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Objective To explore the influence of excessive PTEN expression to fibroblast cycle and collagen secretion induced by LPS.Methods Normal skin fibroblast in the patient with hyperplastic scar were cultured in vitro.When the primary culture was close to 80% fusion,the digestive passage was performed,cultured to the third generation.LPS(0.5 μg/mL) was adopted to stimulate the third generation of normal skin fibroblasts.Defective adenovirus carrying PTEN gene was transfected to the third passage fibroblasts after LPS stimulation.Flow cytometer was adopted to detect the cell cycle.ELISA method was adopted to detect the secreted collagen amont.Results Excessive PTEN expression could inhibit the increase of G2M cell cycle induced by LPS.LPS stimulation could increase the secretion of collagen in skin fibroblasts,yet excessive PTEN expression could inhibit the secretion of collagen induced by LPS.Conclusion LPS could increase the amont of fibroblasts on G2M cell cycle and secretion of collagen,yet excessive PTEN expression can inhibit the effect.
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Objective To review the prevalence of nosocomial infection and the change of drug resistance in a comprehensive intensive care unit (ICU) and to provide theoretical basis for clinical prevention and treatment.Methods The strains of bacteria and fungi were isolated from ICU and their drug resistance was retrospectively analyzed from Jan.1st,2010 to Dec.31th,2011.Results The main pathogen of nosocomial infection were Gram-negative bacteria ( 73.3% ),Gram-positive bacteria ( 17.9% ) and fungi ( 8.7% ).In bacterial infection,Gram-negative and G-positive bacteria accounted for 80.3% and 19.7% respectively.In Gram-negative bacteria,pseudomonas aeruginosa was the major type (21.7%).In Gram-positive bacteria,staphylococcus aureus (31.4%) was most prominent.Drug resistance of bacteria was severe,while that of fungi was mild.Conclusion Bacteria has severe drug resistance and exhibits multi-drug resistance for commonly used antibiotics.The principle of antibiotics application should be mastered and antibiotics should be chosen according to drug-sensitivity tests.
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Objective: To evaluate the protective effect and possible mechanism of ω-3 PUFA supplementation in the early stage of experimental acute bacterial pneumonia in mice.Methods: Mice of Pseudomonas aeruginosa (PA)-induced lung injury were randomly divided into three groups: fish oil supplementation treatment group (n = 8),soybean oil treatment group (n = 8) and normal sodium treatment group (n = 8).Evaluation criteria were inflammatory response 24 h after the injury assessed with TNF-α、IL-1β,IL-10 levels in bronchoalveolar lavage and lung injury evaluation by H-E staining.Results: Compared with the other two groups,the levels of IL-1β,IL-10 in fish oil supplementation treatment group were reduced.No difference was found in the TNF-α level and histological scoring among the groups.Conclusion: The ω-3 PUFA can decrease the elevated mediators of inflammation in bacterial pneumonia,but can't affect the pneumonia progress within 24 hours.
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Objective To explore the changing trend of Ia on monocyte, lymphocyte apoptosis rate, TNF-α and IL-6 in abdominal aorta of burned rats with delayed resuscitation and the influence of application of carbachol on them. Methods Adult male Wistar rats were randomly divided into normal control group(n =8), scald group(n =48) and scald with carbachol treatment group(n =48). In latter two groups, rats were inflicted with 30% total body surface area (TBSA) full-thickness scald and delayed fluid resuscitation. All scald rats were sacrificed at the 6th hours or 1st, 2nd, 3rd, 7th, 14th day after scald, with 8 rats at each time point. Expression of Ia antigen on monocyte and lymphocyte apoptosis rate were determined by direct immunofluorescence on a flow cytometer, and TNF-α and IL-6 was measured by ELISA. Results Expression of la on monocyte was obviously lower than that of controls. The lowest levels were recorded on the 6th hours and 1st day after scald. Subsequently, Ia was elevated gradually, but still lower than that of normal rats(P <0. 01). After administration of carbachol, Ia expression was obviously promoted, compared with the simple scald group (P <0. 01). Lymphocyte apoptosis rate, TNF-α and IL-6 was higher than that of controls(P <0. 01). After administration of cavachol, , lymphocyte apoptosis rate and TNF-α and IL-6 was obviously down-regulated on the 6th hours, 1st day, 2nd day and 3rd day after scald injury, compared with the simple scald group (P < 0. 01 or 0. 05). Conclusion After severe burn with delayed fluid resuscitation, there is a low la expression, high lymphocyte apoptosis rate and increased releasing of proinflammatory cytokine. Immune function was suppressed. Carbacho] could improve the immune function of scald rats with delayed fluid resuscitation.
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Objective To observe the changes of HMGB-1 of burned patient with sepsis and to investigate the effect of carbaehol on production of HMGB-1 from human monocytes stimulated by lipopolysaccharide(LPS) and its mechanism. Methods The peripheral blood samples of burned patients with sepsis and healthy donors were taken for isolation of monoeyte. The subjects were divided into six groups: the menocytes group were added only with 1640 culture medium,LPS group were added only with LPS;nicotine group and carbachol group were with carbachol first or nicotine for 5 rain and then with LPS. The levels of HMGB-1 were tested by EIJSA. α-bungarotoxin + carba-chol group were added with atropine or α-bungarotoxin for 5 min and then were given carbachol and 5 min later were stimulated with LPS. 48 h-incubation later supemate were collected for the detection of HMGB-1 by ELISA. Results HMGB-1 levels of burned patients with sepsis were ( 12.94±6.54)μ/L,which were much higher than that of healthy donors( [ (2.01±0.03 )μ/L), P <0.01 ]. When monoeytes were stimulated by LPS alone, concentrations of HMGB-1 were (9.39±1.37 ) μ/L, which were obviously higher than that of controls [( 1.48±0.69 ) μ/L), P < 0.01]. Mter pretreated by carbaehol or nicotine,concentrations of HMGB-1 were (3.52±1.64)μ/L and (4.01± 1.56) μ/L respectively, which were obviously decreased compared with that of LPS stimulation alone group ( P < 0.01). When pretreatedby atropine before addition of carbachol, there was no significant changes in concentrations of HMGB-1 [ (3.87±2.01 )μ/L]. When pretreated with α-Bungaretoxin before carbachol adminitration, inhibitive effect of carbachol on production of HMGB-1 were blocked [(8.97±1.97 )μ/L ]. Conclusion The release of HMGB-1 in burned patients with sepsis is increased. Carbachol could obviously reduce the production of HMGB-1, which may affect through activating α 7 subunit of cholinergic N receptor.
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Objective: To investigate the immune regulation mechanism and anti-inflammatory effect of ω-3 fatty acids on sepsis patients.Methods: Totle 18 patients were divided into two groups,therapy group and control group. The control group was given routine therapy while the theray group was given routine therapy and fish oil.The expression of HLA-DR on peripheral monocytes was measured by flow cytometry at the beginning and 5 d during therapy, in parallel with serum c-reactive protein(CRP),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and severity of the disease(APACHE Ⅱ score),serum and oxygenation index(PaO2/FiO2)Results: Compared with conventional group,serum concentration of CRP, IL-6 and TNF-α in patients of therapy group were significantly reduced,and oxygenation index improved. There was no difference in the expression of HLA-DR and APACHE Ⅱscore between two groups.Conclusion:Fish oil can decrease the elevated mediators of inflammation on sepsis patients,and can't regulation the expression of HLA-DR on peripheral monocytes.
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Objective:To investigate the immune regulation mechanism and anti-inflammatory effect of ?-3 fatty acids on sepsis patients.Methods: Totle 18 patients were divided into two groups,therapy group and control group.The control group was given routine therapy while the theray group was given routine therapy and fish oil.The expression of HLA-DR on peripheral monocytes was measured by flow cytometry at the beginning and 5 d during therapy,in parallel with serum c-reactive protein(CRP),interleukin-6(IL-6),tumor necrosis factor-?(TNF-?),and severity of the disease(APACHE Ⅱ score),serum and oxygenation index(PaO2/FiO2) Results: Compared with conventional group,serum concentration of CRP,IL-6 and TNF-? in patients of therapy group were significantly reduced,and oxygenation index improved.There was no difference in the expression of HLA-DR and APACHE Ⅱscore between two groups.Conclusion:Fish oil can decrease the elevated mediators of inflammation on sepsis patients,and can't regulation the expression of HLA-DR on peripheral monocytes.
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Objective: To evaluate the protective effect and possible mechanism of ?-3 PUFA supplementation in the early stage of experimental acute bacterial pneumonia in mice.Methods: Mice of Pseudomonas aeruginosa(PA)-induced lung injury were randomly divided into three groups: fish oil supplementation treatment group(n = 8),soybean oil treatment group(n = 8) and normal sodium treatment group(n = 8).Evaluation criteria were inflammatory response 24 h after the injury assessed with TNF-?、IL-1?,IL-10 levels in bronchoalveolar lavage and lung injury evaluation by H-E staining.Results: Compared with the other two groups,the levels of IL-1?,IL-10 in fish oil supplementation treatment group were reduced.No difference was found in the TNF-? level and histological scoring among the groups.Conclusion: The ?-3 PUFA can decrease the elevated mediators of inflammation in bacterial pneumonia,but can't affect the pneumonia progress within 24 hours.