A Hot‐Segment‐Based Approach for the Design of Cross‐Amyloid Interaction Surface Mimics as Inhibitors of Amyloid Self‐Assembly

The design of inhibitors of protein–protein interactions mediating amyloid self‐assembly is a major challenge mainly due to the dynamic nature of the involved structures and interfaces. Interactions of amyloidogenic polypeptides with other proteins are important modulators of self‐assembly. Here we present a hot‐segment‐linking approach to design a series of mimics of the IAPP cross‐amyloid interaction surface with Aβ (ISMs) as nanomolar inhibitors of amyloidogenesis and cytotoxicity of Aβ, IAPP, or both polypeptides. The nature of the linker determines ISM structure and inhibitory function including both potency and target selectivity. Importantly, ISMs effectively suppress both self‐ and cross‐seeded IAPP self‐assembly. Our results provide a novel class of highly potent peptide leads for targeting protein aggregation in Alzheimer’s disease, type 2 diabetes, or both diseases and a chemical approach to inhibit amyloid self‐assembly and pathogenic interactions of other proteins as well.     

A reversed‐phase compatible thin‐layer chromatography autography for the detection of acetylcholinesterase inhibitors

A dual readout autographic assay to detect acetylcholinesterase inhibitors present in complex matrices adsorbed on reversed‐phase or normal‐phase thin‐layer chromatography plates is described. Enzyme gel entrapment with an amphiphilic copolymer was used for assay development. The effects of substrate and enzyme concentrations, pH, incubation time, and incubation temperature on the sensitivity and the detection limit of the assay were evaluated. Experimental design and response surface methodology were used to optimize conditions with a minimum number of experiments. The assay allowed the detection of 0.01% w/w of physostigmine in both a spiked Sonchus oleraceus L. extract chromatographed on normal phase and a spiked Pimenta racemosa (Mill.) J.W. Moore leaf essential oil chromatographed on reversed phase. Finally, the reversed‐phase thin‐layer chromatography assay was applied to reveal the presence of an inhibitor in the Cymbopogon citratus (DC.) Stapf essential oil. The developed assay is able to detect acetylcholinesterase inhibitors present in complex matrixes that were chromatographed in normal phase or reversed‐phase thin‐layer chromatography. The detection limit for physostigmine on both normal and reversed phase was of 1×10^?4 μg. The results can be read by a change in color and/or a change in fluorescence.     

Peptide-derived from Seahorse Exerts a Protective Effect against Cholinergic Neuronal Death in in vitro Model of Alzheimer's Disease

Hippocampus trimaculatus are highly valued and the most heavily traded seahorse species for traditional medicine purposes in many Asian countries. One of the main interesting features in H.trimaculatus is their richness in protein which can be hydrolyzed into bioactive peptides. In the previous study, H. trimaculatus was hydrolyzed using commercial enzymes, the peptides responsible for neuroprotective activity was identified as HTP-1. In the present study, in vitro co-culture system of neuronal (PC12) cells and Aβ₄₂ oligomer-stimulated murine microglia cells (BV2 cells) were used to test neuroprotective effects of HTP-1. The co-culture system showed that HTP-1 protected PC12 cells from BV2 neurotoxic responses. In addition, the activation of PI3K/Akt signaling pathway by HTP-1 were confirmed. The PI3K/Akt activation was found to be mediated through transforming growth factor-β (TGF-β) induction by HTP-1. Furthermore, this signaling pathway was found to up-regulate the expression of pro survival protein expression in PC12 cells. Collectively, HTP-1 has potent protective effect against neuronal cells death in in vitro model of Alzheimer's disease.     

表面解吸常压化学电离质谱法直接分析牙齿微区表面

采用纳升取样表面解吸常压化学电离质谱法(nano-SDAPCI-MS)结合主成分分析(PCA),建立了一种采用具有微米级针尖的金属取样针直接对龋齿不同部位取样并进行快速质谱分析的方法.数据分析结果表明,同一颗龋齿不同部位的质谱指纹谱图之间存在差异;在不需要样品预处理的前提下通过串联质谱快速测定了龋齿中的乳酸、丙酮酸、苯乙酸和丙酸等成分.采用PCA方法可较好地将龋齿病灶位置与邻近正常组织进行区分,也可对不同牙病及健康牙齿进行区分.本方法可方便地对牙齿进行直接微区分析,为鉴别牙齿疾病及观测治疗效果提供了一种快速、简单的方法,为生物体中微细部位的快速取样及直接质谱分析提供了一种可能的解决方案.     

A novel approach for LC-MS/MS-based chiral metabolomics fingerprinting and chiral metabolomics extraction using a pair of enantiomers of chiral derivatization reagents

Chiral metabolites are found in a wide variety of living organisms and some of them are understood to be physiologically active compounds and biomarkers. However, the overall analysis of chiral metabolomics is quite difficult due to the high number of metabolites, the significant diversity in their physicochemical properties, and concentration range from metabolite-to-metabolite. To solve this difficulty, we developed a novel approach for chiral metabolomics fingerprinting and chiral metabolomics extraction, which is based on the labeling of a pair of enantiomers of chiral derivatization reagents (i.e., DMT-(S,R)-Pro-OSu and DMT-3(S,R)-Apy) and precursor ion scan chromatography of the derivatives. The multivariate statistics is also required for this strategy. The proposed procedures were evaluated by the detection of a diagnostic marker (i.e., d-lactic acid) using the saliva of diabetic patients. This method was used for the determination of biomarker candidates of chiral amines and carboxyls in Alzheimer's disease (AD) brain homogenates. As the results, l-phenylalanine (L-Phe) and l-lactic acid (L-LA) were identified as the decreased and increased biomarker candidates in the AD brain, respectively. Therefore, the proposed approach seems to be helpful for the determination of non-target chiral metabolomics possessing amines and carboxyls.     

Fluorescence Polarization Based Nucleic Acid Testing for Rapid and Cost‐Effective Diagnosis of Infectious Disease

A new nucleic acid detection method was developed for a rapid and cost‐effective diagnosis of infectious disease. This approach relies on the three unique elements: 1) detection probes that regulate DNA polymerase activity in response to the complementary target DNA; 2) universal reporters conjugated with a single fluorophore; and 3) fluorescence polarization (FP) detection. As a proof‐of‐concept, the assay was used to detect and sub‐type Salmonella bacteria with sensitivities down to a single bacterium in less than three hours.     

冠心病患者血脂分析与HCY测定的临床应用

目的通过研究并分析出冠心病患者的血脂组成成分,然后通过使用HCY测定的方法测定冠心病患者血清中的同型半胱氨酸含量的实际临床应用。方法 50例患有冠心病的患者和50名健康人员的血清成分中同型半胱氨酸含量,其中治疗组有25例无心肌梗塞的冠心病患者,其余25例研究对象均为有心肌梗塞患者,然后对治疗组患者的血脂组成成分进行测定并分析比较出异常成分。结果通过研究后发现治疗组的研究对象血清中的同型半胱氨酸含量远多于对照组的50例健康人员血清中的同型半胱氨酸含量,其中25例治疗组心肌梗塞患者的同型半胱氨酸含量多于25例治疗组的无心肌梗塞患者的同型半胱氨酸含量,具有统计学意义。冠心病患者的血脂组成成分Apo B100的值增加,冠心病患者的Apo A1的值和Apo A1与Apo B100的比值均减小,经统计学分析得出,具有统计学意义。结论血清同型半胱氨酸的含量改变,以及血脂中Apo B100的值和Apo A1的值与Apo B100的比值发生改变对治疗诊断与预防冠心病都有十分重要的价值。     

维持性血液透析患者血清铁调素水平与心血管疾病的关系研究

目的观察分析维持性血液透析患者血清铁调素水平与心血管疾病的关系。方法选择惠州市第三人民医院2014年12月至2016年12月期间收治的维持性血液透析患者54例作为观察组,包含20例患者出现心血管疾病,另外选择接受体检的健康人25例作为对照组。结果观察组患者的血清铁调素水平、晚期氧化蛋白物及IL-6均显著高于对照组,比较差异具有统计学意义(P0.05);观察组发生心血管疾病患者血清铁调素水平显著高于非心血管疾病患者,数据比较差异具有统计学意义(P0.05)。结论血液透析患者血清铁调素水平与心血管疾病具有相关性,即可作为血透患者发生心血管疾病的新的独立危险因素,但需要进一步验证。     

细菌性阴道病与甲硝唑治疗对妊娠结局的影响探讨

目的探讨细菌性阴道病和甲硝唑治疗对妊娠结局的影响。方法选取吉水县醪桥镇人口与计划生育服务所2014年7月至2015年7月收治的120例中晚期妊娠合并BV者,根据自愿原则分为治疗组和未治疗组(C组)。治疗组中口服用药为A组,阴道用药为B组。结果治疗组中两种不同用药方式都对妊娠合并BV患者有着显著的治疗效果,两组比较无统计学意义(P0.05);治疗组妊娠结局总不良率仅为10%,未治疗组中总不良率高达73.3%,对比差距显著,具备统计学意义(P0.05)。结论对妊娠期合并细菌性阴道病妇女使用甲硝唑治疗,对提升妊娠期妇女生活品质和降低不良妊娠风险有着关键的作用,同时具有十分重要的参考意义。