To investigate the effects of basic fibroblast growth factor (bFGF) on intracellular free Ca 2+ of heating injury, and observe the relationship between mitogen activated protein kinases(MAPKs)signal pathways and Ca 2+ mobilization. Cultured human fibroblasts were heated at 45?C for 10min, then divided into four groups :①basic fibroblast growth factor (10ng/ml) treatment; ②preincubated cells with PD98059 (10?mol/L) for 30 min followed by bFGF (10ng/ml); ③preincubated cells with SB203580 (10?mol/L) for 30min followed by bFGF (10ng/ml); ④preincubated cells with PD98059 (10?mol/L) and SB203580 (10?mol/L) for 30min followed by bFGF (10ng/ml). The cells were incubated with fluorescence Ca 2+ dye fluo 3/AM at 37 C for 30min, then measured by using laser scanning confocal microscope. The results showed fluorescent intensity of fibroblast heating injury was weak, the concentration of intracellular free Ca 2+ increased after stimulation with bFGF treatment. PD98059 and SB203580 could induce calcium oscillation. A rapid decrease of fluorescent intensity was observed after cells were preincubated with PD98059 and SB203580 at the same time. bFGF induced an increase of cytoplasmic and intracellular free Ca 2+ concentrations. It is suggested that MAPKs signaling pathway has a feedback regulation for free Ca 2+ mobilization.

To explore the activities of signal transduction pathway involved in heat-stressed fibroblast in vitro. Human dermal fibroblasts were cultured in Dulbecco′s modified Eagle′s medium with 5% calf serum at 5%CO 2 in a water-saturated atmosphere. Cultured cells were heated to 45℃ for 10min. Western blotting was used to detect ERK1/2 and JNK expression, and the expression of caspase-3 protein was observed by immunoflurescence technique. The results showed that the MAPKs signal transduction pathway was activated in heat-stressed fibroblasts. The expression of JNK reached the peak at 60min, then maintained up to 180min. The expression of ERKs peaked at 30min, then lowered. The expression of caspase-3 was weak at 30min after heat-stress, and became evidently strong at 60min. The signal pathway of ERKs and JNK played important roles in the changes in biologic characteristics of fibroblasts after heat-stress.

The basic concept of gene therapy is to introduce a therapeutic gene into a cell, whose expression can improve to healing of wound. To achieve this goal, the suitable therapeutic gene has been selected and delivered into the reparative cell, which is becoming a focal point works about gene therapy in wound healing. There have been several different therapeutic genes and gene transfer strategies that have been used in models of wound healing. This article discusses several methods that have been used to deliver genes encoding growth factor proteins, stem cells into wounds and the advantages/disadvantages of each approach. We hope a safe vectors system to deliver the effectual transgene in wound healing.

Animals ; Cardiovascular System ; chemistry ; Female ; Group II Phospholipases A2 ; blood ; Group IV Phospholipases A2 ; blood ; Group V Phospholipases A2 ; blood ; Group X Phospholipases A2 ; blood ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

Objective Recent studies suggest that the red blood cell distribution width ( RDW) may play a role in the diag-nosis and treatment of cardiovascular disease.The aim of this study was to investigate the correlation of red blood cell distribution width (RDW) with cardiovascular events in patients with Acute ST-segment elevation myocardial infarction (ASTEMI) in order to provide the basis for improving the diagnosis level and therapeutic effect. Methods Retrospective study was made on the clinical data of 189 patients with ASTEMI enrolled in Sichuan Provincial People′s Hospital from January 2013 to March 2014.The survival rates of patients with ASTEMI with different RDW were estimated by Kaplan-Meier and compared by Log-rank test and the prognosis factors were inves-tigated by Cox proportional hazard regression model. Results 97 patients′RDW levels were more than 13.7% and 92 patients′RDW levels were less than or equal to 13.7% in 189 ASTEMI patients. The hs-CRP, BNP, LVEF and HDL-C were significantly different be-tween two groups (P<0.05).The rates of recurrent myocardial in-farction, heart failure, sudden death cardiac in patients with highRDW were higher than that in patients with low RDW (22.7%vs 8.7%, 27.8%vs 10.9%, 15.5%vs 5.4%)(χ2 =6.915, 8.632, 5.019, P<0.05) .There were 60 patients with high RDW who experienced cardiovascular events and the 1-year cardiovascular event-free survival rate was 38.1%, and 20 patients with low RDW cardiovascular events was 79.3% in follow-up period (χ2 =30.959, P<0.001).Multivariate cox regression showed that age>65 (HR=2.43, 95%CI:1.09~5.44) and RDW>13.7%(HR=2.20, 95% CI:1.10~4.43) were risk prognostic factors. Conclusion There is a certain correlation between RDW and cardiovascular e-vents in patients with ASTEMI.The ASTEMI patients with high RDW hold higher risk of cardiovascular events and poorer prognosis.

Objective To investigate the possible signaling mechanisms by which recombinant human platelet-derived growth factor (rhPDGF) accelerated healing of cutaneous wound in diabetic rats. Methods Four full-thickness skin wounds were incised in the back of 26 male Wistar diabetic rats. The wounded rats were divided into 3 groups (7 or 8 rats each group). One group without treatment was used as a control, and the other 2 groups were treated with rhPDGF at a dose of 7.0 μg/cm2 wound or vehicle ( DMSO/0.9%NaCl, vol/vol 1:1) from 1 to 14 days. The wound healing was evaluated by the measurements of the wound volume and area. Immunofluorescent and immunohistochemical staining were used to examine the phosphorylation of extracellular signal-regulated kinase 1/2(ERK1/2) and the expression of proliferative cell nuclear antigen (PCNA), respectively. Results Granulation tissue appeared in the bed of wound after injury. The number of blood capillary buds and fibroblasts was greater in the rhPDGF-treated group than that in the other 2 groups. A lot of inflammatory cells infiltration and collagen deposition were observed in the wound. The wound-volume in the rhPDGF-treated group was smaller than that in control group ( P ＜ 0.05). The reepithelialization rate in rhPDGF-treated group was higher than that in the other 2 groups at 7 days after injury ( P ＜ 0.05). The expression of PCNA in reparative cells was higher in rhPDGF-treated group than in control group or vehicle-treated group at 3,7 days after injury( P ＜ 0.05). The phosphorylation of ERK1/2 was stronger in rhPDGF-treated group than that in control group or vehicle group at 7 and 14 days after injury( P ＜ 0.05). Conclusion These results suggest that rhPDGF accelerates wound healing and improves healing quality by increasing the phosphorylation of ERK1/2.

Objective To investigate the possible signal transduction pathway during the course of recombinant human platelet derived growth factor (rhPDGF) promoting cutaneous incisional wound healing in diabetic rats. Methods Four full thickness skin wounds were incised in the back of 26 Wistar rats with diabetes mellitus that were randomly divided into two groups, Group A (diabetic rats control) and Group B (treated with rhPDGF at 7.0 ?g/cm2 wound). The granulation, collagen sedimentation, the reepithelialization rate of rhPDGF as well as the inflammatory cell filtration were observed 3,7 and 14 days after wound. Immunofluorescence staining was used to observe the expression of ERKs either around the wound and in the wound center and immunohistochemical method applied to observe the changes of c-fos, proliferation cell nuclear antigen (PCNA) and focal adhesion kinase (FAK). Results The histological investigation showed a lot of granule tissues in the bed of wound, a large of inflammatory cells infiltration and collagen deposition, active growth of granule tissues and significant wound contraction. The number of blood capillary buds and fibroblasts in the Group B were more than that in the Group A. The immunohistochemistry showed that the expressions of ERK1 and c-fos increased significantly three days, strengthened seven days and went weak 14 days after rhPDGF application. In the Group B, at every time point, the expressions of PCNA and FAK were more significant than those in the Group A. Conclusions ERK1/2 signal pathway exerts function in rhPDGF accelerating wound healing of rats with diabetes mellitus and plays an important role in proliferation and migration of reparative cells.

Objective:To observe influence of trimetazidine on cardiac function in patients With chronic heart failure (CHF).Methods:According to number table method,a total of 70 CHF patients accorded With inclusion standards Were randomly and equally divided into trimetazidine group and routine treatment group.According to patient's condition,routine treatment group received digitalis,diuretics,angiotensin converting enzyme inhibitors etc CHF routine freatment.Trimetazidine group additionally received trimetazidine based on routine treatment.The treat-ment period Was four Weeks.Plasma brain natriuretic peptide (BNP)level Was measured and patients received 6min Walking test (6MWT)before and after treatment.Results:Compared With before treatment,there Was significant decrease in plasma BNP level in tWo groups after treatment (P<0.01),compared With routine treatment group, there Was significant increase in decreasing amplitude of BNP level [(655.89±135.61)pg/ml vs.(715.60±181.22) pg/ml,P<0.05]in trimetazidine group;compared With before treatment,there Was significant increase in 6min Walking distance (6MWD)in tWo groups after treatment (P<0.01),compared With routine treatment group,there Was significant increase in increasing amplitude of 6MWD [(181.46±51.16)m vs.(226.06±65.18)m,P<0.01] in trimetazidine group.Conclusion:Treatment trimetazidine can significantly reduce plasma brain natriuretic peptide level and improve heart function based on routine treatment in patients With heart failure.

Objective To study the anti-tumor effect of 5-aza-2′-deoxycytidine(5-aza-dC) on breast carcinoma xenografted in nude mice. Methods The model of breast carcinoma xenografted in nude mice was established.Ten mice were randomized into the treatment group(treated with 5-aza-dC) and control group(treated with PBS).The mass of the tumors before and after treatment were measured in the two groups,and the inhibition rate of the tumor was calculated and the growth curve was drawn.Immunohistochemistry and Western blotting were employed to detect the expression of normal epithelial cell specific-1(NES1)gene. Results The inhibition rate of the tumor in the treatment group was 57.44%,which was significantly different from the control group(P

It designs a way that can easily screen high-pyruvate-producing strain.It is a intelligently selected method which can highly improve the efficiency of strain screening.The principle can be described as the following:On the CaCO-3 medium,a transparent ring can be exhibited based on the reaction of PYR produced by the strain and CaCO-3 in the medium for pyruvate-calcium is a kind of soluble substance,it is obviously that the high-pyruvate-producing strain has a bigger dimension of the transparent ring.On the other hand,color reaction between TTC and ADH indicate the enzyme activities which have a proportional relation with color,our object strain is a weak-ADH-enzyme-activities type with a weak metabolic flux from PYR to alcohol.So the white color strain may be the right choice.