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Objective: To research the functional segments of B-FA molecule binding invariant chain and their characters. Methods:The DNA segments (α1α2, sα1α2 and α3TC ) of B-FA genes were respectively cloned and inserted into prokaryotic or eukaryotic expression plasmids,then they were singly or co-transfected with Ii gene into the engineering bacteria E. coli (BL-21)or 293T cells. After induction of expression,affinity chromatography and SDS-PAGE identification,the binding between B-FA segments and Ii molecule and co-localization in cells were observed with Pull-down and Western blot. Results:First three recombinant prokaryotic expression plasmids and four recombinant eukaryotic expression plasmids were constructed. The single molecules expressed by B-FA segments were observed after an affinity chromatography. Secondly the complexes of Ii/B-FA-α1α2 and Ii/B-FA-sα1α2 were detected by a Pull-down from the co-transfected corresponding prokaryotic expression plasmids,but no complex of Ii andα3TC,also in the western blot it was detected that B-FA-α1α2 or B-FA-sα1α2 as functional segment could bind Ii to form complex. Finally in eukaryotic expression 293T cells B-FA-sα1α2 kept localization, the same as B-FA. Conclusion: Chicken B-FA-α1α2 is function segment to bind with Ii molecule and keeps the location characters same as B-FA. The results of this research first time provide experimental evidence about B-FA functional region binding segment to Ii molecule.
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Objective To evaluate the efficacy of potassium sodium dehydroandroan drographolide succinate (PSDS) combined with routine therapy for rotavirus enteritis in children.MethodsA total of 148 children with rotavirus enteritis were included and divided into an observation group and a control group by random number table method, 74 in each group. The children in the observation group were treated with intravenous PSDS combined with routine therapy, and those in the control group with intravenous ribavirin combined with routine therapy. Serum interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) were determined by enzyme-linked immunosorbent assay, and plasma lactate dehydrogenase (LDH), creatine kinase (CK), and creatine kinase-MB (CK-MB) were determined using standard clinical laboratory procedures. The clinical efficacy was evaluated. Results The total efficacy rate in the observation group was significantly higher than that in the control group (91.9%vs. 78.4%;χ2=2.314,P<0.05). After the treatment, the serum levels of IL-6 (18.24 ± 3.62 ng/mlvs. 25.36 ± 5.25 ng/ml; t=2.425,P<0.05) and TNF-α (20.86 ± 4.28 ng/mlvs. 31.22 ± 7.15 ng/ml;t=2.503,P<0.05), and the plasma levels of LDH (104.25 ± 22.06 U/Lvs. 150.26 ± 37.22 U/L;t=2.316,P<0.05), CK (84.25 ± 13.57 U/Lvs. 107.88 ± 16.28 U/L;t=2.327,P<0.05) and CK-MB (22.30 ± 4.24 U/Lvs. 32.26 ± 7.14 U/L;t=2.426,P<0.05) in the observation group was significantly lower than those in the control group. The time to diarrhea resolution (2.42 ± 0.53 dvs.3.56 ± 0.78 d;t=2.316,P<0.05) and the time to fever resolution(2.11 ± 0.32 dvs.2.63 ± 0.43 d;t=2.472,P<0.05) in the observation group were significantly delayed than those in the control group, and the hospital length of stay longer (6.23 ± 1.42 dvs. 4.35 ± 0.96 d;t=2.413,P<0.05).Conclusions PSDS combined with routine therapy may reduce inflammatory response, protect from myocardial injury, and promote recovery in children with RVE.
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Objective:To study characters of the location in cells and binding activity of chicken Ii and B-L gene expressed products.Methods:The cloned gene segments of chicken Ii,B-LA and B-LB were respectively inserted into prokaryotic or eukaryotic expression plasmids,and then these recombinant plasmids were respectively alone transfected or cotransfected into engineering bacteria, Rosetta(DE3) or 293T cells.All of the recombinant bacteria were induced to express and their products then were renatured.The singly expressed products were detected to their immunogenicity with Western blot, and the co-expressed products were tested their binding with pull-down method and ( SDS-) PAGE.Results:First six of prokaryotic and eukaryotic recombinant expression plasmids were con-structed.The eukaryotic expressed products of Ii, B-LA and B-LB genes located in cellular plasma membrane.The single protein molecules were achieved from prokaryotic expressed products, which were renatured and purified with a Ni-column respectively.Secondly the prokaryotic expression Ii,B-LA and B-LB molecules could respectively induce mouse to product specific anti-bodies, which could recognize the corresponding products in eukaryotic expression.This indicated that they retained their antigenicity.Finally with pull-down from the products in co-expression in engineering bacteria and renaturation the Ii/B-LA or Ii/B-LB complexes were purified and the Ii,B-LA or B-LB monomers were dissociated from these complexes after a SDS treatment.Conclusion:The prokaryotic or eukaryotic expressed products of chicken Ii and B-L genes could retain their antigenicity,and chicken Ii could bind B-L molecules after prokaryotic coexpression and renaturation.These results provide a useful method to study the relation between Ii and MHC molecules.