碱性成纤维细胞生长因子在富集结直肠癌肿瘤干细胞中的作用

目的 研究不同浓度碱性成纤维细胞生长因子（bFGF）作用于结直肠癌细胞不同时间,富集肿 瘤干细胞的效果。方法? 结直肠癌DLD-1细胞分别在含5 ng/mL、10 ng/mL、20 ng/mL bFGF的无血清培 养基中悬浮培养,分为G1、G2、G3组,设置培养时间梯度为10 d、20 d、30 d,获得球细胞。采用流式 细胞术（FCM）检测细胞球中的CD44+、CD133+及CD44+CD133+双阳性的细胞表达比例,Real-time PCR 检测球细胞中的干细胞基因（KLF4、Nanog）;上皮间质转化基因（E-cadherin、Snail）;Wnt/β-catenine通路 基因（Wnt-3a）的 mRNA表达情况,成球实验检测细胞球的自我更新能力。结果 FCM检测结果：CD44+ 阳性表达的细胞表达以G2组20 d最高,CD133+及CD44+CD133+双阳性的细胞表达均以G2组20 d及G3 组10 d最高,差异有统计学意义（F=98.895、147.641、13.321,P<0.05）。 Real-time PCR检测结果：各组中 KLF4、Nanog、Snail以及Wnt-3a mRNA的表达均以G2组20 d表达最高（F=2.424、7.694、2.951、3.771, 均P<0.05）, E-cadherin基因G2组20 d mRNA表达最低（G2组30 d、G1组10 d除外）（F=10.620,P<0.05）。 成球实验结果：各组比较G1组20 d和G2组20 d成球数量明显多于其他组,差异具有统计学意义（F=14.279, P<0.01）;但 G1组20 d和G2组20 d比较,无统计学差异（t=0.605,P=0.578）。 结论 碱性成纤维细胞生长 因子无血清悬浮培养能够有效富集肿瘤干细胞,其中G2组培养20 d较其余组能更有效地富集结直肠癌肿 瘤干细胞。     

基于古文献挖掘与网络药理学的二陈汤治疗肥胖型多囊卵巢综合征的机制研究

目的：探究历代医家治疗肥胖型多囊卵巢综合征的处方规律,归纳核心处方与药物,并利用网络药理学探讨核心处方的潜在物质基础与作用机制。方法：以《中华医典》为资料来源,建立处方数据库,借助 SPSS Modeler以及Gaphi软件,对符合肥胖型多囊卵巢综合征表现的古方进行关联分析。利用TCMSP数据库获取核心处方的成分及靶点,利用GeneCard数据库获取疾病靶点,利用Cytoscape软件构建活性成分-靶点网络,并利用DAVID数据库进行组织富集分析。结果：共得到符合肥胖型多囊卵巢综合征的治疗方剂47首,涉及中药98味,使用频次≥5的中药为茯苓、半夏、甘草、陈皮、川芎等;有强关联的核心药对组合为半夏、陈皮、茯苓、甘草（二陈汤）。利用TCMSP数据库获得药物成分125个,靶基因218个,利用GeneCard数据库获得疾病靶基因2962个,交集靶基因主要富集在胎盘、肝脏、肺、上皮、血液等组织当中。结论：数据挖掘结果提示历代医家治疗肥胖型多囊卵巢综合征时,以燥湿化痰为主要原则,同时佐以养血调经、益气健脾;网络药理学结果在验证了古文献分析结果的同时也为二陈汤治疗肥胖型多囊卵巢综合征的基础研究提供了新的方向。     

CDCA8对肝细胞癌患者预后的影响

目的：肝细胞癌预后相关的分子标志物对提高患者生存质量和改善疗效至关重要,本研究旨在探究细胞分裂周期相关基因8(cell division cycle associated 8,CDCA8)对肝细胞癌预后的影响及潜在分子机制。方法：432例肝细胞癌样本的转录组数据及对应患者的临床信息下载自肿瘤基因表达图谱（The Cancer Genome Atlas,TCGA）数据库。分析肿瘤组织和正常组织中CDCA8 mRNA差异;根据CDCA8 mRNA中位数将肝细胞癌患者分为高表达组和低表达组,绘制生存曲线。采用Kruskal检验分析CDCA8与肿瘤分期、分级和T分期的相关性。采用单因素和多因素Cox回归分析探究CDCA8能否作为肝细胞癌患者的独立预后因子。采用基因集富集分析（gene set enrichment analysis,GSEA）探究CDCA8在肝细胞癌中的潜在作用机制。结果：CDCA8在肿瘤组织中的mRNA水平明显高于正常对照组织（P<0.001）,CDCA8高表达患者的生存率明显低于低表达组（P<0.001）。随着肿瘤分期（P<0.001）、分级（P<0...     

基于生物信息学筛选脓毒症中性粒细胞活化的关键基因

目的基于生物信息学筛选脓毒症中性粒细胞活化的关键基因。 方法从GEO数据库中下载4个成人脓毒症芯片数据集(GSE69528、GSE28750、GSE57065、GSE95233),进行差异表达基因(DEG)筛选。基因本体论(GO)分析DEG,STRING数据库构建蛋白互作网络,Cytoscape软件提取关键基因,CIBERSORT算法估算数据集中的免疫细胞表达水平,ROC曲线评价关键基因对脓毒症患者的诊断价值。取儿童脓毒症相关芯片数据集(GSE66099)进行验证。 结果4个成人脓毒症数据集筛选出299个重叠DEG,其中112个基因上调,187个基因下调。GO主要富集于中性粒细胞活化、中性粒细胞脱颗粒、肽酶调节活性、糖胺聚糖结合、特殊颗粒、囊泡腔等。筛选出脓毒症中性粒细胞活化的关键基因10个,在脓毒症中表达水平均上调,ROC曲线下面积均大于0.7,具有较好的诊断价值。儿童脓毒症数据集验证结果与成人脓毒症数据集结果一致。 结论基于生物信息学方法筛选出脓毒症中性粒细胞活化相关的10个关键基因,为脓毒症的早期诊断、预后判断及治疗提供了潜在新靶点。     

磷酸化蛋白质组学富集策略进展及其在疾病研究中的应用

蛋白质磷酸化是生物体内重要的翻译后修饰,几乎参与调节着细胞增殖、信号转导、新陈代谢、肿瘤发生等过程在内的所有生命活动。对磷酸化蛋白质组的全面分析可以帮助人们深入了解磷酸化蛋白在生命过程中的作用,协助发现生物标志物,辅助疾病的诊疗。然而,磷酸化蛋白丰度低、信号被非磷酸化肽段所掩盖等问题限制了磷酸化蛋白质组学的发展。因此,亟需开发高效的富集策略,如设计新型纳米材料,合并多种分析方法等,以提高检测灵敏度、富集特异性,增大富集容量。本文回顾了近年来磷酸化蛋白质组学研究策略中的富集策略进展及其在疾病研究中的应用。     

环境富集对坐骨神经挤压伤模型小鼠神经再生的影响

目的探讨环境富集对坐骨神经挤压伤模型小鼠神经再生的作用及机制。方法首先对22只C57BL/6小鼠进行坐骨神经挤压建立动物模型, 随后按随机数字表法将模型小鼠分为干预组和对照组, 干预组小鼠置于具备环境富集的鼠笼内进行干预, 对照组置于标准鼠笼中饲养。造模成功2周后, 2组小鼠均行坐骨神经电生理检查和步态分析, 并应用甲苯胺蓝染色观察坐骨神经有髓神经纤维的比例, 采用免疫荧光测定2组坐骨神经中的生长相关蛋白43(GAP43)、髓鞘碱性蛋白(MBP)以及p75神经营养素受体(p75NTR)表达的差异。结果①坐骨神经电生理检测显示, 干预组小鼠坐骨神经复合肌肉动作电位(CMAP)的潜伏期[(1.05±0.04)ms]较对照组[(1.98±0.30)ms]明显缩短(P<0.05), 而其波幅[干预组(10.63±0.90)mV]则明显高于对照组[(6.58±1.25)mV], 且组间差异有统计学意义(P<0.05);②小鼠步态分析显示, 干预组小鼠的平均接触强度[(160.60±20.45)AU]、步幅[(5.11±0.58)cm]和步速[(53.06±7.20)cm/s] 均较对...     

Role of DISC1 Interacting Proteins in Schizophrenia Risk from Genome‐Wide Analysis of Missense SNPs

A balanced translocation affecting DISC1 cosegregates with several psychiatric disorders, including schizophrenia, in a Scottish family. DISC1 is a hub protein of a network of protein–protein interactions involved in multiple developmental pathways within the brain. Gene set‐based analysis has been proposed as an alternative to individual analysis of single nucleotide polymorphisms (SNPs) to get information from genome‐wide association studies. In this work, we tested for an overrepresentation of the DISC1 interacting proteins within the top results of our ranked list of genes based on our previous genome‐wide association study of missense SNPs in schizophrenia. Our data set consisted of 5100 common missense SNPs genotyped in 476 schizophrenic patients and 447 control subjects from Galicia, NW Spain. We used a modification of the Gene Set Enrichment Analysis adapted for SNPs, as implemented in the GenGen software. The analysis detected an overrepresentation of the DISC1 interacting proteins (permuted P‐value = 0.0158), indicative of the role of this gene set in schizophrenia risk. We identified seven leading‐edge genes, MACF1, UTRN, DST, DISC1, KIF3A, SYNE1, and AKAP9, responsible for the overrepresentation. These genes are involved in neuronal cytoskeleton organization and intracellular transport through the microtubule cytoskeleton, suggesting that these processes may be impaired in schizophrenia.     

A Three‐wave Study of Antecedents of Work–Family Enrichment: The Roles of Social Resources and Affect

On the basis of conservation of resources theory (Hobfoll, 1989 ) and the resource‐gain‐development perspective (Wayne, Grzywacz, Carlson, & Kacmar, 2007 ), this paper examines the differential impact of specific social resources (supervisory support and family support) on specific types of affect (job satisfaction and family satisfaction, respectively), which, in turn, influence work‐to‐family enrichment and family‐to‐work enrichment, respectively. A sample of 276 Chinese workers completed questionnaires in a three‐wave survey. The model was tested with structural equation modelling. Job satisfaction at time 2 partially mediated the relationship between time 1 supervisory support and time 3 work‐to‐family enrichment (capital), and the effect of supervisory support on work‐to‐family enrichment (affect) was fully mediated by job satisfaction. Family satisfaction at time 2 fully mediated the relationship between time 1 family support and time 3 family‐to‐work enrichment (affect, efficiency). Implications for theory, practice and future research are discussed. Copyright © 2013 John Wiley & Sons, Ltd.     

Short‐term environmental enrichment,in the absence of exercise,improves memory,and increases NGF concentration,early neuronal survival,and synaptogenesis in the dentate gyrus in a time‐dependent manner

Environmental manipulations can enhance neuroplasticity in the brain, with enrichment‐induced cognitive improvements being linked to increased expression of growth factors, such as neurotrophins, and enhanced hippocampal neurogenesis. There is, however, a great deal of variation in environmental enrichment protocols used in the literature, making it difficult to assess the role of particular aspects of enrichment upon memory and the underlying associated mechanisms. This study sought to evaluate the efficacy of environmental enrichment, in the absence of exercise, as a cognitive enhancer and assess the role of Nerve Growth Factor (NGF), neurogenesis and synaptogenesis in this process. We report that rats housed in an enriched environment for 3 and 6 weeks (wk) displayed improved recognition memory, while rats enriched for 6 wk also displayed improved spatial and working memory. Neurochemical analyses revealed significant increases in NGF concentration and subgranular progenitor cell survival (as measured by BrdU+ nuclei) in the dentate gyrus of rats enriched for 6 wk, suggesting that these cellular changes may mediate the enrichment‐induced memory improvements. Further analysis revealed a significant positive correlation between recognition task performance and BrdU+ nuclei. In addition, rats enriched for 6 wk showed a significant increase in expression of synaptophysin and synapsin I in the dentate gyrus, indicating that environmental enrichment can increase synaptogenesis. These data indicate a time‐dependent cognitive‐enhancing effect of environmental enrichment that is independent of physical activity. These data also support a role for increased concentration of NGF in dentate gyrus, synaptogenesis, and neurogenesis in mediating this effect. © 2013 Wiley Periodicals, Inc.