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Objective·To establish a reliable alcoholic liver disease mouse model (ALDNM) that mimics the drinking pattern of alcoholic liver disease (ALD) patients.Methods·Using the self-designed feeding tubes and liquid diet,ALDNM model was developed through chronic feeding combined with acute gavage of ethanol based on Lieber-DeCarli model and Gao-Binge model.C57BL/6 mice were administered with control liquid diet for adaptation for first 5 d,and then divided into pair-fed group and ethanol-fed group (10 mice each group).Ethanol-fed mice were fed with the liquid diet in which ethanol accounts for 30％ of total energy,while the pair-fed mice were fed with the control diet for 10 d.At the 16th day,ethanol-fed mice and pair-fed mice were respectively gavaged a single dose of 31.5％ ethanol or isocaloric maltose dextrin,and euthanized 9 h later.Sera and livers were collected.The general physiological condition,hepatic tissue pathological changes and serum indexes between Lieber-DeCarli models and ALDNM models were compared.The liver lipids of ALDNM mice were determined by Oil red O (ORO) staining and hepatic triacylglyceride (TAG) test.Meanwhile,the mRNA levels of interleukin-6 (IL-6),tumor necrosis factor α (TNF-α),fatty acid synthase (Fas),long chain fatty acid elongase 6 (Elovl6) and stearyl-CoA desaturase (Scdl) were detected by real-time PCR in ALDNM models.Western blotting was used to detect the changes of phosphorylated signal transduction and transcriptional activator (p-STAT3) in the livers.Results·Lieber-DeCarli model mice were generally in poor condition,and there was no significant change in serum glutamic-pyruvic transaminase (GPT) and glutamic-oxaloacetic transaminase (GOT) compared to pair-fed group.However,in ALDNM models,H-E staining showed that the hepatocytes of ethanol-fed mice were extremely swollen with round volume,increased cytoplasm and filled with large amounts of fat vacuoles.ORO staining analyses showed obvious microsteatosis in the liver cells from all ethanol-fed mice.The hepatosomatic index,liver TAG content,serum GPT and GOT of ALDNM models were significantly higher than those in the pair-fed group,while the serum HDL significantly decreased compared to the pair-fed group.Moreover,the expression levels of both lipid synthesis pathways and inflammatory signaling pathways related genes in livers significantly increased in the ethanol-fed mice of ALDNM model.Conclusion·ALDNM model was successfully constructed.This model is cost-and time-efficient.Moreover,ALDNM model mimics the drinking pattern and pathogenesis of ALD patients with the advantages of stable food intake,good repeatability,and obvious liver damage.
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Objective@#To observe the effects of transient exposure to high glucose on biological behaviors of human dermal microvascular endothelial cells cultured in vitro.@*Methods@#The dividing method and treatment of cells for the detection of all indexes in this study were as follows. Human dermal microvascular endothelial cells of the 4th passage were divided into 3 groups according to the random number table, with 12 wells in each group. Cells in control group (C) were cultured with complete culture solution containing 5 mmol/L D-glucose for 7 d. Cells in transient high glucose group (THG) were cultured with complete culture solution containing 30 mmol/L D-glucose for 2 d and complete culture solution containing 5 mmol/L D-glucose for 5 d. Cells in prolonged high glucose group (PHG) were cultured with complete culture solution containing 30 mmol/L D-glucose for 7 d. (1) The cell morphology in groups C and PHG on culture day 7 and that in group THG on culture day 2 and 7 was observed by inverted optical microscope. (2) On culture day 0, 2, 4, and 7, cell proliferation rate was determined by cell viability analyzing counter. (3) After culture day 2, the scratch experiment was performed, and the cells were further cultured. At post scratch hour (PSH) 0, 24, 48, 72, 96, and 120, the scratch area was measured, and the cell migration rates of the latter 5 time points were calculated. (4) On culture day 0, 2, 4, and 7, the cell apoptosis rate was determined by cell analyzer. (5) Cells were seeded into Matrigel to culture for 24 h after culture day 7. The formation of vessel-like structure was observed by inverted optical microscope. The length and number of branch point of vessel-like structure were calculated. (6) On culture day 2, 4, and 7, mRNA expression of vascularization-related gene tissue inhibitor of matrix metalloproteinase-3 (TIMP-3) was determined with real-time fluorescent quantitative reverse transcription polymerase chain reaction. Data were processed with analysis of variance of factorial design, analysis of variance for repeated measurement, one-way analysis of variance, and LSD test.@*Results@#(1) Cells in group C exhibited ovary shape in cobble stone order on culture day 7. Cells in group THG exhibited long ovary shape and lost cobble stone order on culture day 2 and kept the same changes on culture day 7. Cells in group PHG exhibited long ovary shape and lost cobble stone order on culture day 7. (2) On culture day 0, there was no significant difference in cell proliferation rate among the 3 groups (F=0.23, P>0.05). On culture day 2, cell proliferation rates in groups THG and PHG were similar (P>0.05), which were significantly lower than the cell proliferation rate in group C (with P values below 0.01). On culture day 4 and 7, the cell proliferation rates in groups THG and C were similar (with P values above 0.05), which were significantly higher than those in group PHG (with P values below 0.01). (3) At PSH 24-120, the cell migration rates in groups THG and PHG were similar (with P values above 0.05), which were significantly lower than those in group C (with P values below 0.01). (4) On culture day 0, there was no statistically significant difference in cell apoptosis rate among the 3 groups (F=0.78, P>0.05). On culture day 2, cell apoptosis rates in groups THG and PHG were similar (P>0.05), which were significantly higher than the cell apoptosis rate in group C (with P values below 0.01). On culture day 4 and 7, the cell apoptosis rates in groups THG and C were similar (with P values above 0.05), which were significantly lower than those in group PHG (with P values below 0.01). (5) The length of vessel-like structure of cells in group THG was (1.84±0.10)×105 μm, close to (1.82±0.11)×105 μm in group PHG (P>0.05), both significantly shorter than (2.75±0.23)×105 μm in group C (with P values below 0.01). The numbers of branch point of vessel-like structure of cells in groups THG and PHG were 43±5 and 46±8 respectively, which were close to each other (P>0.05) and both significantly less than 103±21 in group C (with P values below 0.01). (6) On culture day 2, 4, and 7, mRNA expressions of TIMP-3 of cells in groups THG and PHG were similar (with P values above 0.05), which were significantly lower than those in group C (with P values below 0.05).@*Conclusions@#Transient exposure to high glucose can cause metabolic memory of morphology, migration, and angiogenesis in human dermal microvascular endothelial cells cultured in vitro, resulting in sustained changes in biological behaviors. The mechanism may be related to the changes of vascularization-related genes.
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Objective To determine the independent risk factors for sepsis in patients with large area burns.Methods The study enrolled 307 patients with large area burns treated from June 2014 to June 2015.Related variables included age,gender,visit time,emergency eschar open,burn index,tangential excision of eschar during shock stage,delayed resuscitation during shock stage,oxygen pressure(PaO2),carbon dioxide pressure (PaCO2),assisted ventilation mode,mechanical ventilation time,inhalation injury,prophylactic tracheotomy,continuous lactic acid rise,refractory hypernatremia,heart-lung disease history,and diabetes history.Correlation of the variables with the incidence of sepsis was observed.Independent predictors of sepsis in patients with large burns were differentiated using the Logistic regression analysis.Results Delayed resuscitation during shock period (OR =1.747,95％ CI 1.822-7.431,P ＜ 0.05),continuous lactic acid rise (OR =1.758,95％ CI 1.137-4.002,P ＜ 0.05),refractory hypernatremia (OR =2.985,95％ CI 1.074-6.782,P ＜ 0.05),moderate and severe inhalation injury(OR =14.764,95％ CI 0.892-47.323,P ＜ 0.05) and burn index (OR =5.017,95％ CI 1.075-23.572,P ＜ 0.05) were found be independently associated with sepsis in patients with large burns.Conclusions These independent risk factors for sepsis in large area burn patients deserve more attention.Early and timely treatment measures may reduce the incidence of sepsis.
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0.05).The grade of antibiotics tended to decline with the increase in of dressing change freqnency so it could decrease the expenditure of antibiotics.The rate of fungal infection lowered while hospitalization did not appear to be prolonged.CONCLUSIONS After the analysis of the use of antibacterials for burn patients,we find that the proper use of antibiotics and the increase in dressing change would lower the grade of antibiotics and expenditure,at the same time they do not increase the rate of bacterial infection and fungal infection,and do not prolong the duration of patients hospitalization time.
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Des-γ-carboxy prothrombin (DCP)is produced abnormally in hepatocellular carcinoma cells. Vitamin K utilizing deficiency and the decreasing activity of vitamin K-dependent γ-glutamyl carboxylate and coenzyme may involve in production of DCP. The enzymes in bepatocellular carcinoma cells are failed in carboxylating all the ten glutamic acid residues in amino- terminus of prothrombin precursor to Gla and make the difference between DCP and prothrombin. DCP is considered to be the marker for diagnosis of hepatocellular carcinoma. DCP might also be one of the growth-promoting factors to carcinoma cells. Therefore, inhibition of DCP in hepatocellular carcinoma cells is considered to be the therapeutic method of blocking hepatocellular car-cinoma growth. Vitamin K and its analogs are found to have the functions of inhibiting hepatocellular carcinoma cell producing DCP. The mechanism may relate to the increase of γ-glutamyl carboxylase activity.