To find potential biomarkers based on miRNA and their potential targets in splenic monocytes in burn-injured mice. Male Balb/c mice were subjected to sham or scalding injury of 15% total body surface area. Spenic CD11b+ monocytes were purified with magnetic beads. The monocytes were cultured in the presence of lipopolysaccharide. The proliferation of monocytes was detected by MTT assay, and the cytokines in the supernatant were examined by enzyme linked immunosorbent assay. The purified monocytes were also under total RNA extraction. The differential monocytic miRNAs expression between the sham and burn-injured mice was analysed by miRNA microarray. The activity of monocytes was comparable between the two groups (p > 0.05). However, monocytes from burn-injured mice secreted higher levels of tumour necrosis factor (TNF)-α and transforming growth factor-β, but lower level of monocyte chemoattratctant protein-1. A total of 54 miRNAs were differentially expressed in monocytes from burn relative to sham-injured mice (fold >3). Further quantitative reverse transcription polymerase chain reaction confirmed that the expression of miR-146a was significantly down-regulated, while miR-3091-6p was up-regulated after burn injury. Using the combination of Miranda and TargetScan softwares, we found that mir-146a may regulate 180 potential target genes including TNF receptor related factor 6 (TRAF6), interleukin-1 receptor related kinase 1 (IRAK1) and CD28. Mir-3091-6p may regulate 39 potential targets, including SOCS7 (cytokine signal transduction inhibitor 7) and ARRB2 (arrestin, β 2). The miRNAs expressed by monocytes after burn injury may be involved in the regulation of innate immune response in burn injury.

INTRODUCTION: Whether insulin resistance is underlying the deep venous thrombosis (DVT) development in patients with traumatic brain injury (TBI) is elusive. METHODS: A prospective observational study of 73 severe TBI patients who had measurements of plasma insulin, glucose, glucagon-like peptide 1 (GLP-1), inflammatory factors, and hematological profile within 4 preset time periods during 2 weeks after TBI. Ultrasonic surveillance of DVT was tracked weekly. Two-way ANOVA analysis was applied to determine whether the factors could discriminate between patients with and without DVT. Partial correlations of insulin level with other variables were carried out in patients with or without DVT. Factors associated with DVT were analyzed by multivariable logistic regression. Neurological outcome 6 months after TBI was assessed for Glasgow Outcome Scale (GOS). RESULTS: Among patients with an average (SD) age of 53 ± 16 years, DVT developed in 20 patients (27%). The 14-day plasma insulin levels were higher in patients with DVT (P = 0.01). Platelet profile discriminated significantly between patients with and without DVT. None of other factors differed between the two groups. Patients with insulin therapy had significant higher insulin (P = 0.006), glucose (P < 0.001) and GLP-1 (P = 0.01) levels, and were more likely to develop DVT (60% vs. 15%, P < 0.001) with concomitant platelet depletion. Insulin levels correlated with glucose, GLP-1 levels, and platelet exclusively in patients without DVT. Conversely, in patients with DVT, insulin correlated negatively with GLP-1 level (r = -0.297, P = 0.016). Age (P = 0.01) and elevated insulin level at day 4-7 (P = 0.04) were independently associated with DVT. Patients with insulin therapy also showed worse GOS (P = 0.001). CONCLUSIONS: Elevated insulin level in the first 14 days after TBI may present insulin resistance in TBI, which was associated with consistent platelet activation, thus increasing risk of DVT.

Background: Although glucagon-like peptide 1 levels have been closely associated with inflammation and mortality in septic patients, the clinical importance of glucagon-like peptide 1 on hospital-acquired infections and long-term mortality after burn injury remains unexplored. Methods: Plasma samples from 144 burn patients were collected on admission to determine total glucagon-like peptide 1, interleukin 6, and monocyte chemotactic protein-1 levels. Hospital-acquired infections were determined by positive microbial culture. One-year mortality was assessed by telephone interview. Factors associated with glucagon-like peptide 1 were determined by multivariable linear logistic regression. Predicting the clinical importance of glucagon-like peptide 1 on the development of hospital-acquired infections and mortality were determined by Cox proportional hazards models and further by receiver operating characteristic curve analysis. Kaplan-Meier analyses were performed to examine whether the mean glucagon-like peptide 1 level of the cohort could discriminate the hospital-acquired infections-free survival. Results: Median burn size was 41% (19%-70%) of total body surface area. Hospital-acquired infections developed in 36 (25%) patients after a mean of 10 +/- 1 days after injury. Interleukin 6, monocyte chemotactic protein-1, and blood urea nitrogen levels and thrombin time were independently associated with increased glucagon-like peptide 1 levels. Levels of glucagon-like peptide 1 (median, interquartile range) were greater in patients who developed hospital-acquired infections than in those who did not (237 pmol/L, 76-524 vs 80 pmol/L, 51-158; P < .001) and in patients who died (536 pmol/L, interquartile range: 336-891 pmol vs 98 pmol/L, 47-189; P < .001). Although the glucagon-like peptide 1 level could not predict hospital-acquired infections-free survival in individual patients, it could predict 1-year mortality independently (P = .021). Moreover, a glucagon-like peptide 1 level of 200 pmol/L could discriminate hospital-acquired infections-free survival (P < .001). Conclusion: Admission glucagon-like peptide 1 level can discriminate hospital-acquired infections-free survival and predict long-term mortality in a group of patients with burn injury. Our data suggests that glucagon-like peptide 1 may be a predictive biomarker for hospital-acquired infections and mortality in bum patients. (C) 2020 Elsevier Inc. All rights reserved.

Objective: To establish an animal model for posttraumatic stress disorder in burn-injured patients. Methods: Thermal-injured mice with 15% total body surface area were subjected to a series of neurobehavioral tests at 1 and 3 months postburn. Brains were collected for analysis of key molecules expression, spleens for T cell function analysis, and blood for biochemistry and hormones detection. Results: Comparison with sham mice, burn mice showed extremely high locomotion in homecage, open field, and forced swimming tests, indicating a hyper-arousal state. Burn mice exhibited improved spatial memory in Morris Water Maze test and heightened context fear memory in context fear conditioning, suggesting re-experiencing behavior. Although burn mice showed pronounced passive avoidance in the step-through test, their active avoidance capability in response to the conditional stimulus in the shuttle box test was relatively deteriorated. Likewise, the retention of cue-feared memory was impaired in fear conditioning test. The above negative alterations in mood were recapitulated in open-field test, in which the burn mice displayed an anxiety-like behavior with less time spent in the center. However, no sign of depression was found in the forced swimming and sucrose preference tests. The negative mood of burn mice was reinforced by a deficit in sociality and preference for social novelty in social interaction test. These neurobehavioral alterations were associated with an increased expression of brain-derived neurotrophic factor along with a remarkable microgliosis and a moderate astrocytosis in the brain of burn vs. sham mice. Moreover, a prominent Th2 switch and consequent increased nuclear NF-kappa B translocation were seen in the splenic T cells from burn relative to sham mice. Conclusions: We conclude that even mild burn injury could lead to long-lasting cognitive and effective alterations in mice. These findings shed light on the interactions among neuropsychology, neurobiology, and immunology throughout the recovery period of burn injury.

BACKGROUND:Although glucagon-like peptide 1- (GLP-1-) based therapy of hyperglycemia in burn injury has shown great potential in clinical trials, its safety is seldom evaluated. We hypothesize that exendin-4, a GLP-1 analogue, might affect the immune response via the activation of the sympathetic nervous system in burn injury. METHODS:Male Balb/c mice were subjected to sham or thermal injury of 15% total body surface area. Exendin-4 on T cell function in vitro was examined in cultured splenocytes in the presence of β-adrenoceptor antagonist propranolol (1 nmol/L) or GLP-1R antagonist exendin (9-39) (1 μmol/L), whereas its in vivo effect was determined by i.p. injection of exendin-4 (2.4 nmol/kg) in mice. To further elucidate the sympathetic mechanism, propranolol (30 mg/kg) or vehicle was applied 30 min prior to injury. RESULTS:Although the exacerbated burn-induced mortality by exendin-4 was worsened by propranolol pretreatment, the inhibition of T cell proliferation by exendin-4 in vitro could be restored by propranolol instead of exendin (9-39). However, a Th2 switch by exendin-4 in vitro could only be reversed by exendin (9-39). Likewise, the inhibition of splenic T cell function and NFAT activity by exendin-4 in vivo was restored by propranolol. By contrast, the increased splenic NF-κB translocation by exendin-4 in vivo was potentiated by propranolol in sham mice but suppressed in burn mice. Accordingly, propranolol abrogated the heightened inflammatory response in the lung and the accelerated organ injuries by exendin-4 in burn mice. On the contrary, a Th2 switch and higher serum levels of inflammatory mediators by exendin-4 were potentiated by propranolol in burn mice. Lastly, exendin-4 raised serum stress hormones which could be remarkably augmented by propranolol. CONCLUSIONS:Exendin-4 suppresses T cell function and promotes organ inflammation through the activation of the sympathetic nervous system, while elicits Th2 switch via GLP-1R in burn injury.

For the past 60 years of burn therapy in China,the incidence of infection declined gradually,however,the drug-resistant pathogens have become increasingly serious.From the research of the enterogenous infection to the wound coverage by allograft skin,from the endotoxin and exotoxin-targeted therapy of pathogens to empirical usage of antibiotics,these efforts had established the first line of barrier against pathogens and reduced the incidence of sepsis obviously.Recently,many studies have been carried out to investigate the regulatory mechanism underlying the host immunity in defending the pathogens and controlling the excessive inflammation after major burns.The resulting strategies of immunomodulation of burn infection by Chinese and western medicine were contributed to the second immune barrier against the invaded pathogens.With the updated medical technology to treat the infection both symptomatically and etiologically,the incidence and severity of infection were reduced remarkably.In the future,further studies should be focus on applying the new technology to detect the pathogenic microorganism quickly,finding the novel biomarkers for evaluation of immune status,and following up the long-term impact of infection on host immunity in extensively burn patients.

目的 探讨生后发育期不同间隔时间异氟烷处理对大鼠成年后认知功能的影响.方法 选取生后7d的SD大鼠48只,随机分为正常对照组(Naive组),异氟烷处理6h组(G6h组),异氟烷处理3次、每次2h、间隔1d组(G2h-1d组),异氟烷处理3次、每次2h、间隔3d组(G2h-3d组).处理后幼鼠饲养至成年(＞60d),进行旷场实验、八臂迷宫及Morris水迷宫测试并进行组间比较,评价生后发育期异氟烷处理对大鼠成年后空间认知能力的影响.结果 与Naive组比较,各处理组大鼠在旷场实验中的表现均无差异(P＞0.05).雄鼠八臂迷宫测试结果显示,G6h组和G2h-1d组错误次数＞3的百分比明显高于Naive组(P＜0.05),G2h-3d组重复错误次数少于G6h组和G2h-1d组(P＜0.001),入臂总次数明显低于G6h组和G2h-1d组(P=0.003、P=0.008),入臂总耗时明显低于其余3组(P＜0.005).雌鼠八臂迷宫测试结果显示,G6h组错误次数＞3的百分比、重复错误次数、入臂总次数及入臂总耗时均低于Naive、G2h-1d和G2h-3d组(P＜0.005),且G6h组和G2h-3d组重复错误次数低于G2h-1d组(P＜0.001、P=0.024).Naive组和G2h-3d组雌鼠错误次数＞3的百分比和入臂总次数明显高于雄鼠(P＜0.05,P＜0.01),而G6h组雌鼠错误次数＞3的百分比和入臂总次数明显低于雄鼠(P=0.016,P=0.041).Morris水迷宫测试结果显示,与其他3组雄鼠比较,G2h-1d组雄鼠在相反象限耗时最长、潜伏期最长、穿梭平台次数最少,但差异无统计学意义(P＞0.05);与G2h-3d组雌鼠比较,G2h-1d组雌鼠在相反象限耗时增多、穿梭平台次数减少,差异有统计学意义(P=0.033,P=0.040).结论 在生后发育期异氟烷处理总时间一致(6h)的前提下,单次或多次处理对大鼠成年后空间认知行为有一定影响.间隔时间较短的多次异氟烷处理可影响其成年后认知功能;延长处理间隔时间可减弱或消除对认知行为的影响,且具有性别差异.

[Abstract] Objective To investigate the effects of neonatal isoflurane exposures at different interval time on the cognitive function in adult rats. Methods Forty-eight postnatal 7-day SD rats were randomly divided into four groups: normal control group (Naive group), continuous exposure to 1.5% isoflurane for 6h (G6h group), and exposure to isoflurane 3 times, 2h each time, in one day (G2h-1d group) or 3 days interval (G2h-3d group). Open field test (OFT), Radial arm maze (RAM) and Morris water maze (MWM) tests were performed when these rats grew up (>60 days). Behavioral changes were recorded to evaluate the effects of neonatal isoflurane exposure on the cognitive function in adult rats. Results On OFT, no significant difference existed between Naive group and another 3 groups ( P >0.05). The comparison among groups in male rats on RAM test showed: the percentage of error times >3 increased significantly in G6h and G2h-1d group than in Naive group ( P <0.05); the repeated errors were less in G2h-生后不同间隔时间异氟烷处理对大鼠成年后认知功能的影响 G2h-1d were and G2h-1d than in G2h-3d ( for shorter in G2h-3d than in another 3 ( The among groups rats on the of mistake number the repeated the times of and the total time consuming for were decreased significantly in G6h group than G2h-1d and G2h-3d group ( the repeated lower and G2h-3d group than in G2h-1d group ( The and rats mistake number and the times of were higher than of and G2h-3d ( However, with the mistake number and the times of entering arms decreased significantly in rats with the rats in another 3 groups, G2h-1d group spent more time in opposite took the longest incubation period and the least times of shuttling platforms, but the differences were not significant ( with the female rats in G2h-3d group, in G2h-1d group spent increased time in opposite and took decreased frequency for shuttling platforms with statistical significance result in long-term cognitive impairment, but the impairment could be prevented by prolonging the interval time, and in addition, the cognitive impairment has gender difference.

Objective To observe the infiltration of immune cells into the brain and the apoptosis of brain neurons after moderate burn injury. Method Thirty BALB/C male mice were randomly divided into two equal groups: burn group with their backs immersed in water at 94°C for 8s so as to cause second-degree burn in 15% of the total surface area (TBSA),and sham burn group with their backs immersed in water at 37℃..12 brains of mice from each group were removed after intracardiac normal saline perfusion 24 hours after scald.Mononuclear cells from the brain were isolated by discontinuous Percoll gradient centrifugation. For flow cytometry analysis, T cells were labeled with CD3+CD45high,monocytes with CD11b+CD45 high and activated microglia with CD11b+CD45int.The remaining 3 brains of each group were fixed and dehydrated.Then the frozen sections of brain tissue were stained by immunofluorescence for the immune cells,and with TUNEL method for the apoptotic cells.The sections were sealed with mounting medium for fluorescence with DAPI and photographed under confocal microscopy. Results The proportion of T cells in the brain tissue of burn group was[(1.103 ±0.674)%,significantly higher than that of the sham injury group[(0.385 ± 0.109)%,P<0.05]. The proportion of microglia(CD45intCD11b+)of the burn group was(1.017±0.35)%,significantly higher than that of the sham injury group [(0.502 ± 0.358)%,P<0.05],suggesting a higher degree of activation of microglia. Immunofluorescence staining showed an increased distribution of T cells and CD45high cells in the perivascular and choroid plexus of the burn group, compared with the sham injury group. Moreover, more apoptotic neurons were observed in the cerebral cortex and periventricular parenchyma of the brain in the burn mice relative to the sham injury mice. Conclusion As soon as 24 hours after moderate burn,infiltration of peripheral T cells and increasing number of apoptotic neurons can be observed in the brain, which may contribute to the following neuropsychiatric disorders.

Glucagon-like peptide-1 (GLP-1)-based therapy via G protein-coupled receptor (GPCR) GLP-1R, to attenuate hyperglycemia in critical care has attracted great attention. However, the exaggerated inflammation by GLP-1R agonist, Exendin-4, in a mouse model of burn injury was quite unexpected. Recent studies found that GPCR might elicit proinflammatory effects by switching from Gαs to Gαi signaling in the immune system. Thus, we aimed to investigate the possible Gαs to Gαi switch in GLP-1R signaling in monocyte following burn injury.

目的 探讨胰高血糖素样肽(GLP-1)的长效类似物Exendin-4(Ex-4)对烫伤小鼠生存率、炎性反应和器官损伤的影响及其交感神经调控机制.方法 BALB/C小鼠随机分为烫伤组和假伤组,分别行93℃ 15％总体表面积(TBSA)的Ⅲ度烫伤和37℃假伤.烫伤前30min腹腔注射β2肾上腺素能受体阻断剂普萘诺尔(prop,30mg/kg),伤后30min立即腹腔注射Ex-4(2.4nmol/kg).24h后处死动物留取肺脏和血清,生化法检测肝肾功能,ELISA法检测外周血及肺部炎性因子TNF-α、MCP-1和IL-10水平;分离脾脏单个核细胞,观察核蛋白NF-κB p65蛋白表达;此外,烫伤小鼠随机分组观察Ex-4对动物72h存活情况的影响.结果 Ex-4处理的烫伤小鼠病死率有增加趋势.Ex-4对假伤组肺组织炎性细胞因子没有显著影响,但可显著增加烫伤动物肺脏TNF-α、MCP-1和IL-10水平,且此效应可被prop阻断.Ex-4可抑制假伤小鼠血清TNF-α、IL-10水平,而促进烫伤组血清TNF-α、MCP-1、IL-10水平(P＜0.05),且烫伤组prop干预后,反而增强了Ex-4的促细胞因子分泌作用.Ex-4可显著增加烫伤小鼠肝肾功能损伤指标AST/ALT和UR/Cr(P＜0.05),并能被prop阻断;而对假伤组无明显影响.Ex 4能显著上调烫伤小鼠脾脏细胞核NF-κB p65蛋白表达(P＜0.05),且此效应可被prop阻断.结论 Ex-4可通过交感神经机制促进局部组织器官的炎性反应以及器官损伤,Ex-4和激活的肾上腺素能受体共同激活cAMP-PKA-NF-κB信号转导途径,进而调节全身炎性反应.

肠促胰素是一种肠道分泌的降血糖激素,可作用于胰岛β细胞促进胰岛素分泌,降低血糖. 胰高血糖素样肽-1 ( Glucagon-like peptide-1 , GLP-1 )是首先在小肠黏膜L细胞发现的一种肠促胰素,在进食等血糖升高情况下其释放增加. 由于GLP-1的血糖依赖性降血糖特性,糖尿病患者服用后不易发生低血糖事件,因此作为一种新型降血糖药物GLP-1 类胰泌素在临床得到广泛应用[1]. 天然 GLP-1 在体内极易被二肽基肽酶Ⅳ( Dipeptidyl peptidase Ⅳ, DPP-Ⅳ)广泛快速降解,临床上主要使用GLP-1 受体( GLP-1 receptor, GLP-1R )激动剂 Exendin-4 和DPP-Ⅳ活性抑制剂 Liraglutide. 近几年研究发现, GLP-1除具有降血糖作用外,还有心血管保护、神经保护和免疫调节作用,其强大的免疫功能一方面有利于对糖尿病的治疗,另一方面可能引起一定的副作用. 本文重点阐述 GLP-1 的免疫功能及其调节机制,为扩展肠促胰素类药物的临床应用范围,以及规避其免疫学风险方面提供了一定的参考.

在生理或病理状态下,免疫系统与中枢神经系统之间都存在着密切的交流.多种免疫细胞及其相关细胞因子可通过某些途径进入大脑,对神经细胞产生多种神经生物学作用,进而引起认知、情感、社会行为等多个方面的神经行为学改变.本文回顾了几种重要的免疫细胞及细胞因子对神经行为学的影响,阐明其神经免疫学机制,为预防急慢性炎症性疾病导致的神经行为学异常提供预警.

Objective: To investigate the changes of immune function of regulatory T cells (Tregs) in peripheral blood of patients with severe burns, and explore its correlation with the complicated sepsis. Methods: Forty patients with burn area ≥ 30％ total body surface area (TBSA) were enrolled in present study, and divided into sepsis group (n=26) and non-sepsis group (n=14). The peripheral venous blood samples were collected on 1, 3, 7, 14, 21 and 28 days after injury to separate CD4+CD25+Tregs. The expressions of FOXP3 and CTLA-4 on the surface of CD4+CD25+Tregs were measured by flow cytometry, and the levels of mean fluorescence intensity (MFI) of interleukin-10 (IL-10) as well as transforming growth factor-β (TGF-β) in CD4+CD25+Tregs were further determined by flow cytometry after stimulation in vitro. Results: The expressive levels of FOXP3 in CD4+CD25+Treg increased significantly 7 and 14 days after injury, and of CTLA-4 markedly enhanced 14 days after injury in sepsis group than in non-sepsis group (P<0.05). The MFI of IL-10 in CD4+CD25+Tregs at the 1st day after injury was significantly higher than the health controls (P<0.01), and the IL-10 MFI level on day 21 after injury was markedly higher in sepsis group than in non-sepsis group (P<0.05). Likely, TGF-β MFI levels in CD4+CD25+Tregs on day 14 and 21 were significantly higher in sepsis group than in non-sepsis group (P<0.05). Conclusions: The immune response of Tregs shows dramatic enhancement in patients with severe burns, which further down-regulates the cell-mediated immunity by releasing immunosuppressive cytokines including IL-10 and TGF-β. Dynamic monitoring the expression and function of Tregs might be helpful for assessment of the clinical course as well as septic complications in patients following extensive burns.

Objective:To investigate effects of isoflurane anesthesia of different time interval on acute injury of brain function in neonatal rats with consistent total time of isoflurane anesthesia. Methods:Seven-day neonatal Sprague-Dawley (SD) rats were randomly divided into normal control group (breathe the air), continuous anesthesia group (a single 6-hour exposure to 1.5% isoflurane), and intermittent anesthesia 1 day and 3 days groups (three times of 2-hour exposure to anesthesia with an interval of 1 day or 3 days), 12 rats in each group. The ratio of male to female was 5:7. They underwent the test of learning and memory in the radial arm maze (RAM) 21 days after birth, twice a day for 4 days. The number of entry into wrong arms, number of repeated errors, number of total arm entries, and time for completing the task were recorded for evaluation of effect of neonatal isoflurane on cognitive behavior in rats. Results:① Compared with normal control group, the percentage of number of errors > 3 in anesthesia of 3-day interval group was significantly decreased (33.3% vs. 46.9%, P < 0.05), the percentages of repeated errors > 0 and total arm entries > 8 were significantly increased (33.3% vs. 18.8%, 27.1% vs. 13.5%, both P < 0.05), but there were no statistically significant difference in the percentage of mistake number > 3 between continuous anesthesia group, interval anesthesia 1-day group and the normal control group (44.8%, 44.8% vs. 46.9%), the percentages of number of repeated mistake > 0 and total arm entries > 8 in above three groups were slightly increased as compared with those of normal control group (27.1%, 22.9% vs. 18.8%, 20.8%, 21.9% vs. 13.5%, all P > 0.05). No statistical differences in completing the task among normal control group, continuous anesthesia group, interval anesthesia 1 day and 3 days groups were found (minutes: 1.32±0.91, 1.54±1.05, 1.46±0.86, 1.38±0.79, all P > 0.05). ② It was found by gender analysis that the percentages number of repeated errors > 0 and total arm entries > 8 were significantly lower in female rats than those in the male rats only in normal control group (5.0% vs. 28.6%, P < 0.01; 5.0% vs. 19.6%, P < 0.05). There was no obvious gender difference in exposed groups. ③ Compared between groups of female rats, the percentages of repeated mistake > 0 in continuous anesthesia group, interval anesthesia 1 day and 3 days groups (25.0%, 25.0%, 30.0% vs. 5.0%, P < 0.05 or P < 0.01) and the percentage of total arm entries > 8 in interval anesthesia 1 day and 3 days groups were significantly higher than that of normal control group (22.5%, 25.0% vs. 5.0%, both P < 0.05). No significant difference about the RAM task in male rats of all the four groups was found. Conclusions:Different time interval of neonatal isoflurane exposure may develop certain degree of acute brain injury in rats, characterized by cognitive function. Prolongation of the interval time significantly enhanced long-term memory in rats. Multiple neonatal exposures to isoflurane were associated with greater cognitive impairment than a single exposure. In addition, isoflurane can significantly increase cognitional functional disorder in the female, not in the male rats.

OBJECTIVE:To investigate the changes in plasma gelsolin (pGSN) levels in severe burn patients with sepsis, and to evaluate the prognosis of patients when combined with other related clinical indexes.METHODS:Sixty-five severe burn patients with sepsis hospitalized from June 2013 to June 2015 conforming to the study criteria were divided into death group (n=24) and survival group (n=41) according to the clinical outcome on post sepsis diagnosis day (PSD) 28. The pGSN levels of patients were determined on PSD 1, 3, 7, and 14 with double antibody sandwich enzyme-linked immunosorbent assay. The serum level of C-reactive protein (CRP), serum level of procalcitonin, lactate level of arterial blood, Acute Physiology and Chronic Health Evaluation (APACHE) II score, and Sequential Organ Failure Assessment (SOFA) score were determined or recorded on PSD 1. Data were processed with repeated measurement analysis of variance, t test, and chi-square test. On PSD 1, the pGSN level, serum level of CRP, serum level of procalcitonin, lactate level of arterial blood, APACHE II score, and SOFA score of 65 patients were collected to screen the independent risk factors related to death with single factor and multi-factor Logistic regression analysis. Receiver operating characteristic (ROC) curves of the independent risk factors related to death were plotted to evaluate the predictive power for death in 65 patients.RESULTS:(1) The pGSN levels of patients in death group on PSD 1, 3, 7, and 14 were respectively (146±44), (85±24), (28±7), and (19±4) mg/L, obviously lower than those in survival group [(287±82), (179±51), (196±56), and (249±67) mg/L, with t values from 1.735 to 4.304, P<0.05 or P<0.01]. (2) The serum level of CRP, serum level of procalcitonin, lactate level of arterial blood, APACHE II score, and SOFA score of patients in death group on PSD 1 were respectively (56±7) mg/L, (12.54±0.82) μg/L, (2.74±0.27) mmol/L, (24.3±2.4) points, and (11.43±0.57) points, significantly higher than those in survival group [(35±4) mg/L, (2.38±0.16) μg/L, (1.83±0.12) mmol/L, (15.0±1.5) points, and (7.22±0.23) points, with t values from 1.902 to 3.883, P<0.05 or P<0.01]. (3) Multi-factor Logistic regression analysis showed that the pGSN level (odds ratio: 6.83, 95% confidence interval: 4.33-10.25, P<0.01) and APACHE II score (odds ratio: 5.27, 95% confidence interval: 2.28-9.16, P<0.01) were the independent risk factors related to death in 65 patients on PSD 1. (4) The total areas under the ROC curves of pGSN level and APACHE II score for predicting death of 65 patients on PSD 1 were respectively 0.89 and 0.86, and 142 mg/L and 21 points were respectively chosen as the optimal threshold values, with sensitivity of 87% and 83% and specificity of 86% and 89%.CONCLUSIONS:For severe burn patients with sepsis, lowering of pGSN level and elevation of APACHE II score are obviously correlated with increase in case fatality rates. Monitoring the dynamic changes in pGSN level and APACHE II score during the early stage may be useful to predict the prognosis of severe burn patients with sepsis.

Sepsis is defined as severe systemic inflammation in response to invading pathogens, or an uncontrolled hyperinflammatory response, as mediated by the release of various proinflammatory mediators. Although some patients may die rapidly from septic shock accompanied by an overwhelming systemic inflammatory response syndrome (SIRS) triggered by a highly virulent pathogen, most patients survive the initial phase of sepsis, showing multiple organ damage days or weeks later. These patients often demonstrate signs of immune suppression accompanied by enhanced inflammation. Sepsis is a result of a complex process; there is interaction of various pathways, such as inflammation, immunity, coagulation, as well as the neuroendocrine system. This treatise is an attempt to provide a summary of several key regulatory mechanisms and to present the currently recognized molecular pathways that are involved in the pathogenesis of sepsis.

OBJECTIVETo improve the management of the early neurogenic pulmonary edema(NPE)in patients with non-traumatic cerebral hemorrhage.METHODSTotally 140 eligible patients with non-traumatic cerebral hemorrhage who were treated in the emergency department of our hospital from October 2008 to October 2014 were divided into two groups:NPE group(n=25)and non-NPE group(n=115). The clinical data were analyzed and compared.RESULTSAlthough the mean arterial pressure was similar between these two groups,the median pH and the bicarbonate ion(HCO(3)(-))were significantly lower in the NPE group than in the non-NPE group(pH:7.32 vs.7.39,P=0.002;HCO(3)(-),20.6 mmol/L vs.22.7 mmol/L,P=0.01). Multivariate regression analysis indicated that younger age and higher glucose level were significantly correlated with the early onset of NPE in the NPE group than in the non-NPE group(age:50.1 years vs.65.1 years,P=0.0008;glucose,15.4 mmol/L vs.10.78 mmol/L,P=0.001).There were only 3 patients in all with non-traumatic cerebral hemorrhage happened the fulminant NPE in 1 hour. Within 24 hours after patients visited the emergency room,the condition was improved in 20 of 25 patients in the NPE group. However,5 patients died,among whom 3 patients with fulminant NPE(onset within 1 hour)died due to acute respiratory distress syndrome and complicated with multiple organ failure,and 2 died of cerebral hernia.CONCLUSIONSNPE is a rare and severe complication in patients with non traumatic cerebral hemorrhage. The possibility of NPE should be considered in relatively young patients with higher glucose and lower blood pH value. Timely prevention and treatment can improve the outcomes.

With both in vivo and in vitro experiments, the present study was conducted to investigate the effect of regulatory T cell (Treg) on promoting T-lymphocyte apoptosis and its regulatory mechanism through transforming growth factor-beta (TGF-β1) signaling in mice. A murine model of polymicrobial sepsis was reproduced by cecal ligation and puncture (CLP); PC61 and anti-TGF-β antibodies were used to decrease counts of CD4(+)CD25(+) Tregs and inhibit TGF-β activity, respectively. Splenic CD4(+)CD25(+) Tregs and CD4(+)CD25(-) T cells were isolated. Phenotypes, including cytotoxic T-lymphocyte-associated antigen 4 (CTLA-4), forkhead/winged helix transcription factor p3 (Foxp3), and TGFβ1(m+), as well as the apoptotic rate of CD4(+)CD25(-) T cell, were analyzed by flow cytometry. Real-time reverse transcription-polymerase chain reaction was performed to determine mRNA expression of TGF-β1, and the expressions of Smad2/Smad3, Bcl-2 superfamily members of Bcl-2/Bim, cytochrome C, the mitochondrial membrane potential, and caspases in CD4(+)CD25(-) T cells were simultaneously determined. After treatment with PC61 or anti-TGF-β antibody, CTLA-4, Foxp3, and TGFβ1(m+) expressions of CD4(+)CD25(+) Tregs were markedly decreased in comparison to that of the CLP group and the apoptosis rate of CD4(+)CD25(-) T cells was significantly positively correlated with the expression of TGF-β1. Meanwhile, levels of P-Smad2/P-Smad3, proapoptotic protein Bim, cytochrome C, and activity of caspase-3, -8, -9 were downregulated, whereas the mitochondrial membrane potential and antiapoptotic protein Bcl-2 expression were restored. Taken together, our data indicated that the TGF-β1 signal could be partly involved in the apoptosis of CD4(+)CD25(-) T cells promoted by CD4(+)CD25(+) Tregs, therefore inhibition of TGF-β1 expression may provide a novel strategy for the improvement of host immunosuppression following sepsis.

Background Traumatic brain injury (TBI) is one of the leading causes of neurological disability. In this retrospective study, serum total cholinesterase (ChE) activities were analyzed in 188 patients for diagnostic as well as predictive values for mortality. Methods and Findings Within 72 hours after injury, serum ChE activities including both acetylcholinesterase and butyrylcholinesterase were measured. Disease severity was evaluated with Acute Physiology and Chronic Health Evaluation (APACHE) II score, Glasgow Coma Score, length of coma, post-traumatic amnesia and injury feature. Neurocognitive and functional scores were assessed using clinical records. Of 188 patients, 146 (77.7%) survived and 42 (22.3%) died within 90 days. Lower ChE activities were noted in the non-survivors vs. survivors (5.94±2.19 vs. 7.04±2.16 kU/L, p=0.023), in septic vs. non-infected patients (5.93±1.89 vs. 7.31±2.45 kU/L, p=0.0005) and in patients with extremely severe injury vs. mild injury (6.3±1.98 vs. 7.57±2.48 kU/L, p=0.049). The trajectories of serum ChE levels were also different between non-survivors and survivors, septic and non-infected patients, mild and severely injured patients, respectively. Admission ChE activities were closely correlated with blood cell counts, neurocognitive and functional scores both on admission and at discharge. Receiver operating characteristic analysis showed that the area under the curve for ChE was inferior to that for either APACHE II or white blood cell (WBC) count. However, at the optimal cutoff value of 5 kU/L, the sensitivity of ChE for correct prediction of 90-day mortality was 65.5% and the specificity was 86.4%. Kaplan-Meier analysis showed that lower ChE activity (<5 kU/L) was more closely correlated with poor survival than higher ChE activity (>5 kU/L) (p=0.04). After adjusting for other variables, ChE was identified as a borderline independent predictor for mortality as analyzed by Binary logistic regression (P=0.078). Conclusions Lowered ChE activity measured on admission appears to be associated with disease severity and outcome for TBI patients.