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A skin specimen was obtained from each group on day 7, 14, 21 after wounding for histopathological examination. Specimens were immediately fixed in 10% (v/v) neutral buffered formalin, and the solution was replaced every 2 days until the tissues had hardened. Each specimen was embedded in a paraffin block and thin sections (3?��m) were prepared and stained with haematoxylin and eosin (H&E) for general morphological observation. Tissue sections that had been dewaxed and rehydrated routinely were incubated with 3% H2O2 [http://en.wikipedia.org/wiki/PSCD4 CYTH4] for 30?min. The slides were washed with phosphate-buffered saline (PBS, pH 7.4) twice. The sections were blocked with 5% bovine serum albumin in Tris-buffered saline for 20?min. The spent solution was discarded and the sections were incubated with anti-FLAG antibody (1:2000) at 4?��C overnight. The slides were washed with PBS, incubated with rabbit secondary antibody (1:5000) for 1?h, and followed by incubation with streptavidin�Chorseradish peroxidase for 20?min. The antibody binding sites were visualised by incubation with diaminobenzidine�CH2O2 [http://www.selleckchem.com/products/CAL-101.html selleck chemical] solution. All values are expressed as mean?��?standard error of the mean (SEM). One-way analysis of variance followed by Student's unpaired t-test was used to compare parametric data. P?18?mmol/L) after 1 week in the rats that had received STZ. Blood glucose levels were determined when the plasmid treatment was administered, as shown in Table 1; all diabetic rats had average [http://www.selleckchem.com/products/AG-014699.html find more] blood glucose levels that exceeded 18?mmol/L, although there was no statistically significant difference among the six groups. Our data indicate that SPK1 did not directly affect the blood glucose levels of diabetic rats. Wounds in all groups decreased in size gradually over time (Fig. 1). However, there was an obvious delay in the rate of healing in diabetic rats as compared with normal control rats. The wound areas of the diabetic rats were larger than that of the normal control rats after 3�C21 days. Observation of the diabetic rats at 7 days revealed a statistically significant decrease in the wound area following treatment with 125?��g/wound (0.67?��?0.15?cm2, P?