Effect of oleic acid on the proliferation and secretion of pro-inflammatory mediators of human normal fibroblasts and scar fibroblasts / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effect of oleic acid on the proliferation and secretion of pro-inflammatory mediators of human normal fibroblasts and scar fibroblasts.</p><p><b>METHODS</b>Human normal fibroblasts and scar fibroblasts were cultured in vitro and respectively divided into seven groups according to the random number table, with 8 samples in each group. Cells in blank control (BC) group were routinely cultured without addition of other agents. Cells in ethanol-control (EC) group were cultured with medium with the addition of 2% absolute ethanol. Cells in oleic acid groups were cultured with the addition of oleic acid in concentration of 0.25, 0.50, 1.00, 2.00, or 4.00 mmol/L in 2% absolute ethanol. The growth of cells in each group was observed with trypan blue staining on post culture day (PCD) 1-5. On PCD 2, structure of cells in BC, EC, and 1.00 mmol/L oleic acid groups was observed under inverted phase contrast microscope and transmission electron microscope; cell cycle of BC, EC, and 1.00 mmol/L oleic acid groups was measured by flow cytometer; cell proliferation activity in each group was measured by MTT assay; the level of NO in supernatant was assayed by Griess assay; the levels of TNF-α, IL-1β, IL-6, and IL-8 in supernatants in each group were determined by enzyme-linked immunosorbent assay. Data were processed with multifactor and repeated measurement design analysis of variance.</p><p><b>RESULTS</b>(1) There was no significant difference in each index of normal fibroblasts and scar fibroblasts between BC group and EC group. (2) The numbers of normal fibroblasts and scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were significantly lower than those in corresponding BC and EC groups on PCD 2-5 (with F values respectively 13.773 and 11.344, P values all below 0.01). (3) On PCD 2, the numbers of normal fibroblasts and scar fibroblasts in 1.00 mmol/L oleic acid groups decreased, and the cells were aggregating, rounding, and easy to drop off. Cellular membrane disruption, vacuolar degeneration of mitochondrion, pyknosis, and lipid droplets were observed. (4) The percentages of G0/G1 and G2/M phases of normal fibroblasts in 1.00 mmol/L oleic acid group [(93.56 ± 9.98)%, (2.01 ± 0.75)%] were significantly higher than those in BC group [(84.23 ± 10.96)%, (0.37 ± 0.16)%, with F values respectively 3.026, 34.751, P < 0.05 or P < 0.01], while the percentage of normal fibroblasts in S phase [(4.42 ± 0.87)%] was markedly lower than that in BC group [(16.06 ± 1.74)%, F = 136.120, P < 0.01]. The percentages of scar fibroblasts of G0/G1 and G2/M phases in 1.00 mmol/L oleic acid group [(93.86 ± 13.90)%, (1.89 ± 0.66)%] were significantly higher than those in BC group [(83.88 ± 10.42)%, (0.41 ± 0.17)%, with F values respectively 3.529, 32.710, P < 0.05 or P < 0.01], and the percentage of scar fibroblasts in S phase [(3.87 ± 0.63)%] was markedly lower than that in BC group [(15.89 ± 2.02)%, F = 116.508, P < 0.01]. (5) The proliferation rates of normal fibroblasts and scar fibroblasts in 0.50-4.00 mmol/L oleic acid groups were significantly lower than those in corresponding BC and EC groups (with F values respectively 215.945 and 194.555, P < 0.05 or P < 0.01). (6) The content of NO in supernatant of normal fibroblasts in all oleic acid groups was obviously higher than that in BC and EC groups (F = 30.240, P < 0.05 or P < 0.01). The contents of NO in supernatants of scar fibroblasts in 1.00-4.00 mmol/L oleic acid groups were significantly higher than that in BC and EC groups (F = 12.495, P < 0.01). The contents of TNF-α and IL-6 in supernatants of normal fibroblasts and scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were obviously higher than those in corresponding BC and EC groups (with F(TNF-α) values respectively 6.911, 3.818, F(IL-6) values respectively 16.939, 11.600,P < 0.05 or P < 0.01). The contents of IL-1β in supernatants of normal fibroblasts and scar fibroblasts in groups of every concentration of oleic acid were significantly higher than those in corresponding BC and EC groups (with F values respectively 25.117, 9.137, P values all below 0.01). The contents of IL-8 in supernatants of normal fibroblasts in 1.00-4.00 mmol/L oleic acid groups were markedly higher than those in BC and EC groups (F = 2.717, P < 0.05 or P < 0.01). The contents of IL-8 in supernatants of scar fibroblasts in 2.00 and 4.00 mmol/L oleic acid groups were significantly higher than those in BC and EC groups (F = 3.338, P < 0.05). There was no statistically significant difference in above indexes between normal fibroblasts and scar fibroblasts in the same concentration of oleic acid group (with F values from 0.120 to 3.766, P values all above 0.05).</p><p><b>CONCLUSIONS</b>Although oleic acid in high concentration inhibits the proliferation of scar fibroblasts, it also inhibits the proliferation of normal fibroblasts. Oleic acid in high concentration can cause excessive and continued inflammatory reaction by promoting the secretion of pro-inflammatory mediators of normal fibroblasts and scar fibroblasts, and they are detrimental to wound healing.</p>

Investigation and analysis of factors influencing rehabilitation of burn patients / 中华烧伤杂志

<p><b>OBJECTIVE</b>To study the factors influencing health of burn patients in rehabilitation period.</p><p><b>METHODS</b>One hundred and one patients hospitalized in burn department of Xiehe Hospital of Fujian Medical University from February 2008 to October 2008 were investigated by means of General Information Questionnaire, the Eysenck Personality Questionnaire, the Medical Coping Modes Questionnaire, and the Social Support Rating Scale. Their rehabilitation condition was scored according to the Abbreviated Burn-Specific Health Scale. Investigation data were processed by multiple linear regression analysis in order to find out the factors influencing rehabilitation of burn patients.</p><p><b>RESULTS</b>Patients in this group were scored (57 +/- 16) points in physical function, rate [(actual score/possible highest score) x 100%, the same below] 71.1% (the lowest); (97 +/- 19) points in psychological function, rate 80.6%; (53 +/- 8) points in social function, rate 88.4% (the highest); (45 +/- 11) points in general health, rate 74.5%; (251 +/- 44) points in comprehensive health [standard score (314 +/- 55) points], rate 78.5% (upper middle). The factors included in the comprehensive health regression equation (F = 11.602, P < 0.001) were: monthly income, size of burn, number of operations, introverted/extroverted characteristics, living, degree of utilization of support, social support, and resignation. They accounted for 46.6% of the variance of comprehensive health.</p><p><b>CONCLUSIONS</b>Monthly income, size of burn, introverted/extroverted characteristics, living, social support, and resignation are the main factors influencing the rehabilitation level of burn patients.</p>

Changes in the plasma levels of endotoxin in severe burn patients under the treatment of antibiotics / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the changes in the plasma levels of endotoxin in severe burn patients during administration of antibiotics.</p><p><b>METHODS</b>Fifty severe burn patients with burn area larger than 30% TBSA were enrolled in the study, and they were respectively treated with Netilmicin (A group), Cefoperazone (B group), Ceftazidime (C group) and Imipenem/Cilastatin (D group). Venous blood samples were harvested for determination of endotoxins levels before treatment and 1, 2, 3, 5, 7 post-treatment day (PTD).</p><p><b>RESULTS</b>The plasma levels of endotoxin were elevated in different degrees in A, B and C groups. The plasma levels of endotoxin in B group were higher on 1, 2 PTD than on 3, 5, 7 PTD, and they were also higher than that in D group (P < 0.05). The plasma levels of endotoxin in C group reached the peak on 5 PTD [(0.398 +/- 0.172) EU/mL], which were higher than that before treatment [(0.251 +/- 0.142) EU/mL, P < 0.05] and other groups (P < 0.05). The plasma levels of endotoxin in D group were lower on 1, 2 PTD than that before treatment (P < 0.05).</p><p><b>CONCLUSION</b>Different amounts of endotoxins can be released after treatment with antibiotics in severe burn patients. Attention should be paid to the effect of antibiotics on the levels of endotoxin in practice.</p>

The effects of aloe extract on nitric oxide and endothelin levels in deep-partial thickness burn wound tissue in rat / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the effects of polysaccharide extracted from Aloe barbadensis and Aloe barbedensis containing gel on tissue water contents, nitric oxide (NO) and endothelin (ET) levels in wounds of burned rats.</p><p><b>METHODS</b>Four areas of deep-partial thickness burn wounds with 3 cm in diameter were made on each back of 42 male Wistar rats. Single layer gauze impregnated either with 5% (W/W) aloe raw polysaccharide, 10% (W/W) aloe gel, 1% (W/W) sulfadiazine pyridine silver cream (SD-Ag), or normal saline was respectively applied on different wounds. According to different medications, the wounds were divided into aloe raw polysaccharide group, aloe gel group, SD-Ag group and normal saline group. Six rats in each group were sacrificed at 4, 12, 24, 48 post-scald hour (PSH) and on 7, 14, 21 post-scald day (PSD), and the full-thickness skin of wound was harvested for the determination of wound tissues water contents, NO and ET levels, and for calculation of NO/ET ratio. Another 6 normal rats served as normal controls.</p><p><b>RESULTS</b>The water content in the wound tissue in aloe raw polysaccharide group at 12, 24 and 48 PSH [(73.4 +/- 3.8)%, (76.6+/-3.0)%, (70.6+/-3.8)%] and aloe gel group [(74.5+/-2.6)%, (77.1+/-3.6)%, (71.2 +/- 3.1)%] was obviously lower than those in SD-Ag group [(80.1 +/- 4.1)%, (80.5 +/-3.9)%, (76.1 +/-3.8)%, P <0.05]. During 7-21 PSD, all of them returned to the normal level except that in SD-Ag group, as it was still higher than that in normal controls (P < 0.05). The NO content in wound tissue in each group reached the peak at 12 PSH, decreased thereafter, but it was still obviously higher than that of normal controls on 21 PSD (P < 0.05). The ET content in wound tissue of each group reached the peak on 7 or 14 PSD, decreased thereafter, but it was still evidently higher than that in normal controls on 7 or 14 PSD (P < 0.05). The NO content in wound tissue in aloe raw polysaccharide and aloe gel group were markedly lower than those in SD-Ag and normal saline groups at 12 and 24 PSH ( P < 0.05). The NO/ET ratio in each group reached the peak at 12 PSH, decreased thereafter, and it returned to normal value on 14 PSD. On 7 PSD, the NO/ET ratio in aloe gel, SD-Ag and normal saline groups were still significantly higher than that in normal controls, except that returned to normal value in aloe raw polysaccharide group.</p><p><b>CONCLUSION</b>Both aloe raw polysaccharide and aloe gel can decrease wound tissue NO release, optimize NO/ET ratio, lighten vascular inflammatory reaction, and lessen permeability and edema.</p>

Influence of polysaccharide from Aloe vera on the proliferation of the human epithelial cells cultured in vitro / 中华烧伤杂志

<p><b>OBJECTIVE</b>To investigate the influence of polysaccharide from Aloe Vera (AP) on the proliferation of the human epithelial cells cultured in vitro.</p><p><b>METHODS</b>The human epithelial cells undergoing 3 to 4 passages of confluence culture were randomly divided into control and 25, 50, 100, 200 and 400 mg/L AP groups according to different dosage of the polysaccharide (AP) added into the culture medium. In the control group (C), equal volume of DK-SFM medium was added to the culturing cells. The conjugation time of epithelial cells, the changes in the cell morphology and ultrastructure were observed under inverted phase contrast microscope and transmission electron microscope, respectively. The cell proliferation was measured by MTT, cell count analysis and [(3)H]-TdR incorporation. Flow cytometry analysis was employed to detect the cell cycle. The leakage rate of lactate dehydrogenase (LDH) was assayed for the evaluation of the epithelial cell injury.</p><p><b>RESULTS</b>There was no significant difference in the morphology of the epithelial cells among the groups under inverted phase contrast microscope. But under the transmission electron microscope (TEM), the cells in 100 to 400 mg/L AP groups were seen to have proliferated actively, with euchromatin dominant in the nuclei, while heterochromatin was dominant in the cellular nucleus in control and 25 mg/L AP groups. The confluence time of epithelial cells in 50, 100, 200, 400 mg/L AP groups (154 +/- 12, 141 +/- 20, 130 +/- 19, 124 +/- 13) h preceded noticeably than that in control group (182 +/- 8) h, (P < 0.01). The cell proliferation in 100, 200, 400 mg/L groups reached the peak on the 5th day after AP treatment, while that in control and other groups was delayed by 1 to 2 days. The survival rate of the cells in 25 to 400 mg/L AP groups increased dramatically compared with that in control group, with its [(3)H]-TdR incorporation levels significantly increased in a dose dependent manner. The leakage rate of LDH in 200 and 400 mg/L AP groups was lower than that in control group (P < 0.01). The flow cytometric analysis of the cell cycle distribution revealed that the percentage of cell cycle from phase G0/G1 to G2/M and S in 25 to 400 mg/L AP groups increased significantly in a dose dependent manner compared with that in control group (P < 0.01).</p><p><b>CONCLUSION</b>AP might be beneficial to the protection of epithelial cells by promoting cell proliferation through inducing the progression of epidermal cells from phase G0/G1 into G2/M and S phases.</p>

Effect of Aloe coarse polysaccharide on cytokine secretion of keratinocytes in vitro / 中国中药杂志

<p><b>OBJECTIVE</b>To study the effects of Aloe coarse polysaccharide on the levels of growth factors (EGF, TGF-alpha, TGF-beta1) and interleukins (IL-1beta, IL-6, IL-8) and tumor necrosis factor (TNF) in cultured keratinocytes.</p><p><b>METHOD</b>The cultured keratinocytes were treated with Aloe coarse polysaccharide at concentrations of 75, 150, 300, 600, 1 200 mg x L(-1) land the equal volume of media as control group. The levels of EGF, TGF-alpha, TGF-beta1, IL-1beta, IL-6, IL-8 and TNF in the supernatants of cultured keratinocytes were assayed by enzyme-linked immunosorbent assay (ELISA) and radioimmunoassay (RIA).</p><p><b>RESULT</b>Compared with the control group, the levels of EGF, TGF-alpha, IL-1beta, IL-6 and IL-8 were significantly increased by treatment with Aloe coarse polysaccharide (P < 0.05, P < 0.01) and in a dose dependent manner, and the levels of TGF-beta1 and TNF were also increased but no statistical significance.</p><p><b>CONCLUSION</b>Aloe coarse polysaccharide may promote keratinocytes to secrete EGF, TGF-alpha, IL-1beta, IL-6 and IL-8.</p>