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In addition to functioning as an antioxidant, anti-inflammatory and age-defying cellular component, DHA impacts the immune system by facilitating the pathogen invasion. The mechanism through which DHA regulates immune suppression remains obscure. In our study, we postulated that DHA might interact with GPR120 to shape the dendritic cell (DC) differentiation and subsequently drive T cell proliferation during the virus infection. In vitro, the proportion of costimulatory molecules and HLA-DR on DC that generated from exogenous and endogenous (fad3b expression) DHA supplemented mice were significantly lower than wild-type mice. Given the importance of FAs, DHA is not only a critical cellular constituent but also a cell signaling molecule and FA deficiency reduces DC generation; we used GPR120-/- mice to determine whether DHA receptor deficiency disorders DC maturation processing. Novelty, the expression of GPR120 on DC from wild-type (WT) mice was inversely related to DC activation and DC from the GPR120-/- mice maintained a spontaneous maturation status. In vivo, both the excessive activation of GPR120 by DHA and the deletion of GPR120 effectively skewed the balance of Th17/Tregs and reduced the production of VNA and protection of vaccination. Overall, our results revealed a mechanism that the GPR120 self-regulation plays a crucial role in sensing DHA variation, which provides a new prospect for therapeutic manipulation in autoimmune diseases and the design of a vaccine adjuvant.

Previous studies revealed that Japanese encephalitis virus (JEV) infection alters the expression of miRNA in central nervous system (CNS). However, the mechanism of JEV infection contributes to the regulation of miRNAs in CNS remain obscure. Here, we found that a global degradation of mature miRNA in mouse brain and neuroblastoma cells after JEV infection. In additional, the integrative analysis of miRNAs and mRNAs suggests that those down-regulated miRNAs are primarily targeted inflammation genes and the miR-466d-3p target the IL-1β which in the middle of those inflammation genes. Transfection of miR-466d-3p decreased the IL-1β expression and inhibited the JEV replication in NA cells. Interestingly, the miR-466d-3p level increased after JEV infection in the presence of cycloheximide, which indicated that viral protein expression reduces miR-466d-3p. Therefore, we generated all the JEV coding protein and demonstrated that NS3 is a potent miRNA suppressor. Furthermore, the NS3 of ZIKA virus, WNV, DENV1 and DENV2 also decreased the expression of miR-466d-3p. The in vitro unwinding assay demonstrated that the NS3 could unwind the pre-miR-466d and induce the disfunction of miRNA. Using computational models and RNA immunoprecipitation assay, we found that arginine-rich domains of NS3 are critical for pre-miRNA binding and the degradation of host miRNAs. Importantly, site-directed mutagenesis of conserved residues revealed that R226G and R202W significantly reduced the binding affinity and degradation of pre-miR-466d. Together, these results extend the helicase of Flavivirus function beyond unwinding duplex RNA to the decay of pre-miRNAs, which provides a new mechanism of NS3 in regulating miRNA pathways and promoting the neuroinflammation.Author SummaryHost miRNAs had been reported to regulate JEV induced inflammation in central nervous system. We found that the NS3 of JEV can reduce most of host miRNA expression. The helicase region of the NS3 specifically binds to precursors of miRNA and lead to incorrect unwinding of precursors of miRNAs which inhibits the function of miRNAs. This observation leads to two major findings. First, we identified the miR-466d-3p targets to the host IL-1β and E protein of JEV, and NS3 degrades the miR-466d-3p to promote the brain inflammation and viral replication. Second, we proved that the arginine on the helicase of NS3 is the main miRNA binding sites, and the miRNA degradation by NS3 was abolished when the R226 and R202 were mutated on the NS3. These findings were also confirmed with NS3 of ZIKA virus, WNV and DENV which could decrease the expression level of miR-466d-3p to enhance the inflammation. Our study provides new insights into the molecular mechanism of encephalitis caused by JEV, and reveals several amino acid sites to further attenuate the JEV vaccine.

A method for luteoskyrin in Chinese rice wine was developed using high-performance liquid chromatography–ion trap tandem mass spectrometry. The detection limit of luteoskyrin was 0.10 µg/kg. The recovery values of luteoskyrin in fortified Chinese rice wine were between 85.7% and 88.9% at levels between 0.5 µg/kg and 100 µg/kg (R > 0.999). The relative standard deviations were in the range of 1.10% to 3.20%. The method achieved rapid and accurate determination of luteoskyrin in Chinese rice wine while meeting minimum residue measurement requirements.

NLRs are a large family belonging to pattern recognition receptors which could recognize pathogen associated molecular patterns. Class II, major histocompatibility complex, transactivator (CIITA) is a member of NLR family. It is a critical transcription factor which could regulate the expression of MHC class II. In this study, a full-length cDNA of CIITA was cloned from channel catfish according to ten sequenced ESTs. This cDNA contains a 5'-UTR of 71 bp, a 3'-UTR of 238 bp and an ORF of 3,210 bp encoding 1,069 amino acids. Phylogenetic analysis showed that catfish CIITA was conserved with other CIITAs. Quantitative real-time PCR was conducted to detect the expression profiles of CIITA in normal tissues and responding to different pathogens (Edwardsiella tarda, Streptococcus iniae and channel catfish Hemorrhage Reovirus (CCRV)). The expression profile in blood was the highest (53.879-fold) in normal tissues. E. tarda and S. iniae could induce catfish CIITA in head kidney, liver and spleen. CCRV virus could also induce CIITA in head kidney and liver but reduce it in spleen. And S. iniae could induce the expression of CIITA to the highest extent and contrarily CCRV virus to the lowest extent. The expression data showed the tissue-specific and pathogen-specific expression patterns of CIITA responding to different pathogens. These expression data indicated the immune-related functions of CIITA. The data obtained in this study provide a basis for further research aimed at explore the precise immune-related molecular mechanism of CIITA in catfish.