西达本胺增加慢性髓系白血病耐药株K562/ADM细胞对柔红霉素的敏感性

目的 观察西达本胺（CDM）能否影响人慢性髓系白血病耐药株K562/ADM细胞对柔红霉素（DNR）的敏感性,并探讨其可能的分子机制。方法 体外常规培养K562细胞和K562/ADM细胞,给予不同剂量CDM和（或）DNR处理48 h后,采用细胞计数试剂盒8（CCK-8）法检测CDM与DNR对K562和K562/ADM细胞的毒性作用,采用Chou-Talalay中效分析法对两药的联合效应进行评价,采用流式细胞术检测细胞增殖、细胞周期和凋亡,采用Western blotting方法检测组蛋白2AX（H2AX）、γH2AX（Ser139）、共济失调毛细血管扩张征突变基因（ATM）、p-ATM（Ser1981）、乳腺癌易感蛋白l（BRCA1）和p-BRCA1（Ser1524）的蛋白表达水平。 结果 DNR可剂量依赖性地抑制K562/ADM细胞活力（P<0.05）,半数抑制浓度（IC50）为11.76 μmol/L,耐药倍数为18.09;CDM可协同增强DNR对K562/ADM细胞的抑制作用置信区间（CI）（CI<1）,反转倍数为8.11;与对照组相比,DNR组细胞增殖率显著降低（P<0.05）,G2/M期细胞比例和凋亡率明显升高（P<0.05）,而无毒剂量的CDM可协同增强DNR引起的细胞增殖抑制、G2/M期阻滞和细胞凋亡（P<0.05）;耐药株K562/ADM细胞中ATM和BRCA1蛋白表达水平显著高于其亲代K562细胞（P<0.05）;DNR可上调K562/ADM细胞中H2AX、ATM和BRCA1蛋白的磷酸化水平（P<0.05）;CDM与DNR联用可使γH2AX蛋白水平进一步升高,但p-ATM和p BRCA1蛋白水平的变化则相反（P<0.05）。 结论 CMD可反转K562/ADM细胞对DNR的耐药性,这可能与上调H2AX蛋白的磷酸化水平以及下调ATM和BRCA1蛋白的磷酸化水平有关。     

Group A streptococcal M-protein specific antibodies and T-cells drive the pathology observed in the rat autoimmune valvulitis model

Acute rheumatic fever (ARF) and rheumatic heart disease (RHD) are autoimmune mediated diseases triggered by group A streptococcal (GAS) infections. Molecular mimicry between GAS M-proteins and host tissue proteins has been proposed as the mechanism that initiates autoreactive immune responses in ARF/RHD. However, the individual role of antibodies and T-cells specific for GAS M-proteins in the pathogenesis of autoimmune carditis remains under-explored. The current study investigated the role of antibodies and T-cells in the development of carditis in the Lewis rat autoimmune valvultis (RAV) model by transferring serum and/or splenic T-cells from rats previously injected with GAS recombinant M5 protein. Here we report that serum antibodies alone and serum plus in vitro expanded rM5-specific T-cells from hyperimmune rats were capable of transferring carditis to naïve syngeneic animals. Moreover, the rats that received combined serum and T-cells developed more severe carditis. Recipient rats developed mitral valvulitis and myocarditis and showed prolongation of P-R intervals in electrocardiography. GAS M5 protein-specific IgG reactivity and T-cell recall response were also demonstrated in recipient rats indicating long-term persistence of antibodies and T-cells following transfer. The results suggest that both anti-GAS M5 antibodies and T-cells have differential propensity to induce autoimmune mediated carditis in syngeneic rats following transfer. The results highlight that antibodies and effector T-cells generated by GAS M protein injection can also independently home into cardiac tissue to cross-react with tissue proteins causing autoimmune mediated immunopathology.     

血糖、血脂水平与糖尿病视网膜病变患者视力损害的相关性研究

目的探讨血糖、血脂水平与糖尿病视网膜病变患者视力损害的相关性。方法选取2016年1月至2018年1月我院内分泌科收治的250例DR患者进行研究。根据视力分级标准分为三个组:盲组(n=28),低视力组(n=80),低视力以上组(n=142)。比较各组患者FPG、HbAlC、TC、TG、LDL-C、HDL-C、LDL-C/HDL-C、ApoB、ApoA1、ApoB/ApoA1水平,对FPG、HbA1C、LDL-C/HDL-C及ApoB/ApoA1与DR患者视力损害进行Pearman相关性分析和Logistic回归分析。结果三组患者的TC、TG差异均无统计学意义(P>0.05);与低视力以上组相比,盲组和低视力组FPG、HbAlC、LDL-C、LDL-C/HDL-C、ApoB、ApoB/ApoA1更高,HDL-C、ApoA1更低(P<0.05);盲组FPG、HbAlC、LDL-C、LDL-C/HDL-C、ApoB、ApoB/ApoAl显著高于低视力组,HDL-C、ApoA1显著低于低视力组(P<0. 05)。经Spearman相关性分析,FPG、HbAlC、LDL-C/HDL-C、ApoB/ApoA1比值与DR患者视力损害均呈正相关。经Logistic回归分析,FPG、HbAlC、LDL-C/HDL-C、ApoB/ApoA1均是DR患者视力损害的独立危险因素。结论 FPG、HbAlC、LDL-C/HDL-C、ApoB/ApoA1与DR患者的视力损害有密切关系,通过监测这些指标有利于DR患者视力损害的预测,及时采取治疗措施。     

Prion neurotoxicity

Although the mechanisms underlying prion propagation and infectivity are now well established, the processes accounting for prion toxicity and pathogenesis have remained mysterious. These processes are of enormous clinical relevance as they hold the key to identification of new molecular targets for therapeutic intervention. In this review, we will discuss two broad areas of investigation relevant to understanding prion neurotoxicity. The first is the use of in vitro experimental systems that model key events in prion pathogenesis. In this context, we will describe a hippocampal neuronal culture system we developed that reproduces the earliest pathological alterations in synaptic morphology and function in response to PrP^Sc. This system has allowed us to define a core synaptotoxic signaling pathway involving the activation of NMDA and AMPA receptors, stimulation of p38 MAPK phosphorylation and collapse of the actin cytoskeleton in dendritic spines. The second area concerns a striking and unexpected phenomenon in which certain structural manipulations of the PrP^C molecule itself, including introduction of N‐terminal deletion mutations or binding of antibodies to C‐terminal epitopes, unleash powerful toxic effects in cultured cells and transgenic mice. We will describe our studies of this phenomenon, which led to the recognition that it is related to the induction of large, abnormal ionic currents by the structurally altered PrP molecules. Our results suggest a model in which the flexible N‐terminal domain of PrP^C serves as a toxic effector which is regulated by intramolecular interactions with the globular C‐terminal domain. Taken together, these two areas of study have provided important clues to underlying cellular and molecular mechanisms of prion neurotoxicity. Nevertheless, much remains to be done on this next frontier of prion science.     

Overview of Protein‐Based Biopolymers for Biomedical Application

Biopolymers are playing a vital role in biomedical applications. Among them, protein‐based biopolymers are utilized for the fabrication of tissue‐engineering constructs, therapeutic molecule delivery carriers, emulsifiers, and food packaging materials. Wide ranges of proteins are extracted from animal or plant sources and are being utilized for the fabrication of scaffolds for regenerative tissue‐engineering application. Here, an overview about the protein structure, extraction procedure, solubility, and various formulation‐based proteins found in the literature are discussed. Biopolymers display several advantages such as biocompatibility and degradability by enzymes. Methods to overcome the disadvantages of these proteins such as immunogenicity, antigenicity, and solubility are reported. Various crosslinking reagents specific to protein chemistry are discussed as well.     

Emerging hepatitis E virus compared with hepatitis A virus: A new sanitary challenge

Hepatitis A (HAV) and E (HEV) viruses are able to cause liver disease in humans. Among the five classical hepatotropic viruses, they are mainly transmitted via the fecal‐oral route. Historically, many similarities have thus been described between them according to their incidence and their pathogenicity, especially in countries with poor sanitary conditions. However, recent advances have provided new insights, and the gap is widening between them. Indeed, while HAV infection incidence tends to decrease in developed countries along with public health improvement, HEV is currently considered as an underdiagnosed emerging pathogen. HEV autochthonous infections are increasingly observed and are mainly associated with zoonotic transmissions. Extra hepatic signs resulting in neurological or renal impairments have also been reported for HEV, as well as a chronic carrier state in immunocompromised patients, arguing in favor of differential pathogenesis between those two viruses. Recent molecular tools have allowed studies of viral genome variability and investigation of links between viral plasticity and clinical evolution. The identification of key functional mutations in viral genomes may improve the knowledge of their clinical impact and is analyzed in depth in the present review.     

Interaction of antivirals with a heptameric bundle model of the p7 protein of hepatitis C virus

A series of ligands are known experimentally to affect the infectivity cycle of the hepatitis C virus. The target protein for the ligands is proposed to be p7, a 63 amino acid polytopic channel‐forming protein, with possibly two transmembrane domains. Protein p7 is found to assemble into functional oligomers of various sizes, depending on the genotype (GT). Nine ligands are docked to various sites of a computationally derived heptameric bundle of p7 of GT1a. The energy of interaction, here binding energy, is calculated using three different docking programs (Autodock , MOE , LeadIT ). Three protein regions are defined to which the ligands are placed, the loop region and the site with the termini as well as the mid‐region which is supposed to track poses inside the putative pore. A common feature is that the loop sites and poses either within the pore or at the intermonomer space of the bundle are preferred for all ligands with proposed binding energies smaller than −10 kJ/mol. BIT225, benzamine, amantadine, and NN‐DNJ show good overall scoring.     

Satiety and memory enhancing effects of a high-protein meal depend on the source of protein

Objective: High- protein diets have become increasingly popular with various touted benefits. However, the extent to which protein quantity and source affects cognitive functioning through altering postprandial amino acid profiles has not been investigated. Further, whether all protein sources are similarly anorexigenic is uncertain. The objective of this study was to determine the influence of protein level and source on Barnes maze performance, satiety and plasma amino acid levels in male Sprague-Dawley rats.

Methods: Rats were entrained to a meal-feeding schedule consisting of a 30 minutes meal, equivalent to 20% of average daily intake, one hour into the dark phase then ad libitum access to food for 5 h. On test days, rats received one of three isocaloric diets as their first meal, hereafter referred to as Egg White (EW), Wheat Gluten (WG), or Basal, and then were measured for cognitive performance, feeding behavior, or plasma amino acid levels via jugular catheter. Percentage energy from protein was 35% for both EW and WG and 20% for Basal with equal amounts provided by EW and WG proteins.

Results: Rats provided EW performed similarly to Basal on the Barnes maze, whereas WG performed worse. EW increased satiety, whereas WG reduced satiety relative to Basal. Both EW and WG increased postprandial concentrations of large neutral and branched chain amino acids relative to Basal, but in EW, concentrations were slower to peak, and peaked to a higher level than WG.

Discussion: Results demonstrate the importance of protein source for cognition and satiety enhancing effects of a high-protein meal.