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Objective To study the effect of the promoter methylation of coagulation factor Ⅶ (FⅦ) on the coagulation factor Ⅶ activity (FⅦa) in traumatic brain injury (TBI) patients,and the correlation between the promoter methylation in FⅦ and intracranial progressive hemorrhagic injury (PHI).Methods A prospective analysis was conducted on 79 patients with moderate-severe TBI admitted to emergency department from August 2010 to August 2014.The peripheral venous blood samples were collected at admission and then were delivered for measurement of FⅦa.Genomic DNA was isolated from patient blood,and the promoter methylation in FⅦ (CpG2,CpG3,CpG4,CpG5,and CpG6) were analyzed.According to the level of plasma FⅦa,the patients were divided into FⅦa ≥90％ group and FⅦa ＜ 90％ group.Based on the presence of PHI,the patients were divided into PHI group and non-PHI group.The FⅦ promoter methylation,age,gender,systolic blood pressure,Glasgow Coma Scale (GCS),length of stay and mortality between FⅦa≥90％ group and FⅦa ＜ 90％ group,PHI group and non-PHI group were compared.Results There were no significant differences in age,gender,systolic blood pressure,GCS,LOS,and mortality between FⅦa ≥90％ group and FⅦa ＜90％,PHI group and non-PHI group (P ＞ 0.05).The methylation of CpG3 in FⅦa ≥90％ group was less than that in FⅦa ＜90％ group (0.83 ±0.05 vs.0.85 ±0.03) (P＜0.05),while there were no significant differences in other CpG sites between these two groups (P ＞ 0.05).No significant differences in all of methylation levels of the CpG sites between PHI group and non-PHI group were found (P ＞0.05).Conclusions The promoter methylation of FⅦ affects plasma FⅦa concentrations,and higher methylation results in lower FⅦa.The promoter methylation of FⅦ is not associated with PHI in TBI patients.
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Objective To observe the incidence of critical illness-related corticosteroid insufficiency (CIRCI) after severe traumatic brain injury (sTBI) and investigate the relationship between CIRCI and prognosis.Methods This prospective cohort study enrolled 89 sTBI patients (68 males and 21 females;at age range of 15-80 years) hospitalized within 24 hours after sTBI from June 2014 to December 2015.The Glasgow coma scale (GCS) was ≤8 points.The causes of injury included extensive contusion of brain (44 cases),subdural hematoma (21 cases),epidural hematoma (11 cases),primary brain stem injury (8 cases) and diffuse axonal injury (5 cases).Adrenocorticotropic hormone (ACTH) stimulation tests were done within 36 hours after sTBI to identify CIRCI patients.The patients were divided into CIRCI group (50 cases) and non-CIRCI group (39 cases).Moreover,the patients were categorized into survival group (62 cases) and death group (27 cases) based on survival status.The GCS score,mechanical ventilation time,cerebral hernia,survival time and mortality within 28 days were observed in two groups.Results The incidence of CIRCI in sTBI patients was as high as 56％ (50/89).Compared with the non-CIRCI group,the CIRCI group had lower GCS [(5.3 ± 1.7) points vs.(6.1 ± 1.4) points,P ＜ 0.05],and sTBI patients with CIRCI were mechanically ventilated for a longer period of time [(9.9 ± 2.8) days vs.(7.5 ± 1.6) days,P ＜ 0.05].In comparison with non-CIRCI patients,the incidence of brain herniation in sTBI patients with CIRCI was higher (58％ vs.21％,P ＜0.01).The total fatality rate within 28 days was 30％ (27/89).The survival time of CIRCI group was significantly shorter than that of non-CIRCI group (P ＜ 0.05).The fatality rate in the CIRCI group was significantly higher than that of the non-CIRCI group [40％ (20/50) vs.18％ (7/39),P ＜0.05].The incidence of CIRCI in death group was significantly higher than that of the survival group [74％ (20/27) vs.48％ (30/62),P ＜ 0.05].Conclusions The incidence of CIRCI in STBI patients is high.The sTBI patients with CIRCI has significantly higher incidence of brain hernia,longer mechanical ventilation time,higher 28-day mortality and shorter survival time compared with non-CIRCI patients.
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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.
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Objective To observe the correlation between the changes of neural cell apoptosis arid caspase-3 gene expression in brain tissues following acute severe traumatic injury to brain(TIB).Method A total of 120 adult Spraque-Dawley rats were divided into a control group(n=8)，TIB group(n=56)and TIB with administration of caspase-3 inhibitor group(n=56).TIB models of rats were made with Feeney's method.The z-DEVDfmk(5 μg)，caspase-3 inhibitor，was administered by intracerebral infusion，and the rats were sacrificed 1，6，24，48 hours and 3，7，14 days postinjury(n=8 for each interval).The specimens of the injured cerebral cortex，suhcerticai white matter，hippocampus，dentate gyrus and contrahteral corresponding brain tissues were taken for detecting apoptesis of neural cells by the terminal deoxynucleotidyl transferase mediated DUTP nick end labeling (TUNEL)methods and flow cytomeay.Caspase-3 mRNA and protein expression were detected by using RT-PCR，immunohistochemistry and western blot analysis.The caspase-3 activity was detected by using caspase-3 fluorescent assay kit.Student t-test and Spearman correlation analysis were used to analyze the data with SPSS version 10.1 software package.Results Apoptesis indexes(AI)and the apoptesis percentage(AP)of neural cells in the injured brain regions increased quickly after injury，and reached its peak 24 to 48 hours later，then decreased slowly,but it remained at higher level above that of normal till 14 days later(P＜0.01).The levels of caspase-3 mRNA,eastme-3 protein and caspase-3 activity were increased significantly post injury，and reached its peak at 24 to 48 hours，then it gradually decreased.Compared with control group，the levels ofoptical density of caspase-3 proteins in the injured hippocampus and subcortical white matter at 24 and 48 hours post injury increased 1484% and 1690%，caspase-3 mRNA expressiom increased 1043%and 1180%，and the degreas of caspase-3 activity increased 148% and 183%，respectively.The expression of caspase-3 proenzyme and its P17 subarrit increased.After trealment with caspase-3 inhibitor z-DEVD-fmk，the levels of caspase-3 mRNA，protein expression and caspase-3 activity were significantly decreased.and AI and AP were significantly decreased as well.The correlation between caspase-3 mRNA and level of neural apoptesis was positive(r=0.821，P＜0.01)，and it was likewise between caspase-3 protein and level of neural apoptosis(r=0.638.P＜0.01).Interestingly enough，a positive correlation was found between caspase-3 mRNA and easpase-3 proteins(r=0.945，P＜0.01).Conclusions The activation of caspase-3 leads to apoptosis of neural cells after acute TIB.The expression of caspase-3 are consistent with apoptosis of neural cells following TIB.The regulation of caspase-3 induced by TIB occurs at a ceriain critical link before transduction.Caspase-3 inhibitor can efficiently inhibit apoptosis of neural cells following TIB.