1.Construction and expression of eukaryotic expression vector of human IL-37 b gene
Jing YAO ; Jiang CHENG ; Xuefeng PEI ; Jingyu WANG ; Ming YUAN
Acta Laboratorium Animalis Scientia Sinica 2016;24(3):268-272
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.
2.The Dynamic change and clinical significance of C-reactive protein and platelet in patients with acute brain injury
Aifen BI ; Hanbin HU ; Decui PEI ; Xuefeng QIU
International Journal of Laboratory Medicine 2015;(14):2051-2053
Objective To investigate C‐reactive protein in patients with acute brain injury (CRP) ,platelet (PLT) dynamic chan‐ges and clinical significance .Methods A hospital in 2014-2015 120 cases of acute brain injury ,depending on whether surgery di‐vided into :64 cases of surgical group ,56 cases of non‐surgical group .According to Glasgow Outcome Scale (Glasgow Outcome Score ,GOS) divided into 61 cases of poor prognosis group (GOS 1 -3 scores) ,59 patients with good prognosis (GOS 4 -5 scores) .Another choice the same period 50 cases of healthy control group .After the patients were injured 1 d ,3 d ,7 d dynamic tes‐ting CRP ,PLT levels .Results The surgical group and the non‐surgical group patients after injury 1 d serum CRP levels were sig‐nificantly increased ,decreased gradually after all ,the two groups showed a downward trend ,but the surgery group were decreased slowly ,always maintain a high level ,still significantly at 14 d the control group (P<0 .05) .Patients with non‐surgical group de‐creased significantly ,significantly higher than the decline in the extent of the surgery group (P<0 .05) .Surgical and non‐surgical group patients after injury when PLT 1 d showed no increased after 3d began to rise ,increased significantly when 7 d ,PLT levels were elevated degree of surgical group was significantly higher than the non‐surgical group (P<0 .05) .Good prognosis group and a poor prognosis group ,serum CRP on admission were increased ,and reached a peak after 1 d ,after declining in both groups ,with good prognosis group decreased significantly ,while the poor prognosis group ,no significant decline in CRP levels ,continued high levels of state for a long time When postoperative 7d still significantly higher than the normal level (P<0 .05) .At each time point the poor prognosis group ,serum CRP levels were significantly higher than the good prognosis group (P< 0 .05) .When the good prognosis group and a poor prognosis group was mean change 1d PLT little water ,began to increase after the 3 d were poor progno‐sis group of patients was significantly higher than 7 d PLT good prognosis group (P<0 .05) .Conclusion Patients with acute brain injury CRP ,PLT dynamic changes are certain rules ,is to determine the acute phase of the disease of brain injury and prognosis of change .
3.Increased inflammatory reaction in tail-suspension mice infected by K.pneumoniae from spaceflight
Rong LIU ; Jiang CHENG ; Xuefeng PEI ; Mingwen JIA ; Jingyu WANG ; Junfeng WANG ; Changting LIU ; Ming YUAN
Military Medical Sciences 2017;41(5):377-380,389
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.