Negative pressure wound therapy is associated with up-regulation of bFGF and ERK1/2 in human diabetic foot wounds.

Chronic foot wounds are a leading cause of morbidity and hospitalization for patients with diabetes. Negative pressure wound therapy (NPWT) is known to promote healing of diabetic foot wounds, but the underlying molecular mechanisms remain elusive. We propose to gain molecular insights into the wound healing promoting signals underlying the effects of NPWT on diabetic foot wounds in humans. We assessed 30 patients with diabetic foot ulcers. Of these cases, 15 were treated with NPWT, while 15 patients were treated with traditional gauze therapy. Granulated tissue was harvested before and after treatment in both patient groups and histologically analyzed with hematoxylin & eosin as well as Masson's trichrome staining methods. Immunohistochemistry and Western blot analysis was performed to evaluate expression of basic fibroblast growth factor (bFGF) and extracellular signal-regulated kinase (ERK)1/2, previously associated with promoting cellular growth and/or wound healing. Unlike controls, the wounds in the NPWT-treated diabetic patients developed characteristic features of granulated tissue with increased collagen deposition. Immunohistochemical analysis also revealed an increase in bFGF levels in NPWT-treated patients. Western blot analysis further showed a significant up-regulation of bFGF and phosphorylated ERK1/2 protein levels in the NPWT-treated diabetic patients vs. controls. Our studies reveal that NPWT is associated with an up-regulation of bFGF and ERK1/2 signaling, which may be involved in promoting the NPWT-mediated wound healing response.

Relationship between Trp64Arg mutation in the ß3-adrenergic receptor gene and metabolic syndrome: a seven-year follow-up study.

BACKGROUND: It has been shown that the ß3-adrenergic receptor (ß3-AR) gene Trp64Arg mutation was closely related to obesity and insulin resistance, and may be related to the prevalence of metabolic syndrome (MS). The aim of this study was to investigate the relationship between the ß3-AR gene mutation and the prevalence of MS. METHODS: A seven-year follow-up study was initiated in 2000, with 496 samples of simplex obese subjects (body mass index ≥ 25 kg/m(2)) and 248 normal-weight subjects. According to the ß3-AR genotypes, the subjects were classified as Trp64 homozygote group and Arg64 carrier group and after 7 years the prevalence of MS was determined. RESULTS: According to the baseline profile, there were no significant differences in the adiposity, blood pressure, lipid profile, fasting plasma glucose and fasting insulin between Trp64 homozygote group and Arg64 carrier group either in obesity or normal-weight subjects. The results of follow-up study indicated that in obese men the prevalence rate of MS was much higher in Arg64 carrier group than that in Trp64 homozygote group (54.76% vs. 40.85%, P < 0.05), but there was no statistical difference in women of the above groups. The prevalence rate of MS in obese men of both Trp64 homozygote group and Arg64 carrier obese group were obviously higher than that in women of the above groups (40.85% vs. 18.27% and 54.76% vs 21.28%, all P < 0.005). Differences were not statistically significant in the prevalence of MS for normal weight Trp64 homozygote group and normal weight Arg64 carrier group, either between men, between women, or between men and women. Comparison of populations indicated that no matter with the ß3-AR gene mutation or not, the prevalence of MS in obese subjects was significantly higher than normal weight subjects (χ(2) = 28.240 and χ(2) = 15.586, all P < 0.005). Logistic analysis showed that the mutation of ß3-AR gene was associated with the prevalence of MS in men. CONCLUSION: The mutation of ß3-AR gene is the independent risk factor for the prevalence of MS in men.

Virtual screening for Raf-1 kinase inhibitors based on pharmacophore model of substituted ureas.

A three-dimensional (3D) quantitative pharmacophore model was developed from a training set of structurally diverse substituted ureas against Raf-1 kinase, which was well validated to be highly predictive by two methods, namely, test set prediction and Cat-Scramble method. Then a virtual database searching was performed with the pharmacophore model as a 3D query, and the bioactivities of the retrieved hits were predicted by the pharmacophore. Subsequently, molecular docking was carried out on the selected hits whose estimated IC(50) was less than 1 microM. Finally, 29 hits were identified as potential leads against Raf-1 kinase, which exhibited good estimated activities, high docking scores, similar binding mode to experimentally proven compounds and favorable drug-like properties. The study may facilitate the discovery and rational design of novel leads with potent inhibitory activity targeting Raf-1 kinase.

[A prospective study of the relationship between Trp64Arg beta(3)-adrenergic receptor gene polymorphism and metabolic syndrome].

OBJECTIVE: To investigate the relationship between Trp64Arg mutation in beta(3)-adrenergic receptor (beta(3)-AR) gene and the incidence of metabolic syndrome (MS). METHODS: A seven-year follow-up study was conducted in 386 simple obese subjects and 175 normal weight subjects in whom geno-typing of Trp64Arg mutation in beta(3)-AR gene was examined in 2000. RESULTS: There were no differences between a Trp64Trp homozygote group and a Trp64Arg heterozygote group of whether obese or normal weight subjects with respect to adiposity, blood pressure, lipid profile, fasting blood glucose and fasting insulin in the baseline. The results of follow-up indicated that the incidence of MS in the Trp64Arg heterozygote group was higher than that in the Trp64Trp homozygote group of obese males (54.76% vs 40.85%, P < 0.05) but not in the group of obese females. The incidences of MS both in the Trp64Trp homozygote group and Trp64Arg heterozygote group were higher in obese males than in obese females (40.85% vs 18.27% and 54.76% vs 21.28%, all P < 0.01). No significant differences were found in incidences of MS both in the Trp64Trp homozygote group and Trp64Arg heterozygote group of normal weight subjects whether the comparison was made between males and females respectively or between males and females. The overall incidence of MS in the obese subjects were significantly increased than that in the normal weight subjects whether there was gene variant or not (31.30% vs 6.03% and 42.75% vs 12.73%, all P < 0.01). Logistic analysis showed that beta(3)-AR gene variant was associated with increased incidence of MS in males. CONCLUSION: beta(3)-AR gene Trp64Arg mutation is an independent risk factor for the incidence of MS in males.

[Effect of simulated ischemia on activity and heterogeneity of Na+ channel in rabbit ventricular epicardial, midmyocardial and endocardial myocytes].

OBJECTIVE: To study the ionic mechanism of reentrant arrhythmia. METHODS: Single myocytes were enzymatically isolated from the epicardium, midmyocardium and endocardium of the left ventricle free wall of rabbit, followed by whole-cell patch-clamp recording of the Na(+) current (I(Na)) of the 3 cellular subtypes (superfused with normal and then simulated ischemia solution). The currents in the 3 cellular subtypes before and after simulated ischemia lasting for 10, 20, and 30 min, respectively, were compared. RESULTS: No changes was recorded in the configuration of the I-V curves and voltage dependence of I(Na) after simulated ischemia, and the peak I(Na) densities (- 20 mV) were significantly reduced in the 3 cellular subtypes compared with those recorded in normal condition. At the same time, the differences in I(Na) peak current densities in the 3 cellular subtypes underwent variations after simulated ischemia. Simulated ischemia resulted in obvious shift of I(Na) steady-state inactivation curves in the hyperpolarizing direction in the 3 cellular subtypes and inactivation was accelerated, and the differences in the half maximal inactivation voltages (V0.5) between the 3 cellular subtypes were also altered after simulated ischemia. After simulated ischemia, I(Na) recovery from inactivation in the epicardium, endocardium and midmyocardium was all slowed down in comparison with that in normal condition, but without statistical significance. Differences between the recovery curves of three cellular subtypes were noted after ischemia for 30 min. CONCLUSION: Ischemia can affect the activity of Na(+) channel, disrupting the balance of ion channel currents and the heterogeneity of I(Na) among the 3 cellular subtypes, which is responsible for the onset of arrhythmia and partially explains different pharmacological reactions of the 3 cellular subtypes under normal and ischemic conditions.