Effect of recombinant human granulocyte-macrophage colony stimulating factor on wound healing in patients with deep partial thickness burn / 中华烧伤杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy and safety of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel in wound healing in patients with deep partial thickness burn.</p><p><b>METHODS</b>The study was a multicenter, randomized, double-blind, placebo-controlled parallel clinical trial. Three hundred and twenty-one patients (302 cases finally fulfilled the protocol) with deep partial thickness burn were divided into A group (n = 200, with treatment of rhGM-CSF hydrogel, 100 microg/10 g/100 cm2/d), C group (n = 102,with treatment of placebo). Side-effect, systemic condition, wound healing time, wound healing rate, and total effective rate at different time points were observed.</p><p><b>RESULTS</b>There were no obvious differences in vital signs, wound secretion, wound edge reaction, blood and urine routine, liver and kidney function between two groups (P > 0.05). No side-effect was observed. The median wound healing time was 17 days in A group, which was obviously shorter than that in C group (20 days, P < 0.01). The mean wound healing rate in A group was 24.5%, 70.5%, 95.3%, 99.6% respectively on 8th, 14th, 20th, 28th day after treatment, which were obviously higher than that in C group (15.1%, 51.4%, 84.6%, 97.1%, respectively, P < 0.01). The total effective rates in A group on 8th, 14th, 20th day after treatment were also higher than that in C group (P < 0.01).</p><p><b>CONCLUSION</b>rhGM-CSF hydrogel can significantly accelerate wound healing in patients with deep partial thickness burn with certain safety.</p>

Akt gene therapy for cirrhotic rats with portal hypertension / 中南大学学报(医学版)

OBJECTIVE@#To determine whether there is an impaired Akt and eNOS activation in cirrhotic livers, and to investigate the feasibility of transferring adenovirus-mediated Akt gene to the liver for portal hypertension.@*METHODS@#Recombinant adenovirus Ad-myr-HA-Akt and Ad-EGFP were produced by homologoas recombination in 293 cells . The Methods of compound factor, carbon tetrachloride (CCl4), corn flour, and cholesterol plus alcohol were used to construct the hepatic cirrhosis rat models. Ten normal rats were served as a normal control group, and 40 cirrhotic rats were divided into 4 groups randomly: an untreated group, an Ad-myr-HA-Akt treated group, an Ad-EGFP group, and a saline group. Ad-myr-HA-Akt, Ad-EGFP, and saline were transduced into the Ad-myr-HA-Akt treated group, Ad-EGFP group, and saline group via the tail vein respectively. Portal vein pressure, mean arterial pressure, and heart rate were measured in all rats. Protein abundance and phosphorylation status of Akt and eNOS were examined by Western blot. Spectrophotometry was used to measure the NO level. Frozen sections of the liver, heart, lung, kidney, brain, spleen, and testis were made to examine the expression of enhanced green fluorescent protein (EGFP) by fluorescence microscopy on Day 3 in the Ad-EGFP group.@*RESULTS@#The concentration of recombinant adenovirus Ad-myr-HA-Akt after the purification was 5.5 x 10(11)vp/mL and that of Ad-EGFP was 6.0 x 10(11)vp/mL. Akt and eNOS phosphorylations in the liver of cirrhotic rats were obviously impaired. Adenoviral delivery of myr-Akt restored eNOS phosphorylation, increased the NO level and decreased the portal pressure after 3 days of adenoviral infection. In contrast, the livers infected with Ad-EGFP and saline were not changed. The EGFP expression was mainly found under the fluorescence microscopy on the frozen section of liver. Very little fluorescence was detected in the lung and kidney; and there was no detectable EGFP in other organs.@*CONCLUSION@#There is an impaired Akt and eNOS activation in the cirrhotic livers; myr-Akt gene therapy can restore the Akt activation and NO production in the cirrhotic liver, suggesting that this therapy may be helpful in treating portal hypertension.

Increased expression of heat shock protein 70 and heat shock factor 1 in chronic dermal ulcer tissues treated with laser-aided therapy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chronic dermal ulcers are also referred to as refractory ulcers. This study was conducted to elucidate the therapeutic effect of laser on chronic dermal ulcers and the induced expression of heat shock factor 1 (HSF1) and heat shock protein 70 (HSP70) in wound tissues.</p><p><b>METHODS</b>Sixty patients with 84 chronic dermal ulcers were randomly divided into traditional therapy and laser therapy groups. Laser treatment was performed in addition to traditional therapy in the laser therapy group. The treatment efficacy was evaluated after three weeks. Five tissue sections of healing wounds were randomly collected along with five normal skin sections as controls. HSP70-positive cells from HSP70 immunohistochemical staining were counted and the gray scale of positive cells was measured for statistical analysis. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting were performed to determine the mRNA and protein expressions of HSF1 and HSP70.</p><p><b>RESULTS</b>The cure rate of the wounds and the total efficacy in the laser therapy group were significantly higher than those in the traditional therapy group (P < 0.05, P < 0.01, respectively). Immunohistochemical staining revealed that the HSP70-positive cell count was significantly higher in laser therapy group than those in the traditional therapy group and controls (P < 0.01), and the gray scale of the cell signal was obviously lower than traditional therapy group and controls (P < 0.05). By contrast, the traditional therapy group and the control group were not significantly different. The RNA levels of HSF1 and HSP70 were higher in the laser therapy group by RT-PCR, but very low in normal skin and the traditional therapy group. The analysis on the gray scale of the Western blot bands indicated that the expression of HSF1 and HSP70 in the laser therapy group was significantly higher than in the traditional therapy group and the control group (P < 0.01), and the expression in the traditional therapy group was also higher than in the control group (P < 0.05).</p><p><b>CONCLUSION</b>Laser-aided therapy of chronic dermal ulcers plays a facilitating role in healing due to the mechanism of laser-activated endogenous heat shock protection in cells in wound surfaces.</p>

Influence of heat shock factor 1 gene transfection on the expression of inflammatory mediators in macrophages induced by burn serum / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of heat shock factor1 (HSF1) on gene expression of inflammatory mediators in RAW264.7 murine macrophage cells induced by burn serum.</p><p><b>METHODS</b>Sera were separated from blood of normal rats and rats with severe burns, and the recombinant vector pcDNA3. 1/HSF1 was constructed. RAW264.7 macrophages were divided into non-transfection group, vacant vector group (with burn and normal sera stimulation, respectively after vacant vector transfection) and recombinant vector group (with burn and normal sera stimulation, respectively after recombinant vector transfection). Some recombinant vector transfected macrophages without serum stimulation were prepared for the determination of HSF 1 expression with Western blotting. The mRNA expressions of TNF-alpha, HMGB 1 and IL-10 were determined with RT-PCR.</p><p><b>RESULTS</b>The cell line attained after recombinant vector transfection was comparatively stable,with partial activation of HSF 1. Burn sera markedly upregulated TNF-alpha, HMGB1 mRNA expression (0.910 +/- 0.100, 0.860 +/- 0.020, respectively), but downregulated IL-10 expression (0.430 +/- 0.010, respectively) in normal macrophages, while these genes maintained in a very low level in normal macrophages with normal serum stimulation . macrophages with recombinant vector transfection and burn serum stimulation could obviously inhibit the expression of TNF-alpha and HMGB 1, but enhance the IL-10 gene expression (0.130 +/- 0.100, 0.450 +/- 0.020 , 0.450 +/- 0.020, respectively )when compared with that with vacant vector transfection and burn serum stimulation (0.800 +/- 0.050, 0.880 +/- 0.030, 0.420 +/- 0.010, respectively).</p><p><b>CONCLUSION</b>HSF1 can inhibit the expression of some pro-inflammatory mediators in macrophages after a severe burns, indicating that appropriate upregulation of anti-inflammatory mediators might exert protective effects on the organism.</p>

The expression of HIF-1alpha in scars and its relationship to apoptosis / 中华整形外科杂志

<p><b>OBJECTIVE</b>To explore the mechanism of hypoxic environment lessening in the course of scar maturation.</p><p><b>METHODS</b>The expression of hypoxia inducible factor 1alpha (HIF-1alpha) and p53 in granulation of burn wound, burn scars of different stages, and normal skin was detected with immunohistochemical staining. The expressions were quantified by the weight method. The relationship between HIF-1alpha and p53 was investigated.</p><p><b>RESULTS</b>With the scar maturation, the expression of HIF-1alpha gradually decreased while the expression of p53 gradually increased within one year. There was a negative correlation between HIF-1alpha and p53 (P < 0.01).</p><p><b>CONCLUSION</b>p53 plays an important role in scar maturation. Associated with p53, HIF-1alpha may induce apoptosis, which decreases oxygen consumption and lightens hypoxia in the scar maturation.</p>

Human endostatin gene transfected adult skin melanoma cells / 中南大学学报(医学版)

OBJECTIVE@#To study the biological characteristics of human endostatin (hEndo) gene transfected adult skin melanoma cells in vitro and in vivo.@*METHODS@#The plasmid pcDNA3.1 (-)-hEndo was transfected into adult skin melanoma cells by electroporation, and then the stable clones were selected with G418. The transcription and expression of hEndo gene in the transfected melanoma cells were verified by RT-PCR and agarose gel electrophoresis analysis and Western blot. The biological activities of hEndo protein were investigated by MTT in vitro. Stable clones expressing endostatin were subcutaneously injected into the right flank of BALB/c-nu/nu mice of 4 to approximately 6 weeks old. Then the growth of transduced tumors in vivo was investigated.@*RESULTS@#The bands of 624 bp and 5.4 kb were identified from digested plasmid pcDNA3.1 (-)-hEndo. The stable clones were selected with G418 after the eletroporation, the expression of hEndo mRNA was verified by RT-PCR, and Western blot displayed the expression product of hEndo was about 20 kD in the transfected melanoma cells. MTT showed that the conditioned medium of melanoma cells transduced with recombination human endostatin expression vector could inhibit the proliferation of human umbilical vein endothelial cells in vitro. The growth of transduced cells in vivo showed that transfected melanoma cells grew in vivo at a slower rate than the control cells (P < 0.05). RT-PCR showed that endostatin expressed in the transduced tumors.@*CONCLUSION@#Adult skin melanoma cells in vitro transfected with exogenetic hEndo gene can express and secrete active hEndo, and inhibit the growth of transduced tumors in vivo.

Relationship between angiogenesis and expression of HO-1 of scar / 中南大学学报(医学版)

OBJECTIVE@#To investigate the relationshion between the angiogenesis of different kinds of scar and expression of HO-1.@*METHODS@#The expression of heme oxygenase-1 and vessel counted by CD34 of biopsies from different kinds of scars such as hypertrophic scar, keloid, surgical scar and normal skin of 24 cases was valued by immunochemical method, and the relationship was compared between them.@*RESULTS@#The vessel count of hypertrophic scar, keloid was significantly abundant compared with surgical scar or normal skin (P < 0.01). While the expression of HO-1 of hypertrophic scar, keloid was obviously higher than that in surgical scar or normal skin (P < 0.01), decreased from hypertrophic scar, keloid, surgical scar to normal skin. There existed a positive correlation between vessel count and the expression of HO-1 (r = 0. 761, P < 0.01) as well as the number of fibroblastic cells (r = 0. 731, P < 0.01) in the study groups.@*CONCLUSION@#HO-1 might play a important role in the angiogenesis of scar formation. The cause of these changes may be local. Over angiogenesis is one symbol of pathological scar.

Effect of Chinese drugs mixture on immune function of patients with extremely severe burn / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the regulatory effect of Qinghuo Baidu Yin (QHBDY, a mixture prepared with Chinese drugs) on immune function of patients with extremely severe burn (ESB).</p><p><b>METHODS</b>Thirty patients with ESB were divided into two groups, conventional therapy was given to both groups, but QHBDY was given to the treated group additionally. Immunological indices, including peripheral blood T-lymphocyte subsets, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) were determined 3 weeks after treatment to evaluate and compare the therapeutical effect in the two groups.</p><p><b>RESULTS</b>Compared with the control group, CD3, CD4, CD4/CD8, immunoglobin (IgG, IgA and IgM) and complement (C3 and C4) levels were markedly decreased in degree, and recovered earlier and quicker, with CD8 increased mildly (P < 0.01) and turned back more quickly. And so did the parameters of the treated group in comparing with that of the control group at anytime (P < 0.05 or P < 0.01).</p><p><b>CONCLUSION</b>Chinese drugs mixture shows the regulatory effect on both cellular and humoral immune function in patients with ESB.</p>