The E. coli BL21(DE3) strain bearing the plasmid(T_7lac/His6-tag) with the h?TNF?recombinant gene was grown in LB medium containing SO?g/ml kanamycin, at 37 ℃ , up to the late logarithmic phase(OD590 ~ 0.5)then induced with IPTG (final concentration lmmol/L)for 5 hours. SDS-PAGE analysis revealed that expression level of the product(His6-?TNF?was up to 45% of the total bacterial proteins and was mainly as insoluble inclusion bodies ( IBs) . After cell lysis, the IBs was separated by centrifugation, dissolved in 7mol/L urea, then purified by Ni column ( Ni~(2+) -Sepharose 6B) . And the purity of more than 95% and the recovery of about 90% were obtained. The purified product was refolded with the renaturation buffer under low temperafure(

The immune function of spleen after partial splenectomy (PSM) of rabbits was stuided using microwave tissue coagulation (MTC). Streptococcus pneumoniae suspension was injected into the ear veins of rabbits 4 weeks after they were subjected to partial splenectomy using microwave tissue coagulation (PSM) group and to sham operation (SO groups), respectively. India ink was given via portal vein. Residual spleen were resected and microscopic examinations were performed on spleen slices stained using HE to compare phagocytic functions of macrophage. Gradings of phagocytic function in macrophage were of no significant difference between PSM and SO groups (P > 0.05). Weights of spleens showed no significant difference between the two groups (P > 0.05). The phagocytic function of macrophage after PSM can be preserved well. The result of this experiment implies that splenic salvage using MTC is a clinically applicable method.
Animals ; Female ; Macrophages ; immunology ; Male ; Microwaves ; therapeutic use ; Phagocytes ; immunology ; Rabbits ; Regeneration ; immunology ; Spleen ; physiology ; Splenectomy ; methods