1.Cloning and high level expression of gene encoding human sCR1 binding domain in E.coli
Journal of Third Military Medical University 2003;0(15):-
Objective To clone and express the gene encoding the binding domain of human soluble complement receptor type 1 (sCR1). Methods sCR1-SCR15-18 cDNA was amplified using RT-PCR from human monocytes of peripheral blood and sequenced using vector pMD-18T. Recombinant pET32-sCR1-SCR15-18 was constructed using prokaryotic expression vector pET32 and transformed into bacterium BL21. IPTG was used to induce gene expression and the obtained expression product was identified by immunoblotting. Results The gene segment that specifically encodes sCR1 was synthesized, the sequence of which was consistent with that of sCR1-SCR15-18 cDNA as registered at GenBank. A prokaryotic expression recombinant pET32-sCR1-SCR15-18 was constructed. The amount of target protein accounted for 40% of the total bacterial proteins and inclusion bodies were present in the bacteria. Immunoblotting showed a single positive band at the site of 43?10~(3). Conclusion The gene encoding sCR1-SCR15-18 was cloned from human monocytes and efficiently expressed in E.coli.
3.Effect of Qingqihuatan Decoction on pulmonary inflammation in asthmatic mouse
Zhu CHEN ; Yu PENG ; Ping PANG ; Jianhui YANG ; Zhengqing LUO
Chongqing Medicine 2017;46(25):3473-3476
Objective To observe the interventional effect of Qingqihuatan Decoction on airway inflammation and inflammatory reaction of lung tissue in asthmatic mouse.Methods Forty-five BALB/c mice were randomly divided into the normal control group(CON),asthmatic model group(MOD),dexamethasone group(TRE),Qingqihuatan Decoction group(X1) and Qingqihuatan Decoction combined dexamethasone group(X2).The asthmatic mouse model was established by the sensitization and inhalation of OVA and aluminium hydroxide gel.The bronchoalveolar lavage fluid (BALF) was performed the cell counts and eosinophil counts,and the pathological changes of lung tissue were observed.Results Compared with CON group,BALF cell count and eosinophil count in the model group were increased obviously,which in the treatment group were significantly decreased.The effect of the X1 group and X2 group had statistical difference between on 5 d and 15 d.(P<0.05 or P<0.01);the treatment groups could reduce the pathogenical infiltration,lung bullae production and airway epithelial thickening of mouse lung tissue,especially the inflammation damage in X2 group was mild.Conclusion Qingqihuatan Decoction can relieve the airway inflammation and improves the lung tissue inflammatory response in model mouse.