血清降钙素原鉴别病毒性感染、一般性细菌感染和重症细菌性感染的临床应用价值

目的:研究血清中降钙素原在病毒性感染、一般性细菌感染和重症细菌性感染患者中的应用价值。方法:选择2014年6月~2016年12月在我院重症医学科进行诊治的感染患者60例,按照患者感染的严重程度分为病毒性感染患者(A组)15例,一般细菌感染组(B组)22例以及重症细菌性感染组(C组)23例。三组患者均采取纠正水电解质平衡、抗生素抗感染和营养支持等对症治疗,并在入院后第1、3、5、7 d检测患者的血清降钙素原水平,比较三组治疗前后血清降钙素原水平的差异,观察三组血清降钙素原≥0.5μg/L的例数。结果:B组血清降钙素原≥0.5μg/L的例数为20例,占90.91%,C组血清降钙素原≥0.5μg/L的例数为21例,占91.30%,B组和C组之间相比无明显差异(P0.05),但A组血清降钙素原≥0.5μg/L的例数为3例,占20.00%,明显低于B组和C组血清降钙素原≥0.5μg/L的例数,差异均具有统计学意义(P0.05);C组入院后第1、3、5、7 d的血清降钙素原水平均明显高于B组的血清降钙素原水平,差异均具有统计学意义(P0.05);重症细菌性感染患者中,血清降钙素原≥0.5μg/L的患者存活率明显低于血清降钙素原0.5μg/L的患者(P0.05)。结论:血清中降钙素原水平对于重症感染患者的临床诊断和治疗具有较好的应用价值,对于医生判断重症感染患者的预后情况具有很好的效果,值得临床广泛应用推广。     

Microarray Meta-Analysis Identifies Acute Lung Injury Biomarkers in Donor Lungs That Predict Development of Primary Graft Failure in Recipients

Objectives

To perform a meta-analysis of gene expression microarray data from animal studies of lung injury, and to identify an injury-specific gene expression signature capable of predicting the development of lung injury in humans.

Methods

We performed a microarray meta-analysis using 77 microarray chips across six platforms, two species and different animal lung injury models exposed to lung injury with or/and without mechanical ventilation. Individual gene chips were classified and grouped based on the strategy used to induce lung injury. Effect size (change in gene expression) was calculated between non-injurious and injurious conditions comparing two main strategies to pool chips: (1) one-hit and (2) two-hit lung injury models. A random effects model was used to integrate individual effect sizes calculated from each experiment. Classification models were built using the gene expression signatures generated by the meta-analysis to predict the development of lung injury in human lung transplant recipients.

Results

Two injury-specific lists of differentially expressed genes generated from our meta-analysis of lung injury models were validated using external data sets and prospective data from animal models of ventilator-induced lung injury (VILI). Pathway analysis of gene sets revealed that both new and previously implicated VILI-related pathways are enriched with differentially regulated genes. Classification model based on gene expression signatures identified in animal models of lung injury predicted development of primary graft failure (PGF) in lung transplant recipients with larger than 80% accuracy based upon injury profiles from transplant donors. We also found that better classifier performance can be achieved by using meta-analysis to identify differentially-expressed genes than using single study-based differential analysis.

Conclusion

Taken together, our data suggests that microarray analysis of gene expression data allows for the detection of “injury" gene predictors that can classify lung injury samples and identify patients at risk for clinically relevant lung injury complications.     

腹水降钙素原诊断肝硬化自发性腹膜炎的临床价值研究

目的:探讨腹水中降钙素原(PCT)诊断晚期肝硬化腹水并发自发性细菌性腹膜炎(SBP)的临床价值,并确定其参考值水平。方法:选择42例肝硬化失代偿期患者为研究对象(伴SBP22例,非SBP 20例),抽取其住院时、住院后24 h和48 h外周血及腹水标本各一次,进行腹水中有核细胞数计数,并采用免疫荧光层析法同时测定和比较其血清及腹水中PCT的含量。结果:22例伴SBP的患者血清和腹水PCT含量均明显高于20例不伴有SBP的患者(P0.01),而伴SBP的患者腹水PCT含量和同时间点血清PCT含量比较差异均无统计学意义(P0.05)。以入院时腹水PCT含量诊断SBP的ROC曲线的AUC为0.986,而血清PCT、腹水PMN计数的AUC分别为0.942、0.868;入院后24时腹水PCT和血清PCT诊断SBP的ROC曲线的AUC分别为0.998和0.986;入院后48时腹水PCT和血清PCT诊断SBP的ROC曲线的AUC为0.986和0.990。结论:腹水降钙素原可用于晚期肝硬化腹水并发SBP的诊断,且较血清降钙素原和腹水中有核细胞计数具有更高的诊断价值。入院时、入院后24 h和48 h时,腹水PCT大于0.565 ng/m L、0.545 ng/m L和0.410 ng/m L提示患SBP可能性大。     

Procalcitonin Levels Predict Acute Kidney Injury and Prognosis in Acute Pancreatitis: A Prospective Study

Background

Acute kidney injury (AKI) has been proposed as a leading cause of mortality for acute pancreatitis (AP) patients admitted to the intensive care unit (ICU). This study investigated the predictive value of procalcitonin (PCT) for AKI development and relevant prognosis in patients with AP, and compared PCT’s predictive power with that of other inflammation-related variables.

Methods

Between January 2011 and March 2013, we enrolled 305 cases with acute pancreatitis admitted to ICU. Serum levels of PCT, serum amyloid A (SAA), interleukin-6 (IL-6), and C reactive protein (CRP) were determined on admission. Serum PCT was tested in patients who developed AKI on the day of AKI occurrence and on either day 28 after occurrence (for survivors) or on the day of death (for those who died within 28 days).

Results

Serum PCT levels were 100-fold higher in the AKI group than in the non-AKI group on the day of ICU admission (p<0.05). The area under the receiver-operating characteristic (ROC) curve of PCT for predicting AKI was 0.986, which was superior to SAA, CRP, and IL-6 (p<0.05). ROC analysis revealed all variables tested had lower predictive performance for AKI prognosis. The average serum PCT level on day 28 (2.67 (0.89, 7.99) ng/ml) was significantly (p<0.0001) lower than on the day of AKI occurrence (43.71 (19.24,65.69) ng/ml) in survivors, but the serum PCT level on death (63.73 (34.22,94.30) ng/ml) was higher than on the day of AKI occurrence (37.55 (18.70,74.12) ng/ml) in non-survivors, although there was no significant difference between the two days in the latter group (p = 0.1365).

Conclusion

Serum PCT is superior to CRP, IL-6, and SAA for predicting the development of AKI in patients with AP, and also can be used for dynamic evaluation of AKI prognosis.     

Analysis of Serum Inflammatory Mediators Identifies Unique Dynamic Networks Associated with Death and Spontaneous Survival in Pediatric Acute Liver Failure

Background

Tools to predict death or spontaneous survival are necessary to inform liver transplantation (LTx) decisions in pediatric acute liver failure (PALF), but such tools are not available. Recent data suggest that immune/inflammatory dysregulation occurs in the setting of acute liver failure. We hypothesized that specific, dynamic, and measurable patterns of immune/inflammatory dysregulation will correlate with outcomes in PALF.

Methods

We assayed 26 inflammatory mediators on stored serum samples obtained from a convenience sample of 49 children in the PALF study group (PALFSG) collected within 7 days after enrollment. Outcomes were assessed within 21 days of enrollment consisting of spontaneous survivors, non-survivors, and LTx recipients. Data were subjected to statistical analysis, patient-specific Principal Component Analysis (PCA), and Dynamic Bayesian Network (DBN) inference.

Findings

Raw inflammatory mediator levels assessed over time did not distinguish among PALF outcomes. However, DBN analysis did reveal distinct interferon-gamma-related networks that distinguished spontaneous survivors from those who died. The network identified in LTx patients pre-transplant was more like that seen in spontaneous survivors than in those who died, a finding supported by PCA.

Interpretation

The application of DBN analysis of inflammatory mediators in this small patient sample appears to differentiate survivors from non-survivors in PALF. Patterns associated with LTx pre-transplant were more like those seen in spontaneous survivors than in those who died. DBN-based analyses might lead to a better prediction of outcome in PALF, and could also have more general utility in other complex diseases with an inflammatory etiology.     

Acute Liver Failure with Thyrotoxicosis Treated with Liver Transplantation

ObjectiveWe describe a young woman with previously undiagnosed thyrotoxicosis who presented with acute liver failure (ALF).MethodsWe present a case report and review the relevant literature.ResultsAn extensive evaluation excluded possible causes of ALF other than thyrotoxicosis. The management of thyrotoxicosis posed several unique challenges in the setting of ALF, particularly because we did not want to use potentially hepatotoxic thionamides. The patient was treated with prednisone and propranolol and was started on potassium iodide when she was listed for liver transplantation. She underwent an uncomplicated liver transplant and subsequent thyroidectomy and is doing well.ConclusionThis well-characterized case describes thyrotoxicosis as a possible cause of ALF after thoroughly excluding other possible causes and illustrates the challenges of simultaneously managing both disorders. To our knowledge, this is the first report of ALF possibly resulting from untreated thyrotoxicosis that was successfully treated with liver transplantation. (Endocr Pract. 2013;19:e57-e60)     

The Decisive Porlevels in Haloalkane-Mediated Liver Cell Injury

The model hepatotoxine carbon tetrachloride (CC1₄) was used to study haloalkane free radical-induced lipid peroxidation in isolated rat hepatocytes at steady state oxygen partial pressures (pO,) between 0.2 and IOOmmHg. Equilibrium oxygen conditions were achieved by using an oxystat system.

Monitoring of hepatocellular oxygen uptake, malondialdehyde-formation and low-level chemilumine-scence during incubations of CC1₄-supplemented hepatocytes indicated a drastic stimulation of lipid peroxidation at p0₂-levels between 1 and lOmmHg. Above and below this pO₂-region the potency of CC1₄ to induce lipid peroxidation sharply decreased. The evaluation of cellular damages by determining trypan blue exclusion and lactate dehydrogenase leakage revealed that in the presence of CC1₄ hepatocellular injury was significantly increased at those pO₂-levels which were optimal for CC1₄-mediated lipid peroxidation.

The present results demonstrate that CC1₄ is a potent inducer of lipid peroxidation also in the intact hepatocyte, provided that the p0₂ is maintained at distinct low levels. The coincidence of lipid peroxidation and loss of cell viability at the same pO,-range provides further evidence for the assumption that the haloalkane-mediated liver cell injury is due to a peroxidative process which primarily occurs at the hypoxic end of the physiological pO, -levels (1-70 mmHg) in liver.     

Circulating microRNAs Reveal Time Course of Organ Injury in a Porcine Model of Acetaminophen-Induced Acute Liver Failure

ConclusionsMicroRNAs were released passively into the circulation in response to acetaminophen-induced cellular damage. A significant increase in global microRNA was detectable prior to significant increases in miR122, miR192 and miR124-1, which were associated with clinical evidence of liver, kidney and brain injury respectively.     

D-gal致大鼠急性肝损伤过程中肝卵圆细胞增殖及迁延

目的用D-gal建立大鼠急性肝损伤模型,观察肝损伤后再生过程中肝卵圆细胞的增殖和迁延。方法建立大鼠急性肝损伤模型,于第1、3、7和14天分别取肝组织,分别行病理、免疫组织化学,观察卵圆细胞的分布迁移情况,并取第7天肝组织进行组织电镜观察汇管区新增生细胞超微结构。结果病理切片显示肝细胞变性坏死程度以第7天和第14天为主,出现新增生的细胞。免疫组化示随时间阳性细胞明显增多,分布于汇管区,并向小叶中心迁移,形成大量的胆小管,并有部分向坏死区迁移。透射电镜有新生内源性细胞,小于成熟的肝细胞,细胞器较少,有细胞紧密连接,以数个细胞排列成小胆管,与免疫组化一致。结论在大鼠急性肝损伤时HOC被活化、增殖,并向肝小叶中心迁移,全程参与了肝再生过程。     

A Mammalian Cell Regulatory Agent,CeReS-18, Inhibits Yeast Cell Proliferation But Not Bacterial Replication

A cell regulatory sialoglycopeptide, CeReS-18, purified from intact bovine cerebral cortex cells, has exhibited the capability of reversibly inhibiting cellular DNA synthesis and the proliferation of a wide array of mammalian cells. In the present study, the effect of CeReS-18 on the proliferation of bacterial (Bacillus cereus and Escherichia coli) and yeast (Saccharomyces cerevisiae and Schizosaccharomyces pombe) cells was investigated. The results showed that replication and viability of the bacterial cells were not affected by CeReS-18 at any concentration tested, including 15-fold higher than that used for inhibiting mouse 3T3 cell proliferation. In contrast to bacterial cells, CeReS-18 was able to inhibit the replication of yeast cells, in a concentration-dependent, reversible manner, and the addition of calcium to the culture medium could abrogate the inhibitory effect of CeReS-18. A cytotoxic effect of CeReS-18 on both yeast cell species was observed when it was applied at higher concentrations. Received: 13 March 2002 / Accepted: 22 July 2002     

Sake Yeast Suppresses Acute Alcohol-Induced Liver Injury in Mice

Brewer’s and baker’s yeasts appear to have components that protect from liver injury. Whether sake yeast, Saccharomyces cerevisiae Kyokai no. 9, also has a hepatoprotective effect has not been examined. Here we show that sake yeast suppresses acute alcoholic liver injury in mice. Male C57BL/6 mice that had been fed a diet containing 1% sake yeast for two weeks received three doses of ethanol (5 g/kg BW). In the mice fed sake yeast, ethanol-induced increases in triglyceride (TG) and glutamate pyruvate transaminase (GPT) were significantly attenuated and hepatic steatosis was improved. In addition, sake yeast-fed mice showed a smaller decrease in hepatic S-adenosylmethionine (SAM) level and a smaller increase in plasma homocysteine (Hcy) level after ethanol treatment than the control mice, suggesting that a disorder of methonine metabolism in the liver caused by ethanol was relieved by sake yeast. These results indicate that sake yeast protects against alcoholic liver injury through maintenance of methionine metabolism in the liver.     

Liver Cell Calcium Homeostasis in Carbon Tetrachloride Liver Cell Injury: New Data With Fura21

The calcium fluorescent probe fura2 was used to measure concentration of free calcium in the cytosol of isolated rat hepatocytes in suspension. The resting level in untreated hepatocytes was 121 nM. On addition of CCl₄ at a concentration of 0.5 mM, cytosolic free calcium rose sharply and reached a statistically significant (P<0.05) steady plateau level of about 190 nM within five minutes. With a concentration of 1.0 mM CCl₄, cytosolic free calcium rose within ten minutes to a plateau level of about 200 nM. Use of fura2, along with the capacity of Mn²⁺ ions to effectively quench fura2 fluorescence, provided the basis for a simple and decisive method to determine whether the added CCl₄ was permeabilizing the hepatocyte plasma membrane by direct solvent action. It was found that up to a concentration of 1.0 mM, CCl₄ did not permeabilize the plasma membrane, but direct attack on the plasma membrane was unequivocally demonstrated for concentrations of 2 mM CCl₄ and above. Finally, an hypothesis is presented for resolution of the puzzling dilemma that emerged from the observation, reported from two laboratories, that CCl₄ can rapidly mobilize liver mitochondrial calcium despite the well-known relative resistance of these organelles to the damaging effects of this toxic agent.     

Myeloid Cell Sirtuin-1 Expression Does Not Alter Host Immune Responses to Gram-Negative Endotoxemia or Gram-Positive Bacterial Infection

The role of sirtuin-1 (SIRT1) in innate immunity, and in particular the influence of SIRT1 on antimicrobial defense against infection, has yet to be reported but is important to define since SIRT1 inhibitors are being investigated as therapeutic agents in the treatment of cancer, Huntington’s disease, and autoimmune diseases. Given the therapeutic potential of SIRT1 suppression, we sought to characterize the role of SIRT1 in host defense. Utilizing both pharmacologic methods and a genetic knockout, we demonstrate that SIRT1 expression has little influence on macrophage and neutrophil antimicrobial functions. Myeloid SIRT1 expression does not change mortality in gram-negative toxin-induced shock or gram-positive bacteremia, suggesting that therapeutic suppression of SIRT1 may be done safely without suppression of myeloid cell-specific immune responses to severe bacterial infections.     

Mast Cell Stabilization Alleviates Acute Lung Injury after Orthotopic Autologous Liver Transplantation in Rats by Downregulating Inflammation

Background

Acute lung injury (ALI) is one of the most severe complications after orthotopic liver transplantation. Amplified inflammatory response after transplantation contributes to the process of ALI, but the mechanism underlying inflammation activation is not completely understood. We have demonstrated that mast cell stabilization attenuated inflammation and ALI in a rodent intestine ischemia/reperfusion model. We hypothesized that upregulation of inflammation triggered by mast cell activation may be involve in ALI after liver transplantation.

Methods

Adult male Sprague–Dawley rats received orthotopic autologous liver transplantation (OALT) and were executed 4, 8, 16, and 24 h after OALT. The rats were pretreated with the mast cell stabilizers cromolyn sodium or ketotifen 15 min before OALT and executed 8 h after OALT. Lung tissues and arterial blood were collected to evaluate lung injury_. β-hexosaminidase and mast cell tryptase levels were assessed to determine the activation of mast cells. Tumor necrosis factor α (TNF-α), interleukin (IL)-1β and IL-6 in serum and lung tissue were analyzed by enzyme-linked immunosorbent assay. Nuclear factor-kappa B (NF-κB) p65 translocation was assessed by Western blot.

Results

The rats that underwent OALT exhibited severe pulmonary damage with a high wet-to-dry ratio, low partial pressure of oxygen, and low precursor surfactant protein C levels, which corresponded to the significant elevation of pro-inflammatory cytokines, β-hexosaminidase, and tryptase levels in serum and lung tissues. The severity of ALI progressed and maximized 8 h after OALT. Mast cell stabilization significantly inhibited the activation of mast cells, downregulated pro-inflammatory cytokine levels and translocation of NF-κB, and attenuated OALT-induced ALI.

Conclusions

Mast cell activation amplified inflammation and played an important role in the process of post-OALT related ALI.     

Causes and Outcomes of Acute Liver Failure in China

Objectives

No extensive investigation has been performed and thus no representative data are available regarding acute liver failure (ALF) in China. This study aims to investigate the causes and outcomes of ALF in China and establish a prognostic model.

Methods

Patients diagnosed as ALF in seven hospitals in different areas of China from January 2007 to December 2012 were retrospectively selected.

Results

Of the 177 patients included in this study, 112 (63.28%) eventually died. The common causes of ALF were drug toxicity (43.50%), indeterminate etiology (29.38%) and acute viral hepatitis (11.30%). Additionally, traditional Chinese herbs predominated in the causes of drug-induced ALF (30/77). No patients in this study received liver transplantation. In the established model for predicting death in ALF, four variables were finally selected out, including age (P=0.01), the entry hepatic encephalopathy grade (P=0.04), international normalized ratio (P<0.01) and arterial blood ammonia (P=0.02). Using a threshold value of 0.5683, this model had a sensitivity of 95.24% and a specificity of 91.30%.

Conclusions

Traditional Chinese medicine was a major cause of ALF in China. The spontaneous mortality of ALF was high, whereas the rate of liver transplantation was significantly low. The established prognostic model of ALF had superior sensitivity and specificity.     

Oroxylin A Accelerates Liver Regeneration in CCI4-Induced Acute Liver Injury Mice

Introduction

Based on the previous research that oroxylin A can suppress inflammation, we investigated the hepatoprotective role of oroxylin A against CCl₄-induced liver damage in mice and then studied the possible alteration of the activities of cytokine signaling participating in liver regeneration. Wild type (WT) mice were orally administrated with oroxylin A (60 mg/kg) for 4 days after CCl₄ injection, the anti-inflammatory effects of oroxylin A were assessed directly by hepatic histology and indirectly by measuring serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and Albumin. Proliferating cell nuclear antigen (PCNA) staining was performed to evaluate the role of oroxylin A in promoting hepatocyte proliferation. Serum IL-1β, TNF-α, IL-6 and IL-1Ra levels were measured by enzyme-linked immunosorbent assay (ELISA) and liver HGF, EGF, TNF-α, IL-6, IL-1Ra and IL-1β gene expression was determined by quantitative real-time PCR. The data indicated that the IL-6 and TNF-α mRNA of oroxylin A administered group significantly increased higher than the control within 12 hours after CCl₄ treatment. Meanwhile, oroxylin A significantly enhanced the expression of IL-1Ra at the early phase, which indicated that oroxylin A could facilitate the initiating events in liver regeneration by increasing IL-1Ra which acts as an Acute-Phase Protein (APP). In addition, a lethal CCl₄-induced acute liver failure model offers a survival benefit in oroxylin A treated WT mice. However, oroxylin A could not significantly improve the percent survival of IL-1RI^−/− mice with a lethal CCl₄-induced acute liver failure.

Conclusions

Our study confirmed that oroxylin A could strongly promote liver structural remodeling and functional recovery through IL-1Ra/IL-1RI signaling pathway. All these results support the possibility of oroxylin A being a therapeutic candidate for acute liver injury.     

In Vitro Alterations Do Not Reflect a Requirement for Host Cell Cycle Progression during Plasmodium Liver Stage Infection

Prior to invading nonreplicative erythrocytes, Plasmodium parasites undergo their first obligate step in the mammalian host inside hepatocytes, where each sporozoite replicates to generate thousands of merozoites. While normally quiescent, hepatocytes retain proliferative capacity and can readily reenter the cell cycle in response to diverse stimuli. Many intracellular pathogens, including protozoan parasites, manipulate the cell cycle progression of their host cells for their own benefit, but it is not known whether the hepatocyte cell cycle plays a role during Plasmodium liver stage infection. Here, we show that Plasmodium parasites can be observed in mitotic hepatoma cells throughout liver stage development, where they initially reduce the likelihood of mitosis and ultimately lead to significant acquisition of a binucleate phenotype. However, hepatoma cells pharmacologically arrested in S phase still support robust and complete Plasmodium liver stage development, which thus does not require cell cycle progression in the infected cell in vitro. Furthermore, murine hepatocytes remain quiescent throughout in vivo infection with either Plasmodium berghei or Plasmodium yoelii, as do Plasmodium falciparum-infected primary human hepatocytes, demonstrating that the rapid and prodigious growth of liver stage parasites is accomplished independent of host hepatocyte cell cycle progression during natural infection.     

Acute Unilateral Vestibular Failure Does Not Cause Spatial Hemineglect

Objectives

Visuo-spatial neglect and vestibular disorders have common clinical findings and involve the same cortical areas. We questioned (1) whether visuo-spatial hemineglect is not only a disorder of spatial attention but may also reflect a disorder of higher cortical vestibular function and (2) whether a vestibular tone imbalance due to an acute peripheral dysfunction can also cause symptoms of neglect or extinction. Therefore, patients with an acute unilateral peripheral vestibular failure (VF) were tested for symptoms of hemineglect.

Methods

Twenty-eight patients with acute VF were assessed for signs of vestibular deficits and spatial neglect using clinical measures and various common standardized paper-pencil tests. Neglect severity was evaluated further with the Center of Cancellation method. Pathological neglect test scores were correlated with the degree of vestibular dysfunction determined by the subjective visual vertical and caloric testing.

Results

Three patients showed isolated pathological scores in one or the other neglect test, either ipsilesionally or contralesionally to the VF. None of the patients fulfilled the diagnostic criteria of spatial hemineglect or extinction.

Conclusions

A vestibular tone imbalance due to unilateral failure of the vestibular endorgan does not cause spatial hemineglect, but evidence indicates it causes mild attentional deficits in both visual hemifields.