Objective To verify whether the extracellular domain of flt 1 has anti angionesis activity in vivo. Methods A recombinant plasmid pcDNA 3.1/flt 1 n3 was transfected into the human bladder carcinoma EJ cell by lipofectamin, which was constructed by inserting the N terminal first three IgG like domain of VEGF receptor flt 1 (flt 1 n3 ) into eukaryotic expression vector pcDNA 3.1. Results The rflt 1 was functionally expressed in stably transfected EJ and the product secreted in the medium could be specifically bind to rhVEGF165.The result showed that the bladder cancer transfected with rflt 1 had a lower microvessle density than the control. Conclusions It is proved that the expressed rflt 1 n3 can inhibit tumor growth and angiogenesis in nude mice model.

Objective: To investigate the innate immune response of influenza virus-infected glial cells,the transcription levels in chemokines in mouse microglia and astrocytes were detected which pre-infected by human H1N1 or avian H5N1 influenza viruses.Methods: The glial cells isolated from neonatal mice cerebral cortex were cultured and further microglia and astrocytes were purified.The primary mouse microglia and astrocytes were infected in vitro by H1N1 or H5N1 influenza viruses in a multiplicity of infection (MOI) 2.Eight hours post infection,the influenza virus nucleoprotein (NP) was detected by immunofluorescence to identify the proportion of infected cells.The cellular RNA were extracted at 6 h and 24 h to detect the transcriptional level of chemokines by semi-quantitative RT-PCR.Results: More than 95% of the microgha and astrocytes which isolated from mice were infected.The transcription levels of CCL-3,CCL-5,CXCL-2,CXCL-9 and CXCL-10 from infected microglia and astrocytes were upregulated.Futhermore,the mRNA level of CXCL-10 increased much more.In addition,avian H5N1 influenza virus could induce more stronger upregulation of those chemokines than human H1N1 did.Conclusion: The mouse microglia and astro cytes which are infected by H1N1 influenza virus or H5N1 influenza virus could induce upregulation of transcription level of chemokines.

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