Abstract:The object of this study was to investigate the cellulolytic bacterial community in the intestine of the Chinese white pine beetle (Dendroctonus armandi) larvae. A total of 91 cellulolytic bacteria were isolated and assigned to 11 genotypes using amplified ribosomal DNA restriction analysis (ARDRA). Partial 16S rDNA sequence analysis and morphological tests were used to assign the 11 representative isolates. The results showed that the isolates belonged to α-Proteobacteria, γ-Proteobacteria and Firmicutes. Members of γ-Proteobacteria were the most frequently represented species and accounted for 73.6% of all the cellulolytic bacteria. The majority of cellulolytic bacteria in D. armandi larva gut were identified as Serratia and accounted for 49.5%, followed by Pseudomonas, which accounted for 22%. In addition, members of Bacillus, Brevundimonas, Paenibacillus, Pseudoxanthomonas, Methylobacterium and Sphingomonas were found in the D. armandi larva gut. Brevundimonas kwangchunensis, Brevundimonas vesicularis, Methylobacterium populi and Pseudoxanthomonas mexicana were reported to be cellulolytic for the first time in this study. Information generated from the present study might contribute towards understanding the relationship between bark beetle and its gut flora.

Bark beetles oxidize the defensive monoterpenes of their host trees both to detoxify them and to convert them into components of their pheromone system. This oxidation is catalysed by cytochrome P450 (CYP) enzymes and occurs in different stages of the insect. We identified two new CYP4 genes in the Chinese white pine beetle (Dendroctonus armandi), and carried out bioinformatic analysis one the full-length nucleic acid sequences and deduced amino acid sequences. Differential expression of the CYP4 genes was observed between sexes, and within these significant differences amongst development stages, fed on phloem of Pinus armandi and exposed to stimuli((±)- α-pinene, (R)-(+)- α-pinene, (S)-(-)-α-pinene, (S)-(-)-β-pinene and (+)-3-carene) at 8 and 24 h, and their interactions were found upon exposure to host monoterpenes. Increased expression of CYP4 genes suggested that they play a role in the detoxification of monoterpenes released by the host trees. The differential transcript accumulation patterns of these bark beetle CYP4 genes provides insight into the ecological interactions of D. armandi with its host pine.

Objective: The effects of mesenchymal stem cell (MSC)-derived exosomes on brain microvascular endothelial cells under oxygen-glucose deprivation (OGD), which mimics cells in deep hypothermic circulatory arrest (DHCA) in vitro, are yet to be studied. Methods: MSCs were co-cultured with primary rat brain endothelial cells, which were then exposed to OGD. Cell viability, apoptosis, the inflammatory factors (IL-1β, IL-6, and TNF-α), and the activation of inflammation-associated TLR4-mediated pyroptosis and the NF-κB signaling pathway were determined. Furthermore, exosomes derived from MSCs were isolated and incubated with endothelial cells to investigate whether the effect of MSCs is associated with MSC-derived exosomes. Apoptosis, cell viability, and the inflammatory response were also analyzed in OGD-induced endothelial cells incubated with MSC-derived exosomes. Results: OGD treatment promoted endothelial cell apoptosis, induced the release of inflammatory factors IL-1β, IL-6, and TNF-α, and inhibited cell viability. Western blot analysis showed that OGD treatment induced TLR4, and NF-κB p65 subunit phosphorylation and caspase-1 upregulation, while co-culture with MSCs could reduce the effect of OGD treatment on endothelial cells. As expected, the effect of MSC-derived exosomes on OGD-treated endothelial cells was similar to that of MSCs. MSC-derived exosomes alleviated the OGD-induced decrease in the viability of endothelial cells, and increased levels of apoptosis, inflammatory factors, and the activation of inflammatory and inflammatory focal pathways. Conclusion: Both MSCs and MSC-derived exosomes attenuated OGD-induced rat primary brain endothelial cell injury. These findings suggest that at least some of the protective effects of MSCs on endothelial cells are mediated by MSC-derived exosomes.

Disregulation of fatty acid oxidation, one of the major mechanisms for maintaining hepatic lipid homeostasis under fasting conditions, leads to hepatic steatosis. Although obesity and type 2 diabetes-induced endoplasmic reticulum (ER) stress contribute to hepatic steatosis, it is largely unknown how ER stress regulates fatty acid oxidation. Here we show that fasting glucagon stimulates the dephosphorylation and nuclear translocation of histone deacetylase 5 (HDAC5), where it interacts with PPARα and promotes transcriptional activity of PPARα. As a result, overexpression of HDAC5 but not PPARα binding-deficient HDAC5 in liver improves lipid homeostasis, whereas RNAi-mediated knockdown of HDAC5 deteriorates hepatic steatosis. ER stress inhibits fatty acid oxidation gene expression via calcium/calmodulin-dependent protein kinase II-mediated phosphorylation of HDAC5. Most important, hepatic overexpression of a phosphorylation-deficient mutant HDAC5 2SA promotes hepatic fatty acid oxidation gene expression and protects against hepatic steatosis in mice fed a high-fat diet. We have identified HDAC5 as a novel mediator of hepatic fatty acid oxidation by fasting and ER stress signals, and strategies to promote HDAC5 dephosphorylation could serve as new tools for the treatment of obesity-associated hepatic steatosis.