Identification and assignment of function to the genes of Aspergillus fumigatus expressed at 37 degrees C

Aspergillus fumigatus, a fungal pathogen, causes a spectrum of allergic and invasive disorders. In order to rapidly identify genes of this fungus relevant for pathogenesis and as potential antifungal drug targets, 125 expressed sequence tags (ESTs) were generated from 200 phage clones of a non-normalized cDNA library. Out of a novel 68 ESTs, 45 were assigned putative functions based on the sequence similarity. The identities of some of these genes suggest that they may be involved in pathogenesis or autoimmune reactions. Additional genes were identified that are possible targets for the development of antifungal drugs or that may be of use in diagnosing fungal infections.     

Effects of keratin filament disruption on exocrine pancreas-stimulated secretion and susceptibility to injury

Disruption or absence of hepatocyte keratins 8 and 18 is associated with chronic hepatitis, marked hepatocyte fragility, and a significant predisposition to stress-induced liver injury. In contrast, pancreatic keratin disruption in transgenic mice that express keratin 18 Arg89 --> Cys (K18C) is not associated with an obvious pancreatic pathology. We compared the effects of keratin filament disruption on pancreatic acini or acinar cell viability, and on cholecystokinin (CCK)-stimulated secretion, in transgenic mice that overexpress wild-type keratin 18 and harbor normal extended keratin filaments (TG2) and K18C mice. We also compared the response of these mice to pancreatitis induced by a choline-deficient ethionine-supplemented diet or by caerulein. Despite extensive cytoplasmic keratin filament disruption, the apicolateral keratin filament bundles appear intact in the acinar pancreas of K18C mice, as determined ultrastructurally and by light microscopy. No significant pancreatitis-associated histologic, serologic, or F-actin/keratin apicolateral redistribution differences were noted between TG2 and K18C mice. Acinar cell viability and yield after collagenase digestion were lower in K18C than in TG2 mice, but the yields of intact acini and their (125)I-CCK uptake and responses to CCK-stimulated secretion were similar. Our results indicate that keratin filament reorganization is a normal physiologic response to pancreatic cell injury, but an intact keratin cytoplasmic filament network is not as essential in protection from cell injury as in the liver. These findings raise the possibility that the abundant apicolateral acinar keratin filaments, which are not as evident in hepatocytes, may play the cytoprotective role that is seen in liver and other tissues. Alternatively, identical keratins may function differently in different tissues.     

Quantifying macrophage defects in type 1 diabetes

Macrophages from animals prone to autoimmune (type 1) diabetes differ from those of diabetes-resistant animals in processing and clearing apoptotic cells. Using in vitro time-course assays of the number of engulfed apoptotic cells observed within macrophages, we quantified these differences in non-obese diabetic (NOD) versus Balb/c mice. Simple models lead to several elementary parameter estimation techniques. We used these to compute approximate rates of macrophage engulfment and digestion of apoptotic cells from basic features of the data (such as initial rise-times, phagocytic index and percent phagocytosis). Combining these estimates with full fitting of a sequence of model variants to the data, we find that macrophages from normal (Balb/c) mice engulf apoptotic cells up to four times faster than macrophages from the diabetes-prone (NOD) mice. Further, Balb/c macrophages appear to undergo an activation step before achieving their high engulfment rate. In NOD macrophages, we did not see evidence for this activation step. Rates of digestion of engulfed apoptotic cells by macrophages are similar in both types. Since macrophage clearance is an important mechanism of disposal of self-antigen, these macrophage defects could potentially be a factor in predisposition to type 1 diabetes.     

The expression and immunoregulation of immune checkpoint molecule VISTA in autoimmune diseases and cancers

Immune checkpoint inhibitors (ICIs) and immunotherapy have proven to be a transformative therapy for many forms of cancer treatment. While many antibodies targeting the PD-1, PD-L1, and CTLA-4 pathways have been approved for clinical use by the FDA, it is clear that a single ICI is not sufficient to eradicate disease. ICI combination strategies are being extensively investigated to advance cancer treatment to next curative stage. Among the immune checkpoint inhibitors being actively investigated, the potential of VISTA (V-domain Ig suppressor of T cell activation), a unique B7 family member that functions as both ligand and receptor, is being actively pursued. This article summarizes the expression and immunomodulatory effects of VISTA in autoimmune diseases and cancer, and assesses its potential as an additional component of immune checkpoint cancer therapy.     

Mechanisms of Mitochondrial Damage in Keratinocytes by Pemphigus Vulgaris Antibodies

The development of nonhormonal treatment of pemphigus vulgaris (PV) has been hampered by a lack of clear understanding of the mechanisms leading to keratinocyte (KC) detachment and death in pemphigus. In this study, we sought to identify changes in the vital mitochondrial functions in KCs treated with the sera from PV patients and healthy donors. PV sera significantly increased proton leakage from KCs, suggesting that PV IgGs increase production of reactive oxygen species. Indeed, measurement of intracellular reactive oxygen species production showed a drastic increase of cell staining in response to treatment by PV sera, which was confirmed by FACS analysis. Exposure of KCs to PV sera also caused dramatic changes in the mitochondrial membrane potential detected with the JC-1 dye. These changes can trigger the mitochondria-mediated intrinsic apoptosis. Although sera from different PV patients elicited unique patterns of mitochondrial damage, the mitochondria-protecting drugs nicotinamide (also called niacinamide), minocycline, and cyclosporine A exhibited a uniform protective effect. Their therapeutic activity was validated in the passive transfer model of PV in neonatal BALB/c mice. The highest efficacy of mitochondrial protection of the combination of these drugs found in mitochondrial assay was consistent with the ability of the same drug combination to abolish acantholysis in mouse skin. These findings provide a theoretical background for clinical reports of the efficacy of mitochondria-protecting drugs in PV patients. Pharmacological protection of mitochondria and/or compensation of an altered mitochondrial function may therefore become a novel approach to development of personalized nonhormonal therapies of patients with this potentially lethal autoimmune blistering disease.     

弹性成像定量分析对自身免疫性肝病的诊断价值

目的:自身免疫性肝病的发病机理至今尚未明确,与多种疾病之间存在着联系,临床诊断具有一定的难度。本研究利用超声弹性成像技术定量分析自身免疫性肝病的病理特征,探讨该技术的诊断价值,为自身免疫性疾病的治疗提供诊断依据。方法:选取我院2011年3月-2013年6月收治的自身免疫性肝病患者182例,随机分为观察组和对照组。观察组98例患者采用弹性成像定量分析的方法进行诊断,对照组84例则采用常规病理学诊断。观察并比较两组患者的诊断准确率及诊断耗时。结果:观察组检出94例,诊断率为95.92%;对照组检出81例,诊断率为96.43%,两组诊断准确率比较无明显差异(P0.05)。观察组平均诊断耗时为5.6 h;对照组平均诊断耗时为11.6 h,观察组诊断时间比对照组短,差异具有统计学意义(P0.05)。结论:弹性成像定量分析对自身免疫性肝病的诊断准确率与病理活检诊断具有较好的一致性,且诊断耗时短、患者依从性好,值得在临床中进一步推广应用以辅助病理诊断,避免误诊或漏诊现象发生。     

不同营养方式治疗对重症急性胰腺炎患者预后的影响

目的：通过临床试验,}匕较不同营养方式治疗重症急性胰腺炎的效果,并分析不同方法对患者预后的影响。方法：选取我院在2011年11月至2012年11月收治的150例重症急性胰腺炎患者,随机分为三组：肠外营养组（PN）;肠内营养组（EN）;肠外＋肠内营养组（PN＋EN）,每组50例。仔细观察三组患者在住院接受营养治疗期间的情况,分别在第5天和第10天检测三组患者的血淀粉酶和免疫指标,并且对三组患者的住院时间、住院费用、消化道不良反应等指标进行比较。结果：在接受不同的营养支持后,（PN＋EN）组患者的血淀粉酶、免疫指标、住院时间、住院费用、消化道不良反应等指标可见明显下降;与PN组和EN组相比差异显著,有统计学意义（P〈0．05）。结论：重症急性胰腺炎患者接受营养支持对治疗其疾病非常重要,对重症胰腺炎患者实施肠外＋肠内的营养支持更加安全有效,便于患者吸收,有助于保护肠道粘膜屏障,降低感染率,改善患者的身体状况,非常值得临床上广泛推广应用。     

Effect of emodin on long non-coding RNA-mRNA networks in rats with severe acute pancreatitis-induced acute lung injury

Long non-coding RNAs (lncRNAs) contribute to disease pathogenesis and drug treatment effects. Both emodin and dexamethasone (DEX) have been used for treating severe acute pancreatitis-associated acute lung injury (SAP-ALI). However, lncRNA regulation networks related to SAP-ALI pathogenesis and drug treatment are unreported. In this study, lncRNAs and mRNAs in the lung tissue of SAP-ALI and control rats, with or without drug treatment (emodin or DEX), were assessed by RNA sequencing. Results showed both emodin and DEX were therapeutic for SAP-ALI and that mRNA and lncRNA levels differed between untreated and treated SAP-ALI rats. Gene expression profile relationships for emodin-treated and control rats were higher than DEX-treated and -untreated animals. By comparison of control and SAP-ALI animals, more up-regulated than down-regulated mRNAs and lncRNAs were observed with emodin treatment. For DEX treatment, more down-regulated than up-regulated mRNAs and lncRNAs were observed. Functional analysis demonstrated both up-regulated mRNA and co-expressed genes with up-regulated lncRNAs were enriched in inflammatory and immune response pathways. Further, emodin-associated lncRNAs and mRNAs co-expressed modules were different from those associated with DEX. Quantitative polymerase chain reaction demonstrates selected lncRNA and mRNA co-expressed modules were different in the lung tissue of emodin- and DEX-treated rats. Also, emodin had different effects compared with DEX on co-expression network of lncRNAs Rn60_7_1164.1 and AABR07062477.2 for the blue lncRNA module and Nrp1 for the green mRNA module. In conclusion, this study provides evidence that emodin may be a suitable alternative or complementary medicine for treating SAP-ALI.