冠菌素诱导橡胶树次生乳管分化的形成层细胞CUT&Tag文库构建和初步分析

CUT&Tag技术是一种研究蛋白质-DNA互作的新方法,使用超高活性的新型pG-Tn5转座酶,在抗体引导下精准靶向切割目的蛋白附近的DNA序列,从而进行cDNA建库和测序分析。此技术在人类和动物研究中应用广泛,由于植物细胞结构特殊,该技术在植物研究中应用较少。在前期研究中,我们发现组蛋白乙酰化修饰参与茉莉酸诱导橡胶树次生乳管分化的调控,但其分子机制尚未阐明。本研究利用冠菌素(COR)诱导橡胶树萌条维管形成层分化次生乳管的实验系统,通过酶解法获取高质量的形成层区细胞原生质体,使用组蛋白H3乙酰化修饰抗体对次生乳管分化过程中发生组蛋白乙酰化修饰的区域进行原位识别,采用CUT&Tag技术成功构建COR处理橡胶树树皮形成层细胞的cDNA文库。对构建cDNA文库进行质检和测序分析,发现文库质量较好,并通过差异基因的GO和KEGG富集分析,发现生长素、类黄酮代谢和蛋白质泛素化等相关基因得到富集。本研究结果为使用CUT&Tag技术构建植物组织的cDNA文库提供操作方法,为解析组蛋白乙酰化修饰调控橡胶树次生乳管分化的分子机制提供理论基础。

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。

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.

基于改进April Tag技术的吊件空间位置与姿态监测方法

为了解决吊件位置与姿态的监测难题,基于改进的April Tag技术提出了一种吊件位姿视觉测量方法。该方法采用专用磁吸式标签固定方式,降低标签安装难度和要求;通过视觉测量吊件上的April Tag标签位置,结合吊件关键尺寸信息,基于空间坐标变换理论实现了吊件位置和姿态的测量;同时根据标签间的相对关系对测量结果进行数据校验,提高测量精度。对架空输电线路双平臂落地抱杆组塔吊装施工的吊件进行了位置与姿态测量试验。结果表明:本文方法能够有效准确实现吊件空间位置与姿态的测量。

Joint Modeling of Citation Networks and User Preferences for Academic Tagging Recommender System

In the tag recommendation task on academic platforms,existing methods disregard users’customized preferences in favor of extracting tags based just on the content of the articles.Besides,it uses co-occurrence techniques and tries to combine nodes’textual content for modelling.They still do not,however,directly simulate many interactions in network learning.In order to address these issues,we present a novel system that more thoroughly integrates user preferences and citation networks into article labelling recommendations.Specifically,we first employ path similarity to quantify the degree of similarity between user labelling preferences and articles in the citation network.Then,the Commuting Matrix for massive node pair paths is used to improve computational performance.Finally,the two commonalities mentioned above are combined with the interaction paper labels based on the additivity of Poisson distribution.In addition,we also consider solving the model’s parameters by applying variational inference.Experimental results demonstrate that our suggested framework agrees and significantly outperforms the state-of-the-art baseline on two real datasets by efficiently merging the three relational data.Based on the Area Under Curve(AUC)and Mean Average Precision(MAP)analysis,the performance of the suggested task is evaluated,and it is demonstrated to have a greater solving efficiency than current techniques.

Influence of ear tags on the results of body composition analysis in mice

Background:The aim of this study was to investigate the influence of marking meth-ods on the outcomes of body composition analysis and provide guidance for the se-lection of marking methods in mouse body composition analysis.Methods:Male C57BL/6J mice aged 6 weeks were randomly assigned for pre-and post-ear tagging measurements.The body composition of the mice was measured using a small animal body composition analyzer,which provided measurements of the mass of fat,lean,and free fluid.Then,the mass of fat,lean and free fluid to body weight ratio was gained.Further data analysis was conducted to obtain the range and coeffi-cient of variation in body composition measurements for each mouse.The distribution of fat and lean tissue in the mice was also analyzed by comparing the fat-to-lean ratio.Results:(1)The mass of all body composition components in the ear tagging group was significantly lower than that in the control group.(2)There was a significant in-crease in the range and coefficient of variation of body composition measurements between the ear tagging group and the control group.(3)The fat-to-lean ratio in the ear tagging group was significantly lower than that in the control group.Conclusions:Ear tagging significantly lowered the results of body composition analy-sis in mice and higher the results of measurement error.Therefore,ear tagging should be avoided as much as possible when conducting body composition analysis experi-ments in mice.

A Novel Anti-Collision Algorithm for Large Scale of UHF RFID Tags Access Systems

When the radio frequency identification(RFID)system inventories multiple tags,the recognition rate will be seriously affected due to collisions.Based on the existing dynamic frame slotted Aloha(DFSA)algorithm,a sub-frame observation and cyclic redundancy check(CRC)grouping combined dynamic framed slotted Aloha(SUBF-CGDFSA)algorithm is proposed.The algorithm combines the precise estimation method of the quantity of large-scale tags,the large-scale tags grouping mechanism based on CRC pseudo-randomcharacteristics,and the Aloha anti-collision optimization mechanism based on sub-frame observation.By grouping tags and sequentially identifying themwithin subframes,it accurately estimates the number of remaining tags and optimizes frame length accordingly to improve efficiency in large-scale RFID systems.Simulation outcomes demonstrate that this proposed algorithmcan effectively break through the system throughput bottleneck of 36.8%,which is up to 30%higher than the existing DFSA standard scheme,and has more significant advantages,which is suitable for application in largescale RFID tags scenarios.

A U-Shaped Network-Based Grid Tagging Model for Chinese Named Entity Recognition

Chinese named entity recognition(CNER)has received widespread attention as an important task of Chinese information extraction.Most previous research has focused on individually studying flat CNER,overlapped CNER,or discontinuous CNER.However,a unified CNER is often needed in real-world scenarios.Recent studies have shown that grid tagging-based methods based on character-pair relationship classification hold great potential for achieving unified NER.Nevertheless,how to enrich Chinese character-pair grid representations and capture deeper dependencies between character pairs to improve entity recognition performance remains an unresolved challenge.In this study,we enhance the character-pair grid representation by incorporating both local and global information.Significantly,we introduce a new approach by considering the character-pair grid representation matrix as a specialized image,converting the classification of character-pair relationships into a pixel-level semantic segmentation task.We devise a U-shaped network to extract multi-scale and deeper semantic information from the grid image,allowing for a more comprehensive understanding of associative features between character pairs.This approach leads to improved accuracy in predicting their relationships,ultimately enhancing entity recognition performance.We conducted experiments on two public CNER datasets in the biomedical domain,namely CMeEE-V2 and Diakg.The results demonstrate the effectiveness of our approach,which achieves F1-score improvements of 7.29 percentage points and 1.64 percentage points compared to the current state-of-the-art(SOTA)models,respectively.

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。

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.

一种含SNAP-tag TM底物的硫醇荧光探针

合成了一种同时含α,β-不饱和酮活性位点和SNAP-tag TM底物的香豆素母体探针香豆素-烯酮-苯甲氨基鸟嘌呤(CAKBG).通过测试探针CAKBG的紫外-可见吸收光谱和荧光发射光谱,发现探针CAKBG在极性溶剂(例如,甘油、乙二醇、水、DMSO)中有更长的吸收波长,且探针CAKBG对溶液的pH变化不敏感.此外,光谱实验发现生物硫醇Cys和Hcy均能引起探针CAKBG的最大吸收波长发生蓝移,且当激发波长为405 nm时,Cys和Hcy均能导致探针CAKBG在487 nm处的荧光发射峰明显增强.细胞成像结果表明,含有SNAP-tag TM底物的硫醇荧光探针CAKBG在活细胞中可以对Cys、Hcy进行荧光成像.综上,SNAP-Tag TM蛋白标签技术有望今后在硫醇荧光探针的细胞成像中得到推广应用.

TAG HEUER 传奇之作 全面提升

传奇的卡莱拉泛美公路赛以其在墨西哥的严苛挑战而闻名,杰克·豪雅(Jack Heuer)以此为灵感,为TAG HEUER泰格豪雅卡莱拉系列腕表赋予了这一独特且极具影响力的名字。该系列凭借持续挑战边界的精准度,迅速赢得了市场的青睐并占据表迷的腕间,时至今日依旧风靡全球。在对系列进行了无数次的演绎后,日前泰格豪雅再度带来多款全新作品,将竞速传承、前沿材质及创新设计巧妙融合,一方面,是对2016年焕新的备受赞誉的卡莱拉系列运动风格计时码表的大胆演绎,另一方面,标志着该系列的又一次升级演变。

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。

养殖RFID测温定位电子耳标研究——基于智慧农业区块链

首先,介绍了区块链和物联网技术在智慧农业应用的概念;然后,基于结构、温度传感器、RSSI定位和天线等多个模块实现了对测温定位电子耳标的设计;最后,为了验证该RFID测温定位电子耳标的性能,设计了一套放养羊群的养殖信息监测系统,对每只羊的实际位置和体温进行监测。测试结果表明:系统可以准确测量山羊的体温和位置,响应快,稳定性高,符合设计需求。

大西洋中脊TAG热液活动区海底热液沉积物的硫同位素组成及其地质意义

对 1 98 8年取自大西洋中脊TAG热液活动区的海底表层热液沉积物中 36件硫化物样品进行了硫同位素组成分析。结合ODPLeg1 58的近期成果 ,对TAG区热液沉积物的硫同位素组成及其时空演变、硫源、硫同位素偏重的原因和硫的演化进行了探讨 ,得出如下结论 :( 1 )TAG区表层热液沉积物的δ34S值从 3 9‰— 7 6‰变化 ,均值为 5 98‰ ,与其它洋中脊热液活动区相比明显偏重。 ( 2 )从海底表层区到蚀变玄武岩区 ,热液沉积物的硫同位素组成有增大的趋势 ,而时间对热液沉积物的硫同位素组成则没有明显的控制。 ( 3)热液沉积物中硫主要来自玄武岩 ,部分来自海水硫酸盐 ,是海水硫酸盐和玄武岩中硫不同程度混合的结果 ,且各硫源在不同阶段提供硫的比例分配和方式明显不同。 ( 4 )与其它无沉积物覆盖洋中脊中热液活动区相比 ,相对较大比例海水来源硫的加入是导致TAG区硫同位素组成偏重的一个重要原因 ,而TAG区具备有利海水硫酸盐还原作用进行的温度和水 /岩比值条件则是促使该区热液沉积物中海水来源硫相对较多的主要原因。 ( 5)海水的直接混入、流体 -玄武岩相互作用、先期形成硫化物的重溶作用和硫酸盐矿物的还原作用是海水和玄武岩直接或间接提供硫的主要方式 ,也是导致本区硫同位素演化复杂化的主要原因 ,且硫的演化?

6×His-tag在抗菌肽表达纯化中的应用及其对抗菌肽活性的影响

为方便抗菌肽的体外表达和纯化,以蝎子防御素为对象,在其编码基因末端加入6×His-tag,在酵母表达系统中表达,获得有活性的防御素Sd和融合蛋白Sd-His。进一步纯化、体外抑菌活性及热酸稳定性等试验发现,6×His-tag不但没有降低防御素的抑菌活性,反而有稍微的增强趋势。结果表明,6×His-tag可用于抗菌肽的融合表达且不影响其活性,为抗菌肽的表达纯化提供了一个更简便有效的方法。

Tag和现代教育技术

Tag是新一代互联网模式Web2.0的核心应用之一,它实现了分众分类的思想,具有自发性、共享性和动态自适应性的特点。Tag的推广和应用将会增强人们的概括意识,引发评价标准和分类原则的多元化,促进人们的远程交流,有利于社会网络的形成,因此,Tag将在一定程度上影响人们的思维方式和生活方式。本文在分析Tag对网络文化的影响的基础上,指出了Tag在教育技术中诸多方面应用的可行性和优越性。

血清肿瘤标志物MG7AgTAG72CA19－9诊断胃癌的前瞻性对照研究

对胃癌单抗ＭＧ７所识别的胃癌新的相关抗原ＭＧ７Ａｓ、ＴＡＧ７２、ＣＡ１９．９（均为ＩＲＭＡ法）进行了前瞻性规范的诊断试验对照研究，评价ＭＧ７Ａｇ及其联合应用诊断胃癌的价值。结果：ＲＯＣ曲线法选择ＭＧ７Ａｇ界值为７．５ｋＵ／Ｌ。ＭＧ７Ａｇ、ＴＡＧ７２、ＣＡ１９－９诊断胃癌的敏感性分别为４８．８％、４５．３％、３８．５％，无显著性差别（Ｐ＞０．０５）；３者特异性分别为９２．８％、９１．６％、９２．８％（Ｐ＞０．０５）。ＭＧ７Ａｇ与ＴＡＧ７２，ＣＡ１９－９诊断Ⅲ、Ⅳ期胃癌的敏感性显著高于Ⅰ、Ⅱ期，其中ＭＧ７Ａｇ分别为６９．６％和２７．３％。ＭＧ７Ａｇ是一种有价值的胃癌标志物，与ＴＡＧ７２或（和）ＣＡ１９－９联合应用可显著提高诊断胃癌的敏感性。

Tag-TextRank:一种基于Tag的网页关键词抽取方法

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