C4亚型人肠道病毒71型的小鼠肌肉适应株对小鼠具有致死毒力

目的人肠道病毒71型（EV71）感染婴幼儿可导致手足口病,偶尔伴随有严重的并发症。建立EV71的小鼠模型是深入研究病毒感染的致病机制、进行疫苗和药物评价的基础。方法将EV71的临床分离株在小鼠骨骼肌中进行系列传代,得到了小鼠的肌肉适应株MP10。使用MP10经腹腔注射感染10日龄ICR小鼠,对感染小鼠的症状、病毒复制和病理进行了监测。结果与临床分离株相比,MP10对人横纹肌瘤细胞（rhabdomyosarcoma cell,RD细胞）的感染力提高,并且对小鼠的毒力增强,MP10感染10日龄小鼠可导致其瘫痪和死亡。病毒主要在骨骼肌中复制,并引发大面积的坏死性肌炎,同时神经组织未观察到病变。病理分析发现：感染小鼠体内与呼吸运动相关的肌肉均发生严重的坏死性肌炎,推测此类肌肉的坏死会影响小鼠的呼吸功能,从而成为导致小鼠死亡的因素之一。结论建立了C4亚型EV71毒株感染10日龄ICR小鼠的模型,该模型可作为研究EV71感染的致病机制、以及疫苗和药物评价的工具。     

The emerging role of MIF in septic shock and infection

The protein mediator described originally as macrophage migration inhibitory factor (MIF) has been re-discovered recently to be both a novel pituitary hormone and a pro-inflammatory, macrophage-derived cytokine. Emerging studies indicate that MIF plays a pivotal role not only in endotoxic shock but also in the host response to a variety of acute and chronic infections.Abbreviations MIF Macrophage migration Inhibitory Factor     

Mitochondria Redistribution in Enterovirus A71 Infected Cells and Its Effect on Virus Replication

Enterovirus A71(EV-A71) is one of the main causative agents of hand, foot and mouth disease(HFMD) and it also causes severe neurologic complications in infected children. The interactions between some viruses and the host mitochondria are crucial for virus replication and pathogenicity. In this study, it was observed that EV-A71 infection resulted in a perinuclear redistribution of the mitochondria. The mitochondria rearrangement was found to require the microtubule network, the dynein complex and a low cytosolic calcium concentration. Subsequently, the EV-A71 non-structural protein 2 BC was identified as the viral protein capable of inducing mitochondria clustering. The protein was found localized on mitochondria and interacted with the mitochondrial Rho GTPase 1(RHOT1) that is a key protein required for attachment between the mitochondria and the motor proteins, which are responsible for the control of mitochondria movement.Additionally, suppressing mitochondria clustering by treating cells with nocodazole, EHNA, thapsigargin or A23187 consistently inhibited EV-A71 replication, indicating that mitochondria recruitment played a crucial role in the EV-A71 life cycle. This study identified a novel function of the EV-A71 2 BC protein and provided a potential model for the regulation of mitochondrial motility in EV-A71 infection.     

Urine analysis using FTIR spectroscopy: A study on healthy adults and children

Urine spectra from 108 healthy volunteers are studied by attenuated total refraction-Fourier transform infrared (ATR-FTIR) spectroscopy. The spectral features are correlated with observable urine components. The variation of spectra within a healthy population is quantified and a library of reference spectra is constructed. Using the band assignments, these spectra are compared with both age-wise and gender-wise. Children show the least intensity variations compared to both adult groups. Young adults show the highest variation, particularly in the 1650 to 1400 cm⁻¹ and 1200 to 900 cm⁻¹ regions. These results indicate the importance of the size of the control group in comparative studies utilizing FTIR. Age-wise comparisons reveal that phosphate and sulfate excretion decreases with age, and that the variance of phosphate among individuals is higher with adults. As for gender-wise comparisons, females show a slightly higher citrate content at 1390 cm⁻¹ regardless of the age and they show a higher variance in the 1200 to 1000 cm⁻¹ region when compared to men.     

Changes of microcirculation in healthy volunteers and patients with septic shock in Xining

Objective: The purpose of this study is to investigate the characteristic of microcirculation in healthy volunteers and patients with septic shock in both Xining(2 260 m) and Nanjing(10 m). Methods: A total of 62 cases, 33 healthy volunteers, 22 cases in Xining,(2 260 m above sea level) and 11 cases in Nanjing(10 m above sea level); and 29 septic shock, 13 cases in Xining and 16 cases in Nanjing were collected. The total vessel density(TVD), perfused vessel density(PVD), proportion of perfused vessel(PPV) and microcirculation flow index(MFI) of both healthy volunteers and septic shock had been investigated by using sidestream dark field(SDF). Analyzed and managed the image data by using AVA3.0 software. Results: In the healthy volunteers in Xining area(22 cases),the volume of TVD(15.59 ± 2.58 mm/mm~2), PVD(15.58 ± 2.58 mm/mm~2) and PPV(96.60% ± 4.63%) were significant higher than the volume of TVD(10.0 ± 2.10 mm/mm~2), PVD(10.81 ± 2.38 mm/mm~2) and PPV(84.24% ± 8.00%) of the volunteers(11 cases) in Nanjing(11 cases). But the MFI(2.17 ± 0.31) of the healthy volunteers in the Xining was significant lower(P0.05) than the MFI(3.21 ± 0.34) in the healthy volunteers of Nanjing. In the septic shock group(13 cases) in the Xining, the volume of TVD(5.44 ± 1.94 mm/mm~2), PVD(4.18 ± 1.61 mm/mm~2), PPV(42.14%± 5.38%) and MFI(1.05 ± 0.32) compared with the volume of the healthy volunteers in Xining, the TVD(15.59 ± 2.58 mm/mm~2), PVD(5.58 ± 2.58 mm/mm~2), PPV(96.60% ± 4.63%) and MFI(2.17 ± 0.30) were significant lower(P0.05). In the healthy volunteers compare with septic shock group in Nanjing area, the TVD(6.80±1.72 vs 10.00±2.10, P0.05), PVD(5.86±1.58 vs10.81±2.38,P0.05), PPV(45.42±4.86 vs 84.24±4.86, P0.05), MFI(1.28±0.28 vs 3.21±0.34 P0.05), there was significant decreased. In the septic shock group in the Xining compared with the septic shock in Nanjing, there was no significant difference. 10 of 13 patients with septic shock were survived in Xining. 13 of 16 patients with septic shock were survived in Nanjing. Conclusion: The changes of physiological and pathophysiological characteristic in microcirculation induced by hypoxia would be useful for clinical treatment of septic shock at high altitude.     

Analysis of peripheral blood lymphocyte subsets and prognosis in patients with septic shock

The purpose of the study is to study the relationship between peripheral blood lymphocyte subset proportion and prognosis in patients with septic shock. Fifty‐two patients with septic shock, admitted to the intensive care unit between March 2007 and December 2010, were enrolled in this study. Peripheral blood lymphocyte subset proportions were measured using flow cytometry. The percentage of CD3⁺CD4⁺ T lymphocytes and CD19⁺ lymphocytes, CD4⁺/CD8⁺ T cell ratio were substantially lower in patients with septic shock compared to the control group (P < 0.01). The percentage of CD3⁺CD8⁺ T lymphocytes did not differ significantly between the two groups (P > 0.05). The percentage of CD16⁺CD56⁺ lymphocytes was higher in patients with septic shock than in the control group (P < 0.01). Compared with the survivor group, the percentage of CD3⁺CD4⁺ T lymphocytes and CD19⁺ lymphocytes, CD4⁺/CD8⁺ T cell ratio were clearly lower in the non‐survivor group (P < 0.01). There was no difference in the percentage of CD3⁺CD8⁺ T lymphocytes between the non‐survivor and survivor groups (P > 0.05). The percentage of CD16⁺CD56⁺ lymphocytes was higher in the non‐survivor group than in the survivor group (P < 0.05). The total maximum SOFA score and the delta SOFA score were much higher in the non‐survivor group than in the survivor group (P < 0.01). Immune imbalance occurs in patients with septic shock. Peripheral blood lymphocyte subset proportion and SOFA scores can be used to assess the treatment and prognosis of septic shock.     

Cerebrospinal fluid Th1/Th2 cytokine profiles in children with enterovirus 71‐associated meningoencephalitis

Enterovirus 71 (EV71) infection can cause severe neurological complications including meningoencephalitis (ME) in some patients with hand, foot and mouth disease (HFMD). However, to date no studies have reported changes in cytokine concentrations and their correlations with clinical variables in patients with ME following EV71 infection. In this study, responses of Th1/Th2 cytokine, including IL‐2, IL‐4, IL‐6, IL‐10, TNF‐α and IFN‐γ, in cerebrospinal fluid (CSF) from patients with EV71‐related HFMD with ME and patients with febrile convulsions (FC) were analyzed using cytometric bead array technology. It was found that CSF IL‐6 and IFN‐γ concentrations were significantly higher in patients with EV71‐related ME than in those with FC. Additionally, both CSF IL‐6 and IFN‐γ concentrations were correlated with CSF cytology, fever duration and duration of hospital stay. More interestingly, a positive correlation between CSF IL‐6 and IFN‐γ concentrations was observed. Finally, receiver operating characteristic analysis revealed that when a cutoff value of 9.40 pg/mL was set for IL‐6, the sensitivity and specificity were 84.5% and 85.5%, respectively, for discriminating EV71‐related ME from FC. In conclusion, IL‐6 and IFN‐γ may be associated with EV71‐induced neuropathology.     

Differential interferon pathway gene expression patterns in Rhabdomyosarcoma cells during Enterovirus 71 or Coxsackievirus A16 infection

Exposure of cells to type I interferon (IFN) induces an antiviral state that prevents viral infection, but viruses can utilize multiple tactics to antagonize the host immune system. Enterovirus 71 (EV71) and Coxsackievirus A16 (CVA16) are two major pathogens that cause hand, foot, and mouth disease (HFMD), which is prevalent among children. We found that both EV71 and CA16 have different reactions to type I IFN pretreatment and induction patterns of type I IFN on Rhabdomyosarcoma (RD) cells. Further, a human-α and β IFN PCR array was employed to analyze the expressions of 84 genes related to the type I IFN pathway. We found significant up-regulation of multiple genes in the presence of type I IFN and differential regulation patterns during EV71 or CA16 infection in RD cells. For instance, EV71 infection repressed the JAK-STAT signaling pathway and interferon-stimulated gene (ISG) expression, whereas CA16 infection normally triggers the JAK-STAT pathway, leading to the expression of ISGs. Taken together, this study provides a comprehensive view of the differential impacts of EV71 and CA16 infection on 84 genes in the IFN pathway, shedding light on the different resistances of these viruses to type I IFN treatment and cytotoxic effects in RD cells.     

Interleukin-6 and procalcitonin in children with sepsis and septic shock

Objectives. To examine the behavior of interleukin-6 (IL-6) and procalcitonin (PCT) and verify whether they can be used to differentiate children with septic conditions. Methods. Septic children aged between 28 days and 14 years, prospectively enrolled from 01/2004 to 12/2005, were divided into sepsis (SG; n = 47) and septic shock (SSG; n = 43) groups. IL-6 and PCT were measured at admission (T0) and 12 h later (T12h). PCT results were classed as: 0.5 ng/mL = sepsis unlikely; 0.5 to <2 = sepsis possible; 2 to <10 = systemic inflammation; 10 = septic shock. Results. Ninety children were included. At T0, there was a higher frequency of SSG with higher PCT compared with SG [SSG: 30 (69.7%) > SG: 14 (29.8%); p < 0.05]. Similar results were observed at T12h. PRISM was significantly higher for SSG patients with higher PCT than SG patients. At T0, IL-6 levels were higher in SSG [SSG: 213.10 (10.85–396.70) > SG: 63.21 (0.86–409.82); p = 0.001], but not statistically different at T12h. IL-6 levels positively correlated with PRISM score in SSG patients at admission (p = 0.001; r = 0.86). Conclusion. PCT and IL-6 appear to be helpful in early assessment of pediatric sepsis, are of diagnostic value at admission, and are related to disease severity.     

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.     

EV71 infection correlates with viral IgG preexisting at pharyngolaryngeal mucosa in children

Enterovirus 71(EV71) infection causes severe central nervous system damage, particularly for children under the age of 5 years old, which remains a major public health burden worldwide. Clinical data released that children may be repeatedly infected by different members in enterovirus and get even worsen. Mucosa, especially epithelium of alimentary canal, was considered the primary site of EV71 infection. It has been elusive whether the preexsiting viral antibody in mucosa plays a role in EV71 infection. To answer this question, we respectively measured viral antibody response and EV71 RNA copy number of one hundred throat swab specimens from clinically confirmed EV71-infected children. The results released that low-level of mucosal Ig G antibody against EV71 broadly existed in young population. More importantly, it further elucidated that the children with mucosal preexsiting EV71 Ig G were prone to be infected, which suggested a former viral Ig G mediated enhancement of viral infection in vivo.     

Soluble TREM-1 as a diagnostic and prognostic biomarker in patients with septic shock: an observational clinical study

Objectives: The impact of TREM-1-mediated inflammation was investigated in different inflammatory settings.

Methods: Secondary analyses of an observational clinical pilot study, including 60 patients with septic shock, 30 postoperative controls and 30 healthy volunteers.

Results: Plasma levels of sTREM-1 were found to identify patients with septic shock more effectively than procalcitonin and C-reactive protein. Moreover, sTREM-1 was identified to be an early predictor for survival in patients with septic shock.

Conclusion: Due to its diagnostic as well as prognostic value in sepsis syndrome, implementation of sTREM-1 measurements in routine diagnostics should be taken into account.     

Outbreak of central nervous system disease associated with hand,foot, and mouth disease in Japan during the summer of 2000: detection and molecular epidemiology of enterovirus 71

Few outbreaks of the serious enterovirus 71 (EV71) infections, which affect the central nervous system (CNS), had been reported in Japan before 2000. During June through August 2000, a patient died of pulmonary edema caused by brainstem encephalitis accompanied by EV71-induced hand, foot, and mouth disease (HFMD), and many patients complicated by serious CNS disease, including paralysis, were hospitalized in a restricted area in Hyogo Prefecture, Japan (K-area). During the same period, endemics of HFMD were reported in other areas in Hyogo Prefecture, where EV71 was isolated from HFMD patients, but few patients developed aseptic meningitis. The isolations of EV71 from K-area patients were difficult with the use of Vero cells, so the strains were isolated by use of GL37 cells; Vero cells, however, could isolate EV71 strains from other areas in Hyogo Prefecture. We sequenced VP4 coding regions of these EV71 isolates and found that the isolates from K-area had the same sequence, which, except for one isolate, was different from the sequences of EV71 strains isolated from other areas of Hyogo Prefecture. Although these results were not enough to state that EV71 from K-area was a virulent strain, it seemed reasonable to conclude that serious CNS diseases in K-area were caused by EV71 because it was the only infectious agent detected in the inpatients of K-area.     

Comparing the prognostic performance of ASSIST to interleukin-6 and procalcitonin in patients with severe sepsis or septic shock

Abstract

Context: We recently derived and validated a multi-biomarker-based model (ASSIST) to stratify patients with sepsis based on initial mortality risk.

Objective: The objective of this study was to compare the performance of ASSIST to interleukin-6 (IL6) and procalcitonin (PCT).

Methods: The area-under-the-receiver operating characteristic curve for predicting 28-d mortality using ASSIST was compared with that of IL6 (n?=?452) and PCT (n?=?235).

Results: The area under the curve for ASSIST was greater than that of IL6 and PCT.

Conclusions: ASSIST estimated the probability of mortality more reliably than IL6 and PCT in this cohort of patients with sepsis.     

Oxidative stress and septic shock: metabolic aspects of oxygen-derived free radicals generated in the liver during endotoxemia

This review describes the role of oxidative stress caused by endotoxin challenge in sepsis or septic shock symptoms. We observed that endotoxin injection resulted in lipid peroxide formation and membrane damage (near 60-150 kDa) in the livers of experimental animals, causing decreased levels of scavengers or quenchers of free radicals. The administration of alpha-tocopherol completely prevented injury to the liver plasma membrane caused by endotoxin, and suggested that lipid peroxidation by free radicals might occur in a tissue ischemic state, probably by disseminated intravascular coagulation (DIC), in endotoxemia. In mice, depression of Ca(2+)-ATPase activity in the liver plasma membrane may contribute to the membrane damage caused by endotoxin, and the increase of [Ca(2+)](i) in the liver cytoplasm may partially explain the oxidative stress that occurs in endotoxemia. It seems that endotoxin-induced free radical formation is regulated by Ca(2+) mobilization. Moreover, we have suggested that the oxidative stress caused by endotoxin may be due, at least in part, to the changes in endogenous zinc or selenium regulation during endotoxemia. Interestingly, the extent of TNF-alpha-induced oxidative stress may be the result of a synergism between TNF-alpha and gut-derived endotoxin. It is likely that bacterial or endotoxin translocation plays a significant role in TNF-alpha-induced septic shock. On the other hand, although nitric oxide (NO) has been implicated in the pathogenesis of vascular hyporesponsiveness and hypotension in septic shock in our experimental model, it is unlikely that NO plays a significant role in liver injury caused by free radical generation in endotoxemia.     

细胞膜转运分子ABCC3和SLC7A7在肠道病毒A71型感染中作用的初步研究

肠道病毒A71型(enterovirus A71,EV-A71)是导致手足口病(hand-foot-mouth disease,HFMD)的主要病原体之一,目前对其治疗尚无特异高效的抗病毒药物.研究表明,细胞膜转运相关分子参与病毒的入侵、复制以及感染性子代病毒颗粒的释放.为寻找宿主中可有效抑制EV-A71感染的细胞膜转...     

Cold shock response in Lactococcus lactis ssp. diacetylactis: a comparison of the protection generated by brief pre-treatment at less severe temperatures

Acquired freeze–thaw tolerance was investigated for Lactococcus lactis ssp. diacetylactis. Pre-treatment of microorganisms at less severe temperatures to initiate cold tolerance gave L. lactis ssp. diacetylactis improved cell viability after successive freezings and thawings. The ability of cells to survive freezing–thawing was dependent on factors experienced prior to freezing. Factors affecting lactic acid bacteria survival during freezing–thawing cycles include different diluents, growth phase, and cold temperatures. Viability experiments showed that this strain displaying cold shock cryotolerance had an improved survival capacity in stationary phase. The plasmid contents of lactic acid bacteria isolated from different types, strains DRC-2 and DRC-2C, were examined and compared with the plasmid contents of culture collection strains both before and after cold shock treatment. Using agarose gel electrophoresis, no obvious correlation between the cold shock response and the number of plasmids in the cell could be observed.     

Mortality after fluid bolus in children with shock due to sepsis or severe infection: a systematic review and meta-analysis

Introduction

Sepsis is one of the leading causes of childhood mortality, yet controversy surrounds the current treatment approach. We conducted a systematic review to assess the evidence base for fluid resuscitation in the treatment of children with shock due to sepsis or severe infection.

Methods

We searched 3 databases for randomized trials, quasi-randomized trials, and controlled before-after studies assessing children with septic shock in which at least one group was treated with bolus fluids. The primary outcome was mortality at 48 hours. Assessment of methodological quality followed the GRADE criteria. Relative risks (RRs) and 95% confidence intervals (CI) were calculated and data pooled using fixed-effects method.

Results

13 studies met our inclusion criteria. No bolus has significantly better mortality outcomes at 48 hours for children with general septic shock (RR 0.69; 95%CI 0.54–0.89), and children with malaria (RR 0.64; 95%CI 0.45–0.91) when compared to giving any bolus. This result is largely driven by a single, high quality trial (the FEAST trial). There is no evidence investigating bolus vs no bolus in children with Dengue fever or severe malnutrition. Colloid and crystalloid boluses were found to have similar effects on mortality across all sub-groups (general septic shock, malaria, Dengue fever, and severe malnutrition).

Conclusions

The majority of all randomized evidence to date comes from the FEAST trial, which found that fluid boluses were harmful compared to no bolus. Simple algorithms are needed to support health-care providers in the triage of patients to determine who could potentially be harmed by the provision of bolus fluids, and who will benefit.