PLOS One投稿格式

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研究生时经历过的几次论文投稿发表经验汇总

PLoS One（IF>4）：相信很多园友都有投稿该杂志的经历吧，我就说说我所知道的。这个杂志投稿过程中需要填写的要求相对其它杂志要多一些，例如 ethnic assessment 等，另外，因为投稿量巨大的原因，文章在投到该杂志之后要等 20 天左右才能 invited editor，但是在开始审稿后速度则相对快一点，一般主编要求审稿在 10 天内审稿结束，所以文章投到 PLoS One 后如果没有意外，在 40 天内都能收到审稿结果了。我的导师是 PLoS One 的 academic editor，也认识很多 PLoS One 的其它副主编，在投稿时会推荐自己认识的副主编。我在去年有一篇文章投到 PLoS One，是我导师的朋友负责该文章，两个审稿人也是我们推荐的，所以从投稿到接收都非常顺利。PLoS One 的投稿量巨大，academic editor 的数量也很多，也包括很多中国人，即便如此，每个 academic editor 的工作量也是相当大，我登陆我的导师的 PLoS One 账号，发现几乎每天都有新文章的 invitation。
Schizophrenia Research（IF>4）：一个专门研究精神分裂症的杂志，影响因子常年在 4-5 分之间，对文章的质量要求没有那么高。我目前已经在该杂志发表过三篇文章，2011 年第一次在这个杂志投稿，一审 4 个半月，8 个审稿人，均是 positive 意见，不知道主编怎么想的；论文咨询工作QQ2：2035204840 第二篇文章一审 4 个月，1 个审稿人；第三篇文章一审 5 个月，一个审稿人。几乎每一篇文章我们都要催稿，不过催也没用，感觉主编不是特别热情，只是让我们 be patient。如果不是逼不得已，不会投该杂志。
在园子里也混迹了几年时间，大部分时间在看帖，偶尔回帖，求助过园友，请教过园友，也帮助过园友，鼓励过园友，还刺激过园友，嘲讽过园友，当然，一切都无恶意，只是就事论事，还是希望大家能够投稿顺利，多多发文章。

plos ons投稿须知

参考文献图表（tables）
� � � included at the end of the manuscript. 文章最后面 have a concise title Tables occupying more than one printed page should be avoided,
TABLE GUIDE 题目和标注：题目简洁，不超过一句话，卸载图表上方图标上面标注图表序号，例如 Table 1 标注写在图表下面，用来解释缩写等具体要求：可修改，不可为图片类型单独的列有标题或者副标题大小不超过(7 in � � Not exceed 300 words；不应该有引用尽量避免缩写
引言文章主体部分致谢：Funding sources should not be included in the acknowledgments, or anywhere in the
manuscript file. You will provide this information during the manuscript submission process.
作者和单位
作者：名字顺序：名——姓，例如 Kun Zhang 单位信息：associated department, university, or organizational affiliation and its location, including city, state/province (if applicable), and country 备注：作者和单位信息提交后不可修改，请务必仔细通讯作者：One author should be designated as the corresponding author, and his or her email address or other contact information should be included on the manuscript cover page. This information will be published with the article if accepted.

plos one投稿要求

PLOS ONE Manuscript Guidelines1Format RequirementsGuidelines for Standard Sectionso Titleo Authors and Affiliationso Abstracto Introductiono Materials and Methodso Results, Discussion, and Conclusionso Acknowledgmentso Referenceso Tableso Figure Legendso Striking ImagesSpecific Reporting Guidelineso Human Subject Researcho Clinical Trialso Animal Researcho Observational and Field Studieso Cell Line Researcho Systematic Review/Meta-Analysiso Paleontology and Archaeology Researcho Software Paperso Database Paperso New Zoological Taxono New Botanical Taxono New Fungal Taxono Qualitative Research1. Format RequirementsPLOS ONE does not consider presubmission inquiries. All submissions should be prepared with the following files:•Cover letter•Manuscript, including tables and figure legends•Figures (guidelines for preparing figures can be found at the Figure and Table Guidelines) Prior to submission, authors who believe their manuscripts would benefit from professional editing are encouraged to use language-editing and copyediting services. Obtaining this service is the responsibility of the author, and should be done before initial submission. These services can be found on the web using search terms like "scientific editing service" or "manuscript editing service." Submissions are not copyedited before publication.Submissions that do not meet the PLOS ONE Publication Criterion for language standards may berejected.Cover LetterYou should supply an approximately one page cover letter that:•Concisely summarizes why your paper is a valuable addition to the scientific literature •Briefly relates your study to previously published work•Specifies the type of article you are submitting (for example, research article, systematic review, meta-analysis, clinical trial)•Describes any prior interactions with PLOS regarding the submitted manuscript•Suggests appropriate PLOS ONE Academic Editors to handle your manuscript (view a complete listing of our academic editors)•Lists any recommended or opposed reviewersYour cover letter should not include requests to reduce or waive publication fees. Should your manuscript be accepted, you will have the opportunity to include your requests at that time. See PLOS ONE Editorial Policy for more information regarding publication fees.Manuscript OrganizationPLOS ONE considers manuscripts of any length. There are no explicit restrictions for the number of words, figures, or the length of the supporting information, although we encourage a concise and accessible writing style. We will not consider monographs.All manuscripts should be double-spaced and include line numbers and page numbers. Manuscripts should begin with the ordered sections:•Title•Authors•Affiliations•Abstract•Introductionand end with the sections of:•Acknowledgments•References•Figure Legends•TablesFigures should not be included in the main manuscript file. Each figure must be prepared and submitted as an individual file. Find more information about preparing figures here.The title, authors, and affiliations should all be included on a title page as the first page of the manuscript file.There are no explicit requirements for section organization between these beginning and ending sections. Articles may be organized in different ways and with different section titles, according to the authors' preference. In most cases, internal sections include:•Materials and Methods•Results•Discussion•Conclusions (optional)PLOS ONE has no specific requirements for the order of these sections, and in some cases it maybe appropriate to combine sections. Guidelines for individual sections can be found below. Abbreviations should be kept to a minimum and defined upon first use in the text. Non-standard abbreviations should not be used unless they appear at least three times in the text. Standardized nomenclature should be used as appropriate, including appropriate usage of species names and SI units.Manuscript File Type RequirementsAuthors may submit their manuscript files in Word (as .doc or .docx), LaTeX (as .pdf), or RTF format. Only RTF and .doc files can be used during the production process. Word files must not be protected.LaTeX Submissions. If you would like to submit your manuscript using LaTeX, you must author your article using the PLOS ONE LaTeX template and BibTeX style sheet. Articles prepared in LaTeX may be submitted in PDF format for use during the review process. After acceptance, however, .tex files and formatting information will be required as a zipped file. Please consult our LaTeX guidelines for a list of what will be required.Submissions with equations.If your manuscript is or will be in .docx format and contains equations, you must follow the instructions below to make sure that your equations are editable when the file enters production.If you have not yet composed your article, you can ensure that the equations in your .docx file remain editable in .doc by enabling "Compatibility Mode" before you begin. To do this, open a new document and save as Word 97-2003 (*.doc). Several features of Word 2007/10 will now be inactive, including the built-in equation editing tool. You can insert equations in one of the two ways listed below.If you have already composed your article as .docx and used its built-in equation editing tool, your equations will become images when the file is saved down to .doc. To resolve this problem, re-key your equations in one of the two following ways.1. Use MathType to create the equation (recommended)2. Go to Insert > Object > Microsoft Equation3.0 and create the equationIf, when saving your final document, you see a message saying "Equations will be converted to images," your equations are no longer editable and PLoS will not be able to accept your file.Back to top2. Guidelines for Standard SectionsTitleManuscripts must be submitted with both a full title and a short title, which will appear at the top of the PDF upon publication if accepted. Only the full title should be included in the manuscript file; the short title will be entered during the online submission process.The full title must be 250 characters or fewer. It should be specific, descriptive, concise, and comprehensible to readers outside the subject field. Avoid abbreviations if possible. Where appropriate, authors should include the species or model system used (for biological papers) or type of study design (for clinical papers).Examples:•Impact of Cigarette Smoke Exposure on Innate Immunity: A Caenorhabditis elegansModel•Solar Drinking Water Disinfection (SODIS) to Reduce Childhood Diarrhoea in Rural Bolivia: A Cluster-Randomized, Controlled TrialThe short title must be 50 characters or fewer and should state the topic of the paper.Back to top Authors and AffiliationsAll author names should be listed in the following order:•First names (or initials, if used),•Middle names (or initials, if used), and•Last names (surname, family name)Each author should list an associated department, university, or organizational affiliation and its location, including city, state/province (if applicable), and country. If the article has been submitted on behalf of a consortium, all author names and affiliations should be listed at the end of the article.This information cannot be changed after initial submission, so please ensure that it is correct.To qualify for authorship, a researcher should contribute to all of the following: 2Conception and design of the work, acquisition of data, or analysis and interpretation of data3Drafting the article or revising it critically for important intellectual content4Final approval of the version to be publishedAll persons designated as authors should qualify for authorship, and all those who qualify should be listed. Each author must have participated sufficiently in the work to take public responsibility for appropriate portions of the content. Those who contributed to the work but do not qualify for authorship should be listed in the acknowledgments.When a large group or center has conducted the work, the author list should include the individuals whose contributions meet the criteria defined above, as well as the group name.One author should be designated as the corresponding author, and his or her email address or other contact information should be included on the manuscript cover page. This information will be published with the article if accepted.See the PLOS ONE Editorial Policy regarding authorship criteria for more information.Back to top AbstractThe abstract should:•Describe the main objective(s) of the study•Explain how the study was done, including any model organisms used, without methodological detail•Summarize the most important results and their significance•Not exceed 300 wordsAbstracts should not include:•Citations•Abbreviations, if possibleBack to top IntroductionThe introduction should:•Provide background that puts the manuscript into context and allows readers outside the field to understand the purpose and significance of the study•Define the problem addressed and why it is important•Include a brief review of the key literature•Note any relevant controversies or disagreements in the field•Conclude with a brief statement of the overall aim of the work and a comment about whether that aim was achievedBack to top Materials and MethodsThis section should provide enough detail to allow suitably skilled investigators to fully replicate your study. Specific information and/or protocols for new methods should be included in detail. If materials, methods, and protocols are well established, authors may cite articles where those protocols are described in detail, but the submission should include sufficient information to be understood independent of these references.We encourage authors to submit detailed protocols for newer or less well-established methods as Supporting Information. Further information about formatting Supporting Information files, can be found here.Methods sections of papers on research using human or animal subjects and/or tissue or field sampling must include required ethics statements. See the Reporting Guidelines for human research, clinical trials, animal research, and observational and field studies for more information. Methods sections of papers with data that should be deposited in a publicly available database should specify where the data have been deposited and provide the relevant accession numbers and version numbers, if appropriate. Accession numbers should be provided in parentheses after the entity on first use. If the accession numbers have not yet been obtained at the time of submission, please state that they will be provided during review. They must be provided prior to publication. A list of recommended repositories for different types of data can be found here. Methods sections of papers using cell lines must state the origin of the cell lines used. See the Reporting Guidelines for cell line research for more information.Methods sections of papers adding new taxon names to the literature must follow the Reporting Guidelines below for a new zoological taxon, botanical taxon, or fungal taxon.Back to top Results, Discussion, and ConclusionsThese sections may all be separate, or may be combined to create a mixed Results/Discussion section (commonly labeled "Results and Discussion") or a mixed Discussion/Conclusions section (commonly labeled "Discussion"). These sections may be further divided into subsections, each with a concise subheading, as appropriate. These sections have no word limit, but the language should be clear and concise.Together, these sections should describe the results of the experiments, the interpretation of these results, and the conclusions that can be drawn. Authors should explain how the results relate to the hypothesis presented as the basis of the study and provide a succinct explanation of the implications of the findings, particularly in relation to previous related studies and potential future directions for research.PLOS ONE editorial decisions do not rely on perceived significance or impact, so authors shouldavoid overstating their conclusions. See the PLOS ONE Publication Criteria for more information.Back to top AcknowledgmentsPeople who contributed to the work but do not fit the PLOS ONE authorship criteria should be listed in the acknowledgments, along with their contributions. You must ensure that anyone named in the acknowledgments agrees to being so named.Funding sources should not be included in the acknowledgments, or anywhere in the manuscript file. You will provide this information during the manuscript submission process.Back to top ReferencesOnly published or accepted manuscripts should be included in the reference list. Manuscripts that have been submitted but not yet accepted should not be cited. Limited citation of unpublished work shoul d be included in the body of the text only as “unpublished data.”References must be listed at the end of the manuscript and numbered in the order that they appear in the text. In the text, citations should be indicated by the reference number in brackets. Journal name abbreviations should be those found in the NCBI databases. A number of reference software companies supply PLOS style files (e.g., Reference Manager, EndNote).References should be formatted as follows:•Published papers. Hou WR, Hou YL, Wu GF, Song Y, Su XL, et al. (2011) cDNA, genomic sequence cloning and overexpression of ribosomal protein gene L9 (rpL9) of the giant panda (Ailuropoda melanoleuca). Genet Mol Res 10: 1576-1588.Note: Use of a DOI number for the full-text article is acceptable as an alternative to or in addition to traditional volume and page numbers.•Accepted, unpublished papers. Sa me as above, but “In press” appears instead of the page numbers.•Electronic journal articles. Huynen MMTE, Martens P, Hilderlink HBM (2005) The health impacts of globalisation: a conceptual framework. Global Health 1: 14. Available: /content/1/1/14. Accessed 25 January 2012.•Books. Bates B (1992) Bargaining for life: A social history of tuberculosis. Philadelphia: University of Pennsylvania Press. 435 p.•Book chapters Hansen B (1991) New York City epidemics and history for the public. In: Harden VA, Risse GB, editors. AIDS and the historian. Bethesda: National Institutes of Health.pp. 21-28.Back to top TablesTables should be included at the end of the manuscript. All tables should have a concise title. Footnotes can be used to explain abbreviations. Citations should be indicated using the same style as outlined above. Tables occupying more than one printed page should be avoided, if possible. Larger tables can be published as Supporting Information. Please ensure that table formatting conforms to our Guidelines for table preparation.Back to top Figure LegendsFigures should not be included in the manuscript file, but figure legends should be. Guidelines forpreparing figures can be found here.Figure legends should describe the key messages of a figure. Legends should have a short title of 15 words or less. The full legend should have a description of the figure and allow readers to understand the figure without referring to the text. The legend itself should be succinct, avoid lengthy descriptions of methods, and define all non-standard symbols and abbreviations.Further information about figure legends can be found in the Figure Guidelines.Back to top Striking ImagesAuthors are encouraged to upload a "striking image" that may be used to represent their paper online in places like the journal homepage or in search results. The striking image must be derived from a figure or supporting information file from the paper, ie. a cropped portion of an image or the entire image. Striking images should ideally be high resolution, eye-catching, single panel images, and should ideally avoid containing added details such as text, scale bars, and arrows. If no striking image is uploaded, a figure from the paper will be designated as the striking image. Please keep in mind that PLOS's Creative Commons Attribution License applies to striking images. As such, do not submit any figures or photos that have been previously copyrighted unless you have express written permission from the copyright holder to publish under the CCAL license. Note that all published materials in PLOS ONE are freely available online, and any third party is permitted to read, download, copy, distribute, and use these materials in any way, even commercially, with proper attribution.Care should be taken with the following image types in particular:5PLOS ONE is unable to publish any images generated by Google software (Google Maps, Street View, and Earth)6Maps in general are usually copyrighted, especially satellite maps7Photographs8Commercial or government images, slogans, or logos9Images from Facebook or TwitterAuthors must also take special care when submitting manuscripts that contain potentially identifying images of people. Identifying information should not be included in the manuscript unless the information is crucial and the individual has provided written consent by completing the Consent Form for Publication in a PLOS Journal (PDF).For license inquiries, e-mail license [at] .Back to top3. Specific Reporting GuidelinesHuman Subject ResearchMethods sections of papers on research using human subject or samples must include ethics statements that specify:•The name of the approving institutional review board or equivalent committee(s). If approval was not obtained, the authors must provide a detailed statement explaining why it was not needed•Whether informed consent was written or oral. If informed consent was oral, it must be stated in the manuscript:o Why written consent could not be obtainedo That the Institutional Review Board (IRB) approved use of oral consento How oral consent was documentedFor studies involving humans categorized by race/ethnicity, age, disease/disabilities, religion, sex/gender, sexual orientation, or other socially constructed groupings, authors should: •Explicitly describe their methods of categorizing human populations•Define categories in as much detail as the study protocol allows•Justify their choices of definitions and categories, including for example whether any rules of human categorization were required by their funding agency•Explain whether (and if so, how) they controlled for confounding variables such as socioeconomic status, nutrition, environmental exposures, or similar factors in their analysisIn addition, outmoded terms and potentially stigmatizing labels should be changed to more current, acceptable terminology. Examples: "Caucasian" should be changed to "white" or "of [Western] European descent" (as appropriate); "cancer victims" should be changed to "patients with cancer." For papers that include identifying, or potentially identifying, information, authors must download the Consent Form for Publication in a PLOS Journal(PDF), which the individual, parent, or guardian must sign once they have read the paper and been informed about the terms of PLOS open-access license. The signed consent form should not be submitted with the manuscript, but authors should securely file it in the individual's case notes and the methods section of the manuscript should explicitly state that consent authorization for publication is on file, using wording like:The individual in this manuscript has given written informed consent (as outlined in PLOS consent form) to publish these case details.For more information about PLOS ONE policies regarding human subject research, see the Publication Criteria and Editorial Policies.Back to top Clinical TrialsAuthors of manuscripts describing the results of clinical trials must adhere to the CONSORT reporting guidelines appropriate to their trial design, available on the CONSORT Statement website. Before the paper can enter peer review, authors must:10Provide the registry name and number in the methods section of the manuscript11Provide a copy of the trial protocol as approved by the ethics committee and a completed CONSORT checklist as Supporting Information (which will be published alongside the paper, if accepted)12Include the CONSORT flow diagram as the manuscript's "Figure 1"Any deviation from the trial protocol must be explained in the paper. Authors must explicitly discuss informed consent in their paper, and we reserve the right to ask for a copy of the patient consent form.The methods section must include the name of the registry, the registry number, and the URL of your trial in the registry database for each location in which the trial is registered.For more information about PLOS ONE policies regarding clinical trials, see the Editorial Policies.Back to topAnimal ResearchMethods sections of manuscripts reporting results of animal research must include required ethics statements that specify:•The full name of the relevant ethics committee that approved the work, and the associated permit number(s) (where ethical approval is not required, the manuscript should include a clear statement of this and the reason why)•Relevant details for efforts taken to ameliorate animal sufferingFor example:This study was carried out in strict accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. The protocol was approved by the Committee on the Ethics of Animal Experiments of the University of Minnesota (Permit Number: 27-2956). All surgery was performed under sodium pentobarbital anesthesia, and all efforts were made to minimize suffering.The organism(s) studied should always be stated in the abstract. Where research may be confused as pertaining to clinical research, the animal model should also be stated in the title.We ask authors to follow the ARRIVE (Animal Research: Reporting of In Vivo Experiments) guidelines for all submissions describing laboratory-based animal research and to upload a completed ARRIVE Guidelines Checklist to be published as supporting information. Please note that inclusion of a completed ARRIVE Checklist will be a formal requirement for publication at a later date.For more information about PLOS ONE policies regarding animal research, see the Publication Criteria and Editorial Policies.Back to top Observational and Field StudiesMethods sections for submissions reporting on any type of field study must include ethics statements that specify:•Permits and approvals obtained for the work, including the full name of the authority that approved the study; if none were required, authors should explain why•Whether the land accessed is privately owned or protected•Whether any protected species were sampled•Full details of animal husbandry, experimentation, and care/welfare, where relevantFor more information about PLOS ONE policies regarding observational and field studies, see the Publication Criteria and Editorial Policies.Back to top Cell Line ResearchMethods sections for submissions reporting on research with cell lines should state the origin of any cell lines. For established cell lines the provenance should be stated and references must also be given to either a published paper or to a commercial source. If previously unpublished de novo cell lines were used, including those gifted from another laboratory, details of institutional review board or ethics committee approval must be given, and confirmation of written informed consent must be provided if the line is of human origin.Back to top Systematic Review/Meta-AnalysisA systematic review paper, as defined by The Cochrane Collaboration, is a review of a clearly formulated question that uses explicit, systematic methods to identify, select, and critically appraise relevant research, and to collect and analyze data from the studies that are included in the review. These reviews differ substantially from narrative-based reviews or synthesis articles. Statistical methods (meta-analysis) may or may not be used to analyze and summarize the results of the included studies.Reports of systematic reviews and meta-analyses must include a completed PRISMA (Preferred Reporting Items for Systematic Reviews and Meta-Analyses) checklist and flow diagram to accompany the main text. Blank templates are available here:•Checklist: PDF or Word document•Flow diagram: PDF or Word documentAuthors must also state in their "Methods" section whether a protocol exists for their systematic review, and if so, provide a copy of the protocol as Supporting Information and provide the registry number in the abstract.If your article is a Systematic Review or a Meta-Analysis you should:•State this in your cover letter•Select "Research Article" as your article type when submitting•Include the PRISMA flowchart as Figure 1 (required where applicable)•Include the PRISMA checklist as Supporting InformationBack to top Paleontology and Archaeology ResearchManuscripts reporting paleontology and archaeology research must include descriptions of methods and specimens in sufficient detail to allow the work to be reproduced. Data sets supporting statistical and phylogenetic analyses should be provided, preferably in a format that allows easy re-use.Specimen numbers and complete repository information, including museum name and geographic location, are required for publication. Locality information should be provided in the manuscript as legally allowable, or a statement should be included giving details of the availability of such information to qualified researchers.If permits were required for any aspect of the work, details should be given of all permits that were obtained, including the full name of the issuing authority. This should be accompanied by the following statement:All necessary permits were obtained for the described study, which complied with all relevant regulations.If no permits were required, please include the following statement:No permits were required for the described study, which complied with all relevant regulations.See the PLOS ONE Editorial Policies for more information regarding manuscripts describing paleontology and archaeology research.Back to top Software PapersManuscripts describing software should provide full details of the algorithms designed. Describe any dependencies on commercial products or operating system. Include details of the supplied test data and explain how to install and run the software. A brief description of enhancements made inthe major releases of the software may also be given. Authors should provide a direct link to the deposited software from within the paper.See the PLOS ONE Editorial Policies for more information about submitting manuscripts.Back to top Database PapersFor descriptions of databases, provide details about how the data were curated, as well as plans for long-term database maintenance, growth, and stability. Authors should provide a direct link to the database hosting site from within the paper.See the PLOS ONE Editorial Policies for more information about submitting manuscripts describing databases.Back to top New Zoological TaxonFor proper registration of a new zoological taxon, we require two specific statements to be included in your manuscript.In the Results section, the globally unique identifier (GUID), currently in the form of a Life Science Identifier (LSID), should be listed under the new species name, for example: Anochetus boltoni Fisher sp. nov. urn:lsid::act:B6C072CF-1CA6-40C7-8396-534E91EF7FBBYou will need to contact Zoobank to obtain a GUID (LSID). Please do this as early as possible to avoid delay of publication upon acceptance of your manuscript. It is your responsibility to provide us with this information so we can include it in the final published paper.Please also insert the following text into the Methods section, in a sub-section to be called "Nomenclatural Acts":The electronic edition of this article conforms to the requirements of the amended International Code of Zoological Nomenclature, and hence the new names contained herein are available under that Code from the electronic edition of this article. This published work and the nomenclatural acts it contains have been registered in ZooBank, the online registration system for the ICZN. The ZooBank LSIDs (Life Science Identifiers) can be resolved and the associated information viewed through any standard web browser by appending the LSID to the prefix "/". The LSID for this publication is: urn:lsid::pub: XXXXXXX. The electronic edition of this work was published in a journal with an ISSN, and has been archived and is available from the following digital repositories: PubMed Central, LOCKSS [author to insert any additional repositories].All PLOS ONE articles are deposited in PubMed Central and LOCKSS. If your institute, or those of your co-authors, has its own repository, we recommend that you also deposit the published online article there and include the name in your article.Back to top New Botanical TaxonWhen publishing papers that describe a new botanical taxon, PLOS aims to comply with the requirements of the International Code of Nomenclature for algae, fungi, and plants (ICN). In association with the International Plant Names Index(IPNI), the following guidelines for publication in an online-only journal have been agreed such that any scientific botanical name published by us is considered effectively published under the rules of the Code. Please note that。

plos one正文投稿样板

Abstractinterdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc.Materials and MethodsVestibulum adipiscing urna ut lectus gravida, vitae (Fig 1) interdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc. Mauris in mattis sapien. Fusce sodales vulputate auctor. Nam sit amet nulla lacus a, Figs 1 and 2 ultrices tellus. Integer rutrum aliquet sapien, eu fermentum magna pellentesque vitae.Fig 1. This is the Fig 1 Title. This is the Fig 1 legend. Fig 2. This is the Fig 2 Title. This is the Fig 2 legend.interdum. Donec 2pet 2q2221p pq q ++semper viverra mauris vel pulvinar dolor sit amet en 2()1p q +pulvinar et alst.Whole genome RFLP analysisResults and DiscussionLorem ipsum dolor sit amet, consectetur adipiscing elit. Vestibulum adipiscing urna ut lectus gravida, et bland Table 1Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convalli. Fido nemo.Table 1. This is the Table 1 Title.ChemicalW ChemicalX ChemicalY ChemicalZ Chemical 1 Reaction 1W Reaction 1X Reaction 1Y Reaction 1Z Chemical 2 Reaction 2W Reaction 2X Reaction 2Y Reaction 2Z Chemical 3 Reaction 3W a Reaction 3X Reaction 3Y b Reaction 3Z Chemical 4 Reaction 4W Reaction 4X Reaction 4Y Reaction 4Z Chemical 5Reaction 5WReaction 5XReaction 5YReaction 5ZThis is the Table 1 legend. aTable footnotes belong here. bFootnotes should have corresponding symbols in the table.AcknowledgmentsLorem ipsum dolor sit amet, consectetur adipiscing elit. Vestibulum adipiscing urna ut lectus gravida, vitae blandit tortor interdum.References1. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr,McBio GHT, et al. This is the article title. PLoS One 2014 Dec 18; 9(12).2. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr, McBio GHT, et al. Bunny dynamics in cartoon landscapes. PLOS One. Forthcoming 2015.Supporting InformationS1 Fig. This is the S1 Fig Title. This is the S1 Fig legend. S2 Fig. This is the S2 Fig Title. This is the S2 Fig legend. S1 Table. This is the S1 Table Title. This is the S1 Table legend. S2 Table. This is the S2 Table Title. This is the S2 Table legend.。

PLOS_ONE_Formatting_Sample_1_2014

Symbol LegendSymbol Name Definition ¶Pilcrow (paragraph symbol) 1st set of equal contributors & Ampersand 2nd set of equal contributors * Asterisk Corresponding author(s)#a Pound/number sign First Current address#b Pound/number sign Second Current address†Dagger/Cross Deceased^ Caret Consortium/Group AuthorshipThis is the Article TitleJohn Doe1¶, Antonie Data1¶, Johannes van Stats1,#a, Marie Testperson2*, David Ribosome Jr.3,5, Gregory H.T. McBio4,#b, Angela Reviewerson1,2&, Marina Measure1&, on behalf of The Bunny Genome Sequencing Consortium^1 Department, Institution, City, State, Country2 Department of Dermatology, Division of Rabbit Health, Section of Veterinary Medicine, St. Hare Hospital, San Francisco, California, United States of America3 Department of Libraries and Archives, National Contemporary Bunny Museum, Lagomorph, Connecticut, United States of America4 Department of Restoration, National Contemporary Bunny Museum, Lagomorph, Connecticut, United States of America5 Department of Archaeology, Bunny University, Lagomorph, Connecticut, United States of America#a Current Address: Department of Carrot Science, Bunny University, Lagomorph, Connecticut, United States of America#b Current Address: Department of Canine Evasion, Bunny University, Lagomorph, Connecticut, United States of America* Corresponding authorE-mail: testperson@university.ed (MT)¶These authors contributed equally to this work.&These authors also contributed equally to this work.^Membership of the Bunny Genome Sequencing Consortium is provided in the Acknowledgments. Article Title∙Italics, bold type, symbols, and other textformatting will all be reproduced in thepublished article as submitted.∙Capitalization will be automatically formatted in the published article according to PLOS style. Author Byline∙Author names will be published exactly as they appear in the accepted manuscript.∙Indicate affiliations by number only.∙Affiliation footnotes should appear in numerical order at first mention.∙Please use the symbols provided in thisdocument for other designations.∙Numbers and symbols should be insuperscript.∙Do not include titles (Dr., PhD, Professor, etc.).Affiliations∙Affiliations will be published as theyappear in the accepted manuscript.∙Include each component in order ofsmall to large (Department, Division,Section, Institution, City, State,Country).∙Do not include ZIP or Postal Codes,street addresses, or building/officenumbers.∙Do not use abbreviations (e.g. Dept.).∙Do not list positions within aninstitution (e.g. Department Chair,Professor, etc.).∙List each affiliation individually and infull.Corresponding Authorship∙Do not include physical addresses; onlyemail addresses are required.∙List corresponding author’s initials inparentheses after the email address.Contributorship∙Use the symbols provided here toindicate equal contributions.∙If you would like the equal contributionsnotes to read differently, please specifyin your manuscript (e.g., "AR and MMare Joint Senior Authors").Consortia or other Group Authors∙If there is a consortium or group authoron your manuscript, please provide anote that describes where the fullmembership list is available for thereaders.∙The membership list can be listed inthe Acknowledgments, in SupportingInformation, or on the internet.∙Consortia/Group authors can haveaffiliations, but it is not required.。

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plos one的under review

plos one的under review（实用版）目录1.介绍 PLOS ONE 的审稿过程2.PLOS ONE 的审稿标准和要求3.作者在审稿过程中的角色和责任4.PLOS ONE 的审稿周期和结果5.对作者的建议和启示正文PLOS ONE 是一家国际知名的开放获取科学期刊，旨在发布各种学科领域的高质量研究论文。

与其他期刊不同，PLOS ONE 采用严格的同行评审制度，以确保论文的质量和可靠性。

在这里，我们将详细了解 PLOS ONE 的审稿过程，以及作者在审稿过程中的角色和责任。

一、PLOS ONE 的审稿过程PLOS ONE 的审稿过程可以分为以下几个阶段：1.作者提交论文：作者首先需要将论文提交到 PLOS ONE，并完成相关授权和支付手续。

2.编辑部初审：编辑部会对提交的论文进行初步审查，以确保论文符合期刊的范围和格式要求。

如果论文不符合要求，编辑部会要求作者进行修改。

3.送审：经过初步审查，编辑部会将符合要求的论文送交给合适的审稿人进行评审。

PLOS ONE 通常会邀请至少两名审稿人对论文进行评审。

4.审稿人评审：审稿人会根据期刊的审稿标准和要求，对论文进行详细的评审。

评审过程中，审稿人可能会提出修改建议或问题，要求作者进行解答或修改。

5.作者回复：作者需要认真阅读审稿人的意见和建议，并对其进行回应。

如果审稿人要求修改，作者需要对修改后的论文进行重新提交。

6.审稿人复审：在作者提交修改后的论文后，审稿人会对论文进行再次审查，以确保作者已经按照要求进行了修改。

7.编辑部终审：在审稿人完成复审后，编辑部会根据审稿人的意见和建议，对论文进行最终审查。

如果论文符合期刊的要求，编辑部会接受论文并发表。

二、PLOS ONE 的审稿标准和要求PLOS ONE 的审稿标准和要求主要包括以下几点：1.原创性：论文必须是原创研究成果，不能抄袭他人成果。

2.学术价值：论文需要具有较高的学术价值，对相关领域具有贡献。

plos one的under review

PLOS ONE的Under Review：从简单到复杂的全面评估随着科技的飞速发展，学术界对于研究成果的传播和评估要求也越来越高。

PLOS ONE作为开放获取的网络科学期刊，一直以来都是学术界的焦点和关注点之一。

其中，Under Review这一环节更是备受研究者关注。

本文将从浅入深地对PLOS ONE的Under Review进行全面评估，并共享个人见解。

1. 什么是PLOS ONE的Under Review？PLOS ONE是由公共科学图书馆（Public Library of Science）出版的一本跨学科的开放获取科学期刊。

它涵盖了各个科学和医学领域的研究。

而Under Review则是指投稿后经过编辑初审合格后，文章将进入同行评议（Peer Review）阶段。

在这个阶段，专家学者将对文章的学术性、创新性、实验设计和数据解释等方面进行评估，最终决定文章是否适合发表在期刊上。

2. Under Review的意义与作用Under Review是PLOS ONE保证期刊学术质量的一项重要措施。

经过同行评议的文章，更具有权威性和可信度。

通过这一过程，研究者可以接受来自同行专家的建议和意见，有助于提高研究质量，避免发表低质量或有争议的研究成果。

3. Under Review的流程与特点PLOS ONE的Under Review流程十分严谨。

一旦文章进入同行评议阶段，编辑将邀请至少两名同行专家对文章进行评审。

评审专家将通过对文章内容、实验设计、数据分析方法等方面的评估，给出合理的意见和建议。

在此基础上，编辑将根据专家意见作出最终决定，通知作者文章的最终命运。

4. 个人观点与理解对于PLOS ONE的Under Review，我认为这一评审机制是非常有必要的。

它保证了期刊的学术质量和独立性，也促进了学术交流和科研成果的传播。

在这一过程中，作者可以从同行专家的建议中受益，提高自己的研究水平。

也正是由于这一评审机制的存在，PLOS ONE才能成为备受学术界认可和尊重的顶尖期刊之一。

PLosONE

Knocking-Down Cyclin A2by siRNA Suppresses Apoptosis and Switches Differentiation Pathways inK562Cells upon Administration with DoxorubicinXiaohui Wang,Yujun Song,Jinsong Ren,Xiaogang Qu*Division of Biological Inorganic Chemistry,State Key Laboratory of Rare Earth Resources Utilization,Changchun Institute of Applied Chemistry,Graduate School of the Chinese Academy of Sciences,Chinese Academy of Sciences,Changchun,Jilin,ChinaAbstractCyclin A2is critical for the initiation of DNA replication,transcription and cell cycle regulation.Cumulative evidences indicate that the deregulation of cyclin A2is tightly linked to the chromosomal instability,neoplastic transformation and tumor proliferation.Here we report that treatment of chronic myelogenous leukaemia K562cells with doxorubicin results in an accumulation of cyclin A2and follows by induction of apoptotic cell death.To investigate the potential preclinical relevance, K562cells were transiently transfected with the siRNA targeting cyclin A2by functionalized single wall carbon nanotubes.Knocking down the expression of cyclin A2in K562cells suppressed doxorubicin-induced growth arrest and cell apoptosis.Upon administration with doxorubicin,K562cells with reduced cyclin A2showed a significant decrease in erythroid differentiation,and a small fraction of cells were differentiated along megakaryocytic and monocyte-macrophage pathways.The results demonstrate the pro-apoptotic role of cyclin A2and suggest that cyclin A2is a key regulator of cell differentiation.To the best of our knowledge,this is the first report that knocking down expression of one gene switches differentiation pathways of human myeloid leukemia K562cells.Citation:Wang X,Song Y,Ren J,Qu X(2009)Knocking-Down Cyclin A2by siRNA Suppresses Apoptosis and Switches Differentiation Pathways in K562Cells upon Administration with Doxorubicin.PLoS ONE4(8):e6665.doi:10.1371/journal.pone.0006665Editor:Hany A.El-Shemy,Cairo University,EgyptReceived April27,2009;Accepted July7,2009;Published August17,2009Copyright:ß2009Wang et al.This is an open-access article distributed under the terms of the Creative Commons Attribution License,which permits unrestricted use,distribution,and reproduction in any medium,provided the original author and source are credited.Funding:This work was supported by NSFC(20831003,90813001,20833006)and funds from the Chinese Academy of Sciences and Jilin Province.The funders had no role in study design,data collection and analysis,decision to publish,or preparation of the manuscript.Competing Interests:The authors have declared that no competing interests exist.*E-mail:xqu@IntroductionTumor cells are characterized by deregulation of cell cycle checkpoints,leading to uncontrolled cell division and proliferation under conditions where non-transformed cells cannot enter and pass through the cell cycle.All these may come from over-expression of cyclins and the abnormal activation of cyclin-dependent kinases(CDKs)[1].Cyclins are a superfamily of proteins whose levels vary in a cyclical fashion during the cell cycle to activate specific CDK required for the proper progression through the cell cycle.Cyclin A2,which is essential for initiation and progression of DNA replication as well as for cell cycle progression through G1/S and G2/M transitions[2–5],is over-expressed in a variety of human cancers compared with normal cells and tissues[6–15].Deregulated expression of cyclin A2seems to be closely associated with early events in tumor transformation [6,15].In addition,its expression level in many types of cancers appears to be of prognostic value such as prediction of aggressiveness,survival or early relapse[9–14].Until recently it has been held that CDK2,presumed master of the known CDK isoforms,is a promising anticancer target for developing small molecule inhibitors.The first-generation CDK inhibitors,flavopiridol and CY-202,are in late-stage clinical trials, and have only modest activity[16].Recent findings[17],however, suggest that CDK2may not be a key cell cycle player and question whether selective CDK2inhibition is a useful cancer therapy strategy.No cell cycle abnormalities are observed in either a CDK2 null mouse or following acute ablation of CDK2in primary cells, indicating that this gene is not strictly required for cell proliferation [17].In contrast,deletion of cyclin A2in knockout mice is associated with an embryonic lethal phenotype[18].Moreover,Fine et al. demonstrated that cyclin A2and/or cyclin A2-CDK2complex but not CDK2is a promising anticancer target with a high therapeutic index[19].Therefore,inhibitors of CDK2may not be appropriate for cancer therapy and more efforts are focused on inhibition of cyclin A2and/or cyclin A2-CDK2complex activity.We have shown that reduction of cyclin A2in human chronic myelogenous leukaemia K562cells using small interfering RNA significantly inhibits cell proliferation[20],further supporting the notion that cyclin A2can serve as a novel therapeutic target.Apoptosis and differentiation are the predominant two mech-anisms by which chemotherapeutic agents kill tumor cells.Low dose of doxorubicin(DOX)induces erythroid differentiation in K562cells,while high concentration of DOX promotes apoptosis [21].Although many molecular pathways are involved in the apoptosis-regulatory mechanism,evidences suggest that the cell cycle and apoptosis may be interconnected[22–28].Several studies have shown that increased expression of cyclin A2is found in cells in response to several apoptotic stimuli,but very few studies have dealt with this issue directly[29–33].So it is important to clarify whether the decreased expression of cyclin A2is a cause of cell differentiation or a result of differentiation response.Carbon nanotubes possess the unique features of being able to enter a living cell without causing its death or without inflicting other damage and can shuttle biological molecules into mamma-lian cells,indicating their potential application as a vector for the delivery of therapeutic molecules [34–36].Recently we have reported that single wall carbon nanotubes (SWNTs)can induce a sequence-dependent B-A DNA transition [37],selectively induce human telomeric i-motif DNA formation [38],accelerate S1nuclease cleavage rate [39],cause single-stranded poly(rA)to form a duplex structure and bind to human telomeric i-motif DNA under molecular-crowding conditions [40,41].Herein,we explore whether altering the levels of cyclin A 2in K562cells using RNAi delivered by SWNTs can influence cell apoptosis and differenti-ation induced by chemotherapeutic agent DOX.K562cells with reduced cyclin A 2showed a significant decrease in growth suppression,apoptosis and erythroid differentiation,and were differentiated along megakaryocytic and macrophage-monocytic pathways upon administration with DOX.These findings indicate a positive correlation between cyclin A 2and apoptosis induced by DOX and suggest that cyclin A 2is a key regulator of cell differentiation,supporting the notion that cyclin A 2is an important regulator for cell cycle as well as for cell apoptosis and differentiation.To the best of our knowledge,this is the first report that knocking down expression of one gene can switch K562cells differentiation pathways.ResultsUpregulation of cyclin A 2during apoptosis of K562cells induced by DOXDOX can inhibit growth of a variety of cancer cells [42].To measure apoptosis rates induced by varying concentrations of DOX in K562cells,we use acridine orange (AO)/ethidium bromide (EB)staining assay (Fig.1).The percentage of apoptotic cells increased with time in a dose dependent fashion.RT-PCR and western blotting were used for studying the effect of DOX on the expression of cyclin A 2in K562cells.As shown in Fig.2,cyclin A 2expression levels increased with the increase of DOX,and a positive correlation was observed.These results show that expression of cyclin A 2was significantly up-regulated in DOX-treated K562cells at time pointswhen DOX caused a significant amount of apoptosis,indicating that the levels of cyclin A 2are correlated with the ability of DOX to induce apoptosis in K562cells.Suppression of DOX-induced growth inhibition by down-regulation of cyclin A 2with siRNA delivered by SWNTsWe have previously demonstrated that functionalized single wall carbon nanotubes (SWNTs)can efficiently deliver siRNA targeting cyclin A 2into K562cells,resulting in specific suppression of cyclin A 2expression [20].Here,we use SWNTs to transfect cyclin A 2siRNA into K562cells and evaluate the effect of cyclin A 2on growth inhibition induced by DOX.Two hours after transfection,DOX was added.Cell proliferation was examined by trypan blue exclusion method and methylthiazolyldiphenyl-tetrazolium bro-mide (MTT)assay.Fig.3showed cell growth curves of K562cells after various treatments.It can be seen that depletion of cyclin A 2inhibited cell proliferation while carbon nanotubes vector had no apparent effect on growth inhibition as well as no additive or synergetic effect on cell toxicology of DOX.DOX (0.4m M)significantly suppressed cell growth,nevertheless,much less growth inhibition was observed in cells incubated with bothcyclinFigure 1.Dose-and time-dependent apoptosis of K562cells upon administration with DOX.Apoptotic cells were determined by AO/EB staining.Tests were done in triplicate,counting a minimum of 300total cells from at least three random microscope fields each.doi:10.1371/journal.pone.0006665.g001Figure 2.Expression of cyclin A 2in K562cells administered with varying concentrations of DOX.RT-PCR (A)and western blotting (B)were performed 32hours and 60hours after drug administration,respectively.doi:10.1371/journal.pone.0006665.g002Figure 3.Growth curves of K562cells in response to cyclin A 2siRNA delivered by SWNTs and DOX.The viable cells were counted by trypan blue exclusion at indicated time points.The data shown here represent the average of two independent experiments.doi:10.1371/journal.pone.0006665.g003A 2siRNA and DOX,supported by microscopic results (Fig.S1).In order to further quantitively investigate the effect of down-regulation of cyclin A 2on the growth inhibition,MTT assays were carried out 24h after incubation with DOX.Interaction of DOX with MTT was also checked in a cell free system and the results indicated that DOX showed no inference to MTT assay (Fig.S2).As shown in Fig.4,IC 50of non-transfected cells was about 2.5m M.For siRNA transfected K562cells,IC 50was about 5.0m M.The results clearly demonstrate that down-regulation of cellular cyclin A 2level suppress DOX-induced growth inhibition.Suppression of DOX-induced apoptotic cell death by down-regulation of cyclin A 2with siRNA delivered by SWNTsTo investigate whether cyclin A 2participates in cell apoptosis death and determine if reduction of cyclin A 2has an effect on apoptosis induced by DOX,siRNA specific for cyclin A 2was transfected into K562cells by SWNTs,and low dose of DOX (0.4m M)was administered.As shown in Fig.5,carbon nanotubes vector showed no apparent cell toxicology,while numerous larger cells and typical lobular nuclei were observed in cells incubated with DOX.For cells coadministered with cyclin A 2siRNA and DOX,much less apoptotic nuclei were observed,and the size of live cells was much bigger than that of the control cells.Statistics analysis showed that down-regulation of cyclin A 2significantly reduced the apoptosis rate from more than 60%to about 20%.Similar results were obtained by annexin V-PI double staining flow cytometric technique (as shown in Figure S3).These clearly indicated lowering cyclin A 2level in K562cells led to suppression of apoptosis,thus a marked decrease in DOX susceptibility,which provide direct evidence that cyclin A 2is involved in cellular responses to apoptosis.The cytoplasmic subcellular distribution of cyclin A 2correlates with apoptosis of K562cells induced by DOXEarly reports have demonstrated that there is a link between cyclin A 2subcellular localization and its cell function,and the level of cyclin A 2is correlated to cell apoptosis [22,23,26].For clarifyingwhether the subcellular distribution of cyclin A 2in K562cells correlates with apoptosis induced by DOX,indirect immunoflu-orescence detection of cyclin A 2was performed.As shown in Fig.6A,a significant fraction of cells underwent apoptosis and orange nuclei were observed,where DOX was mostly located.The immunofluorescence labeling of cyclin A 2showed its presence predominantly in the nucleus of control K562cells (Fig.S4),whereas in cells administered with DOX,cyclin A 2was mainly located at the cytoplasm of early and late phases of apoptotic cells (Fig.6B).Cyclin A 2labeling was not found in the K562cells incubated with non-immune serum (Fig.6C).The results indicated that translocation of cyclin A 2from the nucleus to cytoplasm was connected with its role in apoptosis.Suppression of cyclin A 2by siRNA can switchdifferentiation pathways of K562cells induced by DOXAs mentioned above,cells treated with both cyclin A 2siRNA and DOX were much bigger than the control.Since enlarged phenotype may suggest cell differentiation,we performed the benzidine staining to assess erythroid differentiation,which was the differentiation pathway of K562cells upon treatment with anthracycline antibiotics including DOX [42,43].Representative microscopy images of the benzidine staining in K562cells after various treatments were shown in Fig.S5.For untreated cultures and cells administered with SWNTs,the percentages of benzidine positive cells were very low (less than 2%).Forty hours after incubation with DOX,around 14%benzidine positive cells were observed.However,down-regulation of cyclin A 2by siRNA in K562cells substantially suppressed erythroid differentiation upon administration with DOX (less than 1%benzidine positive cells,as shown in Fig.7).To determine whether K562cells with reduced cyclin A 2upon treatment with DOX underwent megakaryocytic pathway and monocyte-macrophage differentiation,which are the other two differentiation pathways of K562cells,flow cytometric measure-ment of megakaryocytic specific surface antigen CD61(GPIIIa)and nitro blue tetrazolium (NBT)reduction assay were carried out,respectively.As shown in Fig.S6,in comparison with the control,cells co-administered with cyclin A 2siRNA and DOX showed significant increase in granularity measured by side scatter (side scatter,on Y -axis)and cell size measured by forward scatter (forward scatter,on X -axis),which was in good agreement with our AO/EB staining observation.Cells treated with DOX or SWNTs showed no detectable expression of CD61(GPIIIa)(shown in Fig.S7),whereas in K562cells co-administered with cyclin A 2siRNA and DOX,the fraction of CD61(GPIIIa)positive cells (,10%)was clearly observed (Fig.8).For NBT reduction assay,representative microscopy images of K562cells after various treatments were shown in Fig.S8.In cells treated with DOX or SWNTs,the percentage of NBT positive cells was very low (1%),whereas a small fraction NBT positive cells (,6%)was observed in K562cells co-administered with cyclin A 2siRNA and DOX (Fig.9).Although the fractions of cells undergoing megakaryocytic pathway (,10%)and monocyte-macrophage differentiation (,6%)were small,the difference is statistically significant compared to the control group.Considering that DOX is an erythroid differentiation-inducing agent for K562cells [21,42,43],it is not surprising that only a small fraction of K562cells with reduced cyclin A 2underwent megakaryocytic pathway and monocyte-macrophage differentiation.Taken together,these results indicate that knocking down the expression of cyclin A 2suppressed DOX-induced erythroid differentiation and a small fraction of K562cells with reduced cyclin A 2were differentiated along megakaryocytic andmonocyte-Figure 4.Effect of depletion of cyclin A 2on the survival of K562cells treated with DOX.Cells were transfected with cyclin A 2siRNA (#)or not (N)two hours prior to the addition of drug.The viability was assessed by MTT assay as specified in Materials and Methods.The data shown here were means of two separate experiments performed in triplicate.doi:10.1371/journal.pone.0006665.g004macrophage pathways upon treatment with DOX.These findings suggest that cyclin A 2is an important regulator of cell differentiation.DiscussionCyclin A 2is particularly interesting among the cyclin family because it can activate two different CDKs and functions in both Sphase and mitosis.In S phase,phosphorylated cyclin A 2-CDK2complexes are suggested to play an important role in the initiation of DNA replication.In mitosis,cyclin A 2may contribute to the control of cyclin B stability.Consistent with its role as a key cell cycle regulator,overexpression of cyclin A 2is associated with transformed cells [6–15].However,it is difficult to determine whether elevation of cyclin A 2is a contributing factor or amereFigure 5.Morphological observation of K562cells with a fluorescence microscope.Cells were stained with AO/EB dye mixture as described in Materials and Methods section.Shown here were the representative images of three separate experiments.(A):upper left,untreated control cells;upper right,cells treated with transfection vector SWNTs for 32h;lower left,cells were incubated with 0.4m M DOX for 32h,many cells appeared typical apoptotic bleb phenomenon;lower right,two hours after cyclin A 2siRNA transfection,cells were then administered with 0.4m M DOX for additional 32h.B:quantification of the live,apoptotic,and necrotic cells.Tests were done in triplicate,counting a minimum of 300total cells each.doi:10.1371/journal.pone.0006665.g005consequence of the increased cell proliferation.In haematological malignancies,cyclin A 2is associated with proliferation rate of these disorders and can be used for molecular diagnostics as a proliferation marker [13,14].Single-walled carbon nanotubes (SWNTs)have been considered as the leading candidate for nanodevice applications ranging from gene therapy and novel drug delivery to membrane separations.We have previously showed that siRNA transfection efficiency of lipofectamine 2000in K562cells was low (28%)and some cells underwent apoptosis and necrosis during the process [20].However,SWNTs could efficiently facilitate the coupling of siRNA to form siRNA:SWNTs complexes and carry siRNA into K562cells,significantly knocking down the expression of target gene.No apparent cell toxicology was observed.Hence,in order to directly probe whether cyclin A 2participates in cell apoptosis and differentiation,we employed SWNTs as transfection vector to deliver cyclin A 2siRNA into K562cells to specifically knock down the expression of cyclin A 2and found that carbon nanotubes vector showed no additive or synergetic effect on cell toxicology of DOX,which was consistent with our previous report [20].DOX,a prominent member of anthracycline antibiotics,has been extensively used for treatment of solid tumors and leukemia.It exerts its cytotoxic activity against cancer cells mainly by intercalation into DNA,inhibition of topoisomerase II and helicase activity,leading to cell-cycle arrest at the G2/M phase and apoptosis [42].In clinical applications,doses of DOX are strictly limited by its cardiotoxicity [43].It should be noted that the dose of DOX administrated here (0.4m M)is pharmacological relevant compared to the initial or steady-state plasma concentra-tions observed in patients after standard bolus infusions (5m M and 25–250nM,respectively).It has been reported that there is a link between cyclin A 2and apoptosis [22–28].Hoang et al.showed that in c-myc overexpressing serum deprived rat 1A fibroblasts undergoing apoptosis,cyclin A 2mRNA expression was increased,in contrast to the invariant expression for cyclin B,C,D1and E [24].Moreover,serum-deprived rat 1A fibroblast stably transfected with cyclin A 2exhibitedFigure 6.The sub-cellular distribution of cyclin A 2in K562cells correlated with cell apoptosis.Cells were administered with 0.4m M DOX for 32h.A significant fraction of cells underwent apoptosis.Immunofluorescence detection of cyclin A 2was performed as described in Materials and Methods section.A:representative fluorescence microscopy image of K562cells after treatment.Orange nuclei were observed,where DOX was mostly located;B:representative immuno-fluorescence microscopy image of K562cells.Cyclin A 2,normally located at nucleus,can be observed in cytoplasm of early and late phases of apoptotic cells.C:representative microscopy image of negative control immunofluorescence in K562cells.No significant non-specific signal was observed.doi:10.1371/journal.pone.0006665.g006Figure 7.Knocking–down the expression of cyclin A 2in K562cells significantly inhibited erythroid differentiation induced by low concentration DOX.Cells were transfected with cyclin A 2siRNA or not two hours prior to the addition of 0.4m M DOX.Forty hours later,erythroid differentiation was scored by the benzidine staining method to determine the percentage of hemoglobin-positive K562cells.Tests were done four times,counting a minimum of 300total cells from at least three random microscope fields each.*p ,0.05,**p ,0.001vs.control untreated cells by Student’s t-test.doi:10.1371/journal.pone.0006665.g007increased apoptosis following stimulation of cyclin A 2expression.Meikrantz et al.reported that induction of apoptosis was uniformly associated with activation of cyclin A 2-dependent kinases but not associated with cyclins E or B,and overexpression of the cyclin A 2could circumvent the anti-apoptosis activity of the oncogene BCL-2in human Hela cells [25,26].Furthermore,Hiromura et al.indicated that apoptosis was associated with an increase in cytoplasmic cyclin A 2-CDK2activity following UV irradiation,under these conditions,nuclear cyclin A 2-Cdk2activity decreased significantly [27].Our results showed knocking down the expression of cyclin A 2in K562cells significantly suppressed the apoptosis induced by DOX and a positive correlation between the levels of cyclin A 2and apoptosis was observed.The findings also indicate that the cytoplasmic subcellular distribution of cyclin A 2correlates with its pro-apoptotic role.We speculate that cyclin A 2associated kinases are involved in DOX-induced apoptotic cell death pathways in K562cells,although the exact downstream mechanisms are not known.We have demonstrated that SWNTs could effectively deliver cyclin A 2siRNA into K562cells,significantly suppressing the expression of cyclin A 2with specificity and cell proliferation,and cells with reduced cyclin A 2showed a decrease in the percentage of cells in S phase [20].Several studies have indicated that cancer cells with a high S-phase fraction/high proliferative activity are more sensitive to apoptosis induced by chemotherapy [44,45].Asfor DOX,it is active throughout the cell cycle,but the effect is most pronounced for cells in S phase-G2phase,especially in S phase,where it interferes with the DNA replication and transcription [42].Therefore,it was not surprising that K562cells with reduced cyclin A 2showed a marked decrease in DOX susceptibility.Most of chemotherapeutic agents show significant side effects and not all patients benefit from aggressive chemotherapy.Therefore,searching for tumor biological factors which can predict patient prognosis and chemotherapy response would be of most importance.Several studies have indicated that a high level of cyclin A 2expression may be a marker of poor prognosis in cancers [10–13].Besides,previous studies have shown that cancer patients with high level of cyclin A 2had better chemotherapy response and survival than those with reduced cyclin A 2and low expression of cyclin A 2,indicating that the patients with high expression of cyclin A 2are more suitable for chemotherapy [46–48].Our results demonstrate the pro-apoptotic role of cyclin A 2in human myeloid leukemia K562cells,and indicate that cells with low level of cyclin A 2were more resistant to chemotherapeutic agent DOX.Poon et al.have suggested that a decrease of cyclin A 2,rather than increase,promotes tumorigenesis,and once the tumor has developed,high levels of cyclin A 2simply reflect a high proliferation rate,which can explain this inconsistency [49].Hence,despite its association with transformed cells,evaluating cyclin A 2level in patients will be an important prognostic marker for use of chemotherapy.Patients with high level of cyclin A 2may be more responsive to anticancer drugs through the induction of apoptotic cell death.Moreover,it should be cautious to combine doxorubicin chemotherapy with any small molecule drug targeting cyclin A 2/cyclin A 2associated kinases since it can enhance potential drug resistance.In several systems,it has been reported that down-regulation of cyclin A 2and its associated CDK 2activity are important for successful differentiation [29–33].Ito et al .have suggested cyclin A 2overexpression is directly related with poor differentiation [31].Kiyokawa et al .have indicated that differentiation ofmurineFigure 8.Flow cytometric measurement of megakaryocytic specific surface antigen CD61(GPIIIa)in K562cells co-administered with cyclin A 2siRNA and DOX.Cells were transfected with siRNA by SWNTs two hours prior to the administration of 0.4m M DOX.Sixty hours later,expressions of CD61(GPIIIa)were evaluated by using FITC-conjugated isotype control immunoglobulin and specific anti-CD61(GPIIIa)FITC-conjugated monoclonal antibody.The marker placed to the right of histogram designates positive events.doi:10.1371/journal.pone.0006665.g008Figure 9.Knocking-down expression of cyclin A 2in K562cells induced monocyte-macrophage differentiation upon adminis-tration of DOX.Cells were transfected with siRNA by SWNTs two hours prior to the treatment of 0.4m M DOX.Ninety six hours later,NBT dye reduction was used to qualitatively monitor monocyte-macrophage differentiation.Tests were done twice,counting a minimum of 300total cells from at least three random microscope fields each.**p ,0.001vs.control untreated cells by Student’s t-test.doi:10.1371/journal.pone.0006665.g009erythroleukemia cells induced by hexamethylene is accompanied by a decrease in the level of cyclin A2and CDK2proteins and the persistent suppression of cyclin A2expression may play a role in HMBA-induced commitment to terminal differentiation[30]. Yoshizumi et al.showed down-regulation of cyclin A2gene expression in vivo at both the RNA and protein levels appears to be important in the permanent withdrawal of human and rat cardiomyocytes from the cell cycle during development[32]. Moreover,Rieber et al.demonstrated that the interaction of cyclin A2with E2F is the target for tyrosine induction of B16melanoma terminal differentiation[33].In this work,we found that knocking down the expression of cyclin A2in K562cells significantly suppressed DOX-induced erythroid differentiation and a small fraction of cells with reduced cyclin A2were differentiated along megakaryocytic and monocyte-macrophage pathways upon treat-ment with DOX.To the best of our knowledge,this is the first report that knocking down expression of one gene can switch K562cells differentiation pathways.The results suggested that cyclin A2is directly involved in the checkpoint of cell differentiation pathways and is a key regulator of this process, although the detail downstream mechanisms are not known.For cancer cells with low level of cyclin A2,which are less responsive to chemotherapeutic agents,induction of differentiation might be an alternative bination of cyclin A2siRNA and DOX may provide a novel option of such therapeutic strategy.In conclusion,knocking down the expression of cyclin A2by siRNA delivered by SWNTs suppresses apoptosis and erythroid differentiation,and promotes megakaryocytic and monocyte-macrophage differentiation in human chronic myelogenous leukaemia K562cells upon administration with DOX.The results demonstrate the pro-apoptotic role of cyclin A2and suggest that cyclin A2is a key regulator of cell differentiation,supporting the notion that cyclin A2is an important regulator for cell cycle as well as for cell apoptosis and differentiation.Materials and MethodsEthics statement.The human erythroleukemic cell line K562[20]was used in this study.ChemicalsDoxorubicin(DOX),Acridine Orange(AO),Ethidium Bromide (EB),Methylthiazolyldiphenyl-tetrazolium bromide(MTT),DAPI, Nitro blue tetrazolium(NBT),benzidine and SWNTs(w=1.1nm, purity.90%)were purchased from Sigma-Adrich(St.Louis,MO, USA).Annexin V apoptosis detection kit was obtained from Keygentec(Nanjing,China).The dye mix for the EB/AO staining was100m g/mL acridine orange and100m g/mL ethidium bromide in phosphate buffered saline(PBS).Cell cultureThe human erythroleukemic cell line K562[20]was grown in Iscove’s modified Dulbecco’s medium(Gibco BRL)supplemented with10%fetal calf serum in a humidified37u C incubator with5% CO2.Cells were passed three times per week.DNA fluorochrome staining(DAPI or Hoechst33258)is used as our routine mycoplasma detection and the cells used for experiments are free of contamination.Exponentially growing cells were used for all experiments described below.Cell viability was determined by trypan blue exclusion in a haemocytometer chamber. TransfectionThe siRNA oligonucleotides were synthesized by Genepharma Corporation(Shanghai,China).The sequence used for targeting silencing of cyclin A2was59-CCAUUGGUC CCUCUU-GAUUTT-39.The nonsilencing control siRNA is an irrelevant siRNA with random nucleotides UUCUCCGAACGUGUCAC-GUTT.Functionalized single wall nanotubes(f-SWNTs)were prepared according to method described in our previous work [20].Cyclin A2siRNA:f-SWNTs complexes(w f-SWNTs/w siRNA =40)were added at25n mol/L(siRNA concentration)to culture of cells.Reverse transcriptase-polymerase chain reaction(RT-PCR) Total RNA was extracted using Trizol reagent(Invitrogen) according to the manufacturer’s instructions.The primers and conditions for cyclin A2were TCCATGTCAGTGCTGA-GAGGA(59),GAAGGTCCATGAGACAAGGC(39);94u C for 30seconds,60u C for30seconds,72u C for1minute for25cycles. Three introns are present in this pair of primer so that any contaminating genomic DNA would not be amplified.Primers used for the glyceraldehyde-3-phosphate dehydrogenase (GAPDH)were ACCTGACCTGCCGTCTAGAA(59),TCCAC-CACCCTGTTGCTGTA(39).Two sets of primers were used for each sample,including primers specific for the gene of cyclin A2 and primers for GAPDH as an internal control.All PCR products were visualized on1.5%agarose gel with0.5m g mL21EB. Western blottingFor Western blot analysis,cells were lysed in radio-immuno-precipitation assay(RIPA)buffer(150mmol/L NaCl,50mmol/L Tris-HCl,0.5%sodium deoxycholate,1%NP-40,0.1%SDS, pH7.6)containing protease inhibitors for protein extraction. Protein concentrations were determined using the Bradford assay. 20m g cell samples were denatured by addition of26reducing sample buffer(100mmol/L Tris,4%SDS,25%glycerol,10%b-mercaptoethanol,0.01%bromphenol blue,pH6.8),incubated for 10minutes at95u C,and separated on a12%SDS-PAGE.The proteins were electroblotted to PVDF membrane.After blocking with Tris-buffered saline containing0.05%Tween20(TTBS)and 2%BSA,the membranes were incubated overnight at4u C with appropriate primary antibody diluted in TTBS.Working dilutions were:1/500rabbit polyclonal anti-cyclin A2primary antibody (Lab Vision Corporation,CA,USA);1/400anti-GAPDH primary antibody(Santa Cruz,CA,USA).The membranes were washed three times in TTBS for5min each and then incubated for1h at room temperature with goat anti-rabbit IgG conjugated to horseradish peroxidase(Jackson Immunoresearch,Stratech, UK).After extensive washing in TTBS,the protein–antibody complexes were visualized by CN/DAB substrate according to method described by Yong[50].Images were photographed using a UVP gel documentation system(Ultraviolet Products,Upland, CA,USA).MTT assayK562cells were plated in96-well culture plate at a concentration of0.56104cells/well,and were transfected with cyclin A2siRNA or not by SWNTs.Two hours later,the cells were treated with various concentrations of DOX while the blank control wells were added medium without drug.Cells were then cultured for another24hours and20m L MTT(5mg/mL)was added in each well,followed by additional four hour incubation. The supernatants were then discarded carefully and150m L dimethylsulphoxide was added and shaken vigorously to dissolve the purple precipitation formation.Optical density(OD)of each well was tested using Bio-Rad model-680microplate reader with a wavelength of490nm.。

PLOS ONE 模版

Glucagon-Like Peptide-1Induced Signaling and Insulin Secretion Do Not Drive Fuel and Energy Metabolism in Primary Rodent Pancreatic b-CellsMarie-Line Peyot1,Joshua P.Gray3,Julien Lamontagne1,Peter J.S.Smith2,George G.Holz4,S.R.Murthy Madiraju1,Marc Prentki1.*,Emma Heart2.1Molecular Nutrition Unit and Montreal Diabetes Research Center at the Centre de Recherche du Centre Hospitalier de l’Universite´de Montre´al and Departments of Nutrition and Biochemistry,Universite´de Montre´al,Montre´al,Quebec,Canada,2BioCurrents Research Center(NIH:NCRR),Marine Biological Laboratory,Woods Hole, Massachusetts,United States of America,3Department of Chemistry,United States Coast Guard Academy,New London,Connecticut,United States of America,4State University of New York,Upstate Medical University,Syracuse,New York,United States of AmericaAbstractBackground:Glucagon like peptide-1(GLP-1)and its analogue exendin-4(Ex-4)enhance glucose stimulated insulin secretion(GSIS)and activate various signaling pathways in pancreatic b-cells,in particular cAMP,Ca2+and protein kinase-B (PKB/Akt).In many cells these signals activate intermediary metabolism.However,it is not clear whether the acute amplification of GSIS by GLP-1involves in part metabolic alterations and the production of metabolic coupling factors.Methodology/Prinicipal Findings:GLP-1or Ex-4at high glucose caused release(,20%)of the total rat islet insulin content over1h.While both GLP-1and Ex-4markedly potentiated GSIS in isolated rat and mouse islets,neither had an effect on b-cell fuel and energy metabolism over a5min to3h time period.GLP-1activated PKB without changing glucose usage and oxidation,fatty acid oxidation,lipolysis or esterification into various lipids in rat islets.Ex-4caused a rise in[Ca2+]i and cAMP but did not enhance energy utilization,as neither oxygen consumption nor mitochondrial ATP levels were altered.Conclusions/Significance:The results indicate that GLP-1barely affects b-cell intermediary metabolism and that metabolic signaling does not significantly contribute to GLP-1potentiation of GSIS.The data also indicate that insulin secretion is a minor energy consuming process in the b-cell,and that the b-cell is different from most cell types in that its metabolic activation appears to be primarily governed by a‘‘push’’(fuel substrate driven)process,rather than a‘‘pull’’mechanism secondary to enhanced insulin release as well as to Ca2+,cAMP and PKB signaling.Citation:Peyot M-L,Gray JP,Lamontagne J,Smith PJS,Holz GG,et al.(2009)Glucagon-Like Peptide-1Induced Signaling and Insulin Secretion Do Not Drive Fuel and Energy Metabolism in Primary Rodent Pancreatic b-Cells.PLoS ONE4(7):e6221.doi:10.1371/journal.pone.0006221Editor:Kathrin Maedler,University of Bremen,GermanyReceived April14,2009;Accepted June15,2009;Published July13,2009This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that,once placed in the public domain,this work may be freely reproduced,distributed,transmitted,modified,built upon,or otherwise used by anyone for any lawful purpose.Funding:MP is funded by Merck-Frosst and the Canadian Institute of Health Research.MP is recipient of Canadian chair in diabetes and metabolism.GGH is funded by the National Institutes of Health(DK045817),EH by the American Diabetes Association(7-08-JF-18)and PJSS by the National Science Foundation(P41-RR-001395.JL is a recipient of a fellowship from Fonds de la Recherche en Sante du Quebec.The funders had no role in study design,data collection and analysis, decision to publish,or preparation of the manuscript.Competing Interests:The authors have declared that no competing interests exist.*E-mail:marc.prentki@umontreal.ca.These authors contributed equally to this work.IntroductionThe mechanisms of glucose-stimulated insulin secretion(GSIS) in the b-cell remain to be defined.In addition to the triggering pathway involving a rise in ATP production,K ATP channel closure and a Ca2+rise[1,2,3],fuel signaling is thought to involve additional pathways,in particular anaplerosis/cataplerosis,pyru-vate cycling processes,endogenous lipolysis and enhanced glycerolipid/fatty acid(GL/FFA)cycling[4,5,6,7,8,9].Besides the signals induced by calorigenic nutrients and their associated production of metabolic coupling factors[4],b-cell function is modulated by a variety of neurohormonal agents and glucoincre-tins[10],including glucagon like peptide-1(GLP-1),an incretin hormone secreted by the L-cells of the distal intestine[10,11]. GLP-1levels in the plasma increase rapidly following a meal[12], and this hormone has a profound glucose-lowering effect through both central and peripheral actions[13],the latter effect being particularly at the level of the b-cell[10].GLP-1stimulates insulin gene expression[14],proinsulin biosynthesis[10],and it also potentiates GSIS[10,14].GLP-1also has proliferative[15]and antiapoptotic actions on the b-cell[10].The biologically active form of GLP-1is derived from proglucagon via the action of prohormone convertase enzymes[10,11],and circulating GLP-1is rapidly removed from the circulation following its degradation by dipeptidyl peptidase-4(DPP-4)[16].GLP-1exerts its cellular action by binding to its receptor,a G-protein coupled receptor(GLP-1R),expressed in b-cells,nervous system,heart and kidney[10,11].The activation of the GLP-1R leads to the induction of many signal transduction systems, including cAMP,Ca2+,PI3-Kinase and EGF receptor signaling [10,11,17,18].These multiple actions of GLP-1are also observed upon exposure of b cells to Exendin-4(Ex-4),a peptide that is anincretin mimetic and which lowers levels of blood glucose as a consequence of its ability to activate the GLP-1R[10].GLP-1 induces insulin secretion during short-term exposure to the hormone,or after chronic exposure to the hormone[10,19]. Even though the precise mechanisms of GLP-1action are not fully understood,it is established that the stimulation of GSIS by GLP-1 involves activation of membrane-bound adenylyl cyclase and cAMP production,leading to protein kinase-A(PKA)and Epac [20]activation,and an increase in intracellular Ca2+[10,11,21].A rise in cytoplasmic and mitochondrial Ca2+has been linked to the activation of mitochondrial dehydrogenases,in particular pyruvate dehydrogenase[22],a-ketoglutarate dehydrogenase and isocitrate dehydrogenase[3,22,23].Additionally,islet tissue and the b-cell contain some glycogen[24,25]that might be mobilized following a rise in cellular Ca2+or cAMP[26],thus releasing glucose-1P that may enter the glycolytic pathway following its conversion to glucose-6-P.It is therefore attractive to hypothesize that GLP-1 may indirectly activate b-cell energy metabolism,thereby raising levels of cellular ATP,and possibly influencing other metabolic coupling factors,via its effect on cellular levels of Ca2+and cAMP. Thus,it is generally believed that the cAMP and Ca2+pathways cannot fully account for the complete magnitude of GLP-1 mediated GSIS enhancement[10,11,19].Besides Ca2+signaling,the binding of GLP-1to its receptor also results in the activation of protein kinase-B(PKB/Akt)[27,28,29] and PKB activation in other cell types has been linked to various metabolic effects,including glucose transport in muscle[30] glycogen synthesis[31]and lipolysis[32].It can also be hypothesized that an increase in cellular ATP content might inactivate AMP-activated protein kinase(AMPK)[33],as elevated glucose does[34],which may lead to reduced phosphorylation of hormone sensitive lipase and adipose triglyceride lipase[35],with subsequent enhanced lipolysis[36]and activation of the lipid amplification arm of glucose signaling for insulin secretion[36]. Thus,our previous work has established a role for lipolysis in GSIS [37,38].In addition,it was previously shown that orlistat,a pan-inhibitor of lipases,suppresses the incretin action of GLP-1[39], and that GLP-1enhances lipolysis in HIT(b)cells[40].In the present study we examined whether the acute stimulation of GSIS by GLP-1or Ex-4involves the modulation of glucose, fatty acid and energy metabolism in the b-cells,as studied using isolated islets of both rats and mice.The study was designed to respond to four questions of general interest for b-cell neuropep-tide and fuel signaling.1)Does GLP-1amplify GSIS in part by metabolic signaling?2)Does the activation of major cellular signaling processes(Ca2+,cAMP,PKB etc)in response to a physiological peptide agonist changes b-cell metabolism?3)Does enhanced insulin secretion contribute significantly to total energy consuming processes in the b-cell?4)Is the b-cell similar or different from most tissues in term of metabolic activation, specifically whether it is primarily governed by a‘‘push’’(fuel substrate driven)process,rather than a‘‘pull’’mechanism secondary to enhanced activation of its major cellular function. Materials and MethodsAnimals and dietsEthics Statement.All procedures were performed in accordance with the Institutional Committee for the Protection of Animals at the Centre Hospitalier de l’Universite´de Montre´al or Institutional Guidelines for Animal Care(IACUC)at the Marine Biological Laboratory,in compliance with United States Public Health Service regulations.Wistar rats(200–250g;Charles River)and male Swiss-Webster mice or male CD-1rats were housed under controlled temper-ature(21u C)and light conditions(12-h light/dark cycle)with free access to water and standard chow diet.Isolation and culture of islets and islet cellsPancreatic islets were isolated by collagenase digestion of the pancreas according to Gotoh et al[41].After digestion and washing and separation by histopaque gradient centrifugation, islets were hand-picked and cultured overnight in a humidified incubator with5%CO2.For measurements of insulin secretion, the cell culture medium was RPMI-1640containing11mM glucose and supplemented with10%foetal calf serum,10mM HEPES(pH7.4),1mM sodium pyruvate,100U/ml penicillin and100m g/ml streptomycin(RPMI complete medium).For measurements of oxygen consumption,isolated rat or mouse islets were cultured overnight in RPMI-1640complete medium containing5mM glucose.For single cell studies of mitochondrial ATP levels,the islets were dispersed by incubation in Ca2+/Mg2+ free phosphate buffered saline,3mM EGTA and0.002%trypsin as previously described[42].Islet cells obtained by dispersion of islets were plated on poly-D-lysine coated coverslips(MatTek, Ashland,MA)in35mm Petri dishes.After24h,single islet cells were transduced with Ad-MitoLuc-RFP at50MOI(multiplicity of infection)for12h,after which viral media were replaced with appropriate growth media.Transduction efficiency in single islet cells,determined from RFP fluorescence,reached more then90% under these conditions.Measurement of insulin secretion and insulin content After overnight culture,rat islets were distributed in12-well plates(10islets/well)and incubated for2h in1ml RPMI complete medium containing2.8mM glucose.The islets were then washed and pre-incubated for45min at37u C in KRBH/ 0.07%defatted BSA and2.8mM glucose,followed by incubation for1h in1ml KRBH/0.5%defatted BSA and 2.8,8.3or 16.7mM glucose plus or minus20nM GLP-1-(7–36)-amide (Bachem Americas,Torrance,CA,USA)or20nM Ex-4(Bachem Americas,Torrance,CA,USA),in the presence or absence of 0.3mM palmitate.For whole mouse islets,or populations of single mouse islet cells(plated on the wells of48-well plates),insulin secretion was measured at4,7.5and16.7mM glucose in the presence or absence of10nM Ex-4.At the end of a30min static incubation,media were kept for insulin measurement by radioimmunoassay(Linco research,St.Charles,MO,USA).Islet total insulin content was measured following acid-ethanol(0.2mM HCl in75%ethanol)extraction.Islet fatty acid oxidation and esterificationFatty acid(FA)oxidation and esterification were determined in batches of50islets cultured as described above.After2h incubation in 2.8mM glucose-RPMI complete medium,islets were washed in KRBH/0.25%BSA and pre-incubated for45min at37u C in KRBH/0.25%defatted BSA and2.8mM glucose after which they were incubated for2h(FA oxidation)or4h(FA esterification)in1ml KRBH/0.25%defatted BSA containing2.8, 8.3or16.7mM glucose in the presence or absence of20nM GLP-1,0.1mM(oxidation)or0.2mM(esterification)palmitate, 1m Ci/ml[9,10(n)-3H]palmitate(51Ci/mmol,GE Healthcare, Baie d’Urfe´,QC,Canada),and1mM carnitine.At the end of the incubation,the media were collected for the determination of islet FA oxidation and total lipids were extracted from islets for the measurement of islet FA esterification[38].Glucose oxidation and utilizationGroups of20islets cultured and pre-incubated as described above for the insulin secretion assay,were incubated in a0.6ml Eppendorf tube without capping,in a final volume of70m l KRBH/0.25%defatted BSA containing2.8to16.7mM glucose with D-[U-14C]-glucose for oxidation measurements(250mCi/ mmol,PerkinElmer,Canada)and D-[5-3H]-glucose for utilization measurements(16Ci/mmol,GE Healthcare,Canada)with or without20nM GLP-1[43].For oxidation incubations,this incubation tube was placed upright in an airtight-sealed20ml scintillation vial,which also contained an empty1.5ml Eppendorf tube without capping.The reaction was stopped after90min incubation at37u C with constant agitation by the addition of50m l of a mix consisting of metabolic poisons(400mM citric acid, 10m M rotenone,10m M antimycin and3.5mg KCN,pH4.9). To the empty1.5ml tube in the scintillation vial,250m l of5%(w/ v)KOH was added to trap released14CO2.Incubations were continued for60min at room temperature and glucose oxidation was determined by measuring the KOH-trapped14CO2.For utilization measurements,the scintillation vial also contained 500m l of1mM HCl at the bottom.After stopping the incubations as above,the tightly sealed vials were left at room temperature for 40h and glucose utilization was determined by measuring the amount of3H2O equilibrated into the0.5ml HCl in the vial. LipolysisBatches of60islets,cultured as described above,were washed in KRBH/0.07%BSA and2.8mM glucose and were transferred into0.2mL of KRBH/0.07%BSA medium in a48well plate with2.8,8.3or16.7mM glucose and20nM GLP-1or20nM Ex-4.The plate was incubated for3h at37u C in a humidified atmosphere containing5%CO2,after which the media were collected for glycerol determination by an enzymatic assay[37]. Islet protein content was measured,as previously described[37]. Measurement of[Ca2+]i and cAMP contentMouse islets were dispersed by mild digestion with trypsin-EDTA and the single islet cells were plated on glass cover-slips for [Ca2+]i measurement,or in96-well cell culture plates for cAMP determination.After overnight culture,the islet cells were infected with Ad-MtLuc-RFP(m.o.i.equal to50).Measurements of[Ca2+]i were performed after48h,using fura-2loaded b-cells,imaged at 100X magnification using a dual excitation light source and a ratiometric imaging system(IonOptix Corp.)equipped with filter sets that minimize crossover between fura-2and RFP[44].The cAMP content was determined by immunoassay using a Direct Biotrak EIA kit(Amersham)as described earlier[45].Oxygen ConsumptionOxygen consumption in single rat or mouse islets was measured at37u C in the presence or absence of10nM Ex-4or10m M forskolin by the self-referencing method based on an electrochem-ical oxygen sensor(BioCurrents Center,MBL,Woods Hole,MA) moving between a‘‘near’’and‘‘far’’position at the islet.The magnitude of the amperometric current used for the reduction of oxygen is proportional to the oxygen concentration at that particular point[46].When islet respires,oxygen concentration is lower at near position.Thus,the current used for reduction of oxygen on the sensor will be greater at the far position,and the measured difference in the electric current between far and near position(Difference Current,DC)is greater than zero.When an islet further increases oxygen consumption(in response to a rise in glucose concentration),oxygen concentration at near position decreases even more and causes further increase in the DC. Oxygen consumption was measured in islets incubated in KRBH containing4,7.5,and16.6mM glucose.Mitochondrial ATPChanges of mitochondrial ATP levels(ATPm)were measured in a population of approximately250,000single rat or mouse islet cells infected with Ad-MitoLuc-RFP.This virus was generated using the mt-Luc coding sequence in plasmid VR102.mt-Luc is a fusion protein in which the26amino acid N-terminal signal peptide of cytochrome C oxidase subunit VIII(COX8)is fused to codon-optimized firefly luciferase[47].Ad-MitoLuc-RFP infected islet cells were incubated with KRBH buffer(for72h)containing,either4, 7.5or16.6mM glucose without or with10nM Ex-4.Single islet cells,grown and infected on the PDL-coated glass coverslips inside a 35mm dish(MatTek,Ashland,MA)were placed directly onto the surface of the photocathode optical window of a Hamamatsu R464 photomultiplier tube housed in a37u C heated box.Luciferin was then added to the KRBH at a final concentration of100m M in order to allow the measurement of photoemissions resulting from luciferase-catalyzed oxidation of luciferin[48].PKB/Akt phosphorylationA group of200islets cultured and pre-incubated as described for insulin secretion experiments,was incubated for30min in 1ml KRBH/0.5%defatted BSA containing 2.8or8.3mM glucose in the presence or absence of20nM GLP-1.After30min, islets were lysed in0.1ml of50mM HEPES(pH7.5),2mM sodium orthovanadate,4mM EDTA,100mM sodium fluoride, 10mM sodium pyrophosphate,1mM PMSF,1%(v/v)NP40and protease inhibitors.Total cellular proteins were obtained after sonicating the islets for10s and centrifugation at15,0006g at4u C for12min.The supernatant was collected and the protein content assayed(Pierce).Proteins were resolved by10%sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and elec-trotransferred to nitrocellulose membranes(BioRad,Hercules, CA,USA).After overnight blocking with5%non-fat milk,the membranes were probed with antibodies for pSer473-PKB and total PKB(Cell Signaling Technology,MA,USA)and the proteins were visualized by enhanced chemiluminescence(Pierce). Statistical analysisData are expressed as means6SE.Significance was calculated for multiple comparisons by using one-way analysis of variance (ANOVA)with Bonferroni post-hoc testing.A P-value of,0.05 was considered significant.ResultsGlucose stimulated insulin secretion from isolated rat islets in a dose-dependent manner,and this effect was enhanced by0.3mM palmitate at an intermediate concentration(8.3mM)of glucose (Fig.1A).Both GLP-1and Ex-4markedly potentiated GSIS at8.3 and16.7mM glucose,and palmitate did not further elevate insulin secretion.At16.7mM glucose,GLP-1and Ex-4stimulated insulin secretion3–4fold more than glucose alone.In the presence of GLP-1or Ex-4the amount of insulin released during the 45min time period corresponded to approximately20%of the total islet content of insulin.In mouse islets,Ex-4enhanced GSIS at7.5and16.7mM glucose,but not at4mM glucose(Fig.1B). Overall,Fig.1shows that the islets that were used in the current study were highly responsive to glucose,GLP-1and Ex-4and therefore suitable to be used to respond to the addressed questions. Inasmuch as both GLP-1and Ex-4are agonists of GLP-1receptoron the b-cells,with near equal potency,most studies to date have not found any significant differences between these two agonists in in vitro experiments.In the present study,although most experiments were performed with both GLP-1and Ex-4,because of the similar nature of the data,only either of these is illustrated. PKB has multiple metabolic effects in various cell types[49]and GLP-1acutely activates PKB in INS cells[27]and human islets [29].So far it has not been shown that GLP-1activates PKB in normal rodent b-cells.We examined the effect of GLP-1(20nM) on PKB phosphorylation in rat islets after a30min exposure to the hormone,and observed that GLP-1significantly increased Ser-473phosphorylation of PKB in normal islet cells(Fig.2).It was therefore of interest to determine whether changes in islet intermediate metabolism might be explained by influences of GLP-1on PKB phosphorylation/activation[50,51],in addition to its previously demonstrated stimulatory effects on islet cAMP and Ca2+signaling shown in many studies employing both normal (human,rat,mouse)and tumoral b-cell(for reviews see [3,10,19,20,21]).Tsuboi and co-workers[52]reported that GLP-1receptor activation increased[Ca2+]i,which caused an elevation of mitochondrial ATP in MIN6insulin-secreting cells.In order to ascertain that Ex-4increases[Ca2+]i and cAMP in normal mouse b-cells under our experimental conditions where mito-chondrial ATP and O2consumption were measured,single cell measurements of[Ca2+]i were performed using b-cells loaded with fura-2,infected with Ad-MtLuc-RFP,and equilibrated in KRB containing5.6mM glucose.Under these conditions,Ex-4(10nM) stimulated an increase of[Ca2+]i in these cells(Fig.3A). Importantly,MtLuc expression had no effect on the percentage of cells exhibiting a.100nM increase of[Ca2+]i(Fig.3B).Thus, viral infection did not disrupt the stimulatory action of Ex-4on intracellular Ca2+signaling.Since Ex-4is known to stimulate insulin secretion in a glucose-dependent manner,we examined whether glucose concentration influences Ex-4stimulated intra-cellular Ca2+signaling.This was in fact the case since the action of Ex-4to increase[Ca2+]i was more prominent under conditions in which mouse b-cells were equilibrated in KRB containing7.5mM glucose as compared to5.6mM glucose(Fig.3B).Since it is known that the increase of[Ca2+]i in response to Ex-4 is secondary to b-cell cAMP production[44],it was of interestto Figure1.Acute effects of GLP-1and Ex-4on GSIS in rat(A)and mouse(B)islets.Pancreatic islets were isolated and cultured overnight prior to use as described in Methods.Islets were incubated for1h(A)or30min(B)as described in Methods for examining insulin secretion at indicated concentrations of glucose and GLP-1(20nM)or Ex-4(20nM)in A,or10nM Ex-4in B,in the absence or presence of0.3mM palmitate. Insulin released into the media and the total islet insulin content were measured.Results shown are mean6SE from3independent experiments with quadruplicates(n=12).For A,*p,0.05,**p,0.01,***p,0.001when compared with corresponding2.8mM glucose group.#p,0.05,##p,0.01, ###p,0.001when compared with corresponding groups without GLP-1or Ex-4treatment.For B,{p,0.01when compared to corresponding minus Ex-4group.Insulin secretion in mouse islets at basal glucose levels(4mM)was1.5660.3ng insulin/10islets/30min.doi:10.1371/journal.pone.0006221.g001determine if viral infection altered the ability of Ex-4to increase levels of cAMP in primary mouse islet cells.We found that Ex-4stimulated cAMP production in a dose-dependent manner,in islet cells infected with Ad-MtLuc-RFP (Fig.3C),without any significant difference from cells not infected with Ad-MtLuc-RFP (Fig.3C).Thus,viral infection as described here for Ad-MtLuc-RFP,did not disrupt intracellular Ca 2+signaling and cAMP production in mouse b -cells.We have previously shown [53,54]that GSIS in rat islets and INS cells is accompanied by reduced b -oxidation and increased partitioning of fatty acids into glycerolipids,an event that is thought to be coupled to b -cell activation of insulin release [5].Therefore,we examined whether the acute stimulatory effect of GLP-1on GSIS in islets might result from altered lipid metabolism.There was no significant effect of GLP-1on palmitate oxidation or its incorporation into different classes of glycerolipids or cholesterol esters or phospholipids (Fig.4)at the various tested glucose concentrations.We next assessed whether the acute stimulatory effect of GLP-1on GSIS is associated with changes in islet glucose metabolism.However,rat islet glucose utilization (Fig.5A)or oxidation (Fig.5B)was not significantly affected by GLP-1at all tested glucose concentrations,except for a small increase in utilization at 16.7mM glucose (Fig.5A).Enhanced lipolysis and GL/FFA cycling are thought to play a role in the amplification (K ATP -independent)arm of fuel induced insulin secretion [5,35],and previous work in the HIT cell line showed that GLP-1enhances glycerol release in this tumoral (b )cell [40].However,GLP-1did not enhance lipolysis in rat islets at low,intermediate and high glucose (Fig.5C).We also examined the effect of Ex-4and GLP-1on oxygen consumption at different glucose concentrations in rat and mouse islets since this parameter reflects overall fuel utilization and metabolic activation of a given tissue.Even though oxygen consumption increased with glucose concentration,there was no significant change with Ex-4(Fig.6A &B)or with GLP-1(data not shown),in accordance with the results from glucose oxidation experiments.Also,the adenylate cyclase activator forskolin (10m M)did not affect oxygen consumption in mouse islets (data not shown).Enhanced mitochondrial metabolism and ATP production plays a central role in b -cell fuel signalling [55]and GLP-1was shown to enhance mitochondrial ATP production in the tumoral b -cell line MIN6[52].We further measured mitochondrial ATP levels in isolated rat and mouse islet cells engineered to express mitochondrial-targeted luciferase,after incubation for 5to 30min,with different glucose concentrations and Ex-4.Glucose caused a marked and dose dependent increase in mitochondrial ATP,within 5min of incubation,in both rat and mouse islets cells,but Ex-4did not significantly change mitochondrial ATP at all tested glucose concentrations (Fig.7A &B).Identical results were obtained using 20nM GLP-1(data not shown).We verified that the mitochondrial-luciferase-engineered islet cells respond nor-mally to the respiratory substrates and inhibitors,by examining the effect of methylsuccinate (10mM),a membrane permeable form of succinate and FCCP (10m M),an uncoupler of oxidative phosphorylation.As expected,during a 15min incubation,methylsuccinate enhanced ATP production above the basal (4mM glucose)level,whereas,FCCP reduced ATP levels (data not shown).DiscussionFuel stimulated insulin secretion in the b -cells involves the production of metabolic coupling factors and an elevation in intracellular Ca 2+[1].GLP-1and Ex-4enhance GSIS in the b -cells,cause a rise in cytosolic Ca 2+,elevate cAMP and activate the PKA and PKB signaling pathways [3,10,19,20,21].Because activation of Ca 2+,cAMP and PKB signaling is known to modulate intermediary and energy metabolism in several cell types,we hypothesized that GLP-1signaling to stimulate insulin secretion is in part linked to changes in b -cell metabolism and the production of metabolic coupling factors.The data in fact show that GLP-1or Ex-4barely affect b -cell metabolism at large,and therefore that GLP-1induced insulin secretion may not involve metabolic signaling related to glucose,lipid and energy metabolism.We [1,5]and others [56]provided evidence that enhanced lipolysis in b -cells plays a role in GSIS.However,lipolysis does not appear to be involved in the acute amplification of GSIS by GLP-1in normal islet tissue.Thus,in accordance with a previous study using isolated mouse islets [57],we observed that GLP-1or Ex-4do not affect rat islet lipolysis.Furthermore,islets from hormone sensitive lipase-KO mice exhibited GLP-1stimulation of GSIS similar to that of control mouse islets [37].The previously reported increase in lipolysis by acute treatment with GLP-1in HIT (b )cells [40]is probably due to the inherent differences in established tumoral cell lines from normal islet b -cells.Lipid amplification pathways of GSIS in the b -cell involve reduced fatty acid b -oxidation and concomitant increased esterification of fatty acids into glycerolipids [53,54].We have suggested that enhanced GL/FFA cycling is instrumental in the amplification of GSIS via the generation of lipidsignallingFigure 2.PKB phosphorylation in response to GLP-1in rat islets.Islets were incubated for 30min at 2.8or 8.3mM glucose in the presence or absence of 20nM GLP-1.Activation of PKB by GLP-1was assessed with antibodies specific for Ser 473phospho-PKB and PKB,respectively.(A)Representative immunoblot of phospho-and total PKB in rat islets.(B)Quantitative measurement of PKB phosphorylation after 30min treatment with GLP-1.Results are means 6SE of 4separate experiments.#p ,0.05,when compared to the corresponding ‘minus GLP-1’group.doi:10.1371/journal.pone.0006221.g002。

英文论文投稿信—相关

1.Submit paper1.1 例文1Dear Editor,We would like to submit the enclosed manuscript entitled "GDNF Acutely Modulates Neuronal Excitability and A-type Potassium Channels in Midbrain Dopaminergic Neurons", which we wish to be considered for publication in Nature Neuroscience.GDNF has long been thought to be a potent neurotrophic factor for the survival of midbrain dopaminergic neurons, which are degenerated in Parkinson’s disease. In this paper, we report an unexpected, acute effect of GDNF on A-type potassium channels, leading to a potentiation of neuronal excitability, in the dopaminergic neurons in culture as well as in adult brain slices. Further, we show that GDNF regulates the K+ channels through a mechanism that involves activation of MAP kinase. Thus, this study has revealed, for the first time, an acute modulation of ion channels by GDNF.Our findings challenge the classic view of GDNF as a long-term survival factor for midbrain dopaminergic neurons, and suggest that the normal function of GDNF is to regulate neuronal excitability, and consequently dopamine release. These results may also have implications in the treatment of Parkinson’s disease.Due to a direct competition and conflict of interest, we request that Drs. XXX of Harvard Univ., and YY of Y ale Univ. not be considered as reviewers. With thanks for your consideration, I amSincerely yours,1.2 例文2Dear Editor,We would like to submit the enclosed manuscript entitled "Ca2+-binding protein frequenin mediates GDNF-induced potentiation of Ca2+ channels and transmitter release", which we wish to be considered for publication in Neuron.We believe that two aspects of this manuscript will make it interesting to general readers of Neuron. First, we report that GDNF has a long-term regulatory effect on neurotransmitter release at the neuromuscular synapses. This provides the first physiological evidence for a role of this new family of neurotrophic factors in functional synaptic transmission. Second, we show that the GDNF effect is mediated by enhancing the expression of the Ca2+-binding protein frequenin. Further, GDNF and frequenin facilitate synaptic transmission by enhancing Ca2+ channel activity,leading to an enhancement of Ca2+ influx. Thus, this study has identified, for the first time, a molecular target that mediates the long-term, synaptic action of a neurotrophic factor. Our findings may also have general implications in the cell biology of neurotransmitter release.1.3 例文3Dear Editor:Enclosed are copies of a manuscript entitled "BDNF and NT-4/5 Promote the Development of Long-Term Potentiation in the Hippocampus", which we wish to be considered for publication in Nature.As you know, there is a great deal of interest and excitement recently in understanding the role of neurotrophins in synapse development and plasticity. Our manuscript provides, for the first time, the physiological evidence that neurotrophins regulate long-term potentiation (LTP). The main point of the paper is that the neurotrophins BDNF and NT-4 induce an earlier appearance of LTP in developing hippocampus. In contrast to recent Science article by XX's group, we (and several other LTP groups) did not see that BDNF enhance basal synaptic transmission in adullt hippocampus. However, we found that in adult hippocampus, inhibition of BDNF/TrkB activity attenuated LTP, and weak tetanus that normally cannot induce LTP produced enduring LTP. These findings may have implications in the basic mechanism for regulation of synapse development and long-term modulation of synaptic efficacy.Because of the rather competitive nature of the field and the important implication of our findings, we have not yet presented this work in any public forum. However, confidential discussion with several prominent neuroscientists such as 111 and 222 have generated tremendous excitement. Thus, we feel that this work is of general interest and is suitable for publication in Nature.We would like to suggest Drs. aaa of Y ale Univ., bbb of Harvard Medical School, and ccc of Univ. of California-Berkeley, as reviewers for this manuscript. Due to a direct competition and conflict of interest, we request that Dr. XX and YY. not be considered as reviewers.Thank you very much for your consideration.1．5英文投稿的模版信1发信站: 两全其美BBS (Wed Jun 8 20:57:42 2005), 转信()Dear Sir,We are submitting a manuscript entitled “XXXXX” for your kind consideration for publication in XXXX.In this manuscript, we report a novel approach ..............We would be grateful if the manuscript could be reviewed and considered for publication in this journal.1．6英文投稿的模版信2Dear Professor ×××:One of my paper, which has a title of “××××”, is hoped to be published in your journal . Please take it into your consideration for publication.If you need any more information concerning the manuscript please write to me. I am looking forward to hearing from you.Best regards××××1．7英文投稿的模版信3PLoS GeneticsI submit the accompanying manus cript entitled “Gain and loss of multiple genes during the evolution of Helicobacter pylori” by H. Gressmann et al. for publication in PLoS Genetics.This manuscript presents the global variability within H. pylori in terms of presence or absence of almost all the genes within the two currently available genome sequences. The strains tested are representative of the global genetic diversity within H. pylori and also represent the diverse populations that exist in that organism. Global analyses of genomic content are very rare indeed for bacterial species and are new for H. pylori. By including isolates from the most closely related species, H. acinonychis, as an outgroup, it was possible to deduce that most variable genes were probably present in the last co mmon ancestor of H. pylori and have subsequently been lost in individual isolates. In some cases, the same genes were lost in phylogenetically distinct groupings, reflecting convergent evolution. However, the cag PAI was acquired later, after subpopulations had formed. These conclusions are important for concepts about changes in gene content within species and for the acquisition of pathogenicity islands.We also perform a head-to-head comparison of population structure indicated by whole genome microarrays versus that indicated by sequences of 7 housekeeping gene fragments. The comparison indicates that microarrays yield distorted population structures and are less suitable for this goal than simply sequencing 7 gene fragments. In order to be able to deduce patterns of gain or loss, it was necessary to have an independent population structure based on sequence diversity within core genes. This is an important conclusion because many laboratories are attempting to investigate bacterial population structure on the basis of microarray data on their own and have not realized that an independent population structure is necessary for such efforts..The manuscript will be of great interest to the numerous scientists interested in infections caused by H. pylori as well as to scientists interested in microbial genomic structure and evolution.The following scientists, with whom I have had no contact regarding this manuscript are potentially suitable reviewers:1.Michael McClelland, Sidney Kimmel Cancer Centre, San Diego, CA mmcclelland@2.Al Ivens, Pathogen microarrays, Sanger Center, Hinxten Hall, Cambridge, UKalicat@3.Antonello Covacci, IRIS, Chiron Vaccines, Siena, Italy antonello_covacci@4.Paul O’Toole, University of Cork, Ireland pwotoole@ucc.ie5.Jörg Hacker, University of Würzburg, Germany joerg.hacker@mail.uni-wuerzburg.deI request that Doug Berg not be used as a reviewer due to potential conflict of interest.Sincerely,Mark Achtman, PhD2. When your paper gets rejected — without reviewDear Editor,I would appreciate if you could reconsider to review our manuscript, “111." We feel strongly that this is an important subject that touches one of the central dogmas in neuroscience: xxx. It is also very timely, given the publication of the paper by X a nd Y entitled “222” in the latest issue of Nature Neuroscience. In this paper, the authors xxx. They claimed that xxx. When a paper this provocative has been published by a high profile journal like Nature Neuroscience, we believe that it is worth giving a benefit of doubts. It will be helpful if there are papers that consider other alternative interpretations, or attempt to replicate in the same or different systems.We have observed similar xxx, but we have a completely different interpretation. We found that 1) xxx 2) xxx; 3) xxx. Thus, our paper raises the possibility that xxx reported by X and Y were due to xxx. Specifically, we would like you to consider the following two issues: First, X and Y used aaa, while we used bbb. sssssssss. Second, ccc used by X and Y may not be so specific.In addition to the drastically different opinions regarding xxx, we feel that our findings on xxx is also significant in yyy and will be of interests to general readers of Nature Neuroscience. We therefore did not write our paper to directly challenge the paper by X and Y. However, we willbe willing to re-write the paper in ways you think that will help debate on this important issue.3. When your paper gets rejected — with reviewDear Dr. xx,We received with some surprise your letter of November 4, rejecting this manuscript on the basis of one reviewer’s opinion which you “found persuasive”.We wish to indicate our dissatisfaction with this reviewer’s comments, which appear to ignore the new experiments submitted as part of the revised manuscript.This reviewer states: “111.”This was precisely the point of the xxx experiment which indicated that there were no such deficits.This reviewer further states: “222.” Again, this is a mystifying statement as the detailed rebuttal accompanying this letter described the xxx. Did the reviewer not understand that xxx?Finally, concerning the proposal for a xxx experiment, we believe that you and this reviewer already know that xxx. Thus, it is impossible to do such experiments.While we recognize that the final decision is yours, we feel that reviewer#1 is being unreasonable. We would greatly appreciate it if you would submit this manuscript, reviewer#1’s comments, and our rebuttals, to an additional unbiased reviewer. We would be most surprised if the new reviewer would see the comments of the reviewer#1 as reasonable, but if he/she did so, we would accept a negative decision gracefully.4. 催审稿4.1 例文1(我写的，见笑了！）Dear editor,Thank you for your reading this mail.I submit a paper entitled "…………" in November last year and this paper has been reviewed for more than five months now. I haven't received the Referees' comments on it.I respectfully wish for the comments soon, thank you!Once again, thank you very much for your consideration.Best regards!yours sincerely,………………22-Apr-20054.2 例文2(编辑回信)Editor check on the progress of the paper. The manuscript is in the review process and I anticipate receiving the 2nd reviewer's comment in the next 2 weeks. The first reviewer´s comments are not very encouraging, so I will inform you on the decision as soon as I have received the comments of the second one.Thank you for submitting your research to the ISPRS Journal. I will be happy to keep you informed of how the reviews are progressing.4.3例文3给责任编辑写催问信的. 以下是模板Dear Dr. XXX,We submitted a manuscript (manuscript ID) to “XXXX journal” on X-X-X. Y ou are the execu tive editor. W e have not, yet, received any comment or decision abou t the submission. Please let us know any further information of the reviewing process at your earliest convenience.Many thanks.4.4例文4Dear editor,Thank you for your reading this mail and I am very sorry for sending you this mail again.I want to know if the second comment on my paper (…………) has been received. If it has been received, could you please inform me on the decision?I respectfully wish for your reply, thank you!Once again, Thank you very much for your consideration.Best regards!yours sincerely,………………11-Oct-20055. 催寄会议邀请信Dear Miss. Lee,Many thanks for your hard work.I have received the invitation letter from you by email. You said the original invitation letter would be sent to me on 6 May, but I haven't received it yet.Please send us the original invitation letters for attending the symposium, because the invitation letters are required for us to transact visa. Mr. LI will register when he attend the symposium.It's ok to send the two letters to me together in one mail. Many thanks!Once again, thank you very much for your hard work.Best regards!Yours sincerely,……18-May-20056. Sample Letter to the Editor6.1 on Energy and Global WarmingDear Editor:The presidential race is heating up, but one issue is surprisingly absent from the debate: the environment. With high gas prices and continued tension in the Middle East, now seems to be the perfect time for candidates to start seriously addressing the problems of America's continued dependence on fossil fuels. Air pollution from power plants and vehicles contributes to smog and global warming, and it is estimated that pollution from U.S. power plants kills 30,000 people each year.The Bush Administration has largely neglected these issues; indeed most of its policies seem designed to increase our dependence on fossil fuels. The Administration's national energy policy calls for constructing hundreds of new power plants, increasing coal and nuclear energy, and increasing drilling on public lands. Apparently the plan also includes allowing more pollution from these already-dirty energy sources; the Administration is being sued by several northeast states because of its drastic weakening of the Clean Air Act. The Administration has also been in favor of drilling for oil in the Arctic National Wildlife Refuge, despite the fact that the US Geological Survey estimates that this would yield only six month's worth of oil. A more forward-thinking energy policy would emphasize the development of renewable technologies and increased energy efficiency, especially of our vehicle fleet.The decisions we make now about our energy policy will affect our health, our security, and the environment that we ultimately depend on. Let's hope we hear more serious discussion about these issues before the election.NAMESCHOOLCITYDear Editor:From a student's perspective, I am disturbed by the lack of attention that global warming is receiving in the presidential campaign. In this century, the average global temperature is expected to rise between 2.5 and 10.5 degrees Fahrenheit, which is quite significant if one considers that the temperature only increased 5 –9 degrees between the last ice age and today. The economic effects of such a change will be devastating: Swiss Re estimates that the costs from weather-related disasters will double to $150 billion a year in a decade. And the authors of a recent report from the Pentagon believe that the destabilizing impacts of climate change pose a serious security threat.Unfortunately, our government, despite all its interest in the economy and national security, doesn't seem to be taking notice. Indeed, the Bush Administration's policy on global warming has been to ignore it. Early in his term, Bush withdrew the U.S. from the Kyoto Treaty, the first international treaty to reduce greenhouse gas emissions. Although the treaty hassome flaws, it was an important first step, but unless Russia or the U.S. ratifies it, it cannot go into effect. Domestically the government has opposed any plans to regulate greenhouse gas emissions. It has even gone so far as to manipulate scientific evidence; in 2003, the Administration attempted to make such substantial changes to a section of an Environmental Protection Agency (EPA) report on climate change that the EPA finally deleted the section rather than destroy its scientific credibility.Such short-sighted policies may not have much direct impact today, but it is my generation that is going to have to deal with their consequences. And unless the U.S. government joins the rest of the industrialized world in acting on this problem, those consequences aren't going to be pretty.NAMESCHOOLCITY6.2 on Bush and National ForestsDear Editor:As the presidential election approaches, environmental issues have hardly been discussed, despite its importance to many Americans, especially among young Americans who want to protect our wild lands for fu ture generations. The Bush Administration’s assault on the environment is evident in many areas, but is clearly visible in our National Forests. Since Bush took office more than three years ago, he has created several policies that have rolled back hard earned environmental protections in favor of commercial logging and the oil and industries.Recently the Bush Administration announced their plans to potentially open up the remaining 60 million acres of roadless area in our National Forest to development, oil and gas drilling, and commercial logging. Despite the largest public involvement process in the history of the federal government with more than 600 public hearings and two million public comments in favor of protecting our wild forests, the Bush Administration has now put the Roadless Rule on hold. Under the guise of “fire prevention” and “forest health,” the Bush Administration is chipping away at forest protections policies across the country. Even while many paper and timber corporations are backing away from the logging of old growth forest, the Bush Administration is opening up our most protective areas to commercial loggers and developers. We are headed in a dangerous direction under the Bush Administration.Besides the beauty of our National Forest, these wild areas have some of the highest quality fish and wildlife habitats, backcountry recreation, and clean water supplies in the country. Our treasured wild heritage is being chopped down by the Bush Administration’s relaxed environmental policies, and as young people we want these areas protected for future generations to enjoy. It’s time to stop this assault on our National Forests, and reinstate needed protections from commercial logging, and oil extraction, and development.NAMESCHOOLCITYDear Editor:I was dismayed to hear that the Bush Administration took steps in July to undermine historic protections for our nation’s last remaining wild forests. The Administration announced that it would seek to re-write the Forest Service’s widely popul ar Roadless Rule.These proposed changes will leave countless acres of our country’s most pristine forested areas at risk, and goes against the protections demanded by the American people. To date, an unprecedented 2 million-plus Americans expressed suppor t for the “roadless rule” and for protecting these special places and the wildlife habitat, clean water, and recreational opportunities they provide.Since taking office, the Bush Administration has repeatedly whittled away at the core of the wild forest policy and failed to defend it in court, while simultaneously promoting intensive logging as a remedy for forest fires. There are already more than 440,000 miles of roads that scar our National Forests – roads built for the logging industry and paid for by our tax dollars –and that have destroyed wildlife habitat, caused mudslides and polluted our water.We have enough roads –it is time to safeguard our last wild forests. I hope the Bush administration will begin to listen to the overwhelming majority of Americans rather than to big logging corporations.NAMESCHOOLCITY6.3 on Bush and TradeDear Editor:In May, the Bush Administration signed the Central American Free Trade Agreement (CAFTA). In an election year, this happened without much publicity. The A dministration didn’t seem to want people to know what it was doing, and there are reasons why.CAFTA is NAFTA with a “C.” Under CAFTA, we could expect the same undermining of democracy by corporations, and the same threats to the environment. Under NAFTA, corporations have the right to sue a government directly if they feel that their ability to profit has been undermined by an environmental law or regulation. Examples of such lawsuits include a case where Mexico was forced to pay $16 million after denying a U.S. company a permit to build a toxic waste facility on an environmentally sensitive site, and a current case where a Canadian mining company is suing the U.S. for $50 million over a California law that requires that the holes created by open-pit mining be “backfilled” and that the landscape be brought back to its original form once mining operations have been completed. In a notorious, ongoing case, the Canadian company Methanex is suing the U.S. for $970 million because California enacted a law that banned the harmful gasoline additive MTBE that has severely polluted drinking water in many California communities.CAFTA would replicate this NAFTA provision that undermines state and U.S. law in favor of corporate profits. The threat is all the more serious since U.S. companies would even be able to open a subsidiary in a country like Guatemala in order to turn around and sue the U.S. over laws it doesn’t like.[REP.] XXX SAYS [HE/SHE] [SUPPORTS/ISUNDECIDED ABOUT] CAFTA. [HE/SHE] SHOULD OPPOSE CAFTA. Our democracy and our environment are at stake.NAMESCHOOLCITY7. 邀请某人做审稿人7.1 Reviewer Invitation for MS No. RSE-D-05-00256Dear Dr.----，The following paper has been submitted to …journal…, "Analysis of co-occurrence and wavelets transform textures for classification of vegetation types in very-high resolution imagery." I would greatly appreciate it if you would review the paper.To view the manuscript and the procedure for electronic review, please go to / and log in as a reviewer using the following user name and password:User Name:Password:If possible, we would appreciate receiving your review by 13 Jun 2005. If you cannot review this paper, we would appreciate receiving your recommendation on other well qualified reviewers. Please include their email address.We also invite you to consider RSE for publication of your papers in the remote sensing area. RSE continues to be the #1 remote sensing journal as measured by the Science Citation Index.Reviewers are a critical part of the publication process and, along with authors, are the strength of RSE, so we greatly appreciate your assistance.Sincerely,Marvin BauerEditor-in-ChiefRemote Sensing of Environment7.2 Thank you for agreeing to reviewDear Dr. ------,Thank you for agreeing to review manuscript RSE-D-05-00256, "Analysis of co-occurrence and wavelets transform textures for classification of vegetation types in very-high resolution imagery" for Remote Sensing of Environment. I would appreciate receiving your review by 13 Jun 2005.Y ou may submit your comments online at /.Y our User Name is:and your password is:Y ou will find instructions for completing the review with your comments for the authors, recommendation, and confidential comments to the editor.If you print a copy of the paper and annotate it with suggested edits or other comments, please either mail or fax it to us so that we can provide them to the author. Some reviewers like to do this and it provides additional help to authors in revising and improving their papers. Although Editorial Manager does not directly provide that capability, we do not want to exclude it as an option.A prompt, careful review of this paper will be greatly appreciated by the authors. Thank you for the time and effort that this request entails.Sincerely,Marvin E. BauerEditor-in-ChiefRemote Sensing of EnvironmentUniversity of Minnesota1530 N. Cleveland A venueSt. Paul, Minnesota 55108Fax: 612.625.52128. 接受做审稿人Dear editor,Thank you for inviting me to review the manuscript RSE-D-05-00256.I mainly do research on information extraction from high-resolution remote sensing image. As an author, I really appreciate that the referees review my paper promptly and carefully. So I willreview the paper and give my comments and suggestions as soon as possible.Best regards!Sincerely,9.已接收文章要求改动（如作者信息）Subject: Sorry, some changes of the authors' affiliation ! ( accpeted paper, ID# TRES-PAP-2004-0147.R2)Dear ediror,I am very sorry that I have to make some changes of the authors' information of the accepted manuscript ID # TRES-PAP-2004-0147.R2, entitled "……" .The paper with changes is enclosed in the attached file. The changes are:(1) The first and the third author's affiliation are change.(2) the fourth author's first name and family name was converse and I correct it in the attached file.I am very sorry for it and thank you for your hard work once again!Best regards!yours sincerely,……11-Jul-200511.收到论文接受信的回复发信站: BBS 水木清华站 (Thu Jul 28 23:45:18 2005), 站内请赐教。

PLOS ONE 论文发表被拒稿的实例分析

PLOS ONE 论文发表被拒稿的实例分析医刊汇5018编《PLos ONE》为PLoS系列期刊之一，是为科技人员和医学人员服务的非赢利性机构，致力于使全球范围科技和医学领域文献成为可以免费获取的公共资源，旨在推广世界各地的科学和医学领域的最新研究成果。

PLoS出版了8种生命科学与医学领域的开放获取期刊，可以免费获取全文，比较具有影响力。

PLOS ONE接收并发表的文章快速增加(2010年接近7000篇)，受到各国科研学者越来越多的关注。

然而其对所提交的稿件经过严格的审核，有自行写作稿件的作者面临着多次被拒稿的情况，下面是医刊汇（5018）SCI期刊编辑收到的一位被撤稿作者的求助：撤稿作者投稿PLOS ONE后30天大修，审稿人的意见大略如下：1.写作逻辑需要调整“Many parts of the manuscript are mixed. Some of the introduction belongs to the discussion, some of the results to the methods etc. The discussion should be re-arranged and it should clearly state at the beginning what their findings are. Same for the introduction. It should clearly state the purpose of the article that must be recalled in the discussion.”2.语法问题“Also,they need a language revision. There are too many mistakes especially with articles, concordance, plurals, verbs and wording.”医刊汇（5018）SCI期刊编辑为其提供的修改方案：1.重新整理写作思路，比如将结果中有些介绍性的句子放在Introduction 或Discussion部分。

在PLOSONE上投稿的过程（邮件版）

在PLOSONE上投稿的过程（邮件版）刚刚在PLOS ONE 上发了我读硕期间的第一篇SCI文章，心里挺开心的。

一个虫友曾问我在这上面发文章的流程，希望我能出个技术贴~惭愧的是小弟我记忆力不是太好，投稿的流程忘了差不多了。

其实我也是第一次投，就按杂志给的页面一步步来的，没有遇到什么困难~现在我把我投稿期间杂志给我发的邮件全都整理出来，希望能对大家有点帮助！*************************************************************** ****************Editorial Manager New Account Registration2012年4月22日, 星期日, 下午 5:38Dear Mr Chen,Thank you for registering for the Editorial Manager online submission and peer review tracking system for PLoS ONE.Here is your username and confidential password, which you need to access the Editorial Manager at /.Your username is: ***Your password is: ***Please save this information in a safe place.You can change your password and other personal information by logging into the PLoS ONE website at / and clicking on the Update My Information link on the menu.Best regards,PLoS ONE*************************************************************** ****************这是我注册后网站给我发的邮件~投稿历程正式开始！*************************************************************** ****************PLoS ONE Notification: Your PDF has been built (文章标题)2012年5月28日, 星期日, 下午 10:15Dear Mr Chen,This is an automatic email.The PDF for your submission, "文章标题" has been built. If you have not already done so, please review your manuscript and approve your PDF to complete your submission at /.Username: ****Password: ****Thank you for your time and support.PLoS ONE Staff/*************************************************************** ****************按照网站的要求一项项填好后，上传文件，最后生成了PDF，给我发的邮件。

plos one的参考文献格式

文章标题：PLOS ONE的参考文献格式解析与应用今天我们要讨论的主题是PLOS ONE的参考文献格式。

PLOS ONE是一本开放获取的科学期刊，它要求作者按照特定的参考文献格式来引用已发表的文章，这一点对于保证学术交流的准确性和完整性非常重要。

在本文中，我们将从PLOS ONE的参考文献格式要求开始，逐渐深入探讨其详细规定和实际应用，帮助读者对这一主题有更全面的了解。

1. PLOS ONE关于参考文献格式的要求PLOS ONE要求作者在撰写稿件时按照特定的参考文献格式来引用已发表的文献。

根据PLOS ONE的官方指南，参考文献应按照文中的引用顺序列出，每个引用必须包括作者尊称、文章标题、出版年份、期刊名称、卷号、页码等信息，并使用数字标记。

PLOS ONE还要求在文中标出引用位置并将其标注为上标，以便读者能够方便地找到引用的原始来源。

2. PLOS ONE参考文献格式的详细规定在PLOS ONE的官方指南中，详细规定了不同类型文献的引用格式，包括期刊文章、书籍、章节、网络资源等。

不同类型文献的引用格式略有不同，但都需要包括作者尊称、文章标题、出版年份、期刊名称、卷号、页码等基本信息。

对于网络资源，PLOS ONE还规定了需要包括URL信息和访问日期等信息。

了解这些详细规定对于撰写PLOS ONE稿件的作者至关重要，只有严格按照规定格式列出参考文献，稿件才能通过编辑部的审稿。

3. PLOS ONE参考文献格式的实际应用在实际应用中，撰写PLOS ONE稿件的作者需要仔细核对每一条引用的格式，确保符合官方指南的规定。

特别是对于期刊文章、书籍、网络资源等不同类型的文献，撰写时需要采用不同的格式进行引用，并标注清楚引用位置。

在撰写过程中，作者还需要留意到引用的完整性和准确性，确保每一条引用都能够准确地指向原文。

只有这样，读者才能够便捷地查阅原始文献，理解作者的观点和论据，从而对研究进行深入了解。

结语PLOS ONE的参考文献格式对于保证学术交流的准确性和完整性有着重要的作用。

zotero plosone文献引用格式

一、概述近年来，学术研究对于文献引用格式的要求日益严格，其中Zotero和PLOS ONE是两大广泛应用的文献管理工具，在学术论文的引用中发挥着重要的作用。

正确使用Zotero和PLOS ONE的文献引用格式对于撰写一篇高质量的学术论文至关重要。

本文将就Zotero和PLOS ONE的文献引用格式进行详细介绍和说明，以期对广大学术研究人员有所帮助。

二、Zotero的文献引用格式1. Zotero是一款免费的文献管理软件，能够帮助研究人员方便地管理、整理和引用文献资料。

在使用Zotero进行文献引用时，可以根据所需的引文格式进行设置，包括APA、MLA、Chicago等多种引文格式。

2. 以APA格式为例，当需要引用一篇期刊文章时，可以在Zotero中直接搜索相关的期刊名称或DOI号，然后选择需要引用的文章，点击“Cite”按钮即可生成符合APA格式的引文。

3. 当需要在学术论文中插入引文时，只需在Word文档中选择插入引文位置，然后点击Zotero插件中的“插入引文”按钮，选择相应的文献项即可自动生成文献引用。

4. 需要注意的是，Zotero中的文献引用格式可以根据具体的要求进行自定义设置，确保所生成的引文符合学术期刊或出版社的要求。

三、PLOS ONE的文献引用格式1. PLOS ONE是一家广泛认可的开放获取期刊，其文献引用格式也具有一定的特点。

在PLOS ONE中发表文章时，需要根据其冠方要求使用特定的引文格式。

2. 在PLOS ONE中，文献引用格式一般遵循APA或者PLOS ONE自己的引文格式要求。

在提交论文时，作者需要按照PLOS ONE的格式要求来准备文献引用部分，以确保论文符合期刊的要求。

3. 在PLOS ONE的冠方全球信息站上，也会提供具体的引文格式范例和模板供作者参考，以确保引文格式的准确性和规范性。

四、Zotero和PLOS ONE文献引用格式的结合应用1. 在学术研究中，研究人员通常会使用Zotero来管理和整理文献资料，同时在投稿PLOS ONE等开放获取期刊时，需要按照期刊要求的引文格式来准备文献引用部分。

plos one 伦理应对方法

在科学研究领域，伦理问题一直是一个备受关注的议题。

《PLOS ONE》作为知名的开放获取科学期刊，其伦理应对方法备受关注。

下面将从多个方面分析《PLOS ONE》在伦理应对方面的具体做法。

一、伦理审查在提交论文之前，《PLOS ONE》要求作者必须遵守一定的伦理标准，并进行伦理审查。

对于人体试验和动物试验，作者需要提供相关伦理委员会批准的文件。

这一做法可以有效地保护人和动物的权益，保证研究的道德合法性。

也有利于维护期刊的声誉和学术道德。

二、数据真实性《PLOS ONE》要求作者必须对其研究数据的真实性负责。

任何有关数据的篡改、造假都是不被允许的。

如果有研究者有违规行为，一经发现，将会受到惩处，甚至被开除。

这种严格的数据真实性要求，保证了研究的可信度和学术诚信。

三、作者署名和贡献《PLOS ONE》在论文发表过程中，要求作者必须确保其研究工作的原创性，并对其贡献进行明确署名。

并且，对于引用的其他人的工作，也需要进行恰当的引用和致谢。

这种做法有利于减少学术不端行为，维护学术界的公平竞争环境。

四、披露利益冲突《PLOS ONE》要求作者在论文提交时必须披露任何可能影响其研究结果和结论的潜在利益冲突。

这有利于提高研究结果的客观性和可信度，避免利益驱动的研究行为。

五、审稿人和编辑的道德准则在论文的审稿和编辑过程中，《PLOS ONE》要求审稿人和编辑必须严格遵守道德准则，保护作者的权益，维护审稿的公平和公正。

一旦发现有违反伦理的行为，将会被取消资格，这为保证研究成果的科学性和公正性提供了保障。

六、道德培训和教育《PLOS ONE》为了提高研究人员和编辑的伦理意识，定期组织伦理培训和教育活动，鼓励研究人员和编辑遵守伦理规范，避免学术不端行为的发生。

这种做法有利于加强研究人员和编辑的道德责任感，推动学术的健康发展。

《PLOS ONE》在伦理应对方面采取了一系列切实可行的措施，保证了研究的伦理合法性和学术诚信。

plosone正文投稿样板

plosone正⽂投稿样板Abstractinterdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc.Materials and MethodsVestibulum adipiscing urna ut lectus gravida, vitae (Fig 1) interdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc. Mauris in mattis sapien. Fusce sodales vulputate auctor. Nam sit amet nulla lacus a, Figs 1 and 2 ultrices tellus. Integer rutrum aliquet sapien, eu fermentum magna pellentesque vitae.Fig 1. This is the Fig 1 Title. This is the Fig 1 legend. Fig 2. This is the Fig 2 Title. This is the Fig 2 legend.interdum. Donec 2pet 2q2221p pq q ++semper viverra mauris vel pulvinar dolor sit amet en 2()1p q +pulvinar et alst.Whole genome RFLP analysisResults and DiscussionLorem ipsum dolor sit amet, consectetur adipiscing elit. Vestibulum adipiscing urna ut lectus gravida, et bland Table 1 Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convalli. Fido nemo.Table 1. This is the Table 1 Title.ChemicalW ChemicalX ChemicalY ChemicalZ Chemical 1 Reaction 1W Reaction 1X Reaction 1Y Reaction 1Z Chemical 2 Reaction 2W Reaction 2X Reaction 2Y Reaction 2Z Chemical 3 Reaction 3W a Reaction 3X Reaction 3Y b Reaction 3Z Chemical 4 Reaction 4W Reaction 4X Reaction 4Y Reaction 4Z Chemical 5Reaction 5WReaction 5XReaction 5YReaction 5ZThis is the Table 1 legend. aTable footnotes belong here. bFootnotes should have corresponding symbols in the table.AcknowledgmentsLorem ipsum dolor sit amet, consectetur adipiscing elit. Vestibulum adipiscing urna ut lectus gravida, vitae blandit tortor interdum.References1. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr,McBio GHT, et al. This is the article title. PLoS One 2014 Dec 18; 9(12).2. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr, McBio GHT, et al. Bunny dynamics in cartoon landscapes. PLOS One. Forthcoming 2015.Supporting InformationS1 Fig. This is the S1 Fig Title. This is the S1 Fig legend. S2 Fig. This is the S2 Fig Title. This is the S2 Fig legend. S1 Table. This is the S1 Table Title. This is the S1 Table legend. S2 Table. This is the S2 Table Title. This is the S2 Table legend.。

[专题]PLOSONE期刊发表影响因子

Agricultural Sciences in Chinaimpact factor文章送给2－3个审稿人审阅，under review说明已送审了，而wait for other review说明已有审稿人返回审稿的意见(reviewer comments),但还有审稿人未送回，要等待大家对Plos One的影响因子如何预测？这个期刊的审稿政策决定了论文的平均水平不高。

但同时他们还是发表了少数高水平的论文。

通常一个期刊的影响因子主要取决于引用率高的前20%左右的论文的引用次数，后面大多数贡献不大，所以我的评估应该在1以上，但不会太高，应该小于3.同时，SCI收录与否也没把握。

现在是ISI Web of Knowledge 收录了，但ISI Web of Science没有,后者才包含SCIE.我之所以设置最后一个选项，是担心Plos One和LNCS一样的下场。

PLoS One现在还未收录到SCI中，因此今年ISI可能不会统计它的2008年影响因子了。

不过，有人早已利用Google Scholar对PLoS One在2007年发表的文章进行了引用统计，结果计算出其IF=5.68。

虽然他们没有统计该杂志2006年文章的运用情况，但PLoS One在2006才刊登137篇论文（而2007年为1 229篇），所以对统计结果影响不大。

就算2006年全部文章引用为零（这是不可能的），PLoS One的IF 也达到5.11。

所以说，2008年PLoS One的影响因子应该在5以上见过PLOS one 上有很好的文章，质量也不一定比PLOS系列中其他的期刊的低。

另外老外投稿时和我们对期刊的看法不一样。

我个人觉得PLOS one的引用会比一般的杂志高，但是SCI暂时不会收录，如果收录的话，也是等审稿策略改变以后，要知道，Google scholar有很多重复，去掉了吗。

PLoS One民间统计的影响因子出自这里：http://lampreylinux.wo ... -of-plos-one/另外还有一些关于PLoS One的讨论：http://www.sciencenet. ... spx?id=220746/news/?p=17看看plos one网页上的这句话，PLoS ONE will rigorously peer-review your submissions and publish all papers that are judged to be technically sound. Judgments about the importance of any partic ular paper are then made after publication by the readership (who are the most qualified to dete rmine what is of interest to them). 即使plos one发表一些高水平论文，总的影响因子也不会太高。

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Abstract
interdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc.
Materials and Methods
Vestibulum adipiscing urna ut lectus gravida, vitae (Fig. 1) interdum. Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convallis nunc. Mauris in mattis sapien. Fusce sodales vulputate auctor. Nam sit amet nulla lacus a, Figs. 1 and 2 ultrices tellus. Integer rutrum aliquet sapien, eu fermentum magna pellentesque vitae.
Fig. 1. This is the Fig. 1 Title. This is the Fig. 1 legend. Fig. 2. This is the Fig. 2 Title. This is the Fig. 2 legend.
Lorem
interdum. Donec 2
p
et 2q
22
21p pq q ++semper viverra mauris vel pulvinar dolor sit amet en 2()1p q +
pulvinar et alst.
Whole genome RFLP analysis
Lorem ipsum dolor
Results and Discussion
Lorem
ipsum dolor sit amet, consectetur adipiscing elit.
Vestibulum adipiscing urna ut lectus gravida, et bland Table 1
Donec tincidunt porta sem nec hendrerit. Vestibulum nec pharetra quam, vitae convalli. Fido nemo.
Table 1. This is the Table 1 Title.
Chemical
W Chemical
X Chemical
Y Chemical
Z Chemical 1 Reaction 1W Reaction 1X Reaction 1Y Reaction 1Z Chemical 2 Reaction 2W Reaction 2X Reaction 2Y Reaction 2Z Chemical 3 Reaction 3W a Reaction 3X Reaction 3Y b Reaction 3Z Chemical 4 Reaction 4W Reaction 4X Reaction 4Y Reaction 4Z Chemical 5
Reaction 5W
Reaction 5X
Reaction 5Y
Reaction 5Z
This is the Table 1 legend.
a
Table footnotes belong here. b
Table footnotes belong here.
Acknowledgments
Lorem ipsum dolor sit amet, consectetur adipiscing elit. Vestibulum adipiscing urna ut lectus gravida, vitae blandit tortor interdum.
References
1. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr,
McBio GHT, et al. (2014) This is the article title. PLOS ONE 0(0): e0.
2. Doe J, Data A, van Stats J, Testperson M, Ribosome D Jr, McBio, GHT et al. (In Press) Bunny dynamics in cartoon landscapes. PLOS ONE
Supporting Information
S1 Fig. This is the S1 Fig. Title. This is the S1 Fig. legend. S2 Fig. This is the S2 Fig. Title. This is the S2 Fig. legend. S1 Table. This is the S1 Table Title. This is the S1 Table legend. S2 Table. This is the S2 Table Title. This is the S2 Table legend.。

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