小儿脓毒症休克相关基因的筛选及网络构建

为探究脓毒症休克与SIRS的差异表达基因及网络的构建,筛选潜在的核心基因,从GEO数据库下载相关基因表达谱GSE26378,数据分为脓毒症休克与SIRS各29个样本,通过在线软件GCBI对其进行标准化及差异基因筛选;对差异基因进行GO分析;基于KEGG进行功能通路分析以及基因信号网络分析;差异基因共表达网络分析。结果表明:两组中总共有1 456个基因被识别为差异基因(P0.05),与SIRS组相比,脓毒症休克组中有条859条下调基因,597条上调基因。GO功能富集分析显示差异基因主要参与了细胞周期、细胞免疫、细胞代谢。KEGG功能通路分析显示差异基因主要参与了MAPK信号通路、P53信号通路、wnt信号通路、细胞凋亡信号通路,细胞周期受体信号通路等。共表达分析发现基因CCNB1、NUSAP1、OIP5、SHCBP1、ZWINT、TOP2A、DLGAP5等位于网络中央部位,而基因信号网络分析发现基因PLCB1、PIK3CA、STAT3、CAMK2D、PRKCB、CREB1位于网络核心。基因芯片分析有助于发现脓毒症休克与SIRS患儿外周血单核细胞在转录组学上的改变,而生物信息学网络分析有助于发现潜在的靶点。     

Cytokine and molecular networks in sepsis cases: a network biology approach

Background

Sepsis is a life-threatening condition of organ dysfunction caused by a dysregulated host immune response to infection. We performed network analysis of cytokine molecules and compared network structures between a systematic inflammatory response syndrome (SIRS) or normal control (NC) group and a sepsis group.

Results

We recruited SIRS (n = 33) and sepsis (n = 89) patients from electronic medical records (EMR) according to whether data on PCT, CRP, interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-9, IL-10, IL-12p70, IL-13, IL-17, IL-22, TNF-α, and IFN-γ levels were available. From the public GEO dataset, GSE66099, GSE9960, GSE95233, GSE57065 were downloaded. Genes corresponding to 15 molecules were extracted from an expression array. A correlation matrix was formed for the 15 molecules and statistically significant molecular pairs were used as pairs for network analysis of coexpression. The number of molecular or gene expression pairs significantly correlated among the SIRS or control and sepsis groups are as follows for datasets: EMR, 15 and 15; GEO66099-1, 13 and 15; GEO9960, 13 and 11; GSE95233, 13 and 8; GSE66099-2, 15 and 14; GSE57065, 14 and 13, respectively. Network analysis revealed that network diameter, number of nodes and shortest path were equal to or lower in the sepsis group.

Conclusions

The coexpression network in sepsis patients was relatively small sized and had lower shortest paths compared with the SIRS group or healthy control group. Cytokines with one degree (k = 1) are increased in sepsis group compared with SIRS or healthy control group. IL-9 and IL-2 were not included in network of sepsis group indicating that these cytokines showed no correlation with other cytokines. These data might imply that cytokines tend to be dysregulated in the sepsis group compared to that of SIRS or normal control groups

     

Is the endogenous peroxyl-radical scavenging capacity of plasma protective in systemic inflammatory disorders in humans?

Systemic inflammatory response syndrome (SIRS) in humans is associated with heightened intravascular oxidative stress. The clinical significance of plasma endogenous antioxidative capability in SIRS remains undetermined. Time-sequence changes of plasma total radical-trapping antioxidant parameter (TRAP) and its components were measured in 135 patients with various clinical conditions leading to SIRS. The results were correlated with clinical parameters. Plasma TRAP significantly depressed upon diagnosis of SIRS (SIRS vs. healthy subjects (n = 50), 605.7 +/- 20.4 vs. 803.4 +/- 30.8 microM Trolox equivalent, p <.001). In survivors (n = 86), TRAP declined further during the course of SIRS, followed by a mild recovery at the end of follow-up. General linear mixed model analysis revealed that uric acid, vitamin C, vitamin E and unidentified antioxidants contributed to most of the changes in TRAP (each factor p <.001). In nonsurvivors (n = 49), TRAP increased steadily until death, and the increase was predominantly the result of the increased contribution of bilirubin (p <.01). Higher TRAP levels were not correlated with diminished blood oxidants formation (r = -0.13, p.05), lower intensity of lipid peroxidation (r = 0.261, p <.05) or lesser disease severity of SIRS. The results do not support the hypothesis that the endogenous peroxyl radical scavenging ability of plasma plays a protective role in the course of SIRS.     

Glycyrrhizin inhibits the manifestations of anti-inflammatory responses that appear in association with systemic inflammatory response syndrome (SIRS)-like reactions

In association with the systemic inflammatory response syndrome (SIRS), anti-inflammatory response syndrome is commonly manifested in patients with trauma, burn injury, and after major surgery. These patients are increasingly susceptible to infection with various pathogens due to the excessive release of anti-inflammatory cytokines from anti-inflammatory effector cells. Recently, CC-chemokine ligand 2 (CCL2) found in the sera of mice with pancreatitis was identified as an active molecule for SIRS-associated anti-inflammatory response manifestation. Also, the inhibitory activity of glycyrrhizin (GL) on CCL2 production was reported. Therefore, the effect of GL on SIRS-associated anti-inflammatory response manifestation was investigated in a murine SIRS model. Without any stimulation, splenic T cells from mice 5 days after SIRS induction produced cytokines associated with anti-inflammatory response manifestation. However, these cytokines were not produced by splenic T cells from SIRS mice previously treated with GL. In dual-chamber transwells, IL-4-producing cells were generated from normal T cells cultured with peripheral blood polymorphonuclear neutrophils (PMN) from SIRS mice. However, IL-4-producing cells were not generated from normal T cells in transwell cultures performed with PMN from GL-treated SIRS mice. CCL2 was produced by PMN from SIRS mice, while this chemokine was not demonstrated in cultures of PMN from SIRS mice treated with GL. These results indicate that GL has the capacity to suppress SIRS-associated anti-inflammatory response manifestation through the inhibition of CCL2 production by PMN.     

肾盂灌注冲洗并发全身炎症反应综合征猪模型的建立

目的:建立一种肾镜下肾盂灌注冲洗并发全身炎症反应综合征(SIRS)的猪模型。方法:家猪10头,随机分为假手术组和模型组,每组按时间点分为基础状态(Basic)、术后0h、术后6h、术后12h和术后24h 5个亚组。猪肾穿刺造瘘,利用自体盲肠内容物加入肾盂冲洗液中,模拟经皮肾镜取石术(PCNL)持续肾盂灌注冲洗1h。每个时间点观测肛温(T)、呼吸(RR)、心率(HR);采血行血细胞计数分析(WBC、RBC、PLT)、肾功能(Cr、BUN、β2-MG)测定、血清IL-6、IL-10、TNF-α含量测定;用光镜和电镜观察肾脏组织病理变化。结果:模型组在术后6 h-24 h各项指标与假手术组相比变化明显,差异有统计学意义(P<0.05或<0.01)。模型组肾脏组织光镜和电镜下观察病理切片均较假手术组有明显的形态学改变。结论:本模型较好地模拟了临床病人PCNL并发SIRS的病理生理过程,为进一步研究其发病机制和预防治疗具有重要意义。     

The effect of waning immunity on long-term behaviour of stochastic models for the spread of infection

In stochastic modelling of infectious spread, it is often assumed that infection confers permanent immunity, a susceptible-infective-removed (SIR) model. We show how results concerning long-term (endemic) behaviour may be extended to a susceptible-infective-removed-susceptible (SIRS) model, in which immunity is temporary. Since the full SIRS model with demography is rather intractable, we also consider two simpler models: the susceptible-infective-susceptible (SIS) model with demography, in which there is no immunity; and the SIRS model in a closed population. For each model, we first analyse a deterministic model, then approximate the quasi-stationary distribution (equilibrium distribution conditional upon non-extinction of infection) using a moment closure technique. We look in particular at the effect of the immune period upon infection prevalence and upon time to fade-out of infection. Our main findings are that a shorter average immune period leads to higher infection prevalence in quasi-stationarity, and to longer persistence of infection in the population.     

Site of action of a soluble immune response suppressor (SIRS) produced by concanavalin A-activated spleen cells.

The cellular site of action of SIRS, a soluble immune response suppressor released by Con A-activated spleen cells which suppresses antibody responses to heterologous erythrocytes by murine spleen cells in vitro, was investigated. Exposure of spleen cells to SIRS for 2 hr at 37 degrees C or 1 hr at 4 degrees C was sufficient to suppress 5-day antibody responses in vitro. Similar exposure of splenic or peritoneal exudate macrophages to SIRS also suppressed antibody responses by untreated splenic lymphoid cells; exposure of splenic lymphoid cells to SIRS was without effect. SIRS did not act via T cells which might have contaminated the macrophage preparations. SIRS-mediated suppression could be partially overcome by an excess of normal peritoneal exudate macrophages, but not by an excess of T or B cells. These data indicate that the target cell of SIRS activity is the macrophage. The results are discussed in the context of macrophage functions that could be affected by SIRS.     

Biological expressions of lymphocyte activation. V. Characterization of a soluble immune response suppressor (SIRS) produced by concanavalin A-activated spleen cells.

Supernatant fluids from murine spleen cell cultures incubated with concanavalin A for 48 hr contain a factor(s), soluble immune response suppressor (SIRS), which suppresses plaque-forming cell responses to sheep erythrocytes by murine spleen cells in vitro. In the present studies, some of the biochemical and biophysical properties of SIRS were investigated. SIRS was non-dialysable; the suppressive activity was stable at 56 degrees C for 30 min, but was destroyed by treatment at 70 degrees C for 30 min, 80 degrees C for 10 min, or at pH 2. The suppressive activity was not absorbed by the stimulating antigen, SRBC, or antisera against murine IgG or mu-chain, suggesting that SIRS does not contain immunoglobulin determinants. Murine spleen and thymus, but not kidney cells, however, absorbed SIRS activity. Enzyme treatments revealed that SIRS was resistant to DNase and RNase, but was destroyed by trypsin and chymotrypsin. In gel filtration with Sephadex G-100, SIRS activity eluted in the fraction corresponding to m.w. in the range between 48,000 and 67,000. With polyacrylamide gel electrophoresis, SIRS activity migrated in the region cathodal to albumin. Isopycnic centrifugation in a cesium chloride gradient suggested that SIRS is a glycoprotein. These supernatant fluids with SIRS activity were also found to contain macrophage migration inhibitory factor (MIF). In the experiments using gel filtration, polyacrylamide gel electrophoresis, and isopycnic centrifugation to fractionate supernatant fluids, SIRS and MIF activity were found in the same fractions, and to date we have been unable to dissociate definitively SIRS activity from MIF activity.     

Mathematical Model of BCG Immunotherapy in Superficial Bladder Cancer

Immunotherapy with Bacillus Calmette–Guérin (BCG)—an attenuated strain of Mycobacterium bovis (M. bovis) used for anti tuberculosis immunization—is a clinically established procedure for the treatment of superficial bladder cancer. However, the mode of action has not yet been fully elucidated, despite much extensive biological experience. The purpose of this paper is to develop a first mathematical model that describes tumor-immune interactions in the bladder as a result of BCG therapy. A mathematical analysis of the ODE model identifies multiple equilibrium points, their stability properties, and bifurcation points. Intriguing regimes of bistability are identified in which treatment has potential to result in a tumor-free equilibrium or a full-blown tumor depending only on initial conditions. Attention is given to estimating parameters and validating the model using published data taken from in vitro, mouse and human studies. The model makes clear that intensity of immunotherapy must be kept in limited bounds. While small treatment levels may fail to clear the tumor, a treatment that is too large can lead to an over-stimulated immune system having dangerous side effects for the patient.     

Plasma levels of F-actin and F:G-actin ratio as potential new biomarkers in patients with septic shock

Objective: To compare plasma levels of F-actin, G-actin and thymosin beta 4 (TB4) in humans with septic shock, noninfectious systemic inflammatory response syndrome (SIRS) and healthy controls.

Results: F-actin was significantly elevated in septic shock as compared with noninfectious SIRS and healthy controls. G-actin levels were greatest in the noninfectious SIRS group but significantly elevated in septic shock as compared with healthy controls. TB4 was not detectable in the septic shock or noninfectious SIRS group above the assay’s lowest detection range (78?ng/ml).

Conclusions: F-actin is significantly elevated in patients with septic shock as compared with noninfectious SIRS. F-actin and the F:G-actin ratio are potential biomarkers for the diagnosis of septic shock.     

基于SNP共表达网络肝癌分子分型及预后分析

基于SNP突变数据与mRNA表达谱关联分析,构建一种肝癌分子分型方法并对比不同分型预后的差异,并对不同分型肝癌的发生发展机制进一步研究。首先通过TCGA数据库收集359例肝细胞癌患者的SNP突变数据和mRNA表达数据,采用Wilcoxon秩和检验,筛选突变后差异表达基因,并通过生物信息学工具String和Cytoscape构建差异表达基因的蛋白互作网络,筛选连接度最高的10个Hub基因。利用Consensus Cluster Plus软件包,基于Hub基因mRNA表达水平构建NMF分子分型模型,再结合生存数据评估各分型患者的预后。最后利用加权基因共表达网络分析(WGCNA),识别与肝癌分子分型相关的模块,并针对关键模块的基因进行通路富集,从而对不同分型肝癌的基因表达谱进行比较。结果：NMF模型将肝癌分为高危、低危2个分型,其中CDKN2A和FOXO1基因对分型贡献度高。生存分析显示低危组患者的生存情况显著优于高危组,高危组富集多个与肿瘤细胞侵蚀、转移、复发过程相关的信号通路,低危组则与细胞周期和胰液分泌相关。本研究在无先验性信息的前提下,基于突变后显著差异表达的Hub基因表达水平构建的...     

Effects of a disease affecting a predator on the dynamics of a predator-prey system

We study the effects of a disease affecting a predator on the dynamics of a predator-prey system. We couple an SIRS model applied to the predator population, to a Lotka-Volterra model. The SIRS model describes the spread of the disease in a predator population subdivided into susceptible, infected and removed individuals. The Lotka-Volterra model describes the predator-prey interactions. We consider two time scales, a fast one for the disease and a comparatively slow one for predator-prey interactions and for predator mortality. We use the classical “aggregation method” in order to obtain a reduced equivalent model. We show that there are two possible asymptotic behaviors: either the predator population dies out and the prey tends to its carrying capacity, or the predator and prey coexist. In this latter case, the predator population tends either to a “disease-free” or to a “disease-endemic” state. Moreover, the total predator density in the disease-endemic state is greater than the predator density in the “disease-free” equilibrium (DFE).     

Targeted metabolomics for discrimination of systemic inflammatory disorders in critically ill patients

The occurrence of systemic inflammatory response syndrome (SIRS) remains a major problem in intensive care units with high morbidity and mortality. The differentiation between noninfectious and infectious etiologies of this disorder is challenging in routine clinical practice. Many biomarkers have been suggested for this purpose; however, sensitivity and specificity even of high-ranking biomarkers remain insufficient. Recently, metabolic profiling has attracted interest for biomarker discovery. The objective of this study was to identify metabolic biomarkers for differentiation of SIRS/sepsis. A total of 186 meta-bolites comprising six analyte classes were determined in 143 patients (74 SIRS, 69 sepsis) by LC-MS/MS. Two markers (C10:1 and PCaaC32:0) revealed significantly higher concentrations in sepsis. A classification model comprising these markers resulted in 80% and 70% correct classifications in a training set and a test set, respectively.This study demonstrates that acylcarnitines and glycerophosphatidylcholines may be helpful for differentiation of infectious from noninfectious systemic inflammation due to their significantly higher concentration in sepsis patients. Considering the well known pathophysiological relevance of lipid induction by bacterial components, metabolites as identified in this study are promising biomarker candidates in the differential diagnosis of SIRS and sepsis.     

脓毒症患儿内生吗啡动态变化及临床意义

目的:检测脓毒症患儿血清内生吗啡(EM)动态变化,探讨脓毒症患儿血清EM水平的临床意义。方法:选择2017年6月-2018年2月华中科技大学同济医学院附属同济医院收治的30例伴有严重脓毒症或脓毒症休克的患儿作为脓毒症组,30例全身炎症反应综合征(SIRS)患儿作为SIRS组,30例健康儿童作为对照组。脓毒症组、SIRS组及对照组分别于入组后第1、3、6、9天抽取外周静脉血,采用免疫荧光法检测血清降钙素原(PCT)水平,采用酶联免疫吸附试验(ELISA)测定血清EM水平,采用流式细胞术(FCM)检测细胞免疫功能。结果:SIRS组、脓毒症组患儿的第1、3、6天血清PCT水平显著高于对照组,且随时间延长PCT水平逐渐降低,至第9天降至正常水平(P0.05),而脓毒症组与SIRS组间血清PCT水平无统计学差异(P0.05)。脓毒症组第1、3、6天血清EM水平均高于SIRS组(P0.05),第9天两组血清EM水平比较差异无统计学意义(P0.05)。脓毒症组EM水平随时间延长而降低,至第9天降至SIRS组的水平(P0.05)。与SIRS组相比,第1天脓毒症组的CD3~+T细胞数量增多(P0.05),两组CD4~+、CD8~+T细胞数量、CD4~+/CD8~+比例比较无统计学差异(P0.05)。结论:脓毒症患儿中血清EM水平较高,有可能作为诊断脓毒症特异性较高的生物标志物。     

A role for histamine type II (H-2) receptor binding in production of the lymphokine, soluble immune response suppressor (SIRS)

Soluble immune response suppressor (SIRS) is an immunosuppressive protein produced by human and murine suppressor cells activated by a variety of agents. Because histamine has been reported to activate suppressor cells, the possibility that it also induced SIRS production was investigated. Human lymphocytes treated with 10(-4) M histamine for less than 1 hr released a suppressive substance into culture supernatants that was physically, functionally and antigenically similar to human SIRS. Cimetidine and ranitidine, structurally distinct histamine type II (H-2) receptor antagonists, prevented histamine-induced SIRS production. In further experiments, suppression of human polyclonal IgM PFC responses by Con A and interferons, substances that activate the SIRS pathway, was inhibited by H-2 receptor antagonists. Activation of lymphocytes to produce SIRS by Con A or interferons was blocked by cimetidine or ranitidine. These data demonstrate that production of SIRS is induced by histamine, and raise the possibility that H-2 receptor binding may play a role in the SIRS pathway.     

Fundamental-applied aspects of systemic inflammation in terms of physiologic and typical pathological process

The concept of systemic inflammatory response syndrome (SIRS) by sepsis as well as quality of SIRS criteria, classification, and PIRO system has been a subject to analytical criticism in terms of theory of physiologic and typical pathological process. It has been disclosed SIRS can be considered only as the syndrome, that solves particular clinical tasks, but not as a basic model of pathogenesis of critical states. In authors' opinion it is more correctly to discuss systemic inflammation as a typical pathologic process that appears as a complex of one or another "resuscitation" syndrome in a clinical course.     

A Time Since Recovery Model with Varying Rates of Loss of Immunity

For many infectious diseases, immunity wanes over time. The majority of SIRS models assume that this loss of immunity takes place at a constant rate. We study temporary immunity within a SIRS model structure if the rate of loss of immunity can depend on the time since recovery from disease. We determine the conditions under which the endemic steady state becomes unstable and periodic oscillations set in, showing that a fairly rapid change between slow and rapid immunity loss is necessary to produce oscillations.     

Biomarkers in critically ill patients with systemic inflammatory response syndrome or sepsis supplemented with high-dose selenium

ObjectiveLow levels of selenium (Se) and glutathione peroxidase (GSHPx), a key selenoenzyme, were documented in systemic inflammatory response syndrome (SIRS) and sepsis, both associated with high mortality. Se supplementation had mixed effects on outcome. We hypothesized that Se supplementation could have a different impact on biomarkers and 28-day mortality in patients with SIRS vs. sepsis.MethodsAdult patients with SIRS or sepsis were randomized to either high-dose (Se+, n = 75) or standard-dose (Se−, n = 75) Se supplementation. Plasma Se, whole blood GSHPx activity, C-reactive protein (CRP), procalcitonin (PCT), prealbumin, albumin and cholesterol levels were measured serially up to day 14.ResultsThere was no difference in mortality between Se− (24/75) vs. Se+ group (19/75; p = 0.367) or between SIRS and septic patients (8/26 vs. 35/124; p = 0.794). There was a trend to reduced mortality in SIRS patients in the Se+ vs. Se− group (p = 0.084). Plasma Se levels increased in the Se+ group only in patients with sepsis but not in patients with SIRS. Plasma Se levels correlated with GSHPx. In SIRS/Se+ group, Se correlated only with GSHPx. In SIRS/Se− group, Se correlated with cholesterol but not with other biomarkers. In sepsis patients, Se levels correlated with cholesterol, GSHPx and prealbumin. Cholesterol levels were higher in survivors in the Se− group.ConclusionsSe levels correlated with GSHPx activity and other nutritional biomarkers with significant differences between SIRS and sepsis groups. High-dose Se supplementation did not affect mortality but a strong trend to decreased mortality in SIRS patients warrants further studies in this population.     

Inference of Type-Specific HPV Transmissibility,Progression and Clearance Rates: A Mathematical Modelling Approach

Quantifying rates governing the clearance of Human Papillomavirus (HPV) and its progression to clinical disease, together with viral transmissibility and the duration of naturally-acquired immunity, is essential in estimating the impact of vaccination programmes and screening or testing regimes. However, the complex natural history of HPV makes this difficult. We infer the viral transmissibility, rate of waning natural immunity and rates of progression and clearance of infection of 13 high-risk and 2 non-oncogenic HPV types, making use of a number of rich datasets from Sweden. Estimates of viral transmissibility, clearance of initial infection and waning immunity were derived in a Bayesian framework by fitting a susceptible-infectious-recovered-susceptible (SIRS) transmission model to age- and type-specific HPV prevalence data from both a cross-sectional study and a randomised controlled trial (RCT) of primary HPV screening. The models fitted well, but over-estimated the prevalence of four high-risk types with respect to the data. Three of these types (HPV-33, -35 and -58) are among the most closely related phylogenetically to the most prevalent HPV-16. The fourth (HPV-45) is the most closely related to HPV-18; the second most prevalent type. We suggest that this may be an indicator of cross-immunity. Rates of progression and clearance of clinical lesions were additionally estimated from longitudinal data gathered as part of the same RCT. Our estimates of progression and clearance rates are consistent with the findings of survival analysis studies and we extend the literature by estimating progression and clearance rates for non-16 and non-18 high-risk types. We anticipate that such type-specific estimates will be useful in the parameterisation of further models and in developing our understanding of HPV natural history.