Results:The miR-24 overexpression group exhibited decreased cell proliferation and migration,and expressions of VEGF and Sp1 compared with the control group (P ＜ 0.01).No tube like network structure was formed in the miR-24 overexpression group.However,inhibition of miR 24 in HUVECs markedly in creased cell proliferation and migration,enhanced tube for mation and expressions of VEGF and Sp1 (P＜0.05 or P＜ 0.01).Conclusion:MiR-24 suppressed the proliferation,migration and tube formation of HUVECs,and the mechanism might be related to the down-regulation of VEGF expression.Sp1 might participate in this regulation process.

Mechanisms of inhibition by aspirin of endometrial carcinoma cell proliferation,migration and invasion

Objective:To investigate the effects and mechanisms of aspirin on the proliferation,migration and invasion of endometrial carcinoma HEC-1A cell lines.Methods:HEC-1A cells were cultured to the exponential phase and treated with different concentrations of aspirin (0.625 mmol/ L,1.25 mmol/L,2.5 mmol/L,5 mmol/L and 10 mmol/L) for 24 to 120 hours.Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT) assay.The migration and invasion of HEC-1A cells were detected by transwell assay.The protein expressions of vascular endothelial growth factor (VEGF) and ascular endothelial growth factor receptor 2 (VEGFR-2) in HEC-1A cells were determined by western blotting.Results:MTT results showed that aspirin inhibited the growth and proliferation of endometrial cancer cells in concentration and time-dependent manner.Aspirin had a significant inhibitory effect on the migration and invasion of HEC-1A cells (P＜0.05).In addition,aspirin obviously suppressed concentration-dependently the expression levels of VEGF and VEGFR-2 (P ＜ 0.05).Conclusion:Aspirin could inhibit the proliferation,migration and invasion of endometrial cancer cells.The anti-tumor mechanism of aspirin might be related to the inhibition of tumor angiogenesis via blocking the VEGF/VEGFR-2 signaling pathway.

Role of vascular endothelial growth factor in reconstructive surgery after surgical excision of malignant tumor

As a key mediator of normal physiological angiogenesis,vascular endothelial growth factor(VEGF)has been regarded as an emancipator to plastic surgeon,and yet a misfortune to oncology surgeon,due to its sin-gular biological effect.Therefore in some clinical cases,especially for some malignant tumor patients having en-dured radical surgery and being craving for a reconstructive surgery,VEGF plays a role full of paradoxes.To make a clinical balance,we should find a point to inhibit tumor cell from utilizing VEGF and make a permission to normal tissues to employ it.

Objective:To discuss the clinical value of Serum TSH combined with 3 kinds of VEGF (VEGF-C, VEGF-D and VEGFR-3) determination in the early diagnosis of papillary thyroid carcinoma (PTC). Method:Selected 37 cases of patients with thyroid benign tumor (Benign group) and 37 cases of patients with PTC (PTC group), then collected the serum of these both groups, to determine the TSH, VEGF-C, VEGF-D and VEGFR-3 levels of all cases by chemiluminescence immunoassay and enzyme linked immunosorbent assay respectively. Through Logistic model, to calculate the curve area of TSH combined with 3 kinds of VEGF.Results:PTC group: VEGF-C, VEGFR-3 and TSH levels were obviously higher than that in Benign group (P<0.05); and VEGF-C, VEGFR-3 and TSH levels inⅢ-Ⅳ period patients were obviously higher than that in I-Ⅱ period patients (P<0.05); AUC area of VEGF-C, VEGFR-3 and TSH were respectively 0.805, 0.736 and 0.710, reached to significance level (P<0.05); AUC area of combined diagnosis was 0.859.Conclusion:VEGF-C, VEGFR-3 and TSH between papillary thyroid carcinoma and thyroid benign tumor had significant difference. Combined determination could improve the early diagnose rate of PTC, and could be regarded as one of the important auxiliary index of PTC early diagnosis.

Background: Clinical trials have revealed that the antivascular endothelial growth factor (VEGF) therapies are effective in retinopathy of prematurity (ROP). But the low level of VEGF was necessary as a survival signal in healthy conditions, and endogenous placental growth factor (PIGF) is redundant for development. The purpose of this study was to elucidate the PIGF expression under hypoxia as well as the infl uence of anti-VEGF therapy on PIGF.
Methods: CoCl2-induced hypoxic human umbilical vein endothelial cells (HUVECs) were used for an in vitro study, and oxygen-induced retinopathy (OIR) mice models were used for an in vivo study. The expression patterns of PIGF under hypoxic conditions and the infl uence of anti- VEGF therapy on PIGF were evaluated by quantitative reverse transcription-polymerase chain reaction (RTPCR). The retinal avascular areas and neovascularization (NV) areas of anti-VEGF, anti-PIGF and combination treatments were calculated. Retina PIGF concentration was evaluated by ELISA after treatment. The vasoactive effects of exogenous PIGF on HUVECs were investigated by proliferation and migration studies.
Results: PIGF mRNA expression was reduced by hypoxia in OIR mice, in HUVECs under hypoxia and anti-VEGF treatment. However, PIGF expression was reversed by anti-VEGF therapy in the OIR model and in HUVECs under hypoxia. Exogenous PIGF significantly inhibited HUVECs proliferation and migration under normal conditions, but it stimulated cell proliferation and migration under hypoxia. Anti-PIGF treatment was effective for neovascular tufts in OIR mice (P<0.05).
Conclusion: The finding that PIGF expression is iatrogenically up-regulated by anti-VEGF therapy provides a consideration to combine it with anti-PIGF therapy.