RESUMEN

BACKGROUND: Recent seminal studies have revealed that endosomal reactive oxygen species (ROS) promote rather than inhibit viral infection. Some ROS generators, including shikonin and H2O2, have the potential to enhance recombinant adeno-associated virus (rAAV) transduction. However, the impact of ROS on rAAV intracellular trafficking remains unclear. METHODS: To understand the effects of ROS on the transduction of rAAV vectors, especially the rAAV subcellular distribution profiles, this study systematically explored the effect of ROS on each step of rAAV intracellular trafficking pathway using fluorescently-labeled rAAV and qPCR quantification determination. RESULTS: The results showed promoted in-vivo and in-vitro rAAV transduction by ROS exposure, regardless of vector serotype or cell type. ROS treatment directed rAAV intracellular trafficking towards a more productive pathway by upregulating the expression of cathepsins B and L, accelerating the rAAV transit in late endosomes, and increasing the rAAV nucleus entry. CONCLUSIONS: These data support that ROS generative drugs, such as shikonin, have the potential to promote rAAV vector transduction by promoting rAAV's escape from late endosomes, and enhancing its productive trafficking to the nucleus.

Asunto(s)

Dependovirus , Peróxido de Hidrógeno , Especies Reactivas de Oxígeno/metabolismo , Transducción Genética , Dependovirus/genética , Peróxido de Hidrógeno/metabolismo , Endosomas , Vectores Genéticos

RESUMEN

Cartilage degeneration and inflammation are important features of rheumatoid arthritis (RA). Chondrocyte inflammation and apoptosis have been increasingly demonstrated to be related to cartilage decomposition. In this study, we analyzed the protective role of kallistatin against RA and its associated mechanisms. We obtained in vitro and in vivo RA models using IL-1ß and heat-inactivated Mycobacterium tuberculosis, respectively. Our results showed that kallistatin mitigated IL-1ß-mediated chondrocyte apoptosis and inhibited the synthesis of ECM-degrading generation, like matrix metalloproteinase (MMP)-3/13 and a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS)-4/5, in IL-1ß-mediated chondrocytes. Furthermore, kallistatin markedly suppressed IL-1ß-mediated inflammation while decreasing the levels of inflammatory factors and mediators via the NF-κB pathway. Daily administration of kallistatin reduced the expression levels of PGE2, TNF-α, IL-1ß, and IL-6. Histochemical analysis revealed that the kallistatin-treated rats exhibited reduced RA severity compared with control mice. In summary, kallistatin suppressed IL-1ß-mediated inflammation in chondrocytes via the NF-κB pathway. Administration of kallistatin remarkably inhibited RA development, accompanied by reduced inflammation and apoptosis. Therefore, kallistatin administration can be used as a candidate therapeutic strategy for RA.

Asunto(s)

Artritis Reumatoide , FN-kappa B , Ratas , Ratones , Animales , FN-kappa B/metabolismo , Células Cultivadas , Transducción de Señal , Artritis Reumatoide/tratamiento farmacológico , Artritis Reumatoide/metabolismo , Inflamación/tratamiento farmacológico , Inflamación/metabolismo , Condrocitos/metabolismo , Interleucina-1beta/metabolismo

RESUMEN

The decomposition of surface EMG signals can provide valuable information about the recruitment and firing of motor units from surface EMG recordings. According to the physiologic characteristic of the surface EMG signals generation, a method of the decomposition of SEMG signals based on the technique of convolved mixing blind source separation was proposed. Using simulated SEMG signals, the performance of the decomposition algorithm was analyzed and compared with that of the decomposition technique adopting Independent Component Analysis (ICA). The experiment results show that the proposed method could decompose SEMG signals effectively, and it's performance is better than the ICA decomposition method, no matter for the simulated or recorded SEMG signals.