Objective To investigate the effect of topical external administration of recombinant human epidermal growth factor (rhEGF) when controll ing blood sugar on expression of epidermal growth factor receptor (EGFR) and EGFR mRNA of wound in diabetes mell itus (DM) combined with scald. Methods A total of 136 male Wistar rats weighing (188.57 ± 6.59) g were randamly divided into 4 groups (groups A, B, C, and D, n=34). The rats was made DM model by intraperitoneal injected 60 mg/kg streptozocin in groups A, B, and C; rats were injected buffer alone in group D as control group. After 8 weeks, the rats of 4 groups were placed in 80℃ hot water for 6 seconds for preparation of the back deep II degree scald model. In group A, the blood sugar level was controlled at the level of group D 1 week before scald model; within 24 hours after models preparation, rhEGF was sprayed on wound at 150 U/cm2 . In group B, the rats were given the same treatment as group A except not controll ing blood sugar. In group C, the blood sugar was controlled as group A and wound was suture fixation with 1% silver sulfadiazine cream at 24 hours after the model. In group D, the same treatment as group A was given after injury. The heal ing rate of the wound was detected at 3, 7, 11, 15, and 21 days after injury; the EGFR mRNA expression was determined by mRNA hybridization in situ, and the EGFR protein expression was deterimined by immunohistochemistry and Western blot at 1, 3, 5, 7, 11, 15, and 21 days. Results All the rats survived at the end of experiment. There was no significant difference in the heal ing rate of the wound among the 4 groups at 3 days (P gt; 0.05). The heal ing rate of the wound was significantly higher in groups A and D than in groups B and C (P lt; 0.05) at 7, 11, 15, and 21 days. The expession of EGFR mRNA in 4 groups was observed by hybridization in situ, which mainly distributed in the dermal fibroblasts, capillary endothel ial cells and remnants of skin and wound edge epithel ium of the subsidiary; the expessions reached the peak at 5 days in group A, at 7 days in groups B and C, and at 11 days in group D; and the peak level was significantly higher in groups A and D than in groups B and C (P lt; 0.05). Immunohistochemistry and Western blot showed that the expession of EGFR protein was observed in 4 groups and reached the peak level at 7 days in groups A and B, and at 11 days in groups C and D; showing significant difference between groups B, C and groups A, D (P lt; 0.05). Conclusion External appl ication of rhEGF when controll ing blood sugar can accelerate obviously the wound heal ing in DM combined with scald. After controll ing blood sugar, external appl ication of rhEGF can boost obviously the expressions of EGFR mRNA, EGFR, and the extending process of signal conduction.
Objective Epidermal stem cells (ESCs) can actively partici pate in wound heal ing and enhance reepithel ial ization. To establ ish ideal diabetes mell itus (DM) rat models and to investigate the expression of keratin 19 (K19),β1-integrin, β-catenin, and prol iferating cell nuclear antigen (PCNA) in ESCs of DM rat model, then to study the potential mechanism of difficult recovering wounds of diabetic skin. Methods Twenty male SD rats (weighing 250-300 g) were dividedinto DM group and normal control group randomly (n=10). The DM rat model was made by intraperitoneal injected 65 mg/kg streptozocin (STZ), the normal control group was not treated. At 4 weeks after injection, pancreatic tissue was harvested for HE staining in two groups. The ESCs isolated from full-thickness skins of the back of two group rats were culutured and identified. The 2nd passage of ESCs were obtained for immunocytochemical staining of K19, β1-integrin, β-catenin, and PCNA. Meanwhile, the cell cycle were measured by flow cytometry. The cell colony formation rates were detected after 1 week. Results The achievement ratio of DM rat model was 90% with good stabil ity. HE staining showed that the number of islet cells significantly decreased with degeneration and necrosis in DM group; the structure of islet cell was clear without degeneration and necrosis in normal control group. The integral absorbance values of positive expression for K19, β1-integrin, β-catenin, and PCNA in ESCs of DM group (82.63 ± 14.77, 21.59 ± 4.71, 6.49 ± 6.58, and 90.77 ± 12.44, respectively) were significantly lower than those in the normal control group (151.24 ± 42.83, 54.48 ± 17.43, 116.39 ± 9.26, and 110.62 ± 20.67, respectively) (P lt; 0.01). The clone forming efficiency of ESCs in DM group (6.43% ± 1.01% ) was significantly lower than that in the normal control group (11.37% ± 1.62%) (P lt; 0.01). Flow cytometry indicated that 88.89% of cultured ESCs in the DM group were in resting state/ pre-DNA-synthetic gap (G0/G1), and the apoptosis rate was 3.98%; 91.50% in the normal control group and the apoptosis rate was 0. Conclusion The DM rat model can be effectively induced by intraperitoneal injected 65 mg/ kg STZ. The decreased amount and the low prol iferation and differentiation capacity of ESCs may be one of the important mechanisms of difficult recovering wounds of DM rats.
Objective To investigate the perioperative management and the results of surgical treatment of spinal tuberculosis associated with diabetes mellitus. Methods The cl inical data were analysed retrospectively from 42 patients with spinal tuberculosis associated with type 2 diabetes mell itus who were surgically treated between July 2001 and January 2009.There were 22 males and 20 females with an average age of 56.5 years (range, 41-78 years). The disease duration was 4-18 months (mean, 7.5 months). The involved vertebrae included 2 cervical vertebrae, 13 thoracic vertebrae, 17 thoracolumbar vertebrae, and 10 lumbar vertebrae. Of them, 18 patients compl icated by paraplegia, and 25 patients had more than one of concomitant diseases. Anterior debridement and bony grafting with anterior instrumentation fixation were performed in 16 patients; anterior debridement and bony grafting with posterior instrumentation fixation in 4 patients; posterolateral costotransversectomy debridement and interbody fusion with posterior instrumentation fixation in 8 patients; posterior debridement and bony grafting with posterior fixation in 7 patients; sinus resectomy and focus debridement in 2 patients; anterior debridement and bony grafting in 3 patients; and CT guided percutaneous catheter drainage in 2 patients. Postoperative anti-tuberculosis treatment was given for 12-24 months. Results The patients were followed up 1.5-5.0 years, with a mean period of 3.5 years. One patient died of pulmonary infection after 1 week of operation; 1 patient died of myocardial infarction after 2 years of operation; and other patients survived without tuberculosis recurrence. Among 38 patients who received bony grafting, 34 patients achieved bony fusion, 3 suffered bony grafting failure without kyphotic deformity or instabil ity except 1 patient who died from pulmonary infection. Among 18 cases compl icated by paraplegia, nerve function improved to a certain extent. The intraoperative and postoperativecompl ications occured in 28 cases. The systemic compl ications mainly included heart insufficiency in 5, heart rrhythmia in 3, pulmonary infection in 2, urinary tract infection in 2, and stree ulcer in 1; they were cured after medical treatment. The local complications mainly included sinus tract formation in 5, pleural tear in 2, neurologic injury in 2, intraoperative tear of inferior vena cava in 2, and the loosening of transpedicular screws in 4; they all were cured effectively. Conclusion Surgical treatment of spinal tuberculosis associated with diabetes mell itus appears to be a beneficial procedure on the condition that the blood glucose is controlled and the associated disorders and postoperative complications are properly handled, and reasonably selection of surgical procedures is very important. Instrumentation fixation provides adequate stabil ity to allow early mobilization.
To study the influence of maggot secretion on expression of bFGF and connective tissue growth factor(CTGF) in ulcer tissue of diabetes mell itus(DM)rats and its antibacterial function. Methods There were 40 3-month-old SD male rats (weighing 300-350 g) which were randomly divided into 2 groups: control group and experimental maggot secretion group. The model of ulcer wound of DM rats was made. The ulcer wound of DM rats in maggot secretiongroup spread maggot secretions, but no secretion on ulcer wound was found in control group. The morphological and tissue changes of ulcer wound were observed at different times, and the conditions of bacterial infection on ulcer wound in the two groups were checked. Tissue sl ices were prepared on 7, 14 and 21 days, respectively; immunohistological detection of bFGF and CTFG in ulcer wound of the two groups was done; and the cell number of positive expression of bFGF and CTFG was counted. Results It was found that the heal ing of ulcer was dominant in experimental group; the wound was clean; the tissue regenerated and no Staphylococcus aureus infection was seen. Bad heal ing was obtained in control group; tissue necrosis was found and the rate of Staphylococcus aureus infection was 60%. Positive expression cell number of bFGF in ulcer wound was detected on 7 and 14 days after operation with 23.76 ± 3.34 and 52.76 ± 4.84 in experimental group, and 18.88 ± 2.16 and 46.04 ± 4.00 in control group. Positive expression cell number of CTGF in ulcer wound was detected on 7 and 14 days after operation with 18.76 ± 3.24 and 46.52 ± 4.07 in experimental group, and 12.52 ± 3.03 and 40.52 ± 3.96 in control group. There was significant difference between positive expressions of bFGF and CTFG in the two groups (P﹤0.05). Conclusion The maggot secretion can elevate the expressions of bFGF and CTFG in ulcers, promote heal ing and prevent bacterial infection.