Objective To establish a mouse model of biliogenic severe acute pancreatitis (SAP) by using a self-made device for retrograde injection of sodium taurocholate into common bile duct,and to investigate the improvement of the device on retrograde injection of sodium taurocholate into common bile duct and its safety.Methods Thirty-six adult male ICR mice were randomly divided into biliogenic SAP model group and sham group,with 18 mice in each group.A 40 U disposable insulin syringe,a 200 μL tips and a 25 μL micro-syringer were used as basic materials for making the mouse common bile duct injection device [National Utility Model Patent (ZL 2014 2 0694365.4)].In model group,3.5％ sodium taurocholate (1 mL/kg) was injected retrogradely into the common bile duct of mice,whilst in sham group,the mice underwent the injection of equal amount of normal saline instead.Six mice in each group were sacrificed at 6,24 and 48 hours after operation,and the abdominal aortic blood was collected.Serum amylase (AMY),alanine aminotransferase (ALT),creatine kinase-MB (CK-MB),serum creatinine (SCr),oxygenation index (PaO2/FiO2) as well as serum Ca2+ were.determined.Pathological change in pancreas was observed under conventional light microscopy after hematoxylin and eosiu (HE) staining,and the impairment was evaluated by a widely used score system.Results The injection device was easily placed into mouse common bile duct under macroscopic observation.Six hours after operation,the levels of serum AMY,ALT and SCr in model group were significantly higher than those in sham group,and peaked at 24 hours,and they slightly decreased at 48 hours,which were still significantly higher than those of the sham group [24-hour AMY (U/L):7 325 ± 1 154 vs.1 737 ± 197,24-hour ALT (U/L):176.0±5.0 vs.38.3 ± 2.0,24-hour SCr (tmol/L):46.3 ± 1.5 vs.17.8 ±0.6,all P ＜ 0.01].The level of CK-MB at 6 hours in the model group was significantly higher than that of the sham group,and peaked at 48 hours (U/L:749.8±42.2 vs.383.3±35.5 at 6 hours,3 340.1 ± 203.6 vs.704.6 ± 63.5 at 48 hours,both P ＜ 0.01).PaO2/FiO2 at 6 hours after the operation in model group was significantly lower than that of sham group,then it began to rise at the similar level in sham group at 48 hours [mmHg (1 mmHg =0.133 kPa):327.5±33.8 vs.424.8±31.0 at 6 hours,P ＜ 0.01;429.8 ±41.8 vs.464.7±43.3 at 48 hours,P ＞ 0.05].Ca2+ level in model group was continuously decreased after operation,and it was significantly lower than that of sham group at 48 hours (mmol/L:1.58 ± 0.14 vs.2.45 ± 0.21,P ＜ 0.01).The pancreatic edema was obvious after operation in sham group,with the observation time prolongation,the changes were gradually improved;pancreatic focal necrosis was found at 6 hours after operation in model group,and it was secondary aggravated,and pancreatic lobule structure disappearance and inflammatory cells extensive infiltration was found at 48 hours.Pathological score of the model group was significantly higher than that of sham group at each time point,and peaked at 48 hours (13.3 ±0.3 vs.3.0±0.1,P ＜ 0.01).Conclusion It is a highly efficient and low-cost way to induce biliogenic SAP in mice by retrograde injection of 3.5％ sodium deoxycholate into common bile duct via the self-made injection device,and the model conformed to the clinical characteristics of biliogenic SAP.

Objective To investigate the effects of lentivirus-mediated heat shock protein 70 (HSP70) gene on voltage-gated calcium channels on the cell membrane of pheochromocytoma cell 12 (PC 12 cells) induced by ischemic/hypoxia and its mechanisms.Methods PC12 cells at logarithmic phase were collected,which were challenged with hypoxia for 6 hours and reoxygenation for 12 hours to stimulate the nerve cells suffered ischemia/reperfusion pathological process in vitro.PC12 cells were divided into non-infection group,infected by lentivirus containing green fluorescent protein (GFP) without HSP70 gene lentivirus control group (GFP lentivirus control group),and infected by lentivirus containing HSP70 and GFP gene recombined lentiviral infection group (HSP70 recombined lentiviral infection group).Real-time fluorescence quantitative reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot were used to determine mRNA and protein expressions of L-type voltage-gated Ca2+ channel subunits cav1.2 and cav1.3,receptor gated channel subunits NR1 and NR2,and Na+/Ca2+ exchange (NCX) in PC12 cells.Results After being challenged with hypoxia/reoxygenation,the mRNA and protein expressions of cav1.2,NR1 and NR2 in the PC12 cells were significantly lower in HSP70 recombined lentiviral infection group than those of GFP lentivirus control group and non-infection group [cav1.2 mRNA (2-△△Ct):3.13 ± 0.46 vs.5.12 ± 0.52,5.13 ± 0.66;NR1 mRNA (2-△△△Ct):1.61 ± 0.44 vs.3.23 ±0.82,3.31 ±0.78;NR2 mRNA (2-△△Ct):2.09±0.41 vs.3.91 ±0.64,3.88±0.62;cav1.2 protein (gray value):2.82±0.39 vs.3.98±0.23,3.96±0.24;NR1 protein (gray value):1.84±0.35 vs.2.79±0.21,2.86±0.23;NR2 protein (gray value):0.87±0.24 vs.1.57±0.31,1.33±0.44;all P ＜ 0.01].But there were no statistical differences in the mRNA and protein expressions of cav1.2,NR1 and NR2 between GFP lentivirus control group and non-infection group (all P ＞ 0.01).There were no statistical differences in the mRNA and protein expressions of cav1.3 and NCX among non-infection group,GFP lentivirus control group and HSP70 recombined lentiviral infection group [cav1.3 mRNA (2-△△Ct):4.82 ± 0.32,4.72 ± 0.36,4.82 ± 0.29;NCX mRNA (2-△△Ct):3.49 ± 0.78,3.47 ± 0.71,3.56 ± 0.65;cav 1.3 protein (gray value):2.63±0.40,2.64±0.39,2.68±0.39;NCX protein (gray value):3.27±0.48,3.34±0.48,3.31 ±0.42;all P ＞ 0.01].Conclusion Exogenous HSP70 affects the expression of L-type voltage-gated Ca2+ channel subunit cav1.2 and receptor gated channel subunits NR1 and NR2,which may protect PC12 cells from the injury caused by ischemic/hypoxia.

Objective To screen the potential biomarkers in plasma of rats with acute paraquat (PQ) poisoning using gas chromatography-mass spectrometry (GC-MS) based metabonomics technology,and to provide concrete evidence for early diagnosis.Methods Eight Sprague-Dawley (SD) rats were randomly divided into PQ poisoning group (intragastricly administrated with PQ solution 100 mg/kg) and control group (intragastricly administrated with the same volume of normal saline) according to the random number table,with 4 rats in each group.The general situation of rats was observed at 2,24 and 48 hours after administration.The blood of eye sockets was collected,the endogenous small molecule metabolites in plasma were determined with GC-MS method,and metabolic profile analysis and random forest analysis were performed to filter the potential biomarkers.Results ① The rats in PQ poisoning group gradually appeared lack movement,tachypnea,abdominal seizure and other symptoms of poisoning.In control group,the vital signs were stable.② The metabolites in plasma of rat were analyzed with GC-MS analysis,and the diagrammatic figure was plot as combined with principal component analysis (PCA) and partial least squares-discriminated analysis (PLS-DA) model,which showed that the distribution of plasma metabolism in PQ poisoning group was more diffuse but in the control group was more intensive,indicating that the metabolic patterns in two groups were different.From 2 hours after PQ administration,the metabolic trajectory in PQ poisoning group was significantly deflected compared with that of the control group,which was similar to control group until 48 hours,indicating that the metabolites in plasma of rat showed obvious difference in the early period.Five kinds of potential biomarkers with large weights were selected by random forest method which were serine,L-asparagine,hexadecanoic acid,octadecanoic acid,and arachidonic acid,the retention time was 15.259,24.345,33.334,37.695,and 40.254 minutes,respectively.The levels of serine,L-asparagine,arachidonic acid in PQ poisoning group were significantly higher than those of the control group,peaked at 48,48 and 24 hours,respectively (40.884-5.38 vs.28.85±2.32,6.61±1.31 vs.0.76±0.65,14.21±4.28 vs.4.42±1.19,all P ＜ 0.01),and the levels of hexadecanoic acid and octadecanoic acid were significantly lowered,reached tough at 48 hours (39.09 ± 10.23 vs.83.99 ± 20.49,44.03 ± 3.60 vs.140.76 ± 73.91,P ＜ 0.05 and P ＜ 0.01).The changes in these biomarkers were related to the toxicity of PQ,indicating that PQ could interfere the energy and lipid metabolism in rats.Conclusion Combine with the metabonomics analysis,screened plasma serine,L-asparagine,arachidonic acid content in PQ poisoning rats increased significantly,and hexadecanoic acid and octadecanoic acid content decreased significantly,which can preliminary diagnose acute PQ poisoning with animal general performance.

Objective To investigate the effects of stress hyperglycemia on prognosis in patients with severe cerebral vascular diseases.Methods A retrospective analysis was conducted.416 patients with severe cerebral vascular diseases confirmed by radiological imaging admitted to intensive care unit (ICU) of Guangdong General Hospital from December 2013 to June 2015 were enrolled.According to the values of randomise blood glucose (RBG) and glycosylated hemoglobin (HbA1c) and diabetes history,the patients were divided into euglycemia group (RBG ＜ 11.1 mmol/L,HbA1c ＜ 0.065,without diabetes history),diabetes group (RBG ≥ 11.1 mmol/L,HbA1c ≥ 0.065,with diabetes history),and stress hyperglycemia group (RBG ≥ 11.1 mmol/L,HbA1c ＜ 0.065,without diabetes history).The nosocomial infection rate,the length of ICU stay and 28-day mortality were compared among the three groups.Survival analysis was performed using Kaplan-Meier method,and multivariate Cox proportional hazard model was used to estimate the risk of death.Results Among 416 patients,there were 40 cases with stress hyperglycemia,46 with diabetes and 330 with euglycemia,with the incidence of stress hyperglycemia of 10.81％ (40/370).The nosocomial infection rates in the stress hyperglycemia group and diabetes group were significantly higher than those of the euglycemia group [55.00％ (22/40),52.17％ (24/46) vs.18.79％ (62/330),both P ＜ 0.01],and the length of ICU stay was significantly longer than that of the euglycemia group (days:16.53 ± 6.26,15.79 ± 8.51 vs.9.23 ± 4.29,both P ＜ 0.01).No significant differences in nosocomial intection rate and length of ICU stay were found between stress hyperglycemia group and diabetes group (both P ＞ 0.05).The 28-day mortality rate in stress hyperglycemia group was significantly higher than that of diabetes group and euglycemia group [47.50％ (19/40) vs.26.09％ (12/46),10.30％ (34/330),P ＜ 0.05 and P ＜ 0.01].It was showed by Kaplan-Meier survival analysis that 28-day cumulative survival rate in stress hyperglycemia group was significantly lower than that of euglycemia group and diabetes group (log-rank =6.148,P =0.043).It was showed by Cox death risk analysis that stress hyperglycemia was the risk factor of death in patients with severe cerebral vascular disease [hazard ratio (HR) =1.53,95％ confidence interval (95％CI) =1.04-1.26,P =0.001].Conclusion The patients with stress hyperglycemia may have a higher 28-day mortality and a poorer prognosis compared with those with diabetes and normal blood glucose in severe cerebral vascular diseases.

Objective To observe the effect of agmatine (AGM) on inflammatory factor in Kupffer cells of liver,and to investigate the protective effects of AGM on severe trauma-induced liver injury in mice and its possible mechanism.Methods Forty-two adult male BALB/c mice were randomly divided into sham group,model group,and AGM treatment group,with 14 mice in each group.The mice model of trauma-hemorrhage was reproduced by hindlimbs fracture combined with 35％ of orbital bleeding.The mice in the sham group were only anesthetized without other treatments.The mice in AGM treatment group were given intraperitoneal injection of 200 mg/kg AGM when limited recovery was performed,and the mice in model group were given the equal amount of normal saline.Seven mice in each group were sacrificed at 12 hours and 24 hours,respectively,after modeling,and blood samples and liver tissue were harvested,and liver Kupffer cells were isolated.Serum alanine aminotransferase (ALT),aspartate transaminase (AST)and lactic dehydrogenase (LDH) were determined with automatic biochemistry analyzer.Hepatic pathological changes were observed with light microscope using hematoxylin and eosin (HE) staining.The levels of tumor necrosis factor-α(TNF-o) and interleukin-6 (IL-6) in serum,hepatic homogenate and Kupffer cell supernatant were determined with enzyme linked immunosorbent assay (ELISA).The mRNA expressions of pro-inflammatory cytokines TNF-α and IL-6 in the Kupffer cell were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR).Results ① The normal liver tissue structure was found in sham group.At 24 hours after modeling in the model group,the changes in pathobiology were found as following:neutrophil infiltration,hepatocytes swelling,hyperemia,and necrosis,as well as the abnormality of parameters reflecting liver function.AGM could significantly improve the pathological changes in liver tissue caused by severe trauma,and ameliorate the liver function.② There were no significant differences in the levels of TNF-α and IL-6 in serum and hepatic tissue at 12 hours after modeling,and the parameters at 24 hours in model group were higher than those at 12 hours,which were significantly higher than those of the sham group [serum TNF-α (ng/L):80.8±4.7 vs.34.7±4.7,IL-6 (ng/L):104.0±9.0 vs.55.4±3.3;liver TNF-α (ng/mg):405.2± 19.6 vs.57.2±10.0,IL-6 (ng/mg):58.4±7.7 vs.14.3±2.1,all P ＜ 0.01].AGM could effectively reduce the levels of TNF-o and IL-6 in serum and hepatic tissue [serum TNF-α (ng/L):58.2 ± 3.1 vs.80.8 ± 4.7,IL-6 (ng/L):74.1 ± 6.6 vs.104.0± 9.0;liver TNF-α (ng/mg):248.7 ± 22.5 vs.405.2 ± 19.6,IL-6 (ng/mg):22.5 ± 3.1 vs.58.4 ± 7.7,all P ＜ 0.01].③ The levels of TNF-o and IL-6 in Kupffer cells supernatant were significantly higher than those of the sham group,and they were further increased after lipopolysaccharide (LPS) stimulation for 24 hours.AGM could effectively reduce the levels of TNF-α and IL-6 in Kupffer cells [TNF-α (ng/L):256.6 ± 5.6 vs.465.5 ± 5.2,IL-6 (ng/L):1 185.5 ± 64.4 vs.2 018.8 ± 53.2,both P ＜ 0.01],and also decreased the mRNA expressions of TNF-α and IL-6 [TNF-α mRNA (2-△△Ct):7.2±0.4 vs.13.5±0.4,IL-6 mRNA (2-△△Ct):13.2±0.7 vs.21.3 ± 1.6,both P ＜ 0.01].Conclusion Agmatine can reduce trauma-induced acute hepatic injury via suppression of cytokines release in Kupffer cells,and can ameliorate the liver function.

Objective To investigate the role of micro-RNAs (miR-101 and miR-125a-5p) in autophagy of lipopolysaccharide (LPS) derived human THP-1 macrophages.Methods Human monocytic leukemia cell line THP-1 was cultured in vitro,and it was differentiated into macrophages after being induced with phorbol (50 μg/L) for 48 hours.THP-1 macrophages were stimulated with LPS in 0,250,500,1 000 μg/L respectively for 12 hours,miR-mimic was transfected into THP-1 macrophages as induced by Lipofectamine RNAiMAX,and the transfection efficiency of miRNA was determined with fluorescence microscopy.Enzyme linked immunosorbent assay (ELISA) was used to determine the levels of tumor necrosis factor-α (TNF-α) and monocyte chemotaxis protein-1 (MCP-1) in the supernatants of culture.Western Blot was used to detect the protein expressions of autophagy proteins ATG4D,Beclin1,and LC3 Ⅱ.The expression levels of miR-101 and miR-125a-5p were determined by quantitative reverse transcription-quantitative polymerase chain reaction (RT-qPCR).Results ① The releasing levels of TNF-α and MCP-1 induced by LPS with 250,500,1 000 μg/L were significantly increased as compared the cells without LPS stimulation [TNF-α (ng/L):1 336.1 ± 18.5,1 340.6±24.8,1 364.5± 14.9 vs.47.6±4.4;MCP-1 (ng/L):996.3 ±51.3,934.6±84.3,974.2±69.5 vs.21.3±6.5,all P ＜ 0.01],but no significant differences were found among the three LPS stimulation groups.The protein expressions of ATG4D,Beclin1 and LC3 1Ⅱ were up-regulated in the presence of different LPS concentrations (0,250,500,1 000 μg/L) for 12 hours in THP-1 macrophages (when compared with the cells without LPS stimulation,t value of ATG4D was 8.103,38.410,52.020,P value was 0.015,0.001,＜ 0.001;t value of Beclin1 was 3.026,5.328,3.482,P value was 0.047,0.034,0.037;t value of LC3 Ⅱ was 3.836,6.200,4.665,P value was 0.018,0.003,0.010),and the optimal concentration was 500 tg/L LPS.When THP-1 macrophages were stimulated with 500 μg/L LPS for 12 hours,the expression levels of miR-101 and miR-125a-5p were down-regulated significantly as compared with the cells without LPS stimulation [miR-101 (2-△ △Ct):0.68 ± 0.08 vs.1.95 ±0.26,t =8.047,P =0.001;miR-125a-5p (2-△ △Ct):0.23 ± 0.06 vs.1.69± 0.42,t =5.975,P =0.004].② The higher transfection efficiency was showed under fluorescence microscope.Westem Blot results showed the protein expressions of ATG4D,Beclin1 and LC3 Ⅱ were down-regulated as induced by an over-expression of miR-101 or miR-125a-5p in THP-1 macrophages,and more obviously down regulated by co-transfected with miR-101 and miR-125a-5p (compared with negative control group,t value was 14.550,5.855,14.180,P value was 0.005,0.014,＜ 0.001).Conclusion miR-101 and miR-125a-5p can inhibit the autophagy in LPS challenged THP-1 macrophages,and the potential mechanism might be related to target regulation of ATG4D.

Objective To observe the features of the changes in the whole blood N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in pregnant patients with complication of hypertensive disorders,its correlation to the severity of the illness,and to investigate the diagnostic value of point-of-care testing of NT-proBNP in patients with hypertensive disorders complicating pregnancy.Methods A prospective observation was conducted.Sixty-nine patients with hypertensive disorders complicating pregnancy admitted to Department of Critical Care Medicine of Liaocheng People's Hospital in Shandong Province from April 2013 to April 2015 were enrolled.All patients were divided into gestational hypertension group (n =16),preeclampsia group (n =30) and eclampsia group (n =23).At the same time,30 age-matched normal pregnant women were enrolled as the control group.The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score of all patients with hypertensive disorders complicating pregnancy were calculated within 24 hours after intensive care unit (ICU) admission.NT-proBNP in venous blood at 1,3,5 days after ICU admission was determined with point-of-care testing,in order to analyze the correlation of changes in NT-proBNP value in each group and the severity of the disorder.Receiver operating characteristic (ROC) curve was plotted to evaluate the diagnostic value of NT-proBNP in the whole blood in hypertensive disorder complicating pregnancy.Results The APACHE Ⅱ score of the eclampsia group was significantly higher than that of the preeclampsia group,and it was higher than that of gestational hypertension group (15.91 ± 1.06,13.73 ± 1.09,10.31 ± 1.10,all P ＜ 0.01).The NT-proBNP in normal pregnancy group was lower than 125.00 ng/L,with a mean of 90.00 (79.75,100.00) ng/L.With the aggravation of the disease,NT-proBNP was gradually increased.On the first day in ICU,the NT-proBNP of the eclampsia gronp was significantly higher than that of the preeclampsia group,and it was higher in preeclampsia group than that of gestational hypertension group [ng/L:1960.00 (1 226.00,3 229.00),859.50 (626.75,2439.00),505.00 (171.25,604.05),P ＜ 0.05 or P ＜ 0.01].With the extension of duration of treatment,the levels of NT-proBNP (ng/L) in the eclampsia group,preeclampsia group,and gestational hypertension group was gradually decreased,and had a statistically significant difference on the fifth day as compared with that of the first day [310.00 (210.00,430.00) vs.1 960.00 (1 226.00,3 229.00) in eclampsia group,265.00 (229.50,333.25) vs.859.50 (626.75,2439.00) in preeclampsia group,and 203.00 (115.50,259.25) vs.505.00 (171.25,604.05) in gestational hypertension group,all P ＜ 0.01].APACHE Ⅱ score of the patient with hypertensive disorders complicating pregnancy was positively correlated with the level of NT-proBNP on the first day in ICU (r =0.795,P =0.000).It was shown by ROC curve analysis that the area under the ROC curve (AUC) of NT-proBNP in the whole blood for the diagnosis of the patient with hypertensive disorder complicating pregnancy was 0.986 [95％ confidence interval (95％C/) =0.753-0.924].When the cutoff value was 122.50 ng/L,the sensitivity was 97.1％,and the specificity was 100.0％.No patient died,all the 69 patients recovered and discharged.Conclusions The levels of NT-proBNP in the whole blood in the patients with hypertensive disorder complicating pregnancy,especially those with eclampsia,were significantly higher,and it was correlated with the severity of illness.After treatment,the levels were gradually lowered with the improvement of the disease.Therefore,it is concluded that the point-of-care testing of NT-proBNP in the whole blood has an excellent value for the diagnosis and evaluation of hypertensive disorder complication pregnancy.

Objective To investigate the clinical effect of early use of enteral nutrition therapy in elderly diabetic patients suffering from severe lower respiratory tract infection.Methods A prospective,randomized,open,controlled trial was conducted.Patients aged ≥ 60 years old with diabetes mellitus complicated by severe lower respiratory tract infection admitted to Department of Geriatrics of the Ninth People's Hospital Affiliated to Shanghai Jiaotong University School of Medicine from July 2013 to June 2015 were enrolled,and they were divided into the observation group (nasogastric tube infusion of fresubin) and control group (ordinary liquid diet) according to the random number table method,with 60 patients in each group.Nutritional status,inflammation state and immunological indexes before treatment (0 day) and 4,7,14 days after treatment,the outcome of the disease and the nutrition related complications were compared between two groups.Results After treatment,serum albumin (ALB),pro-albumin (PA),immunoglobulin A and G (IgA,IgG) were significantly increased compared with those before treatment in both groups [at 7 days,ALB (g/L):28.37 ± 0.40 vs.26.72 ± 0.37 in control group,29.12 ± 0.25 vs.26.86 ± 0.26 in observation group;PA (mg/L):53.80 ± 6.28 vs.43.76 ± 6.93 in control group,58.46 ± 8.70 vs.44.68 ± 7.33 in observation group;IgG (g/L):11.62±4.72 vs.9.98±3.71 in control group,13.36±4.58 vs.9.88±3.27 in observation group;IgA (g/L):2.31 ±0.35 vs.1.50±0.39 in control group,3.07±0.48 vs.1.37±0.29 in observation group;all P ＜ 0.05].Compared with the control group,the level of PA in observation group was significantly increased from 7 days on (mg/L:58.46 ± 8.70 vs.53.80 ± 6.28,P ＜ 0.05),while ALB,IgG,IgA levels in observation group increased at 14 days [ALB (g/L):33.24 ± 0.45 vs.30.76±0.79,IgG (g/L):15.03 ±3.73 vs.11.45 ±2.83,IgA (g/L):3.56±0.32 vs.2.50±0.16,all P ＜ 0.05].The levels of C-reactive protein (CRP) and procalcitonin (PCT) in both groups gradually lowered,but they were significantly lower in observation group than those in the control group from 4 days on [CRP (mg/L):17.72±4.23 vs.20.96±5.83,PCT (ng/L):123±37 vs.257±88,both P ＜ 0.05],up to 14 days.The hospital mortality rate of the observation group was lowered compared with that of the control group (6.67％ vs.8.89％),and the duration of mechanical ventilation was significantly shortened (hours:145.00±19.39 vs.193.00± 18.97,P ＜ 0.05),insulin dosage was also significantly decreased (U:33.52 ± 5.74 vs.49.71 ± 6.99,P ＜ 0.05).There was no significant difference in the incidence of abdominal distension,diarrhea and reflux of gastric contents between the two groups,and they were relieved after treatment and had no influence on further enteral nutrition therapy.Conclusion Early administration of the complicated enteral nutrition in elderly diabetic patients with severe lower respiratory tract infection cannot only decrease the levels of pro-inflammatory factors in patients,but also shorten the duration of mechanical ventilation,enhance immunity,improve the curative effect with little influence on the blood glucose level.

Objective To explore the mechanism of high mobility group protein 1 (HMGB1) involved in endoplasmic reticulum stress (ERS) induced by brain ischemia/reperfusion (I/R),based on I/R-HMGB1-ERS as the breakthrough point.Methods The brain of rats birthed 1-3 days was harvested,and the brain cells were cultured in vitro,which were used in the experiment when the cells were in the third passage.The cells were divided into two groups:cells in blank control group were cultured under the normal conditions without any treatment,and the cells in hypoxia/reoxygenation group were cultured with 99.9％ nitrogen for 60 minutes (hypoxia) followed by opening the bottle neck for reoxygenation 120 minutes to simulate I/R model.The HMGB1 gene was silenced by using small interfering RNA (siRNA,siRNA and transfection reagent Lipofectamine 2000 mixture gradient was transfected into the cultured cells) as HMGB1-siRNA transfection group,and blank control (without any treatment) and negative control group (transfected with control siRNA) served as controls.The mRNA and protein expressions of HMGB1 and ERS related molecules were determined by real-time reverse transcription-polymerase chain reaction (RT-PCR) and Western Blot.Results ① In cells of hypoxia/reoxygenation group,the mRNA and protein expressions of HMGB1 and ESR related proteins,including glucose regulating protein 78 (GRP78),C/EBP homologous protein (CHOP) and caspase-12,were significantly higher than those of blank control group with statistical difference (the value in blank control group was served as baseline 1,HMGB1 mRNA:3.19±0.48 vs.1,t =2.183,P =0.008;GRP78 mRNA:2.07±0.33 vs.1,t =3.292,P =0.016;CHOP mRNA:1.93±0.28 vs.1,t =2.573,P =0.021;caspase-12 mRNA:2.42±0.42 vs.1,t =2.261,P =0.027:HMGB1 protein:2.28±0.36 vs.1,t =2.042,P =0.009;GRP78 protein:1.33±0.24 vs.1,t =2.781,P =0.016;CHOP protein:1.67±0.34 vs.1,t =2.174,P =0.021;easpase-12 protein:1.36±0.44 vs.1,t =3.192,P =0.008).It was indicated that ERS related molecules involved in cell hypoxia/reoxygenation process.2② After HMGB1 gene was silenced by siRNA,the cells after hypoxia/reoxygenation showed a decrease in the mRNA and protein expressions of HMGB1 and ERS related moleculars as compared with those of blank control group and negative control group (served the value in blank control group as baseline 1,HMGB1 mRNA:0.27±0.12 vs.1,1.02 ± 0.04;GRP78 mRNA:0.16 ± 0.13 vs.1,0.96 ± 0.04;CHOP mRNA:0.47 ± 0.09 vs.1,0.98 ± 0.07;caspase-12 mRNA:0.31 ±0.11 vs.1,1.05±0.02;HMGBI protein:0.23±0.04 vs.1,1.08±0.01;GRP78 protein:0.14±0.09 vs.1,1.35±0.03;CHOP protein:0.32±0.10 vs.1,0.93±0.06;caspase-12 protein:0.27±0.09 vs.1,0.97±0.08;P ＜ 0.05 or P ＜ 0.01).It was indicated that HMGB1 involved in ERS related with GPR7,CHOP,caspase-12.Conclusion Hypoxia/reoxygenation brain intracellular HMGB1 and ERS related molecules expression levels were significantly up-regulated,and silencing HMGB1 gene can significantly inhibit the expression levels of these molecules,and I/R-HMGB 1-ERS pathway may participate in the mechanism of brain I/R injury.

The metabolic response to stress plays a key role in the adaptive response during critical illness.Multiple mechanisms including the stimulation of the sympathetic nervous system,inflammation,and immune responses were involved.Insulin resistance is one of the main features in metabolic response to stress.Metabolic response to stress was manifested by disorders of energy consumption,such as glucose,lactic acid,lipids,and proteins.The decrease in fat-free body mass and cell mass,relative excess of adipose tissues,and increased extracellular fluid volumes were also involved.Therapeutic interventions,including hormone supplementation,enhanced protein intake,and early mobilization,are considered for prevention and therapy of metabolic response to stress.The review aims to summarize the pathophysiological mechanisms,clinical consequences,and therapeutic implications of metabolic response to stress in critical illness.