Objective To study the effect of adenoviral-delivered short hairpin RNA (shRNA) target against permeability glycoprotein (Pgp) as a new drug in anti-epileptic drug resistance epilepsy treatment and to evaluate its efficiency. Methods MDR Sprague-Dawley (SD) rat estrocyte model was induced by Coriaria Lactone (CL), mainly over-expressing mdrlb. To reverse the drug resistance, astrecytes were treated with constructed replication deficient adencvirus AdS-EGFP-shRNAI-U6 delivering short hairpin (shRNA) target agianst mdrlb gene. Total RNA and protein were extracted from the infected cells, mdr1 b level was detected by Quantitative Real-time PCR whereas Pgp by Western blot, Rhodamine123 (Rho123) efflux ratio by Flow Cytometry. Results AdS-EGFP-shRNA1-U6 was succesfully constucted with high virus titer of 6×1010 pfu/ml. The interference efficency of AdS-EGFP-shRNA1-U6 agianst mdrlb in rat astrecyte model was about 94%. The Rho123 efllux ratio was about 15. 8%, significiently lower than control group which was 56. 2% (F = 127.5, P < 0. 05). Conclusions Pgp over-expression has been successfully suppressed and MDR has been reversed, which may provide a premising approach for refractory epilepsy remedy.

To study the effect of batroxobin(DF-521) on atherosclerosis, we divided 50 Japanese big ear rabbits into control group and high-lipid group. After the atherosclerosis model was successfully established, the high-lipid rabbits were divided into 3 groups(placebo group, treatment group 1 and treatment group 2). Batroxobin was injected in the treatment groups, and saline was injected in placebo group and control group. Getting the aorta before, inter and after treatment, dyeing the lipid, endothelium, smooth muscle, collagen fibers of the vascular plaque(the elastic fibers are of autofluorescence), we observed them with the light microscope and laser scanning confocal microscope. From the results, we found that the atherosclerotic plaque in the treatment groups, tended to be static four weeks later, but there was no obvious difference between treatment group 1 and treatment group 2. These implied that batroxobin possessed the action of stabilizing the atherosclerotic plaque, but the dosage-effect was not clear and the principle needed more study.
Animals ; Arteriosclerosis ; drug therapy ; pathology ; Batroxobin ; therapeutic use ; Disease Models, Animal ; Female ; Fibrinolytic Agents ; therapeutic use ; Male ; Rabbits

A DNA fragment encoding human interleukin 4 was obtained by PCR from pORF-hIL4 plasmid. The amplified fragment was inserted into prokaryotic expression vector PQE60 and recombinant protein was expressed in E. Coli M15 by adding isopropyl-beta-D-thiogalactoside (IPTG). The hIL-4 protein was present as insoluble inclusion bodies in the bacterial extract. After denaturation of inclusion bodies with 5 mol/L guanidine hydrochloride, the supernate was diluted to get renaturized. Then dialysis and Ni chelating chromatography were used for purification. TF-1 proliferation assay of recombinant human interleukin 4 was performed, and then rhIL-4 was fit to be used for proliferation of human dendritic cells from monocyte in vitro.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Humans ; Inclusion Bodies ; metabolism ; Interleukin-4 ; biosynthesis ; genetics ; Protein Folding ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification