Objective To discuss the levels of inhibitory protein Ga-2 (Gαi2) in hippocampus of brain and the effects of Gαi2 on neuronal intracellular Ca2+ level in cerebral ischemia reperfusion injury model of rats.Methods Ninety SD rats were randomly (random number) assigned to sham group (n =30),ischemic-reperfusion (IR) group (n =30),Pertussis toxin (PT) group (n =30).The blood flow of right common carotid artery of rat was blocked for 90 min to make ischemia reperfusion model.The levels of Gαi2 in hippocampus was assayed by immunohistochemistry and Western blotting after ischemia reperfusion for 6,12,24 h in each group.The average fluorescence values of intracellular Ca2+ levels in hippocampus of rats in three groups were detected by using Flow CytoMeter (FCM).Neuronal cell apoptosis was measured by TUNEL.Results After restoration of middle cerebral artery blood flow for different lengths of time,the levels of hippocampus Gαi2 and Ca2+ levels in IR group were significantly higher than those in Sham group (P ＜0.01).The hippocampus Ca2+ levels in PT group were higher than those in IR group (P ＜ 0.01).The apoptotic rates of neurons in PT group were lower than those in IR group (P ＜0.05).Conclusions The level of Gαi2 in cerebral ischemia reperfusion injury model was increased.Gαi2 might reduce the calcium ion concentration of neurons after cerebral ischemia and rcduce the neuronal cell apoptosis in this model.Gαi2 might play a role in protecting neuron from cerebral ischemia reperfusion injury.

Objective To study the role of src-suppressed C kinase substrate (SSeCKS) in the secretion of tumor necrosis factor (TNF-α) in rat pulmonary micro-vascular endothelial cells (PMVEC) induced by lipopolysaccharide (LPS).Methods Wistar rat PMVEC cultured in vitro were randomly (random number) divided into several groups (n =4) as per exposure to given dosage of LPS for different lengths of time and to different dosages of LPS for given length of time.After PMVEC exposed to 10 mg/L LPS for 1 hour (h),3 h,6 h,12 h and 24 h or 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 h,the levels of TNF-αin the supernatant of culture medium were examined by the method of enzyme linked immunosorbent assay (ELISA).Another PMVEC was pre-treated by protein kinase C (PKC) inhibitor bis-indolylmaleimide (BIM) for 0.5 h or had the transfection of SSeCKS-specific small interfering RNA (siRNA) for 48 h before 10 mg/L LPS challenge for 24 h,and subsequently the supernatant was also examined by ELISA.One-way analysis of variance (ANOVA) was employed for statistical analysis by SPSS version 10.0 to compare values among all groups.A significant difference was presumed as a probability value ＜ 0.05.Results After PMVEC incubated with 0.1 mg/L,1 mg/L and 10 mg/L LPS for 24 hours,the levels of TNF-αsecreted were (253.70 ± 23.55),(327.88 ± 37.25),(403.20 ± 36.22),respectively,which were higher than that in un-stimulated PMVEC (82.28 ± 22.56,all P =0.000).After 10 mg/L LPS challenge for one hour,the level of TNF-αin the supernatant of PMVEC raised substantially (170.11 ±49.22),peaked at the time of 6 h (404.82 ± 13.78),then persisted at a higher level until 24 h (395.67 ± 36.23) than that in un-stimulated PMVEC (84.60 ± 23.61,P =0.001,0.000,0.000,respectively).After PMVEC pre-incubated with BIM,the level of LPS-induced TNF-αdecreased obviously (200.44 ± 27.39 vs.402.28 ± 31.07,P =0.000).Compared with LPS challenged PMVEC (407.28 ± 32.64),depletion of endogenous SSeCKS in PMVEC after inhibited by SSeCKS-siRNA significantly attenuated increase in the level of LPS-induced TNF-α (195.20 ± 13.28,P =0.000).Conclusions Down-activation of SSeCKS and PKC can inhibit the secretion of TNF-αin PMVEC induced by LPS,relieving the inflammatory response of PMVEC.

Objective To study the clinical significance of the injury and functional change of hypothalamic-pituitary-adrenal (HPA) axis after acute severe traumatic brain injury (TBI) in the rats.Methods A total of 60 adult healthy male Spraque-Dawley rats were randomly (random number) divided into 3 groups (n =20 in each group):sham operation group,model group and treatment group.The TBI models of rats were established by Feeney' s method.A low dose of dexamethasone (0.6 mg/kg) was injected into the abdominal cavity 20 minutes,24 hours and 48 hours after injury in treatment group,while rats of sham operation group and model group received equal volume of normal saline instead.All the rats were injected 1 μg adrenocorticotropic hormone (ACTH) into the abdominal cavity.The related parameters were detected at four time points,3 hours,12 hours,24 hours and 72 hours after cerebral contusion.The plasma corticosterone (CORT) and ACTH levels were measured by chemiluminescence.The hypothalamic,pituitary and adrenal of the rats were taken out for observing interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) expression detecting by immunohistochemical techniques at 72nd hour after TBI.One-way ANOVA and SNK-q test were used to analyze the results with SPSS 17.0 software package.Results The levels of ACTH and CORT on 3rd hour of model group raised remarkably compared with that of sham operation group,then they reduced gradually.The levels of CORT were lower than that of sham operation group at every time points after ACTH stimulation test (P ＜0.05 or P ＜0.01).The levels of CORT at all time points of treatment group were changed remarkably compared with that of model group.However,the ACTH levels of treatment group on 24 h increased slightly than that of model group.And the tendency of them was similar to model group (P ＜ 0.05 or P ＜ 0.01).The number of the hypothalamus and pituitary cells which express IL-6 and TNF-α in model group was more significantly increased when compared with that in sham operation group (P ＜ 0.01),while the number of this kind of cell in treatment group was significantly decreased than that in model group (P ＜ 0.01).The number of the adrenal cortex cells which express IL-6 in treatment group was more significantly decreased when compared with that in model group (P＜ 0.01),while the number of this kind of cell in model group was significantly increased than that in sham operation group (P ＜ 0.01).However,there was no significant difference of the TNF-α between all the groups (P ＞ 0.05).Conclusions Functional change of adrenal occurs early in the severe acute traumatic brain injury rats,and the response of adrenal to ACTH decreased as time goes by.Low-dose,short-course dexamethasone can delay the pathological changes,reduce the inflammatory response of HPA axis and increase the sensitivity of adrenal response to ACTH.

Objective To Explored the relative factors which caused the extubation failure in neurological intensive care unit (NICU).Methods It was a retrospective study.40 cases of patients who met the criteria,were brought into statistical analysis.They were admitted in NICU in Nan Fang Hospital from December 2008 to February 2011.The name,sex,age,diagnosis,respiratory parameters,24 hours discrepancy quantity,sputum,and Glasgow Coma Scale,Full Outline of UnResponsiveness Scale were recorded.SPSS 13.0 was used as statistic software.P ＜ 0.05 was considered statistically significant.Results Both in extubation successful and failure groups,GCS and Four were significantly different (all P ＜ 0.05).Howerer,there were no statistically significant in the other factors.There were significantly differences between GCS and Four in predicting extubation results (P =0.012).Logistic multiple regression showed that Four and GCS grade were predictive factor of extubation failure (P =0.041).Conclusions The result suggests that it is statistically significant to use GCS and Four as factors to predict extubation results.It can be widely used to help medical personnels monitoring the changes of patients'clinical conditions,judging prognosis,and making treatment plan in NICU.Wether other factors would effect the extubation results,more prospective,randomized controlled studies were needed.

Objective To investigate the cell survival of the combination of fibrin glue and adiposederived stem cells (ADSCs) in rats when implanted into ischemic myocardium and the improvement of heart function.Methods The rat ADSCs were isolated from the subcutaneous adipose tissues.The surface phenotype of these cells was analyzed by flow cytometry.Myocardial infarction was induced in female rats using coronary artery ligation.One week after MI,surviving rats were randomized (random nuber) into 4 groups,control group (n =10),fibrin group (n =10),cell group (n =10) and combination group (n =10).100 μl of PBS was injected into the ischemic myocardium in control group.100 μl of Fibrin glue were injected into ischemic myocardium in fibrin group.100 μl of ADSCs labeled with DAPI were injected into the infract along the border zone in cell group.ADSCs in 100 μl of fibrin glue were injected into the infract in combination group.Four weeks after the injection the surviving rats underwent examination of heart functions by the Hemodynamics.The rats were killed and their hearts were taken out to undergo immunohistochemistry with 4,6-diamidino-2-phenylindole (DAPI) and actin and factor Ⅶ to measure the area of cardiac infarction and the capillary density.The heart infarcted size was calculated by masson trichrome staining.All data was analyzed by software SPSS 15.0,ANOVA comparison tests and the student t test were used,and P ＜ 0.05 was considered as statistically significant.Results Four weeks after the cells were transplanted,LVSP and + dp/dtmax of combination group were highest among all groups.The heart infarcted size of the combination group was (28.5 ± 3.6) ％,significantly less than those of the cell group (33.33 ± 2.3) ％ and fibrin group (35.96 ± 2.11) ％,both P ＜ 0.05.The capillary density of the combination group was (108.7 ± 11.38) /mm2,significantly greater than those of the cell group and that of the fibrin group,and greater than that of the control group.DAPI and actin double staining detected a varied increase in the number of surviving cardiomyoctyes at the heart infarcted area.Conclusions Transplantation of ADSCs with fibrin glue brings better improvement in cell survival and in restoration of heart function than either cellular or fibrin therapy alone.

Objective To investigate the effects of Tanshinone Ⅱ A (extracted from Chinese herb medicine Salvia miltiorrhiza Bge.) on ventricular arrhythmias of rabbit hearts induced by ischemia in order to illuminate its mechanism of anti-arrhythmia.Methods Thirty rabbits were randomly (random number)divided into normal group,ischemic group and Tanshinone Ⅱ A group.Model of wedge shaped mass of rabbit left ventricular myocardium with coronary perfusion was prepared,and then by using floating glassy microelectrode,the trans-mural ECG,QT interval,the trans-mural dispersion of re-polarization (TDR) and trans-membrane action potentials from both endocardium and epicardium were simultaneously and wholly recorded.The incidence of ventricular arrhythmia in myocardium was observed after ischemia for thirty min.Results Under the condition of acute ischemia,compared with normal group,the incidence of ventricular arrhythmia and TDR were significantly increased in ischemia group (P ＜ 0.01),while incidence of ventricular arrhythmia and TDR were significantly reduced in tanshinone ⅡA group compared with ischemia group (P ＜ 0.05).The incidences of ventricular arrhythmia in normal,ischemia and Tanshinone Ⅱ A groups were 0/10,9/10 and 2/10 respectively.Conclusions Tanshinone Ⅱ A prevents ventricular arrhythmia and reduces TDR significantly in ischemic rabbit hearts.

Objective To compare the point-of-care testing (POCT) and laboratory testing of myocardial damage markers in the diagnosis and prognosis of acute coronary syndrome (ACS).Methods A total of 3467 patients with ACS who were treated in the Emergency Department Beijing Chaoyang Hospital Affiliated to Capital Medical University between January 1,2006 and June 30,2010 were retrospectively reviewed.The patient demographics (age,sex,past medical history and smoking history) and laboratory testing results (myocardial damage markers,D-dimer,NTproBNP,and ejection fraction [EF]) were analyzed.The patients who received POCT or laboratory testing of myocardial damage markers were compared with regard to emergency department stay (i.e.,the time from the emergency visit to interventional or conservative treatment),cardiovascular events during hospitalization (congestive heart failure,ventricular fibrillation,and cerebrovascular disease),and 28-day mortality rate.Results The emergency department stay,incidence of a cardiovascular event,and 28-day mortality in the POCT group were all lower than that in the laboratory testing group (P =0.000).A prolonged emergency department stay result in an increased incidence of 28-day mortality.The higher level of D-dimer and decreased EF prompted an increased incidence of 28-day mortality.Conclusions Compared with conventional laboratory testing,POCT can significantly shorten the length of an emergency department stay for an ACS patient,decrease the incidence of a cardiovascular event,and improve the prognosis.

Objective To explore the effects of pyrrolidine dithiocarbamate (PDTC) on the acute lung injury and the activation of Nrf2 pathway after Paraquat (PQ) induced lung injury.Methods Fortyeight adult male SD rats with lung injury induced by PQ were randomly (random number) divided into control group and PDTC group.Three animals were sacrificed at every 1-week interval,7d,14d and 21 days after PQ intoxication,and the lungs of rats were removed for acute lung injury score after HE staining,and for lung fibrosis assessment after Masson staining,and the levels of reduced glutathione (GSH) and malondialdehyde (MDA) in the lung tissue homogenate were assayed and the phosphorylation of Nrf2 (nuclear-E2-related factor 2) was detected by Weston blot.The mean values of detected variables between two groups were compared by t test,and survival curve was tested by Wilcoxon (Gehan) test.Results The intoxication symptoms of rats were obvious,and 4 rats in control group and 9 rats in PDTC group survived until 21days.The survival time of animals in PDTC group was longer than that in control group (Wilcoxon (Gehan) =10.17023,P =0.001).The levels of MDA in control group were higher than those in PDTC group,while the levels of GSH in control group were lower than those in PDTC group.The levels of phosphorylation of Nrf2 in PDTC group were higher than those in control group at 1-week intervals,1-week:(0.32±0.04) vs.(0.23±0.05),P=0.003; 2-week:(0.62±0.06) vs.(0.33±0.03),P＜0.001; 3-week:(0.61 ±0.04) vs.(0.33±0.05),P＜0.001.The acute lung injury (ALI) scores in PDTC group were lower than those in control group,1-week:(5 ± 0.95) vs.(8 ± 1.23),P =0.002 ; 2-week:(9±1.18) vs.(11±1.02),P=0.019; 3-week:(11±1.33) vs.(12±1.42),P=0.002.The percentages of lung fibrosis at 1-week intervals after PQ intoxication were (40.87 ± 7.25) ％,(43.38 ±5.71)％ and (45.91 ± 3.97)％ in control group,and they were higher than those in PDTC group (32.92±2.34)％,(33.45 ±3.04)％ and (35.27 ±3.81)％ in PDTC group,P=0.017,0.001 and 0.001 respectively.Conclusions Attenuation of acute lung injury and lung fibrosis,and prolongation of survival time of SD rats by PDTC were associated with activation of Nrf2 pathway.

Objective To observe the effect of cardiac massage by subdiaphragmatic compression (D-CPR) on the length of time required from cardiac arrest (CA) to restoration of spontaneous circulation (ROSC),hemodynamics,rate of ROSC,survival rate of 6 h,level of Caspase3 in myocardial cells and apoptosis index (AI) of myocardial cells and compare the effect of standard cardiac massage by chest compression (S-CPR) on those variables in order to choose the more effective resuscitation method for the patient with CA during abdomen operations.Methods A total of 32 healthy New Zealand rabbit were randomly (random number) divided into two groups,namely S-CPR group and D-CPR group (n =16 in each group).All of rabbits were anesthetized with ketamine and Shumianxin (a kind of hypnotics) by intraperitoneal injection,subsequently tracheotomy was made for endotracheal intubation,and right internal jugular vein was catheterized for monitoring central venous pressure (CVP) and left common carotid artery was for indwelling cannula to monitor arterial blood pressure.Lead-2 of ECG was placed.After laparotomy and vital signs of rabbits stabilized for 5 minutes,the endotracheal tube was clamped at the end expiration for 8 minutes to make asphyxial cardiac arrest model.The effects of two different methods were observed and compared in respects of changes in hemodynamics、length of time elapsed from CA to ROSC、ROSC rate and the survival rate in 6 h.The level of Caspase3 in myocardial cells and AI of myocardial cells were detected by using immunohistochemistry staining method and TUNEL,respectively 6 hours after successful resuscitation.Results ①The length of time consumed fiom CA to ROSC in D-CPR group was shorter than that in S-CPR group (P ＜0.05) ②Coronary perfusion pressure (CPP) and MAP 15 minutes after CPR were higher in D-CPR group than those in S-CPR group (P ＜ 0.05).③SBP and DBP after ROSC were higher in D-CPR group than those in S-CPR group.④ROSC rate in D-CPR group was significantly higher than that in S-CPR group (81％ vs.43％,P ＜0.05).⑤Survival rate in 6h in D-CPR group was substantially higher than that in S-CPR group (75％ vs.25％,P ＜ 0.05).⑥HE staining showed that severe myocardial damage manifesting in edema of myocardial cell,indistinguishable cell boundary,and patchy necrosis with infiltration of scanty inflammatory cells were found in S-CPR group.While in D-CPR group,mild myocardial damage in form of slight cellular edema and distinctive cell boundary was observed.⑦Level of Caspase3 in myocardial cells in terms of integrated optical density (IOD) of postive Caspase3 cells was substantially lower in D-CPR group than that in S-CPR group (P ＜ 0.05).⑧Apoptosis index (AI) of cells was lower in D-CPR group than that in S-CPR group (P ＜ 0.05).Conclusions ①The hemodynamics in D-CPR group was more stable than that in S-CPR.group,and D-CPR increased CPP,MAP,ROSC rate and survival rate in 6h,improving achievement of successful resuscitation.②D-CPR was more effective in terms of shortening the length of time for restoration of spontaneous circulation、decreasing level of Caspase3 in myocardial cells、decreasing apoptosis index of myocardial cells and ameliotating myocardial damage from ischemic repeffusion injury.

Objective To investigate the coagulant function of Thrombomodulin-Protein C-EPCR System in rats with hemorrhagic shock in virto study.Methods In this study,30 rats were divided into three groups,with each group 10 rats:control group,the sham shock group,hemorrhagic shock group.Hemorrhagic shock model was subjected to computer-controlled arterial hemorrhage to 40 mm Hg for 60 min The rat blood serum samples were obtained from rats in sham shock group and shock group after maintaining 3 hours,and fetal bovine serum (FBS) was used as control group.Then medium contained fore-mentioned serums cultured with SV40-transformed aortic rat endothelial cells for 6 hours,12 hours and 18hours.The messenger RNA (mRNA) expression of thrombmodulin (TM),Endothelial cell protein C receptor in the SV40-transformed aortic rat endothelial cells which were treated with serums were detected by reverse-transcription polymerase chain reaction.Results Compared to control group,SVARECs were incubated for 6hrs with shock serum,the mRNA expression of TF,TM and t-PA was significantly higher,and continued elevating at the time of 18 hours (P ＜ 0.01).Conclusions Serums from hemorrhagic shock rats upregulate TMmRNA EPCRmRNA expression on the endothelial cells surface.It is suggested that Thrombomodulin-Protein C-EPCR System pay a key role of anticoagulation,the protection of endothelial function.