Expression of coxsackie-adenovirus receptor in keratinocytes of mouse skin after heat stimulation and the effect of coxsackie-adenovirus receptor on dendritic epidermal T lymphocytes / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the influence of heat stimulation on expression of coxsackie-adenovirus receptor (CAR) in keratinocytes (KCs) of mouse skin and the effect of CAR on production of cell growth factors by dendritic epidermal T lymphocytes (DETCs).</p><p><b>METHODS</b>(1) Twenty BALB/c mice were divided into heat stimulation group (HS) and control group (C) according to the random number table, with 10 mice in each group. Mice in group HS were inflicted with scald milder than superficial-thickness by dressing wet hot gauze, which had been soaked in 100°C hot water for 3 min, in the hair removed area on the back for 1 to 3 s, while mice in group C were sham injured by dressing a wet gauze which had been soaked in water of room temperature for 3 min in the hair removed area on the back for 1 to 3 s. Square full-thickness skin specimens measuring 2.0 cm × 2.0 cm in size were obtained from the center of the bare skin. The expression of CAR in skin tissue sections were detected by immunohistochemistry staining. The mRNA and protein expression levels of CAR in skin tissue sections were respectively determined by real-time fluorescent quantitation RT-PCR and Western blotting. (2) KCs were isolated and cultured from full-thickness skin obtained from the trunk of 2 fetal BALB/c mice, and they were divided into 2 groups according to the random number table, with 5 wells in each group. The cells in group HS and group C were respectively cultured in 42°C and 37°C, 5% CO2 incubator for 1 h, and then all the cells were cultured in 37 °, 5% CO2 incubator for 6 h. The apoptosis of the cells and their expression of CAR were detected by flow cytometer. (3) Five BALB/c mice were sacrificed, and full-thickness skin was obtained from the trunk. The DETCs were divided into 7 groups according to the random number table after being isolated and purified from the skin specimens. Cells in group C were cultured without any stimulation, and cells in the 0.5, 1.0, 2.0, 4.0, 8.0, and 16.0 mg/L CAR groups were respectively cultured with corresponding concentration of recombinant mice CAR nutrient solutions, with 5 wells in each group. The contents of insulin-like growth factor I (IGF-I) and keratinocyte growth factor (KGF) were determined with ELISA. Data were processed with independent samples t test and one-way analysis of variance.</p><p><b>RESULTS</b>(1) The immunohistochemistry staining showed that there was mild positive staining in the skin tissue sections of mice in group C, while the positive staining was more obvious in group HS. The positive staining was mainly located in KCs, hair follicles, and sweat gland epithelial cells, while no positive staining was observed in fibroblasts. The mRNA expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.157 ± 0.027 and 0.773 ± 0.029. There was statistically significant difference between them (t = 3.052, P < 0.01). The protein expression levels of CAR in skin tissue sections in group C and group HS were respectively 0.23 ± 0.09 and 0.89 ± 0.14. There was statistically significant difference between them (t = 2.556, P < 0.05). (2) The apoptosis rates of KCs in group C and group HS were respectively (5.7 ± 1.3)% and (7.4 ± 1.7)% (t = 0.464, P > 0.05). The expression rates of CAR in KCs in group C and group HS were respectively (48 ± 6)% and (80 ± 8)%. There was statistically significant difference between them (t = 2.585, P < 0.05). (3) The contents of IGF-Iin culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (23.1 ± 1.8), (22.5 ± 2.1), (31.2 ± 2.5), (39.7 ± 2.3), (61.8 ± 3.5), (45.1 ± 2.8), and (29.0 ± 2.0) µg/L. There was statistically significant difference among 7 groups (F = 3.414, P < 0.05). The contents of KGF in culture supernatants in group C and 0.5, 1.0, 2.0, 4.0, 8.0, 16.0 mg/L CAR groups were respectively (131 ± 9), (217 ± 12), (355 ± 21), (563 ± 21), (535 ± 34), (292 ± 20), and (245 ± 10) ng/L. There was statistically significant difference among 7 groups (F = 5.063, P < 0.01).</p><p><b>CONCLUSIONS</b>The expression of CAR in KCs would rise after HS. The optimum CAR concentration to increase IGF-I and KGF production in DETCs is low.</p>

Methodology of BMSCs in repairing ureteral injury in mice via renal artery transplantation / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of bone marrow mesenchymal stem cells (BMSCs) on ureteral injury repair via renal artery transplantation.@*METHODS@#The left ureteral obstruction model was set up in 49 Balb/c mice by micro vascular clamp. The microscopic vascular clamp was taken out to lift the left ureteral obstruction after 10 days. The mice were randomly divided into an experimental group (n=25) and a control group (n=24). Balb/c mice BMSCs transfected by luciferase (Luc) were transplanted immediately through the renal artery after removing the microscopic vascular clamp from the experimental group; while mice in the control group was closed the incision after the microscopic vascular clamp was removed immediately and without BMSCs transplant. Magnetic resonance imaging (MRI) was used to scan the experimental mice. By measuring the left renal pelvis volume of the experimental mice at different time points and comparing the left ureter recanalization rate after removing left ureteral obstruction of the experimental group and the control group, we evaluated the repair effect of BMSCs on ureteral injury.@*RESULTS@#The volume of the left renal pelvis in experimental mice became bigger obviously after the left ureter was obstructed (P<0.01). The left renal pelvis volume of the experimental group and the control group had no statistical significance 10 days after the left ureteral obstruction was set up (P=0.693). In the experimental group, the left ureter recanalization rate was higher than that in the control group, after removing the left ureteral obstruction (P=0.012).@*CONCLUSION@#Transplantation through the renal artery can promote the restoration of ureteral injury in mice.

Determination of ephedrine hydrochloride in Juyuanzhike Tablets by HPLC / 中成药

AIM: To establish a method for the determination of ephedrine hydrochloride in Juyuanzhike Tablets (Radix Platycodonis,Radix Polygalae,ephedrinelydrochloride,etc.). METHODS :Ephedrine hydrochloride in Juyuanzhike Tablets were determined by HPLC. RESULTS : The linear range was 0.2～1.6 ?g,r =0.9999.The average recovery was 97.7% and RSD was 0.58%,respectively. CONCLUSION : The method is simple,feasible and reproducible. It can be used for the quality control of Juyuanzhike Tablets.