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Objective To investigate the expression of thymosin ?4 in the foreskin,normal skin(except the foreskin),hypertrophic scar,and keloid,and its relationship to pathological scars.Methods In situ hybridization was applied to detect the expression of thymosin ?4 mRNA [With the probe of BM005698(EST) gene marked by digoxin] in four groups: foreskin,normal skin(except the foreskin),hypertrophic scar,and keloid.Both qualitative and semi-quantitative analysis were performed.Results Thymosin ?4 mRNA was expressed in all the four groups.The proportion of thymosin ?4 mRNA expression in the keloid was 2/10,which was 25% of that in the foreskin(8/10),and was 28.6% of that in the normal skin(7/10).The positive rate of thymosin ?4 mRNA expression in the dermis was higher than that in the epidermis;and was high in fibroblasts,intravascular endothelial cells,and macrophages.The mean OSD of the positive samples was 0.03?0.01 in the keloids,which was significantly lower than that in the normal skins(0.09?0.03) and hypertrophic scars(0.09?0.02)(P=0.000 for both).Compared with the normal skins,the expression increased by 231.5%(P
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Objective To construct a recombinant lentiviral vector for human thymosin ?4 (TMSB4) and to test the mRNA and protein expression of TMSB4 in 293T cells after being infected by shRNA lentivirus. Methods We designed a specific sequence of small hair RNA targeting TMSB4 gene; the complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was cloned into the pGCSIL-GFP vector to construct a lentiviral vector. The vector was converted into the competent DH5a coli,which confirmed by PCR and sequencing. Then the viral vector and the packed systemic vector were cotransfected 293T cells to get the lentivirus. The virus titer was determined. Afterwards,in the 293T cells infected with the lentivirus,the expression of TMSB4 was detected by real time PCR and immunocytochemistry. Similarly,a primary fibroblast was also infected with the lentivirus. Results Compared with negative control cells,the mRNA and protein levels of TMSB4 in 293T cells infected with the lentivirus were reduced by 44% (0.56?0.11 vs 1.00?0.06,F=89.673,P
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Objective:To explore the mechanism of action of thymosin ? 4 (T? 4) in septic shock by observing the 72h survival rate of septic shock mice treated with thymosin ? 4 and exploring the changes of TNF-? and IL-1?.Methods:LPS(lipopolysaccharide) was intraperitoneally injected into clean Kunming mice to establish the animal model.①The T? 4,between normal and septic shock mice were determined.②The survival rate of septic shock mice was studied by intraperitoneal injection of T? 4.Animals were divided into four groups of 30 and each received LPS(25mg/kg)intraperitoneal injection.Group Ⅰ was LPS control group.GroupⅡ,Ⅲ and Ⅳ were individually injected with T? 4 at 0h,0h,2h;0h,2h,4h post LPS through caudal vein.③TNF-? and IL-1? were examined in septic shock mice by ELISA.Results:T? 4 was obviously decreased in the blood of septic shock mice.T? 4 could raise the 72h survival rate of septic shock mice and down-regulate inflammatory mediators.Conclusion:The optimal protective methods in the mouse appears to be 5mg/kg T? 4 given immediately following(0h) and at 2h and 4h successively post LPS.T? 4 down-regulates the inflammatory mediators,which may be the mechanism of T? 4 in curing the septic shock.
