表达Strep-tag II的重组塞内卡病毒的构建与鉴定

为构建表达Strep-tag II的重组塞内卡病毒(SVA),本研究在感染性cDNA克隆p SVA-I212V/S460L的基础上,采用重叠延伸PCR扩增Strep-tag II并采用无缝克隆法将其插入SVA VP1基因C端,构建感染性cDNA克隆,命名为p SVA-Strep,并采用双酶切和测序鉴定正确后,将p SVA-Strep转染BHK-21细胞拯救重组病毒,收集病变细胞上清,并将含有病毒的细胞上清和野生型SVA(SVA-WT)分别感染BHK-21细胞,以未感染病毒的BHK-21细胞作为阴性对照,感染后24 h分别以鼠源SVA VP2蛋白单克隆抗体(MAb)5D10(1∶1000)、鼠源Strep-tag II MAb(1∶2000)作为一抗,以FITC标记的山羊抗小鼠Ig G(1∶2000)为二抗,采用间接免疫荧光试验(IFA)对重组病毒鉴定。IFA结果显示,以鼠源SVA VP2蛋白MAb 5D10为一抗,SVA-WT和重组病毒感染的细胞均出现绿色荧光,以Strep-tag II MAb作为一抗时,仅有重组病毒感染的细胞出现绿色荧光,经SVA-WT感染的BHK-21细胞及阴性对照细胞均无绿色荧光,表明拯救了Strep-tag II标记的重组病毒,并将其命名为r SVA-Strep。将r SVA-Strep在BHK-21细胞中连续传10代,每隔2代采用引物VP1-F/R经PCR鉴定并对第10代重组病毒的VP1基因测序以鉴定r SVA-Strep的遗传稳定性。结果显示,每隔两代的重组病毒经PCR扩增后均出现约780 bp的目的条带,第10代重组病毒扩增的VP1基因测序结果显示Strep-tag II基因仍在重组病毒中,且无基因突变,表明r SVA-Strep的遗传稳定性较强;将SVA-WT和r SVA-Strep分别以MOI 0.01感染BHK-21细胞,在感染后4 h、8 h、12 h、24 h、36 h、48 h和72 h收获病毒测定病毒滴度并绘制生长曲线,结果显示在病毒感染36 h内r SVA-Strep和SVA-WT生长动力学基本一致,表明Strep-tag II的引入在病毒感染36 h内对SVA的复制基本无影响。采用0.2%甲醛充分灭活r SVA-Strep,并利用Strep-TactinRXT Purification纯化试剂盒纯化该灭活病毒,将r SVA-Strep灭活病毒上清加至Strep-Tactin亲和层析柱,分别用不同体积洗脱缓冲液依次洗脱病毒并通过western blot鉴定病毒的纯化效果。对纯化过程中不同洗脱液洗脱病毒的western blot结果显示,不同体积的洗脱液中E1、E2和E3均出现VP0和VP2两条带,且E1和E2的条带最明显,表明洗脱缓冲液的最佳洗脱体积约为2.2 CV;以上结果表明r SVA-Strep可通过Strep-Tactin亲和层析柱一步法纯化。本研究在SVA中插入Strep-tag II标签,为SVA灭活疫苗的快速纯化提供技术手段,也为其他病原的快速纯化方法的建立提供参考。

Differential Expression Analysis of Proteins Regulated by Rare Earth Cerium in Soybean Leaves at Seedling Stage

This study was to explore the functional mechanism of rare earth regulating soybean leaves and the characteristics and functions of differentially expressed proteins under the regulation of rare earth. In this study, Dongnong 42 was used as material, and 30 mg·L^(-1) CeCl_(3) solution was sprayed on soybean leaves at the seedling stage. Tandem mass tag(TMT) quantitative proteomics technique and bioinformatics analysis were used to identify soybean leaf proteins. A total of 8 510 proteins were identified, and 127 differentially expressed proteins(DEPs) in response to rare earth cerium regulation were identified, among which 64 were upregulated and 63 were down-regulated. The gene ontology(GO) annotation indicated that DEPs were mainly involved in metabolic process, cellular process, response to stimulus, biological regulation, and response to a stimulus;DEPs in cell module categories were mainly involved in cells, cell part, organelle, membrane, membrane part, organelle par, and protein-containing complex;DEPs in molecular functional categories were mainly involved in catalytic activity, binding and antioxidant activity. Kyoto encyclopedia of genes and genomes(KEGG) pathway significantly enriched starch and sucrose metabolism, glycolysis/gluconeogenesis, galactose metabolism, pentose phosphate pathway, and MAPK signaling pathway-plant. These DEPs were mainly involved in photosynthesis, glucose metabolism and stress response. Forty-six differential protein interaction networks were identified by protein interaction network analysis. This experiment provided a reference for studies of the mechanism of rare earth cerium regulating soybean leaf function from the proteomic perspective.

Serum proteins differentially expressed in gestational diabetes mellitus assessed using isobaric tag for relative and absolute quantitation proteomics

BACKGROUND As a well-known fact to the public,gestational diabetes mellitus(GDM)could bring serious risks for both pregnant women and infants.During this important investigation into the linkage between GDM patients and their altered expression in the serum,proteomics techniques were deployed to detect the differentially expressed proteins(DEPs)of in the serum of GDM patients to further explore its pathogenesis,and find out possible biomarkers to forecast GDM occurrence.METHODS Subjects were divided into GDM and normal control groups according to the IADPSG diagnostic criteria.Serum samples were randomly selected from four cases in each group at 24-28 wk of gestation,and the blood samples were identified by applying iTRAQ technology combined with liquid chromatography-tandem mass spectrometry.Key proteins and signaling pathways associated with GDM were identified by bioinformatics analysis,and the expression of key proteins in serum from 12 wk to 16 wk of gestation was further verified using enzyme-linked immunosorbent assay (ELISA).RESULTS Forty-seven proteins were significantly differentially expressed by analyzing the serum samples between the GDMgravidas as well as the healthy ones. Among them, 31 proteins were found to be upregulated notably and the rest16 proteins were downregulated remarkably. Bioinformatic data report revealed abnormal expression of proteinsassociated with lipid metabolism, coagulation cascade activation, complement system and inflammatory responsein the GDM group. ELISA results showed that the contents of RBP4, as well as ANGPTL8, increased in the serumof GDM gravidas compared with the healthy ones, and this change was found to initiate from 12 wk to 16 wk ofgestation.CONCLUSION GDM symptoms may involve abnormalities in lipid metabolism, coagulation cascade activation, complementsystem and inflammatory response. RBP4 and ANGPTL8 are expected to be early predictors of GDM.

34.TAG·西海美术馆:面朝大海的艺术长廊

设计:让·努维尔事务所地点:山东青岛西海岸新区时间:2012—2021年规模:17000 m^(2)TAG·西海美术馆位于一座半岛的海边,远离市区^([1,2])。从规划意图看,整个项目由美术馆和艺术工坊两部分组成,但由于工坊部分处于停工状态,艺术社区氛围尚未形成,作为主体建筑的美术馆便成了荒野中的孤独存在,公众可达性较差(图1)。尽管如此,从设计研究角度,这座美术馆仍有许多精彩看点值得关注。

TAG HEUER 竞速激情

继去年以卡莱拉系列Skipper计时码表的精钢版本标志着品牌重返帆船运动与航海领域之后,TAG Heuer泰格豪雅如今欣然发布一款以18K 5N玫瑰金打造的全新版本。金质的Skipper计时码表与精钢版本类似,同样基于2023年前期为纪念卡莱拉系列计时码表60周年推出且热销的“玻璃盒”(glassbox)设计,并搭载品牌自制机心Heuer 02(型号:TH20-06),这是一款为帆船赛计时而特别调整的Heuer 02机心。

Connotea中Social Tagging机制研究

选取Connotea为研究对象,统计分析标签、被标引资源、标引者两两关联关系,发现标签覆盖资源范围较广,标签共现现象突出,部分用户标引活跃,资源平均标签数较低,用户对内容关注度规律性转移,科研领域的用户比较多地使用规范词。提出增加标引词结构关联和细粒内容定位标引。

国外Social Tagging研究群体分析

基于抽取的国外知名数据库的学术文献,分析探讨Social Tagging研究群体的学科特征和研究视角。结果表明当前Social Tagging的研究群体主要局限于信息科学与计算机科学,且未形成居于统治权威地位的研究团体和研究人员。自从Social Tagging产生以来,"Social Tagging的组成功能"的研究一直是Social Tagging的基础性研究维度,不过随着应用深入发展,研究维度逐步转而融合信息检索和人机交互这两个相关领域知识分析"Tagger的行为及Social Tagging的界面设计"和"Social Tagging自身的组织和使用"这两个主要研究维度。

利用MRI Tagging分析β受体阻滞剂对左室心肌运动的影响

目的 探讨不同剂量β受体阻滞剂对左室心肌运动特征的影响及其作用机制.方法 13名志愿者注射不同剂量的短效β受体阻滞剂Esmolol,每个注射剂量起效4 min后利用MRI Tagging对左室心肌圆周向收缩运动、放射向收缩运动及长轴收缩运动进行磁化标记追踪,对获得的心肌运动结果进行心肌最大收缩程度、达到最大收缩时间、收缩速度及心肌做功分析.结果 注射小剂量Esmolol（5～25 μg·kg-1·min^-1）时,心肌运动程度增加,所需时间缩短,心肌做功增加;而当剂量增加至50～150 μg·kg-1·min^-1时,心肌运动明显较未用药时减弱,可以看到β受体阻滞剂Esmolol的负性肌力作用开始显现.结论 小剂量β受体阻滞剂与大剂量相比,更易对心肌运动产生正性肌力作用,这可能与不同种类心肌细胞表面β受体分布差异有关.

Molecular tagging techniques and their applications to the study of complex thermal flow phenomena1,211

This review article reports the recent progress in the development of a new group of molecule-based flow diagnostic techniques, which include molecular tag- ging velocimetry （MTV） and molecular tagging thermometry （MTT）, for both qualitative flow visualization of thermally induced flow structures and quantitative whole-field mea- surements of flow velocity and temperature distributions. The MTV and MTT techniques can also be easily combined to result in a so-called molecular tagging velocimetry and ther- mometry （MTV＆T） technique, which is capble of achieving simultaneous measurements of flow velocity and temperature distribution in fluid flows. Instead of using tiny particles, the molecular tagging techniques （MTV, MTT, and MTV＆T） use phosphorescent molecules, which can be turned into long-lasting glowing marks upon excitation by photons of appropriate wavelength, as the tracers for the flow veloc- ity and temperature measurements. The unique attraction and implementation of the molecular tagging techniques are demonstrated by three application examples, which include： （1） to quantify the unsteady heat transfer process from a heated cylinder to the surrounding fluid flow in order to exam- ine the thermal effects on the wake instabilities behind the heated cylinder operating in mixed and forced heat convec- tion regimes, （2） to reveal the time evolution of unsteady heat transfer and phase changing process inside micro-sized, icing water droplets in order to elucidate the underlying physics pertinent to aircraft icing phenomena, and （3） to achievesimultaneous droplet size, velocity and temperature measure- ments of ＂in-flight＂ droplets to characterize the dynamic and thermodynamic behaviors of flying droplets in spray flows.

The Detectability for the Myocardial Fibrosis by Tagging Imaging on Cardiovascular Magnetic Resonance

Purpose: Myocardial fibrosis causes cardiac dysfunction, arrhythmias, and sudden death. Tagging imaging on cardiovascular MR can measure the intra-myocardial motion from the dynamic deformation of lines superimposed on the myocardium. The purpose of this study was to evaluate the detectability of myocardial fibrosis using tagging imaging and to compare this with conventional cine imaging. Materials and Methods: We reviewed 4 normal control (NML) subjects, 4 patients with myocarditis (MYO), and 4 patients with old myocardial infarction (ICM). We measured circumferential strain (Ecc) from tagging imaging, and regional wall thickening (rWT) from cine imaging. Fibrosis was determined from a late gadolinium enhancement (LGE) image. We evaluate diagnostic performance by comparing values of the area under curve (AUC) using ROC analysis. Results: Mean values of Ecc and rWT decreased in the area of LGE both in MYO and ICM patients. AUC values of Ecc and rWT in all subjects were 0.98 and 0.84, respectively (p < 0.0001). These values in MYO patients were 0.95 and 0.72 (p = 0.007), respectively, and 0.99 and 0.75, respectively, in ICM patients (p = 0.0008). Conclusions: Both Ecc and rWT decreased in the area with fibrosis in the patients with MYO and ICM. Tagging imaging showed better detectability of myocardial fibrosis than did cine imaging.

Unified Framework of Performing Chinese Word Segmentation and Part-of-Speech Tagging

The paper proposes a unified framework to combine the advantages of the fast one-at-a-time approach and the high-performance all-at-once approach to perform Chinese Word Segmentation(CWS) and Part-of-Speech(PoS) tagging.In this framework,the input of the PoS tagger is a candidate set of several CWS results provided by the CWS model.The widely used one-at-a-time approach and all-at-once approach are two extreme cases of the proposed candidate-based approaches.Experiments on Penn Chinese Treebank 5 and Tsinghua Chinese Treebank show that the generalized candidate-based approach outperforms one-at-a-time approach and even the all-at-once approach.The candidate-based approach is also faster than the time-consuming all-at-once approach.The authors compare three different methods based on sentence,words and character-intervals to generate the candidate set.It turns out that the word-based method has the best performance.

铜麦6号苗期耐高铵与耐盐特性的鉴定及TaGS基因的表达模式分析

为鉴定小麦品种铜麦6号的耐高铵及耐盐特性,对铜麦6号、中国春及抗逆性较强的烟农19、三抗10号、矮优王、烟农21、山农25的发芽势和发芽率进行测定,发现铜麦6号的耐铵指数位列第一,耐盐指数位列第二。以中国春为对照品种,在高铵、高盐及其双重胁迫处理下,测定其苗期的叶片鲜重、根鲜重、主根和第一叶叶片长度以及根冠比,发现高铵、高盐及其双重胁迫处理7 d后,铜麦6号的上述指标均不同程度高于中国春(除铜麦6号的根冠比在高盐处理1 d后低于中国春外),说明铜麦6号具有较强的耐高铵和耐高盐特性。进一步采用qRT-PCR检测小麦苗期谷氨酰胺合成酶TaGS基因家族成员(根中TaGS1;1、TaGS1;2和TaGS1;3基因以及叶片中TaGS2基因)的表达模式,发现高铵、高盐及其双重处理均可诱导铜麦6号苗期TaGS基因上调表达,且高盐处理下上调表达更明显,其中除高铵与高盐双重处理7 d后的TaGS1;2基因以及高铵和高盐处理1 d后的TaGS2基因在铜麦6号中的表达量低于中国春外,其他三种处理不同时间点铜麦6号中这两个基因的相对表达量均不同程度高于中国春。这些结果说明,铜麦6号苗期TaGS基因家族与其耐高铵和耐高盐特性相关,其中TaGS1;2和TaGS2基因的贡献较大。

Improved hidden Markov model for speech recognition and POS tagging

In order to overcome defects of the classical hidden Markov model (HMM), Markov family model (MFM), a new statistical model was proposed. Markov family model was applied to speech recognition and natural language processing. The speaker independently continuous speech recognition experiments and the part-of-speech tagging experiments show that Markov family model has higher performance than hidden Markov model. The precision is enhanced from 94.642% to 96.214% in the part-of-speech tagging experiments, and the work rate is reduced by 11.9% in the speech recognition experiments with respect to HMM baseline system.

A New Part-of-Speech Tagging System Based on Closed-words,Word Form and Rules

A method of part-of-speech tagging of English text based on closed-words, wold-form and rules, its abstract model and formal description of its realizing procedure are presented. Finally, an experimental example is givento illustrate the application of this method.

A lexicalized second-order-HMM for ambiguity resolution in Chinese segmentation and POS tagging

Hidden Markov Model(HMM) is a main solution to ambiguities in Chinese segmentation and POS (part-of-speech) tagging. While most previous works for HMM-based Chinese segmentation and POS tagging consult POS information in contexts, they do not utilize lexical information which is crucial for resolving certain morphological ambiguity. This paper proposes a method which incorporates lexical information and wider context information into HMM. Model induction and related smoothing technique are presented in detail. Experiments indicate that this technique improves the segmentation and tagging accuracy by nearly 1%.

Tag与Tagging

Tag,直译标签,是一种由用户自定义的、用于描述信息的关键词。Tagging是用户为信息赋予Tag的行为。Delicious,Flickr等Web2.0网站的发展促进了它的流行,使之成为社会化书签、相册服务、博客等网站的常见功能。从目前网络发展形势而言——个人自生成内容(博文、图片、视频等)迅速增加,各类信息海量涌来——Tag可谓信息管理的强大利器。

Jointly Part-of-Speech Tagging and Semantic Role Labeling Using Auxiliary Deep Neural Network Model

Previous studies have shown that there is potential semantic dependency between part-of-speech and semantic roles.At the same time,the predicate-argument structure in a sentence is important information for semantic role labeling task.In this work,we introduce the auxiliary deep neural network model,which models semantic dependency between part-of-speech and semantic roles and incorporates the information of predicate-argument into semantic role labeling.Based on the framework of joint learning,part-of-speech tagging is used as an auxiliary task to improve the result of the semantic role labeling.In addition,we introduce the argument recognition layer in the training process of the main task-semantic role labeling,so the argument-related structural information selected by the predicate through the attention mechanism is used to assist the main task.Because the model makes full use of the semantic dependency between part-of-speech and semantic roles and the structural information of predicate-argument,our model achieved the F1 value of 89.0%on the WSJ test set of CoNLL2005,which is superior to existing state-of-the-art model about 0.8%.

Tandem Mass Tag-based proteomics analysis reveals the vital role of inflammation in traumatic brain injury in a mouse model

Proteomics is a powerful tool that can be used to elucidate the underlying mechanisms of diseases and identify new biomarkers.Therefore,it may also be helpful for understanding the detailed pathological mechanism of traumatic brain injury(TBI).In this study,we performed Tandem Mass Tag-based quantitative analysis of cortical proteome profiles in a mouse model of TBI.Our results showed that there were 302 differentially expressed proteins in TBI mice compared with normal mice 7 days after injury.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses showed that these differentially expressed proteins were predominantly involved in inflammatory responses,including complement and coagulation cascades,as well as chemokine signaling pathways.Subsequent transcription factor analysis revealed that the inflammation-related transcription factors NF-κB1,RelA,IRF1,STAT1,and Spi1 play pivotal roles in the secondary injury that occurs after TBI,which further corroborates the functional enrichment for inflammatory factors.Our results suggest that inflammation-related proteins and inflammatory responses are promising targets for the treatment of TBI.

Morpho-Syntactic Tagging of Text in “Baoule” Language Based on Hidden Markov Models (HMM)

The label text is a very important tool for the automatic processing of language. It is used in several applications such as morphological and syntactic text analysis, index-ing, retrieval, finished networks deterministic (in which all combinations of words that are accepted by the grammar are listed) or by statistical grammars (e.g., an n-gram in which the probabilities of sequences of n words in a specific order are given), etc. In this article, we developed a morphosyntactic labeling system language “Baoule” using hidden Markov models. This will allow us to build a tagged reference corpus and rep-resent major grammatical rules faced “Baoule” language in general. To estimate the parameters of this model, we used a training corpus manually labeled using a set of morpho-syntactic labels. We then proceed to an improvement of the system through the re-estimation procedure parameters of this model.

Gene Tagging by Activating DNA Transposon nDart in Indica Rice

As International Rice Genome sequencing Project (2005) demonstrated, the rice genome contains various transposons and about 13% of the genome is occupied by DNA transposons. So far, only a few DNA