Coptidis alkaloids extracted from Coptis chinensis Franch attenuate IFN-γ-induced destruction of bone marrow cells

Jinyu Li; Xiaoying Meng; Changzhi Wang; Huijie Zhang; Hening Chen; Peiying Deng; Juan Liu; Meiyier Huandike; Jie Wei; Limin Chai

doi:10.1371/journal.pone.0236433

Peer Review History

Original SubmissionMarch 4, 2020
1 May 2020 Decision Letter - Xing-Ming Shi, Editor PONE-D-20-06039 The Coptidis alkaloids extracted from Coptis chinensis Franch attenuates the IFN-γ-induced immune destruction in bone marrow cells PLOS ONE Dear Dr. Chai, Thank you for submitting your manuscript to PLOS ONE. After careful consideration, we feel that it has merit but does not fully meet PLOS ONE’s publication criteria as it currently stands. Therefore, we invite you to submit a revised version of the manuscript that addresses the points raised during the review process. In addition to address the concerns raised by the reviewers, please have the English polished so that readers can better understand the study. We would appreciate receiving your revised manuscript by Jun 15 2020 11:59PM. When you are ready to submit your revision, log on to https://www.editorialmanager.com/pone/ and select the 'Submissions Needing Revision' folder to locate your manuscript file. 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PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. 2, In the Methods, please provide the precise part of Coptis chinensis Franch used in this work (roots, stems, leaves, etc). - We note that Coptis chinensis Franch was obtained from Beijing Xidan Pharmaceutical Co, Ltd, China. As such, in the Methods, please provide the product number and lot number. - We noticed minor instances of text overlap with the following previous publications from your group, which need to be addressed: https://bmccomplementmedtherapies.biomedcentral.com/articles/10.1186/s12906-019-2781-4 https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0180417 In your revision please ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed. 3. Thank you for including the following funding information within your acknowledgements; "This work was supported by the State Administration of Traditional Chinese Medicine, National Chinese Medicine Clinical Research Base, Research special [grant number: JDZX2015191]." We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." [Note: HTML markup is below. Please do not edit.] Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ******** 2. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 3. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 4. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: This is a well-written study in which the effect of coptidis alkaloids on IFN-gamma induced destruction in mice bone marrow cells. All experiments and analyses are performed on high technical standards; data are adequately presented and discussed. I have only two minor points: 1. What are bone marrow cells? Which type of cells are present there? This should be shown by Authors, because several cell types could mediate the effect of alkaloids and IFN-gamma. Authors used ANOVA statistics, but how they proved the normal distribution of data, which is a pre-requisite for this statistical method? Reviewer #2: Dear Limin Chai, M.D. Thank you for your manuscript submission. Your manuscript titled “The Coptidis alkaloids extracted from Coptis chinensis Franch attenuates the IFN-γinduced immune destruction in bone marrow cells ” has been reviewed and here we put forward some suggestions so that you can make essential modifications before you submit it next time. 1)Some typos or grammatical errors should be corrected. (e.g. The fifth line of the Cell treatment part, ‘...... cells were harvest’. The first line of Discussion part, ‘Coptis chinensis are known for its antibacterial ......’.) We would like you to read your whole manuscript again to make sure there are no mistakes like these. 2)We would like you to provide more reliable information in figure 5 about your immunofluorescence test. Pictures of DAPI staining should be provided to confirm the total number and the location of cells in each visual field. 3)We would like you to provide the all the pictures of the whole membranes of your WB tests (including protein markers) in the supplementary figure to ensure the results are reliable. 4)We suggest that animal experiments in vivo should be added to further demonstrate your point of view. For example, IHC or HE staining might be applied to analyze changes of the IFN-γ-induced bone marrow tissues after being treated with coptis or not in different time. These issues have to be dealt with before this manuscript get published. We hope our suggestions help you to modify your article to address these issues. Yours sincerely Dr Huang ****** 6. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No [NOTE: If reviewer comments were submitted as an attachment file, they will be attached to this email and accessible via the submission site. Please log into your account, locate the manuscript record, and check for the action link "View Attachments". If this link does not appear, there are no attachment files to be viewed.] While revising your submission, please upload your figure files to the Preflight Analysis and Conversion Engine (PACE) digital diagnostic tool, https://pacev2.apexcovantage.com/. PACE helps ensure that figures meet PLOS requirements. To use PACE, you must first register as a user. Registration is free. Then, login and navigate to the UPLOAD tab, where you will find detailed instructions on how to use the tool. If you encounter any issues or have any questions when using PACE, please email us at figures@plos.org. Please note that Supporting Information files do not need this step. https://doi.org/10.1371/journal.pone.0236433.r001
Revision 1
23 Jun 2020 Author Response Journal Requirements: 1. PLOS ONE now requires that authors provide the original uncropped and unadjusted images underlying all blot or gel results reported in a submission’s figures or Supporting Information files. This policy and the journal’s other requirements for blot/gel reporting and figure preparation are described in detail at https://journals.plos.org/plosone/s/figures#loc-blot-and-gel-reporting-requirements and https://journals.plos.org/plosone/s/figures#loc-preparing-figures-from-image-files. When you submit your revised manuscript, please ensure that your figures adhere fully to these guidelines and provide the original underlying images for all blot or gel data reported in your submission. See the following link for instructions on providing the original image data: https://journals.plos.org/plosone/s/figures#loc-original-images-for-blots-and-gels. In your cover letter, please note whether your blot/gel image data are in Supporting Information or posted at a public data repository, provide the repository URL if relevant, and provide specific details as to which raw blot/gel images, if any, are not available. Email us at plosone@plos.org if you have any questions. Response: We have provided the original uncropped gel image in the uploaded files. 2, In the Methods, please provide the precise part of Coptis chinensis Franch used in this work (roots, stems, leaves, etc). - We note that Coptis chinensis Franch was obtained from Beijing Xidan Pharmaceutical Co, Ltd, China. As such, in the Methods, please provide the product number and lot number. - We noticed minor instances of text overlap with the following previous publications from your group, which need to be addressed: https://bmccomplementmedtherapies.biomedcentral.com/articles/10.1186/s12906-019-2781-4 https://journals.plos.org/plosone/article?id=10.1371%2Fjournal.pone.0180417 In your revision please ensure you cite all your sources (including your own works), and quote or rephrase any duplicated text outside the methods section. Further consideration is dependent on these concerns being addressed. Response: Thanks for this comment. We have provided the precise part lot number of Coptis chinensis Franch in the Methods of revised manuscript. We also have removed and re-described the relevant contents which overlap with our previously published works in the revised manuscript. 3. Thank you for including the following funding information within your acknowledgements; "This work was supported by the State Administration of Traditional Chinese Medicine, National Chinese Medicine Clinical Research Base, Research special [grant number: JDZX2015191]." We note that you have provided funding information that is not currently declared in your Funding Statement. However, funding information should not appear in the Acknowledgments section or other areas of your manuscript. We will only publish funding information present in the Funding Statement section of the online submission form. Please remove any funding-related text from the manuscript and let us know how you would like to update your Funding Statement. Currently, your Funding Statement reads as follows: "The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript." Response: We have removed the funding-related text and Funding Statement in the revised manuscript. This work was supported by Research special for National Chinese Medicine Clinical Research Base of State Administration of Traditional Chinese Medicine of the People’s Republic of China (Grant number: JDZX2015191, http://www.satcm.gov.cn/). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. We have uploaded the information of funding on the revised files. Reviewers' comments: Reviewer #1: This is a well-written study in which the effect of coptidis alkaloids on IFN-gamma induced destruction in mice bone marrow cells. All experiments and analyses are performed on high technical standards; data are adequately presented and discussed. I have only two minor points: 1. What are bone marrow cells? Which type of cells are present there? This should be shown by Authors, because several cell types could mediate the effect of alkaloids and IFN-gamma. Response: We obtained bone marrow cells from the tibias, femurs and humeris of BALB/c mice. We focused on the apoptotic BMCs and the proliferation and differentiation of helper T cells and regulatory T cells induced by IFN-γ in the bone marrow microenvironment treated by alkaloids extracted from Coptis chinensis Franch. From the above consideration, we did not separate and identify the cellular constituent of bone marrow cells in this study. 2. Authors used ANOVA statistics, but how they proved the normal distribution of data, which is a pre-requisite for this statistical method? Response: Thank you for this comment. We used the Shapiro-Wilk test method for data normality testing. We have added instructions about data normality testing in the statistical method section of the revised manuscript. Reviewer #2: Your manuscript titled “The Coptidis alkaloids extracted from Coptis chinensis Franch attenuates the IFN-γinduced immune destruction in bone marrow cells ” has been reviewed and here we put forward some suggestions so that you can make essential modifications before you submit it next time. 1. Some typos or grammatical errors should be corrected. (e.g. The fifth line of the Cell treatment part, ‘...... cells were harvest’. The first line of Discussion part, ‘Coptis chinensis are known for its antibacterial ......’.) We would like you to read your whole manuscript again to make sure there are no mistakes like these. Response: According to the reviewer’s comment, we have corrected the grammatical errors and the unclear or awkwardly worded sentences. And then, the manuscript has been edited by American Journal experts. 2. We would like you to provide more reliable information in figure 5 about your immunofluorescence test. Pictures of DAPI staining should be provided to confirm the total number and the location of cells in each visual field. Response: Thank you for comment on this important concern. According to your comment, we added the pictures obtained in bright field corresponding to the immunofluorescence pictures in Figure 5. The pictures in bright field also could confirm the total number and the location of cells in the visual field. Because of the coVID-19 outbreak, we cannot supplement the relevant experiments of DAPI staining. We hope that this supplement will meet with approval. 3. We would like you to provide the all the pictures of the whole membranes of your WB tests (including protein markers) in the supplementary figure to ensure the results are reliable. Response: We have provided the original uncropped gel image in the uploaded files. We used a pre-dyed marker in the WB tests. The marker could not be displayed on the films. For this reason, we provided the original uncropped gel image without protein markers. 4. We suggest that animal experiments in vivo should be added to further demonstrate your point of view. For example, IHC or HE staining might be applied to analyze changes of the IFN-γ-induced bone marrow tissues after being treated with coptis or not in different time. Response: Thank you for this comment. The animal experiments for modified Danggui Buxue Tang (Radix astragali, Radix Angelicae sinensis, and Coptis chinensis Franch) have been done in our previous work. The experimental results have been published in our papers (1 Jingwei Zhou, Xue Li, Peiying Deng, et al. Chinese Herbal Formula, modified Danggui Buxue Tang, attenuates apoptosis of hematopoietic stem cells in immune mediate aplastic anemia mouse model, Journal of Immunology Research, 2017, 2017(2017): 9786972; 2 Peiying Deng, Xue Li,Yi Wei, et al. The herbal decoction modified Danggui Buxue Tang attenuates immune-mediated bone marrow failure by regulating the differentiation of T lymphocytes in an immune-induced aplastic anemia mouse model, PLOS ONE, 2017, 12(7): e0180417). In this study, we focus on the exact mechanism of Coptis chinensis Franch for regulating the apoptosis of BMCs and the proliferation and differentiation of helper T cells and regulatory T cells induced by IFN-γ in the bone marrow microenvironment in vitro. https://doi.org/10.1371/journal.pone.0236433.r002
8 Jul 2020 Decision Letter - Xing-Ming Shi, Editor Coptidis alkaloids extracted from Coptis chinensis Franch attenuate IFN-γ-induced destruction of bone marrow cells PONE-D-20-06039R1 Dear Dr. Chai, We’re pleased to inform you that your manuscript has been judged scientifically suitable for publication and will be formally accepted for publication once it meets all outstanding technical requirements. Within one week, you’ll receive an e-mail detailing the required amendments. When these have been addressed, you’ll receive a formal acceptance letter and your manuscript will be scheduled for publication. An invoice for payment will follow shortly after the formal acceptance. To ensure an efficient process, please log into Editorial Manager at http://www.editorialmanager.com/pone/, click the 'Update My Information' link at the top of the page, and double check that your user information is up-to-date. If you have any billing related questions, please contact our Author Billing department directly at authorbilling@plos.org. If your institution or institutions have a press office, please notify them about your upcoming paper to help maximize its impact. If they’ll be preparing press materials, please inform our press team as soon as possible -- no later than 48 hours after receiving the formal acceptance. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information, please contact onepress@plos.org. Kind regards, Xing-Ming Shi, Ph.D Academic Editor PLOS ONE Additional Editor Comments (optional): Reviewers' comments: Reviewer's Responses to Questions Comments to the Author 1. If the authors have adequately addressed your comments raised in a previous round of review and you feel that this manuscript is now acceptable for publication, you may indicate that here to bypass the “Comments to the Author” section, enter your conflict of interest statement in the “Confidential to Editor” section, and submit your "Accept" recommendation. Reviewer #1: All comments have been addressed Reviewer #2: (No Response) ******** 2. Is the manuscript technically sound, and do the data support the conclusions? The manuscript must describe a technically sound piece of scientific research with data that supports the conclusions. Experiments must have been conducted rigorously, with appropriate controls, replication, and sample sizes. The conclusions must be drawn appropriately based on the data presented. Reviewer #1: Yes Reviewer #2: Yes ****** 3. Has the statistical analysis been performed appropriately and rigorously? Reviewer #1: Yes Reviewer #2: Yes ****** 4. Have the authors made all data underlying the findings in their manuscript fully available? The PLOS Data policy requires authors to make all data underlying the findings described in their manuscript fully available without restriction, with rare exception (please refer to the Data Availability Statement in the manuscript PDF file). The data should be provided as part of the manuscript or its supporting information, or deposited to a public repository. For example, in addition to summary statistics, the data points behind means, medians and variance measures should be available. If there are restrictions on publicly sharing data—e.g. participant privacy or use of data from a third party—those must be specified. Reviewer #1: Yes Reviewer #2: Yes ****** 5. Is the manuscript presented in an intelligible fashion and written in standard English? PLOS ONE does not copyedit accepted manuscripts, so the language in submitted articles must be clear, correct, and unambiguous. Any typographical or grammatical errors should be corrected at revision, so please note any specific errors here. Reviewer #1: Yes Reviewer #2: Yes ****** 6. Review Comments to the Author Please use the space provided to explain your answers to the questions above. You may also include additional comments for the author, including concerns about dual publication, research ethics, or publication ethics. (Please upload your review as an attachment if it exceeds 20,000 characters) Reviewer #1: (No Response) Reviewer #2: We are so glad that you and your team made some modification to your manuscript according to the comments. We believe that your manuscript can now be accepted and published in the journal of PLOS ONE ****** 7. PLOS authors have the option to publish the peer review history of their article (what does this mean?). If published, this will include your full peer review and any attached files. If you choose “no”, your identity will remain anonymous but your review may still be made public. Do you want your identity to be public for this peer review?** For information about this choice, including consent withdrawal, please see our Privacy Policy. Reviewer #1: No Reviewer #2: No https://doi.org/10.1371/journal.pone.0236433.r003
Formally Accepted
13 Jul 2020 Acceptance Letter - Xing-Ming Shi, Editor PONE-D-20-06039R1 Coptidis alkaloids extracted from Coptis chinensis Franch attenuate IFN-γ-induced destruction of bone marrow cells Dear Dr. Chai: I'm pleased to inform you that your manuscript has been deemed suitable for publication in PLOS ONE. Congratulations! Your manuscript is now with our production department. If your institution or institutions have a press office, please let them know about your upcoming paper now to help maximize its impact. If they'll be preparing press materials, please inform our press team within the next 48 hours. Your manuscript will remain under strict press embargo until 2 pm Eastern Time on the date of publication. For more information please contact onepress@plos.org. If we can help with anything else, please email us at plosone@plos.org. Thank you for submitting your work to PLOS ONE and supporting open access. Kind regards, PLOS ONE Editorial Office Staff on behalf of Dr Xing-Ming Shi Academic Editor PLOS ONE https://doi.org/10.1371/journal.pone.0236433.r004

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