Increased inflammatory reaction in tail-suspension mice infected by K.pneumoniae from spaceflight / 军事医学

Objective To explore the changes in inflammatory reactions in tail-suspension mice infected by Klebsiella pneumoniae from spaceflight.Methods Tail suspension was used to simulate the physiological effects of microgravity.C57BL/6 mice were randomly divided into control (Con),control+K.pneumoniae T16-169 (Con+T16-169),tail suspension (TS) and tail suspension+K.pneumoniae T16-169 (TS+T16-169) groups.The level of inflammatory cytokines TNF-α,IL-6 and IL-1β mRNA in lung tissue and the plasma cytokine concentration were detected by RT-qPCR and xMAP technology,and HE staining was used to represent the morphological changes in lung tissue.Results Compared with the control group,the expression of inflammatory cytokines in lung tissue and plasma concentrations of all experimental groups were increased,and the difference in TS+T16-169 group was the most significant (P<0.01 or P<0.001).HE staining showed that the lung tissues in Con+T16-169 and TS+T16-169 groups were damaged in different degrees,and the damage of TS+T16-169 group was the most serious.Conclusion The K.pneumoniae from spaceflight significantly increases the expression of inflammatory cytokines in lung tissue and plasma concentrations after infecting tail-suspension mice,and induces more serious damages to the lung tissue,which suggests that inflammatory reactions can be increased in tail suspension mice infected by K.pneumoniae from spaceflight.

Construction and expression of eukaryotic expression vector of human IL-37 b gene / 中国实验动物学报

Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells ( PB-MCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eu-karyotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detec-ted by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P<0. 01) after transfection in THP-1 cells. Conclusions We successful-ly constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foun-dation for further study on IL-37 functions and its association with related diseases.