Yeast-based automated high-throughput screens to identify anti-parasitic lead compounds

We have developed a robust, fully automated anti-parasitic drug-screening method that selects compounds specifically targeting parasite enzymes and not their host counterparts, thus allowing the early elimination of compounds with potential side effects. Our yeast system permits multiple parasite targets to be assayed in parallel owing to the strains’ expression of different fluorescent proteins. A strain expressing the human target is included in the multiplexed screen to exclude compounds that do not discriminate between host and parasite enzymes. This form of assay has the advantages of using known targets and not requiring the in vitro culture of parasites. We performed automated screens for inhibitors of parasite dihydrofolate reductases, N-myristoyltransferases and phosphoglycerate kinases, finding specific inhibitors of parasite targets. We found that our ‘hits’ have significant structural similarities to compounds with in vitro anti-parasitic activity, validating our screens and suggesting targets for hits identified in parasite-based assays. Finally, we demonstrate a 60 per cent success rate for our hit compounds in killing or severely inhibiting the growth of Trypanosoma brucei, the causative agent of African sleeping sickness.     

Pseudomonas fluorescens,a potential bacterial antagonist to control plant diseases

Abstract

Fluorescent Pseudomonads belong to plant Growth Promoting Rhizobacteria (PGPR), the important group of bacteria that play a major role in the plant growth promotion, induced systemic resistance, biological control of pathogens etc. Many strains of Pseudomonas fluorescens are known to enhance plant growth promotion and reduce severity of various diseases. The efficacy of bacterial antagonists in controlling fungal diseases was often better as alone, and sometimes in combination with fungicides. The present review refers to occurrence, distribution, mechanism, growth requirements of P. fluorescens and diseases controlled by the bacterial antagonist in different agricultural and horticultural crops were discussed. The literature in this review helps in future research programmes that aim to promote P. fluorescens as a potential bio-pesticide for augmentative biological control of many diseases of agriculture and horticultural importance.     

Quantitative analysis of site-specific N-glycans on sera haptoglobin βchain in liver diseases

The site-specific characterization of N-glycans in glycopro- teins with the potential of clinical application is important. In our previous report, the overall N-glycans of sera haptoglobin （Hp） β chain were found to be different in liver diseases. Hp β chain contains four potential sites of N-glycosylation. In this study, we investigated the potential change of N-glycans on Hp β chain in a site-specific fashion. Sera Hp β chain in healthy individuals as well as patients with hepatitis B virus （HBV）, liver cirrhosis （LC） and hepatocellular carcinoma （HCC） were purified, digested and subjected to liquid chromatography-electro- spray ionization-higher energy collision dissociation mass spectrometry, which allowed identification and structure determination of the glycopeptide, as well as the relative quantification of glycans present on each glycopeptide. The quantitative results revealed that the sialylation of NLFLN207HSEN211 ATAK and the fucosylated structure at all glycopeptides increased significantly in LC and HCC patients compared with those in HBV patients and healthy individuals. A set of different N-glycan patterns of Hp β chain in various liver diseases has been determined. Thus, the sialylated and fucosylated glycoforms of Hp β chain might be related to early hepatocarcinogenesis and also might be useful as novel differential markers for LC and HCC patients.     

单气囊小肠镜在诊断胶囊内镜检查阴性的可疑小肠疾病患者中的应用价值

目的：探讨单气囊小肠镜（single-balloonenteroscop,SBE）在胶囊内镜检查阴性的可疑小肠疾病患者中的应用价值。方法：选取在我院行胶囊内镜检查无异常发现,后行单气囊小肠镜检查的可疑小肠疾病患者24例,分析后者对胶囊内镜检查阴性可疑小肠疾病患者的阳性发现率和病因的分布特点。结果：24例行SBE检查者有22例（91．7％）获得成功,2例失败,10例被检出阳性病变,其中间质瘤5例,小肠息肉2例,过敏性紫癜1例,血管畸形2例,SBE对胶囊内镜检查阴性的患者小肠疾病的再检出率为41．7％。结论：胶囊内镜和SBE在小肠疾病的诊断上有着各自的优缺点,对于胶囊内镜检查阴性的可疑小肠疾病患者进一步行SBE检查有助于确诊。     

不同剂量丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性变的临床研究

目的：探讨采用不同剂量的丹参注射液联合波尼松龙治疗口腔粘膜下纤维性病的治疗效果,为今后的治疗提供更多的依据。方法：选择从2010年1月至2013年1月期间在我院口腔科治疗的100例口腔粘膜下纤维性病患者,根据门诊号,随机将患者分为低剂量组、次低剂量组、中剂量组、高剂量组和对照组,每组各20例,低剂量组、次低剂量组、中剂量组、高剂量组,分别使用不同剂量丹参注射液联合波尼松龙治疗,对照组单纯使用波尼松龙治疗,观察治疗一个疗程后患者口腔粘膜情况及张口度。结果：中剂量组和高剂量组情况改善要明显好于低剂量、次低剂量组、对照组,差异具有显著性（P〈0．05）。结论：丹参注射液联合泼尼松龙治疗口腔粘膜下纤维性病疗效令人满意,其中低剂量丹参注射液便有效果,一定范围内剂量越高,疗效越好,值得在临床推f,     

过氧化物酶体增殖物激活受体α的研究进展

过氧化物酶体增殖物激活受体（Peroxisome proliferator activated receptors,PPARs）作为核受体超家族的一员,其作用广泛,可调节脂肪细胞因子表达、抑制炎症因子、改善胰岛素抵抗等。PPARs有三种亚型,分别是：PPARα、PPARβ／δ和PPARγ。其中PPARα是PPARs最主要的亚型,主要分布在肝脏中。PPARα由不饱和脂肪酸或贝特类降脂药物等配体活化后形成异二聚体,调控靶基因的表达,发挥生物学功能。PPARα参与调节肝脏脂质吸收、脂肪酸氧化、酮体生成、胆固醇代谢等脂代谢过程,以及糖代谢、炎症反应和细胞增殖等,与脂肪性肝病、肝脏炎症反应、乙肝病毒复制和肝癌等肝脏疾病密切相关。本文对PPARα的结构、作用机制、生物学功能及其与肝脏疾病的关系进行综述。PPARα作为肝脏疾病一个新的治疗靶点,阐明其与肝脏疾病发生机制之间的关系,有助于为肝脏疾病的治疗提供新的途径。     

1991-2010年宁夏水稻病虫害发生特征与经济损失分析

水稻病虫害发生种类繁多、暴发频繁,是威胁宁夏水稻稳产、高产的重要因素,但对其变化趋势与实际危害损失不清楚.基于宁夏水稻统计数据、稻田覆盖类型遥感数据和水稻产量数据,分析1991年至2010年宁夏水稻病虫害发生特征与经济损失情况.结果表明:1991年到2010年期间,宁夏水稻病害、虫害年均发生面积分别为5.3万hm2和2.0万hm2,其年均防治面积分别为8.4万hm2和1.6万hm2;水稻病害的发生程度下降了23.10％,防治程度上升了77.30％.20年间宁夏水稻虫害的发生面积、发生程度、防治面积和防治程度均显著增加.防治水稻病害、虫害后,分别挽回稻谷为2.67万吨、0.28万吨,其挽回损失量在20年期间分别增加了55.15％、2775.0％,表明水稻病虫害防治意义重大.由于气候变化等诸多因子,导致1991年到2010年宁夏水稻病害、虫害年均造成的实际稻谷损失量分别为0.71万吨与0.13万吨,水稻病害实际损失量在0值附件波动、虫害呈现波动增加趋势,说明水稻病害防控效果好、虫害的防控还有提升的空间.从全区各市县分布来看,水稻病虫害主要分布在宁夏的银北地区.为有效地防止或减少病虫害对水稻产量的损失,应加强农田景观变化和气候变化等对水稻病虫害发生与灾变的风险评估和监测预警,开展区域性水稻病虫害综合治理研究,并建立相应的防控新对策与技术体系.     

孝感城区中老年对慢性病及社区管理现状认知情况的调查研究

目的:调查孝感城区中老年对慢性病及其社区管理现状认知情况。方法:选择2010年7月至2015年7月在孝感城区3个社区的500名中老年人作为研究对象,利用自拟调查问卷对所有受试者进行数据调查,分析孝感城区中老年对慢性病的认知情况,及对慢性病社区管理现状的认知情况,分析影响慢性病认知的因素。结果:在慢性病种类中,孝感城区中老年人对于高血压和糖尿病,及风湿性关节炎与慢性支气管炎的认知率较高,而对于癌症、偏瘫及血栓的认知率较低,差异有统计学意义(P0.05)。在慢性病的危险因素中,孝感城区中老年人对于缺乏锻炼、吸烟及膳食不合理的认知率明显较高,而对于酗酒、缺乏保健知识及悲观情绪的认知率明显较低,差异有统计学意义(P0.05)。在慢性病社区管理现状上,孝感城区中老年对于指导其改变生活习惯的认知率较高,为62.00%,而在进行三级预防、强化慢性病自我管理、定期举办慢性病专题讲座以及建立社区卫生的信息服务平台等方面的认知率明显较低,分别为22.40%、36.80%、42.20%及21.80%,差异有统计学意义(P0.05)。Logistic分析显示,文化程度低、经济收入低、未患慢性病及年龄≥60岁均为慢性病认知的危险因素。结论:孝感城区中老年对慢性病及其社区管理现状认知仍存在一定的不足,相关社区卫生管理部门应针对问题的影响因素,积极采取措施,改善社区管理现状。     

Discovery and characterization of COVA322, a clinical-stage bispecific TNF/IL-17A inhibitor for the treatment of inflammatory diseases

Biologic treatment options such as tumor necrosis factor (TNF) inhibitors have revolutionized the treatment of inflammatory diseases, including rheumatoid arthritis. Recent data suggest, however, that full and long-lasting responses to TNF inhibitors are limited because of the activation of the pro-inflammatory T_H17/interleukin (IL)-17 pathway in patients. Therefore, dual TNF/IL-17A inhibition is an attractive avenue to achieve superior efficacy levels in such diseases. Based on the marketed anti-TNF antibody adalimumab, we generated the bispecific TNF/IL-17A-binding FynomAb COVA322. FynomAbs are fusion proteins of an antibody and a Fyn SH3-derived binding protein. COVA322 was characterized in detail and showed a remarkable ability to inhibit TNF and IL-17A in vitro and in vivo. Through its unique mode-of-action of inhibiting simultaneously TNF and the IL-17A homodimer, COVA322 represents a promising drug candidate for the treatment of inflammatory diseases. COVA322 is currently being tested in a Phase 1b/2a study in psoriasis (ClinicalTrials.gov Identifier: NCT02243787).     

Effect of EGCG On Fe(III)‐induced conformational transition of silk fibroin,a model of protein related to neurodegenerative diseases

The abnormal aggregation of amyloid proteins is reported to play a critical role in the etiology of neurodegenerative disorders. Studies have shown that excessive ferric irons are associated with the misfolding of amyloid proteins, and that (‐)‐epigallocatechin gallate (EGCG) is a good metallic ion chelator with inhibitory effect on the aggregation of amyloid proteins. EGCG has been thus considered as a potential drug candidate for the treatment of neurodegenerative diseases. However, the mechanism of action for EGCG in inhibition of aggregation of amyloid proteins is still remaining unclear. Silk fibroin (SF) shares similarities with amyloid proteins in some amino acid sequences and fibrillation kinetics. In this work, therefore, we used SF as a model of protein to investigate the effects of Fe(III) and EGCG on conformational transition by using turbidity assay, thioflavin T (ThT) fluorescence spectroscopy, Raman spectroscopy, and atomic force microscope (AFM). We demonstrated that low concentration of Fe(III) ions promoted the formation of β‐sheet conformers, while high concentration of Fe(III) ions inhibited further aggregation of SF. EGCG could significantly inhibit the conformational transition of SF when induced by Fe(III), and decrease the amount of β‐sheet conformers dose‐dependently. The findings provide important information regarding to EGCG as a potential agent for the prevention and treatment of neurodegenerative diseases. Fe(III) can accelerate the conformation transition of silk fibrion (SF) from random coil into β‐sheet, while (‐)‐epigallocatechin gallate (EGCG) inhibits Fe(III)‐induced β‐sheet aggregation of SF., 2016. © 2015 Wiley Periodicals, Inc. Biopolymers 105: 100–107, 2016