Silencing of VEGF inhibits human osteosarcoma angiogenesis and promotes cell apoptosis via VEGF/PI3K/AKT signaling pathway.

BACKGROUND: Osteosarcoma is a kind of highly malignant tumor and the growth and metastasis is closely related to angiogenesis. Vascular endothelial growth factor (VEGF) is an important angiogenesis-promoting factor. In the current study, we investigated the effects of suppressed VEGF on osteosarcoma and its molecular mechanism provided for a basis by targeting angiogenesis. MATERIAL/METHODS: We established bearing human osteosarcoma Wistar rats model by subcutaneous inoculation of human SaOS-2 cells and the adenovirus vector Ad-VEGF-siRNA was constructed for further study. We assessed the efficiency of VEGF silencing and its influence on SaOS-2 cells. The expression of mRNA and protein were detected by RT-PCR and western blotting, respectively. Intratumoral microvessel density (MVD), VEGF and CD31 were evaluated by immunohistochemistry. We detected the cell apoptotic rates by flow cytometry. RESULTS: Our results indicated that Ad-VEGF-siRNA could effectively suppressed the expression of VEGF expression, inhibited the proliferation capability and promoted apoptosis of SaOS-2 cells in vitro. Silencing of VEGF expression also suppress osteosarcoma tumor growth and reduce osteosarcoma angiogenesis in the Wistar rats model in vivo. Furthermore, We found that phosphoinositide 3-kinase (PI3K) and protein kinase B (AKT) activation were considerably reduced while inhibition VEGF expression in SaOS-2 cells. CONCLUSION: Our data demonstrated that VEGF silencing could suppress cells proliferation, promote cells apoptosis and reduce osteosarcoma angiogenesis through inactivation of VEGF/PI3K/AKT signaling pathway.

IGF1 3'UTR functions as a ceRNA in promoting angiogenesis by sponging miR-29 family in osteosarcoma.

Osteosarcoma is one of the most common malignant bone tumors in human worldwide. Angiogenesis is a pivotal process during osteosarcoma development. Insulin-like growth factor 1 (IGF1) has been reported to promote angiogenesis. However, the role of 3' untranslational region (3'UTR) of IGF1 mRNA in angiogenic activity in osteosarcomas is still unknown. In the present study, we performed gain-of-function assays to investigate the role of IGF1-3'UTR in angiogenesis. For the first time, we demonstrated that IGF1 3'UTR increased VEGF expression and promotes angiogenesis in osteosarcoma cells. In addition, RNA-immunoprecipitation and luciferase reporter assays showed that IGF1 3'UTR was a direct target of miR-29s. Our data also demonstrated that there existed a competition of miR-29s between IGF1-3'UTR and VEGF mRNA, and IGF1-3'UTR promoted angiogenesis at least in part via sponging miR-29s. Taken together, our study suggests that IGF1-3'UTR functions as a ceRNA in promoting angiogenesis by sponging miR-29s in osteosarcoma.

Epidemiological features of viral encephalitis in Cangzhou of China with use of multiplex RT-PCR for five RNA viruses.

OBJECTIVE: With use of multiplex RT-PCR testing the five RNA viruses associated with viral encephalitis, the aim of research is to find out the epidemiological features of viral encephalitis in Cangzhou of China. METHODS: Patients hospitalized in Cangzhou central hospital with the diagnosis of viral encephalitis from January 2010 to December 2012 were retrospectively analyzed. The sensitivity and specificity of multiplex RT-PCR was compared with ELISA through testing CSF samples of enterovirus (EVs), Japanese encephalitis virus (JEV), mumps virus (MUV), measles virus (MV) and rubella virus (RV). RESULTS: Disease incidence of viral encephalitis in Cangzhou of China was 18.6 per 100 thousand, and the main pathogen focused on EVs, MUV, JEV, MV and RV, which positive rate were 27.8%, 14.4%, 12.2%, 6.7% and 3.3% respectively. The sensitivity and specificity were all higher than ELISA. CONCLUSION: The most common pathogens responsible for viral encephalitis in Cangzhou, Hebei province, China, were EVs, and the multiplex RT-PCR was a rapid, sensitive, accurate method of testing the viruses responsible for causing these illnesses.

Association of polymorphisms in Toll-like receptors 4 and 9 with risk of pulmonary tuberculosis: a meta-analysis.

BACKGROUND Findings regarding the association of the single-nucleotide polymorphisms (SNPs) rs4986790 and rs4986791 in Toll-like receptor 4 and rs187084, rs574386, and rs352139 in Toll-like receptor 9 (TLR9) with pulmonary tuberculosis (PTB) susceptibility are inconsistent. We conducted a meta-analysis to systematically summarize and clarify the association between these SNPs and PTB susceptibility. MATERIAL AND METHODS A systematic literature search for relevant studies up to December, 2014 was performed in PubMed, EMBASE, Web of Science, Chinese National Knowledge Infrastructure (CNKI), and Wanfang databases. Information was gathered from each eligible study. Odds ratios (ORs) and 95% confidence intervals (95% CIs) were used to pool the effect size. RESULTS Finally, a total of 16 case-control studies on these polymorphisms were enrolled in this meta-analysis. The meta-analysis results suggest there was no association between these polymorphisms and PTB risk PTB risk in all the genetic models overall. However, for TLR4 rs4986791, a significant increased PTB risk was found in Africans, and for TLR9 rs352139 a significant increased PTB risk was found in Asians after subgroup analysis by ethnicity, although the enrolled studies were limited. CONCLUSIONS There was no association between the polymorphisms in TLR4 and 9 and PTB risk overall, but TLR4 rs4986791 and TLR9 rs352139 might be associated with increased PTB risk in Africans and Asians, respectively. Additional well-designed, larger-scale epidemiological studies are needed to validate our results.