ABSTRACT
Objective To study the expression and change of neuropeptide substance P (SP) during the wound healing of deep partial thickness scalding in diabetic rats. Methods Eighty-four Wistar rats were randomly divided into diabetes mellitus group (n=42) and control group (n=42). Diabetic rat models were established by intraperitoneal injection of streptozotocin (STZ) in diabetes mellitus group, and those in control group were intraperitoneally injected with aseptic citrate buffer solution. Deep partial thickness scalding with diameter of 2 cm on the back were prepared in all the rats. The pre-scalding and post-scalding wound specimens of different time points were obtained, and the percentages of wound closure were calculated. The wound specimens were also obtained for immunohistological staining to compare the areas with positive staining of SP, and ELISA was employed to detect the expression of SP in the wound tissues. Results The percentage of wound closure was significantly lower in diabetes mellitus group than that in control group from 7 days post-scalding (P< 0.01). The areas with positive staining of SP in diabetes mellitus group were much smaller than those in control group at different time points, which was most significant on the seventh day post-scalding[(1 350.93±99.28) μm2 vs(1 715.86± 103.41) μm2](P < 0.01). The expression of SP in the wound tissues was significantly lower in diabetes mellitus group than that in control group at different time points, which was most significant on the seventh day post-scalding[(114.04±9.96) vs(143.39±8.94)](P<0.01). Conclusion The significantly lower expression of SP in wound site may be one of the causes of delayed wound healing in diabetic rats.
ABSTRACT
Objective To construct the lentivirus carrying human ?-catenin-EGFP(enhanced green fluorescent protein)and observe its expression in human follicle stem cells.Methods The ?-catenin gene sequence was amplified by RT-PCR from extraction of total RNA of human vascular endothelial cells.TA cloning technique was utilized to acquire gene subcloned pUCm-T-?-catenin.After transformation reaction,candidate clone was further analyzed by PCR and gene sequencing.Then the plasmid was transfected into FT293 cells.After identification by Western blotting,the plasmid was transfected into FT293 cells again for packaging.Infection titer was monitored by green EGFP expression.The expression of ?-catenin-lentivirus in human follicle stem cells were observed under inverted fluorescence microscope.Results The ?-catenin gene was cloned into the lentivirus successfully.The high expression of green fluorescence protein in FT293 cell line was found under fluorescent microscope.Viral titer checked by real-time PCR was about 2.0?108 TU/mL.When the multiplicity of infection(MOI)was 10,the infection efficiency of ?-catenin-lentivirus in human follicle stem cells was nearly 80% after infection 48 h around.After 3 weeks of continuous observation,we found the infection efficiency still keeping in the range of 80%-90%.Conclusion The lentivirus expression vector for ?-catenin was successfully constructed.It can steadily infect human follicle stem cells and the infection efficiency is considerable high.