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      • Ficolin2 defends against virulent Mycobacteria tuberculosis infections in vivo and its insufficiency is associated with infections in human

        Fengling Luo,Yubin Wang,Xiaoming Sun,Qilong Wang,Yunfeng Ma,Qin Pan,Xiao-Lian Zhang 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1

        Tuberculosis (TB) caused by Mycobacterium tuberculosis (M.tb) remains a major global health problem. Human ficolin2 (L-ficolin/P35) is a recently identified lectin complement pathway activator present in normal plasma associated with infectious diseases, however, little is known about the roles and mechanisms of ficolin2 during M.tb infection.We describe in this study for the first time L-ficolin expression levels in tuberculosis (TB) patients. We found that serum levels of ficolin2 of 107 pulmonary TB patients were much lower compared with 107 healthy controls, and recurrence TB patients have even lower levels than onset TB patients. In vitro analysis showed that Ficolin2 bound to virulent M.tb H37Rv strain much stronger than to non-virulent M. bovis BCG and M. smegmatis. Ficolin2/ficolin A bound to the H37Rv surface glycolipid portion and blocked H37Rv infection to human lung A549 cells. We further determined that the extrogenous ficolin2 had remarkable protecting effects against virulent H37Rv strain infections in C57BL/6J mice. FicolinA (ficolin2 like molecule) knockout mice had greatly increased susceptibility to the H37Rv infection. Ficolin2 activated macrophages via phosphorylation of JNK and stimulated IFN-□, IL-17, TNF-□ and nitric oxide (NO) secretions. Ficolin2 also stimulated IFN-□ production of CD8+T cells, but not CD4+T cells. The opsono-phagocytosis was promoted by ficolin2 as well. Our data demonstrate that ficolin2 can defend against virulent M.tb H37Rv infection both in vitro and in vivo mainly via activating macrophage proinflammatory cytokines IFN-□ and NO production and opsonophagocytosis, and its insufficiency is associated with more susceptible infections in human.

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        Amygdalin inhibits HSC-T6 cell proliferation and fibrosis through the regulation of TGF-β/CTGF

        Huanhuan Luo,Liang Li,Jianbang Tang,Fengxue Zhang,Fang Zhao,Da Sun,Fengling Zheng,Xinhua Wang,Xinhua Wang 대한독성 유전단백체 학회 2016 Molecular & cellular toxicology Vol.12 No.3

        Amygdalin is one of the nitrilosides that was widely used to treat cancer, inhibit liver fibrosis. In the present study, the aim was to determine the antifibrotic potential of amygdalin and examine its mechanisms of action in vitro. Firstly, we found amygdalin significantly inhibited HSC-T6 cells proliferation. Both mRNA and protein of transforming growth factor-β (TGF-β) were decreased in HSC-T6 cells during amygdalin treatment. Secondly, to investigate functional role of TGF-β, both TGF-β over-expression vector and siRNA against TGF-β were transfected into HSC-T6 cells respectively. The results showed that over-expression of TGF-β promoted proliferation of HSC-T6 cells, whereas TGF-β knockdown inhibited cell viability. Moreover, our data even indicated that TGF-β could promote cell proliferation independent of amygdalin treatment. Finally, we found amygdalin could inhibit expression of the classical fibrotic factor αSMA, which suggested an antifibrotic effect of amygdalin. While the TGF-β antagonized anti-fibrotic effect of amygdalin. To assess the mechanisms, we examined expression of CTGF in cultured HSC-T6 cells. Our results showed that CTGF was down-regulated in HSCT6 cell treated by amygdalin, but was up-regulated when exogenous TGF-β introduced. As CTGF was one of the downstream factors in the TGF-β pathway. These might suggest that amygdalin inhibited HSC-T6 cells proliferation and fibrosis via TGF-β/CTGF pathway.

      • Aptamers against ManLAM of Mycobacterium tuberculosis inhibits virulent Mycobacterium tuberculosis infection in mice and rhesus monkeys

        Qin Pan,Qilong Wang,Fengling Luo,Min Li,Xianru Xia Dongdong Shi,Xiao-Lian Zhang 한국당과학회 2012 한국당과학회 학술대회 Vol.2012 No.1

        Tuberculosis (TB) has been a major world-wide cause of death for centuries. One-third of the world’s population is infected with Mycobacterium tuberculosis (M.tb), the etiologic agent of TB. The development of potent new anti-TB drugs is urgently needed. The major M.tb surface lipoglycans, mannose-capped Lipoarabinomannan (ManLAM) had immunosuppressive effects during M.tb infection. In this study, aptamer ZXL1 which specifically bound to ManLAM from the virulent strain M.tb H37Rv was screened out by Systematic Evolution of Ligands by EXponential enrichment (SELEX). The binding affinity of ZXL1 to ManLAM was measured as 8.907X10-8 M of quilibrium dissociation constant (Kd) value by surface plasmon resonance (SPR) analysis. We found that aptamer ZXL1 prevented the ManLAM-induced immunosuppressive effects on DCs and inhibited M. tb entry into macrophages. More importantly, we found that single injection of aptamer ZXL1 significantly prolonged the survival rate of infected mice, and prevent the infected rhesus monkeys from weight loss. The Bacterial numbers were significantly lower in the lungs and spleens in ZXL1-treated groups. These results suggest that aptamer ZXL1 can be used as antimycobacterial agent or as TB vaccine adjuvent.

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