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1.
Objective: To study the effects of the generation 4 polyamidoamine/vascular endothelial growth factor antisense oligodeoxynucleotide (G4PAMAMNEGFASODN) compound on the expressions of vascular endothelial growth factor (VEGF) and its mRNA of breast cancer cells and on the inhibition of vascular endothelial cells. Methods: We examined the morphology of G4PAMAM/VEGFASODN compound and its pH stability, in vitro transfection efficiency and toxicity, and the expressions of VEGF and its mRNA. Methyl thiazolyl tetrazolium assay was used to detect the inhibitory function of the compound on vascular endothelial cells. Results: The compound was about 10 nm in diameter and was homogeneously netlike. From pH 5 to 10, it showed quite a buffered ability. The 48-h transfection rate in the charge ratio of 1:40 was 98.76%, significantly higher than that of the liposome group (P<0.05). None of the transfection products showed obvious toxicity on the cells. The expressions of both VEGF protein and its mRNA after G4PAMAM/VEGFASODN transfection decreased markedly. Conclusion: With a low toxicity, high safety, and high transfection rate, G4PAMAMNEGFASODN could be a promising gene vector. Specifically, it inhibits VEGF gene expression efficiently, laying a basis for further in vivo animal studies.  相似文献   

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3.
1IntroductionRibosome-inactivating proteins(RIPs)occur natu-rally in a variety of higher plant species,and theyfunction by catalytic depurination of a specific aden-osine residue located near the3*terminus of eukary-otic large ribosomal subunit rRNA,preventing EF-2/GTP binding and thereby blocking peptidyl-tRNAtranslocation during protein synthesis[1].Many RIPsare potent antiviral and antifungal proteins in vitro[2,3],but it may beinsufficient for field application,as com-pared to the ac…  相似文献   

4.
Objective: To investigate the relationship between the expression of endothelial nitric oxide synthase (eNOS), vascular endothelial growth factor (VEGF) and angiogenesis in primary astrocytoma. Methods: Thirty-seven primary astrocytomas and 4 astrocytic hyperplasia samples were collected and divided into three groups according to histological grade. The expression of eNOS, VEGF and factorⅧrelated antigen (FⅧRAg) were assayed by immunohistochemistry. Microvascular density was assessed by FⅧRAg immunoreactivity. The intensity of immunoreactivity was graded according to the percentage of positive tumor cells. Results: No eNOS and VEGF were expressed in the astrocytes and vascular endothelium in astrocytic hyperplasia. The expression of eNOS or VEGF was light in low-grade astrocytoma and strong in glioblastoma. eNOS expression in astrocytoma was very positively correlated with VEGF. eNOS and VEGF expression in anaplastic astrocytoma was median in contrast to the low grade astrocytoma and glioblastoma. Lower microvascular density was found in low grade astrocytoma than that in higher grade malignant ones. The expressions of eNOS and VEGF were correlated with microvascular density and tumor malignancy. Conclusion: This finding suggests that eNOS and VEGF may have cooperative effect in tumor angiogenesis and play an important role in the pathogenesis of primary astrocytoma.  相似文献   

5.
为了提高血管内皮生长因子(VEGF)121的免疫原型,文章用戊二醛一步偶联法将VEGF121与HSP65偶联形成HSP65-VEGF121复合物,并研究其免疫学效果,结果发现HSP65可以提高VEGF121的免疫原性,但抗肿瘤效果并未显著增强.  相似文献   

6.
Mucoepidermoid carcinoma undergoes uniquely vigorous angiogenic and neovascularization processes, possibly due to proliferation of vascular endothelial cells (ECs) induced by mucoepidermoid carcinoma cells (MCCs) in their three-dimensional (3D) microenvironment. To date, no studies have dealt with tumor cells and vascular ECs from the same origin of mucoepidermoid carcinoma using the in vitro 3D microenvironment model. In this context, the current research aims to observe neovascularization with mucoepidermoid carcinoma microvascular ECs (MCMECs) conditioned by the microenvironment in the 3D collagen matrix model. We observed the growth of MCMECs purified by immunomagnetic beads and induced by MCCs, and characteristics of tubule-like structures (TLSs) formed by induced MCMECs or non-induced MCMECs. The assessment parameters involved the growth curve, the length, the outer and inner diameters, and the wall thickness of the TLSs, and the cell cycle. Results showed that MCCs induced formation of the TLSs in the 3D collagen matrix model. A statistically significant difference was noted regarding the count of TLSs between the control group and the induction group on the 4th day of culture (t=5.00, P=0.001). The outer and inner diameters (t 1=5.549, P 1=0.000; t 2=10.663, P 2=0.000) and lengths (t=18.035, P=0.000) of the TLSs in the induction group were statistically significant larger than those in the control group. The TLSs were formed at the earlier time in the induction group compared with the control group. It is concluded that MCCs promote growth and migration of MCMECs, and formation of the TLSs. The 3D collagen matrix model with MCMECs induced by MCCs in the current research may be a favorable choice for research on pro-angiogenic factors in progression of mucoepidermoid carcinoma.  相似文献   

7.
目的：通过康脑液干预观察其对脑缺血再灌注损伤大鼠血管内皮生长因子（vascular endothelial growth factor,VEGF）、脑源性神经生长因子（brain derived neurotrophic factor,BDNF）、基质金属蛋白酶（matrix metalloproteinase-9,MMP-9）表达的影响,探讨康脑液对脑缺血再灌注损伤的保护机制.方法：将雄性SD大鼠随机分为假手术组、脑缺血再灌注模型组及康脑液28.6、14.3、7.15 g·kg-1·d-1剂量组（灌胃给药7 d）,改进Longa等线栓法制备大鼠右侧大脑中动脉阻塞（middle cerebral artery occlusion,MCAO）再灌注模型.于缺血2h后再灌注（分别于再灌注后6h、12h、24 h、72 h、7d处死大鼠）;采用TTC染色法观察大鼠的脑梗死面积;免疫组织化学法观察大鼠脑组织VEGF、BDNF、MMP-9的表达.结果：比较各组缺血再灌注24 h大鼠脑梗死灶面积,发现28.6、14.3 g·kg--1·d-1剂量组较脑缺血再灌注模型组明显减小（P＜0.05）;与脑缺血再灌注模型组相比,28.6、14.3 g·kg--1·d-1剂量组各时间点的VEGF、BDNF表达量明显上调（P＜0.01）,MMP-9表达的阳性细胞明显减少（P＜0.01）,差异有统计学意义.结论：康脑液可促进大鼠局灶性脑缺血后脑组织中VEGF,BDNF的表达,同时抑制脑内MMP-9的表达,缩小脑梗死面积,发挥对神经血管单元（neurovascular unit,NVU）的保护作用,减轻脑缺血再灌注损伤.  相似文献   

8.
应用免疫组化方法,比较性地研究了大肠癌组织及其同一患者形态正常大肠粘膜组织中血管内皮细胞生长因子的表达,分析血管内皮细胞生长因子表达与大肠癌发生的年龄、性别、病理分期等因素的关系。结果显示,癌组织与正常大肠粘膜的血管内皮细胞生长因子表达存在明显差异,血管内皮细胞生长因子的表达与大肠癌患者年龄、性别、分化程度等因素不存在相关性,而与Ducks分期存在明显相关性。表明肿瘤血管的生成在大肠癌的生长发展方面起着至关重要的作用,血管内皮细胞生长因子的检测可以作为大肠癌诊断的重要参考指标。  相似文献   

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10.

Objective

To construct a novel non-viral vector loaded with growth and differentiation factor-5 (GDF-5) plasmid using chitosan, hyaluronic acid, and chondroitin sulfate for osteoarthritis (OA) gene therapy.

Methods

Nano-microspheres (NMPs) were prepared by mixing chitosan, hyaluronic acid, and chondroitin sulfate. GDF-5 plasmid was encapsulated in the NMPs through electrostatic adsorption. The basic characteristics of the NMPs were observed, and then they were co-cultured with chondrocytes to observe their effects on extracellular matrix (ECM) protein expression. Finally, NMPs loaded with GDF-5 were injected into the articular cavities of rabbits to observe their therapeutic effects on OA in vivo.

Results

NMPs exhibited good physicochemical properties and low cytotoxicity. Their average diameter was (0.61±0.20) μm, and encapsulation efficiency was (38.19±0.36)%. According to Cell Counting Kit-8 (CCK-8) assay, relative cell viability was 75%–99% when the total weight of NMPs was less than 560 μg. Transfection efficiency was (62.0±2.1)% in a liposome group, and (60.0±1.8)% in the NMP group. There was no significant difference between the two groups (P>0.05). Immunohistochemical staining results suggested that NMPs can successfully transfect chondrocytes and stimulate ECM protein expression in vitro. Compared with the control groups, the NMP group significantly promoted the expression of chondrocyte ECM in vivo (P<0.05), as shown by analysis of the biochemical composition of chondrocyte ECM. When NMPs were injected into OA model rabbits, the expression of ECM proteins in chondrocytes was significantly promoted and the progression of OA was slowed down.

Conclusions

Based on these data, we think that these NMPs with excellent physicochemical and biological properties could be promising non-viral vectors for OA gene therapy.
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11.
Objective:To explore the feasibility of using regenerated silk fibroin membrane to construct artificial skin substitutes for wound healing, it is necessary to evaluate its cytocompatibility. Methods:The effects of regenerated silk fibroin film on cytotoxicity, adhesion, cell cycle, and apoptosis of L929 cells, growth and vascular endothelial growth factor (VEGF) expression of ECV304 cells, and VEGF, angiopoietin-1 (Ang-1), platelet-derived growth factor (PDGF) and fibroblast growth factor 2 (FGF2) expressio...  相似文献   

12.
Angiogenesis is required for solid tumor growth and facilitates tumor progression and metastasis. The inhibition effects of O-（chloroacetyl-carbamoyl） fumagillol （TNP-470）, an angiogenesis inhibitor, and gemcitabine, a chemotherapeutic agent, on expression of growth factors were investigated using human pulmonary adenocarcinoma cell line, A549. The A549 cells were divided into four groups： control group, 10^-6 mg/ml gemcitabine treated group, 10^-4 mg/ml TNP-470 treated group and gemcitabine＋TNP-470 treated group. The mRNA and protein expression of vascular endothelial growth factor （VEGF） and its receptors, FMS-like tyrosine kinase-l （FLT-1） and kinase insert domain-containing receptor （KDR）, in different groups were measured. The growth of A549 cell cultured with gemcitabine or TNP-470 was inhibited in an almost dose-dependent manner. Although gemcitabine （10^-6 mg/ml） alone and TNP-470 （10^-4 mg/ml） alone had no effect on the mRNA and protein expression of VEGF and its receptors （FLT-1, KDR） in A549 cells compared to the control （P〉0.05）, 10^-6 mg/ml gemcitabine in combination with 10^-4 mg/ml TNP-470 had significant effect （P〈0.01）. Moreover, combination of the two drugs significantly inhibited the mRNA expression of VEGF, FLT-1 and KDR compared to either drug alone （P〈0.05）. This study suggests that combined treatment with TNP-470 plus gemcitabine may augment the antiangiogenic and antineoplastic effects in lung cancer cells in vitro.  相似文献   

13.
Objective: To evaluate the transduction efficiency of a recombinant adenovirus carrying the gene for green fluorescent protein (Ad-GFP) into the primary cultures of fetal neural stem cells (NSCs) by the expression of GFP. Methods: The Ad-GFP was constructed by homologous recombination in bacteria with the AdEasy system; NSCs were isolated from rat fetal hippocampus and cultured as neurosphere suspensions. After infection with the recombinant Ad-GFP, NSCs were examined with a fluorescent microscopy and a flow cytometry for their expression of GFP. Results: After the viral infection, flow cytometry analysis revealed that the percentage of GFP-positive cells was as high as 97.05%. The infected NSCs sustained the GFP expression for above 4 weeks. After differentiated into astrocytes or neurons, they continued to express GFP efficiently. Conclusion: We have successfully constructed a viral vector Ad-GFP that can efficiently infect the primary NSCs. The reporter gene was showed fully and sustained expression in the infected cells as well as their differentiated progenies.  相似文献   

14.
Homoharringtonine (HHT) has currently been used successfully in the treatment of acute and chronic myeloid leukemias and has been shown to induce apoptosis of different types of leukemic cells in vitro. Emerging evidence suggests that angiogenesis may play an important role in hematological malignancies, such as leukemia. How ever, whether HHT can relieve leukemia by anti-angiogenesis is still unknown. We investigated the anti-angiogenesis potential of HHT with the human umbilical vein endothelial cell line (ECV304) and leukemic cell line (K562) in vitro. Cellular proliferation was determined by MTT assay and apoptosis was analyzed by flow cytometry. The mRNA expression of vascular endothelial growth factor (VEGF) was assessed by RT-PCR and VEGF protein production was detected by Western blot. Inhibition of cell proliferation and induction of apoptosis by HHT were discovered in ECV304 cells, and appeared in a dose- and time-dependent manner. Also, treatment with HHT caused down-regulation of VEGF mRNA expression in K562 cells in similar dose- and time-dependent manner and inhibition of VEGF protein production in K562 cells in response to the enhancing concentration of HHT. The results demonstrated that HHT could also induce apoptosis in endothelium and down-regulate VEGF expression in K562 cells. In conclusion, we believe HHT has anti-angiogenesis potential and speculate that HHT might exert its anti-leukemia effects via reduction of angiogenesis.  相似文献   

15.
INTRODUCTION Homoharringtonine (HHT) is a cephalotoxin alkaloid with anti-leukemic activity and had been used successfully in the treatment of acute and chronic myeloid leukemias (O払rien et al., 1995; 1999; Feldman et al., 1992). The principal mecha-nism of action by HHT is the inhibition of protein synthesis in a dose- and time-dependent manner by binding to ribosome and inhibiting polypeptide chain elongation (Tujebajeva et al., 1989; Zhou et al., 1995). HHT had been shown to indu…  相似文献   

16.
INTRODUCTION Soil-borne pathogens, including Pythium spp. and Fusarium spp., cause significant yield losses in horticulture and agriculture crops (Mao et al., 1997). Current practices for controlling plant diseases are based largely on disease resistant crops, cultivation management in fields and application of synthetic pesticides (Elizabeth and Emmert, 1999). Biological control using antagonistic microbes to reduce the use of chemical pesticides in a system of integrated plantdisease …  相似文献   

17.
对虾白斑综合症病毒（WSSV）编码与核酸合成代谢相关的核糖核苷酸还原酶（Ribonucleotide Reductases,RR）,与病毒DNA的复制有关．利用克隆技术将RR基因克隆到L4440载体,构建体内合成dsRNA的大肠杆菌HTn5工程菌．诱导该工程菌合成了RR基因的特异双链RNA（RR—dsRNA）和非特异双链RNA（gfP—dsRNA）,分别与WSSV混合共注射凡纳滨对虾,在感染72h后用病毒检测试剂盒提取DNA模板用于荧光定量PCR分析,结果显示RR—dsRNA能有效抑制WSSV病毒粒子的增值．  相似文献   

18.
Ds-echinoside A (DSEA), a non-sulfated triterpene glycoside, was isolated from the sea cucumber Pearsonothuria graeffei. In vitro and in vivo investigations were conducted on the effects of DSEA on tumor cell adhesion, migration, invasion, and angiogenesis. In this study, we found that DSEA inhibited the proliferation of human hepatocellular liver carcinoma cells Hep G2, with a half-maximal inhibitory concentration (IC50) of 2.65 μmol/L, and suppressed Hep G2 cell adhesion, migration, and invasion in a dose-dependent manner. DSEA also reduced tube formation of human endothelial cells ECV-304 on matrigel in vitro and attenuated neovascularization in the chick embryo chorioallantoic membrane (CAM) assay in vivo. Immunocytochemical analysis revealed that DSEA significantly decreased the expression of matrix metalloproteinase-9 (MMP-9), which plays an important role in the degradation of basement membrane in tumor metastasis and angiogenesis. DSEA also increased the protein expression level of tissue inhibitor of metalloproteinase-1 (TIMP-1), an important regulator of MMP-9 activation. From the results of Western blotting, the expressions of nuclear factor-kappa B (NF-κB) and vascular endothelial growth factor (VEGF) were found to be remarkably reduced by DSEA. These findings suggest that DSEA exhibits a significant antimetastatic activity through the specific inhibition of NF-κB-dependent MMP-9 and VEGF expressions.  相似文献   

19.
Objective： Study blood vessel injury and gene expression indicating vascular endothelial cell apoptosis induced by mannitol with and without administration of anti-oxidative vitamins. Methods： Healthy rabbits were randomly divided into four groups. Mannitol was injected into the vein of the rabbit ear in each animal. Pre-treatment prior to mannitol injection was per- formed with normal saline （group B）, vitamin C （group C） and vitamin E （group D）. Blood vessel injury was assessed under electron and light microscopy. In a second experiment, cell culture specimen of human umbilical vein endothelial cells were treated with mannitol. Pre-treatment was done with normal saline （sample B）, vitamin C （sample C） and vitamin E （sample D）. Total RNA was extracted with the original single step procedure, followed by hybridisation and analysis of gene expression. Results： In the animal experiment, serious blood vessel injury was seen in group A and group B. Group D showed light injury only, and normal tissue without pathological changes was seen in group C. Of all 330 apoptosis-related genes analysed in human cell culture specimen, no significant difference was seen after pre-treatment with normal saline, compared with the gene chip without pre-treatment. On the gene chip pre-treated with vitamin C, 45 apoptosis genes were down-regulated and 34 anti-apoptosis genes were up-regulated. Pre-treatment with vitamin E resulted in the down-regulation of 3 apoptosis genes. Conclusion： Vitamin C can protect vascular endothelial cells from mannitol-induced injury.  相似文献   

20.
Objective: To test the influence of homocysteine on the production and activation of matrix metalloproteinase-2 (MMP-2) and tissue inhibitors of matrix metalloproteinase-2 (TIMP-2) and on cell migration of cultured rat vascular smooth muscle cells (VSMCs). Also, to explore whether rosuvastatin can alter the abnormal secretion and activation of MMP-2 and TIMP-2 and migration of VSMCs induced by homocysteine. Methods: Rat VSMCs were incubated with different concentrations of homocysteine (50–5 000 μmol/L). Western blotting and gelatin zymography were used to investigate the expressions and activities of MMP-2 and TIMP-2 in VSMCs in culture medium when induced with homocysteine for 24, 48, and 72 h. Transwell chambers were employed to test the migratory ability of VSMCs when incubated with homocysteine for 48 h. Different concentrations of rosuvastatin (10−9–10−5 mol/L) were added when VSMCs were induced with 1 000 μmol/L homocysteine. The expressions and activities of MMP-2 and TIMP-2 were examined after incubating for 24, 48, and 72 h, and the migration of VSMCs was also examined after incubating for 48 h. Results: Homocysteine (50–1 000 μmol/L) increased the production and activation of MMP-2 and expression of TIMP-2 in a dose-dependent manner. However, when incubated with 5 000 μmol/L homocysteine, the expression of MMP-2 was up-regulated, but its activity was down-regulated. Increased homocysteine-induced production and activation of MMP-2 were reduced by rosuvastatin in a dose-dependent manner whereas secretion of TIMP-2 was not significantly altered by rosuvastatin. Homocysteine (50–5 000 μmol/L) stimulated the migration of VSMCs in a dose-dependent manner, but this effect was eliminated by rosuvastatin. Conclusions: Homocysteine (50–1 000 μmol/L) significantly increased the production and activation of MMP-2, the expression of TIMP-2, and the migration of VSMCs in a dose-dependent manner. Additional extracellular rosuvastatin can decrease the excessive expression and activation of MMP-2 and abnormal migration of VSMCs induced by homocysteine.  相似文献   

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