Mechanism of action of Orthosiphon stamineus against non-alcoholic fatty liver disease: Insights from systems pharmacology and molecular docking approaches

Non-alcoholic fatty liver disease (NAFLD) is one of the most common complications of a metabolic syndrome caused by excessive accumulation of fat in the liver. Orthosiphon stamineus also known as Orthosiphon aristatus is a medicinal plant with possible potential beneficial effects on various metabolic disorders. This study aims to investigate the in vitro inhibitory effects of O. stamineus on hepatic fat accumulation and to further use the computational systems pharmacology approach to identify the pharmacokinetic properties of the bioactive compounds of O. stamineus and to predict their molecular mechanisms against NAFLD. Methods: The effects of an ethanolic extract of O. stamineus leaves on cytotoxicity, fat accumulation and antioxidant activity were assessed using HepG2 cells. The bioactive compounds of O. stamineus were identified using LC/MS and two bioinformatics databases, namely the Traditional Chinese Medicine Integrated Database (TCMID) and the Bioinformatics Analysis Tool for the Molecular Mechanism of Traditional Chinese Medicine (BATMAN-TCM). Pathway enrichment analysis was performed on the predicted targets of the bioactive compounds to provide a systematic overview of the molecular mechanism of action, while molecular docking was used to validate the predicted targets. Results: A total of 27 bioactive compounds corresponding to 50 potential NAFLD-related targets were identified. O. stamineus exerts its anti-NAFLD effects by modulating a variety of cellular processes, including oxidative stress, mitochondrial β-oxidation, inflammatory signalling pathways, insulin signalling, and fatty acid homeostasis pathways. O. stamineus is significantly targeting many oxidative stress regulators, including JNK, mammalian target of rapamycin (mTOR), NFKB1, PPAR, and AKT1. Molecular docking analysis confirmed the expected high affinity for the potential targets, while the in vitro assay indicates the ability of O. stamineus to inhibit hepatic fat accumulation. Conclusion: Using the computational systems pharmacology approach, the potentially beneficial effect of O. stamineus in NAFLD was indicated through the combination of multiple compounds, multiple targets, and multicellular components.     

基于TCGA数据初步探查肝内胆管癌预后相关基因

目的 分析tCGA数据库中肝内胆管癌（ICC）高通量测序数据，寻找其预后相关基因，构建风险模型，并研究其在ICC组织中表达及作用通路。方法 下载tCGA数据库中33例ICC组织和8例癌旁组织中的RNA-seq表达矩阵数据和患者临床资料信息，利用edgeR软件包进行基因差异表达分析，通过单因素Cox回归分析筛选出预后相关差异基因，对差异基因绘制生存曲线，筛选出具有临床意义的基因，经多因素Cox回归分析并构建风险模型，通过京都基因与基因组百科全书（KEGG）通路富集分析了解预后相关基因的作用通路。结果 通过edgeR分析后得到6 617个差异基因（筛选标准为|log2 Fold Change|＞1，P＜0.05），其中高表达组4 094个，低表达组2 523个。通过功能富集发现，这些基因主要集中在化学物致癌作用、药物代谢-细胞色素P450系统、细胞色素P450对异生物质的代谢影响以及视黄醇代谢通路。经单因素Cox回归、R软件“survival”包生存曲线分析显示，UCN2、CST1、PROS1、SLC35E4、PEMT五个基因对ICC患者预后存在显著性影响。通过多因素Cox回归分析，CST1、PEMT、PROS1构建的风险模型对ICC患者预后具有判断作用。结论 UCN2、CST1、PROS1、SLC35E4、PEMT基因可能成为ICC预后判断指标，为后续临床试验提供数据支持。     

Chemical shift differences between free and Fab-bound peptide correlate with a two-stage selection of peptide sequences from a random phage display library to delineate critical and non-critical residues for antibody recognition

Epitope libraries provide a method to identify peptide ligands for antibodies, receptors or other binding proteins. As such, they provide a powerful tool to rapidly identify lead ligands in the drug discovery process. In an attempt to correlate structural information with the results from peptide screening, we have used NMR spectroscopy of peptide/antibody complexes to demonstrate that core residues identified through a two-stage selection process undergo a larger structural change upon binding antibody than do positions in the peptide amenable to a variety of side chains. The model system used was the M2 monoclonal antibody/Flag? octapeptide epitope system. We have analyzed two peptides: Ac-Asp-Tyr-Lys-Leu-Gly-Asp-Asp-Leu-NH₂ (peptide l), which contains several non-core positions randomized, and Ac-Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Leu-NH₂ (peptide 2), which closely corresponds to the original Flag? sequence. Enrichment of the peptides with ¹⁵N facilitated the investigation by permitting spectral editing of the peptide resonances in the presence of antibody. For peptide 1 the absolute shifts for the free vs. Fab-bound peptide were found to be largest for the amide groups of Asp-1 and Asp-6, in agreement with classification of these residues as critical by the phage display library selection process. For peptide 2 the largest absolute shifts were observed for Asp-1 and Asp-4, with the other aspartic acid residues also showing significant but smaller changes. © Munksgaard 1995.     

富氧对海拔3700m高原人体运动心力储备的影响

目的　探讨富氧对高原人体运动心力储备方面的影响。方法　对海拔 3 70 0m高原的 1 2名健康青年富氧 (氧浓度为 2 4%～ 2 5 %)前后分别进行踏阶运动 ,采用心力监护仪采集和记录心动周期和心力信息 ,把完成规定运动量运动后第一心音 (S1 )幅值对安静时S1 幅值增加的相应倍数评估心肌收缩能力储备指数 (CCRI) ;利用舒张期和收缩期时限数据计算舒张期 /收缩期比值 (D/S比 )。结果　运动后较安静时HR ,D/S ,S1 幅值均增高 ,有非常显著性差异 (P <0 .0 1 ) ;富氧运动较未富氧运动CCRI,D/S ,S1 幅值增高 ,有显著性差异 (P <0 .0 5 ) ,HR无统计学意义 (P >0 .0 5 )。结论　高原低氧环境下心脏储备主要是心肌收缩能力储备而不是心率储备。富氧对增强机体心力储备具有重要作用     

Literacy for early childhood: Learning from the learners

The originator of LETTERLAND describes a unique teaching model which blends a structured phonics approach with whole language teaching. Children are introduced to a language about language which enables them to acquire a confident mastery of print based on parallel learning. Its pictogrammatic approach creates a stimulus to inquiry and a learning environment which is imaginative and friendly.     

Differential effects of two types of environmental novelty on activity and sleep in BALB/cJ and C57BL/6J mice

Change in the sleeping environment can produce significant alterations in sleep. To determine how these alterations may vary with the amount of change and the relative reactivity of the sleeper, we examined the influences of environmental novelty on sleep in two mouse strains that differ in behavioral anxiety. Mice [BALB/cJ (n=7) and C57BL/6J (n=8)] were implanted for recording EEG and activity via telemetry. Following baseline data collection, activity and sleep were examined over 46 h after routine cage change, after placing a simple novel object (PVC Tee) in the home cage, and after handling controls. Mice of both strains showed immediate increases in activity and decreases in rapid eye movement sleep (REM) and non-REM (NREM) after cage change and novel object. Within strain, changes in activity and sleep were greater after cage change than after novel object. Changes in activity and sleep time were significantly correlated in each strain. Compared to C57BL/6J mice, BALB/cJ mice exhibited greater and longer duration initial reductions in sleep time, and greater increases in EEG slow wave activity power after cage change and novel object, but these changes were not followed with subsequent increases in sleep time. In contrast, C57BL/6J mice showed significantly greater subsequent increases in sleep time following the initial reductions induced by both manipulations. The results suggest that initial decreases and subsequent increases in sleep time are related to putative differences in the intensity of environmental novelty (cage change>novel object) and to previously described strain differences in anxiety (BALB/cJ>C57BL/6J).     

全外显子组测序和目标序列靶向捕获测序在遗传性视网膜变性基因诊断中的差异

目的:对比外显子组测序(whole exome sequencing,WES)和目标序列靶向捕获测序检测中国遗传性视网膜变性(inherited retinal dystrophies,IRDs)患者致病基因变异的差异。方法:收集182例IRDs家系,所有先证者均接受系统的眼科检查和必要的全身检查,采集患者及家属血样并提取基因组DNA。按照就诊的时间顺序将患者平均分为两组,一组91例接受WES,另一组91例应用本课题组设计并定制的“遗传性眼病基因诊断芯片” (hereditary eye disease enrichment panel,HEDEP)进行IRDs致病基因外显子区域靶向捕获测序。对候选致病基因用Sanger测序进行验证,并对家系成员进行共分离分析,使用多重连接依赖的探针扩增技术对拷贝数变异进行验证,针对二代测序捕获效率低的区域如RPGR ORF15区,应用Sanger 测序补充检测。根据美国医学遗传学与基因组学学会和分子病理学协会(American College of Medical Genetics and Genomics and the Association for Molecular Pathology,ACMG/AMP)制定的《ACMG/AMP基因变异分类标准与指南》将检测到的所有基因变异进行分类,本文只包含“致病的”、“可能致病的”的基因变异,不包含“意义不明确的”、“可能良性的”和“良性的”基因变异。结果:应用HEDEP确诊的家系共51例,阳性率为56.04%(51/91);应用WES确诊的家系共30例,阳性率为33.00%(30/91);总阳性率44.51%(81/182)。平均测序深度以及测序覆盖度方面,HEDEP优于WES,此外HEDEP具有检测拷贝数变异潜力。本研究共检测到29个IRDs基因的致病突变,最常见的致病基因为USH2A、ABCA4和RPGR,基因突变频率分别为11.54%(21/182)、6.59%(12/182)、3.85%(7/182);共发现43个新的致病突变,并检测到6例家系携带RPGR ORF15区的突变。结论:针对临床确诊的IRDs病例,HEDEP较WES能获得更高的基因诊断阳性率和更精确的诊断结果,可作为IRDs基因诊断的首选方法,WES可作为其他基因诊断方法的补充手段。同时,本研究丰富了IRDs致病基因的突变频谱,为将来IRDs基因诊断、遗传咨询和基因治疗奠定了基础。     

大孔吸附树脂富集茵陈蒿汤中蒿属香豆素的工艺研究

目的 研究大孔吸附树脂富集茵陈蒿汤中蒿属香豆素的工艺条件及参数。方法 以蒿属香豆素为指标，考察大孔吸附树脂富集茵陈蒿汤中蒿属香豆素的最佳工艺条件。结果提取液上大孔吸附树脂柱，吸附30min后，以5倍树脂量的蒸馏水洗去杂质，再以3～4倍树脂量的70％乙醇洗脱蒿属香豆素为最佳工艺。结论 此法可较好地富集茵陈蒿汤中的主要有效成分蒿属香豆素。     

大孔吸附树脂富集小蓟中咖啡酸酯类成分的研究

目的通过对23种大孔吸附树脂筛选,寻找适用于纯化小蓟中咖啡酸酯类成分的大孔吸附树脂,为从小蓟中工业化生产咖啡酸酯类成分提供依据。方法采用23种大孔吸附树脂对小蓟提取物进行吸附纯化,以总咖啡酸酯收率和纯度作为指标综合评价。结果23种吸附树脂中,以树脂HPD-100吸附洗脱的总咖啡酸酯的收率最高为87.6%、纯度为52.2%。结论HPD-100树脂综合性能良好,适用于小蓟中咖啡酸酯类成分的纯化。     

用VEF值评价地铁车站内挥发性有机物的污染水平

[目的 ]评价地铁车站内站台和站厅挥发性有机物 (VOCs)的污染水平 ,并判断来源。 [方法 ]用气相色谱质谱联用仪对地铁车站站台和站厅挥发性有机物浓度进行测定 ,并用挥发性有机物富集指数 (VEF)进行分析。 [结果 ]两车站站台和站厅监测点白天及夜晚的CO2 浓度差异存在显著性 (P <0 .0 1) ,监测点白天和夜晚的VOCs浓度差异无显著性 (P >0 .0 5 )。地铁车站内VOCs的主要来源来自其自身室内排放源 ,室外污染物浓度及通风状况等可能对室内VOCs有影响。 [结论 ]地铁车站内存在VOCs污染 ,用VEF评价地铁车站内VOCs污染状况 ,能较简便、准确地反映其污染物来源 ,并可比较不同车站之间VOCs的浓度水平