Abstract We have recently demonstrated that dendritic cell (DC)-mediated immune modulation and deviation can be accomplished through RNA interference (RNAi), highlighting the therapeutic potential of RNAi-modified DC as antigen-specific tolerogenic vaccines. To date, an RNAi-based vaccine has not been reported. The current study was designed to develop siRNA-modified DC as antigen-specific, tolerogenic vaccines for prevention and intervention of autoimmune arthritis. Using small interfering RNA (siRNA) that specifically targets IL-12p35 gene (IL-12 siRNA), we have generated a type of DC that exhibits multiple tolerogenic characteristics. Immunization with type II collagen (CII)-pulsed and IL-12 gene-silenced DC (CII-pulsed/gene-silenced DC) resulted in antigen-specific nonresponsiveness in T cell responses. Vaccination with CII-pulsed/gene-silenced DC prevented collagen-induced arthritis (CIA) onset in a murine rheumatoid arthritis model. Furthermore, administration of CII-pulsed/gene-silenced DC was sufficient to inhibit progression of CIA. The therapeutic effects were further evidenced by decreased clinical scores, inhibited inflammatory infiltrates, and suppressed T cell and B cell responses to CII. In conclusion, this study is the first to demonstrate the therapeutic utilization of RNAi-modified DC as antigen-specific tolerogenic vaccines for autoimmune arthritis.
Abstract The assembly of snRNP cores, in which seven Sm proteins, D1/D2/F/E/G/D3/B, form a ring around the nonameric Sm site of snRNAs, is the early step of spliceosome formation and essential to eukaryotes. It is mediated by the PMRT5 and SMN complexes sequentially in vivo. SMN deficiency causes neurodegenerative disease spinal muscular atrophy (SMA). How the SMN complex assembles snRNP cores is largely unknown, especially how the SMN complex achieves high RNA assembly specificity and how it is released. Here we show, using crystallographic and biochemical approaches, that Gemin2 of the SMN complex enhances RNA specificity of SmD1/D2/F/E/G via a negative cooperativity between Gemin2 and RNA in binding SmD1/D2/F/E/G. Gemin2, independent of its N-tail, constrains the horseshoe-shaped SmD1/D2/F/E/G from outside in a physiologically relevant, narrow state, enabling high RNA specificity. Moreover, the assembly of RNAs inside widens SmD1/D2/F/E/G, causes the release of Gemin2/SMN allosterically and allows SmD3/B to join. The assembly of SmD3/B further facilitates the release of Gemin2/SMN. This is the first to show negative cooperativity in snRNP assembly, which provides insights into RNA selection and the SMN complex's release. These findings reveal a basic mechanism of snRNP core assembly and facilitate pathogenesis studies of SMA.
Background: Activating transcription factor 2 (ATF2) regulates the expression of downstream target genes and is phosphorylated by the Ras-extracellular-signal-regulated kinase (ERK) pathway. Acetylation of ATF2 is necessary for this type of regulation. However, the molecular mechanism by which the Ras-ERK pathway mediates the regulation of acetylated ATF2 is unknown. This study investigates the mechanism of Ras-ERK pathway-mediated regulation of acetylated ATF2 in maintaining the characteristic phenotype of pancreatic cancer cells. Methods: This study was carried out using ASPC-1 and BXPC-3 pancreatic cancer cell lines transfected with the double mutant RasG12V/T35S. The levels of phosphorylated ERK1/2 were measured to establish the activated Ras-ERK pathway. The regulation of acetylated ATF2 was examined by detecting the protein level using western blotting, and the effects on cancer cell phenotype were measured using cell viability, proliferation, migration, and apoptosis assays. Also, chromatin immunoprecipitation (ChIP) assays were used to measure the effect on respective downstream target genes. Results: The results showed that RasG12V/T35S reduced the level of acetylated ATF2 in ASPC-1 and BXPC-3 cells. Compared to wild-type ATF2, the mutant ATF2K357Q (which mimics the irreversible acetylated form of ATF2) reduced the cancer cell phenotype and showed decreased enrichment on target genes upon transfection with Ras. Moreover, the level of acetylated ATF2 was regulated by the degradation of p300 through E3 ubiquitin ligase mouse double minute 2 homolog (MDM2). Conclusions: Activation of the Ras-ERK pathway regulates acetylated ATF2 through degradation of p300 via a proteasome-dependent pathway, which alters the transcription of downstream target genes responsible for the cancer cell phenotype.
Aiming at the fault early warning of transformer oil chromatographic commonly used the methods in the national standard.In this paper, a method is proposed to obtain alarm thresholds of each component gas within transformer oil chromatogram based on statistical analysis of numerous data.Compared with the value specified in national standard, the threshold obtained there is relatively conservative and can effectively prevent the potential failure from developing.Followed by that, on the basis of determining each gas threshold, through Pearson correlation analysis, a combination alarm strategy with decisionsupporting gas group is proposed.Such method can effectively avoid false alarm due to chromatography measurement error caused by equipment, human and environment factors, so that the alarm accuracy can be improved.The application of the theoretical results in this paper will provide a refined fault early warning method of transformer oil chromatogram to the substation operation and maintenance staffs.
Integrated gasification fuel cell (IGFC) systems combining coal gasification and solid oxide fuel cells (SOFC) are promising for highly efficient and environmentally friendly utilization of coal for energy production. Most IGFC system analyses performed to date have used non-dimensional thermodynamic SOFC models that do not resolve the intrinsic constraints of SOFC operation. In this work, a one-dimensional finite volume model for planar SOFC is developed and verified using literature data. Special attention is paid to making the model capable of supporting recent SOFC technology improvements, including the use of anode-supported configurations, metallic interconnects, and reduced polarization losses. Results are presented for SOFC operation on humidified hydrogen and methane-containing syngas, under co-flow and counter-flow configurations; detailed internal profiles of species mole fractions, temperature, current density and electrochemical performance are obtained. The effects of performance, fuel composition and flow configuration on SOFC performance and thermal profiles are evaluated, and the implications of these results for system design and analysis are discussed.
The demethylation of methylmercury has received substantial attention. Here, a novel chemical method for the demethylation of methylmercury is proposed. The low-toxicity graphene-fulvic acid (FA, a ubiquitous material in the environment) was synthesized without the use of a chemical reagent. The hybridized graphene-FA presented an indirect open band gap of 2.25-2.87 eV as well as adequate aqueous dispersion. More importantly, the hybridized graphene-FA exhibited 6- and 10-fold higher photocatalytic efficiencies for the demethylation of methylmercury than FA and free FA with graphene, respectively. This result implies that immobilized, rather than free, FA accelerated the catalysis. Furthermore, inorganic mercuric ion, elemental mercury, and mercuric oxide were identified as the primary demethylation products. For free FA with graphene, graphene quenches the excited-state FA, inhibiting the demethylation by electron transfer. In contrast, the graphene of the self-assembled graphene-FA serves as an electron reservoir, causing electron-hole pair separation. Graphene-FA showed a negligible toxicity toward microalgae compared to graphene. The above results reveal that the green synthesis of graphene and organic molecules is a convenient strategy for obtaining effective cocatalysts.
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