Reviewer’s report
Title: Uric acid induced hepatocytes lipid accumulation through regulation of miR-149-5p/FGF21
axis
Version: 0 Date: 06 Nov 2019
Reviewer: Reviewer 2
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses  a testable research
question(s) (brief or other article types: is there a clear objective)?
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for the
objective?
No - there are major issues
EXECUTION - Are the experiments and analyses performed with technical rigor to allow confidence
in the results?
No - there are major issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
No - there are issues with the statistics in the study
INTERPRETATION  -  Is  the  current  interpretation/discussion  of  the  results  reasonable  and  not
overstated?
No - there are major issues
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically sound? If
not, can revisions be made to make the work technically sound?
Maybe - with major revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: 
The proposed study provides insight view and novel strategy potential applicable into the clinical
setting for the treatment of NAFLD and hyperuricemia in particular. The study is well-described but
important limitations are related to the sole use of cell lines.
REQUESTED REVISIONS: 
The proposed study is interesting but requiring additional refinements and supportive data to reach
(clinically) relevant conclusions. Both mouse (AML-12) and human (HepG2) cell lines have been
tested, while more accurate and clinically relevant results would be collected by the use of primary
(human and murine) hepatocytes. The technique to isolate primary liver cells is quite well-established,
and cryopreserved primary cells are commercially available. Variability, in particular in human samples
need to be considered during such analysis and it would potentially provide a much better insight view
and mechanistic outcome. Regarding the in vivo experiments: Interestingly, the authors did not find
important to analysis or comment difference between control group (SCD) and the allopurinol-treated
group. Moreover, Introduction section is not covering the allopurinol use and the  reasons to design the
study around such pharmacological treatment only. Large parts  of the Results section is actually
introductive or commentary to the results. Conclusions and comments on the provided results should be
avoided in the Results sections and only address in the final part of the manuscript. miRNA profile and
analysis required supportive description and motivation for the cut-off and statistical analysis. Once
again, comparison between ctrl and HFD+A group is required and motivated. Transfection experiments
in primary liver cells would be the better choice and implemented to give strength to the study. Cell
line may be used a preliminary approach to set up technique or only when primary cells have been
proved inefficiently in such approach. Long-term expression/effects should also be monitored. Finally,
statistical analysis should be motivated and reconsidered. Histological images need to be uploaded to
higher resolutions to be evaluated
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
No
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
No
Are you able to assess any statistics in the manuscript or would you recommend an
additional statistical review?
If an additional statistical review is recommended, please specify what aspects require further
assessment in your comments to the editors.
I am able to assess the statistics
Quality of written English
Please indicate the quality of language in the manuscript:
Acceptable
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below.
If your reply is yes to any, please give details below.
This reviewer has been recruited by a partner organization, Research Square. Reviewers with declared
or  apparent  competing  interests  are  not  utilized  for  these  reviews.  This  reviewer  has  agreed  to
publication of their comments online under a Creative Commons Attribution License attributed to
Research Square and was paid a small honorarium for completing the review  within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer recommendation.
I agree to the open peer review policy of the journal. I understand that my name will be included
on my report to the authors and, if the manuscript is accepted for publication, my named report
including any attachments I upload will be posted on the website along with the authors'
responses. I agree for my report to be made available under an Open Access Creative Commons
CC-BY license (http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments
which I do not wish to be included in my named report can be included as confidential comments
to the editors, which will not be published.
I agree to the open peer review policy of the journal
Reviewer’s report
Title: Uric acid induced hepatocytes lipid accumulation through regulation of miR-149-5p/FGF21
axis
Version: 0 Date: 06 Nov 2019
Reviewer: Reviewer 2
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses  a testable research
question(s) (brief or other article types: is there a clear objective)?
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for the
objective?
No - there are major issues
EXECUTION - Are the experiments and analyses performed with technical rigor to allow confidence
in the results?
No - there are major issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
No - there are issues with the statistics in the study
INTERPRETATION  -  Is  the  current  interpretation/discussion  of  the  results  reasonable  and  not
overstated?
No - there are major issues
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically sound? If
not, can revisions be made to make the work technically sound?
Maybe - with major revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: 
The proposed study provides insight view and novel strategy potential applicable into the clinical
setting for the treatment of NAFLD and hyperuricemia in particular. The study is well-described but
important limitations are related to the sole use of cell lines.
REQUESTED REVISIONS: 
The proposed study is interesting but requiring additional refinements and supportive data to reach
(clinically) relevant conclusions. Both mouse (AML-12) and human (HepG2) cell lines have been
tested, while more accurate and clinically relevant results would be collected by the use of primary
(human and murine) hepatocytes. The technique to isolate primary liver cells is quite well-established,
and cryopreserved primary cells are commercially available. Variability, in particular in human samples
need to be considered during such analysis and it would potentially provide a much better insight view
and mechanistic outcome. Regarding the in vivo experiments: Interestingly, the authors did not find
important to analysis or comment difference between control group (SCD) and the allopurinol-treated
group. Moreover, Introduction section is not covering the allopurinol use and the  reasons to design the
study around such pharmacological treatment only. Large parts  of the Results section is actually
introductive or commentary to the results. Conclusions and comments on the provided results should be
avoided in the Results sections and only address in the final part of the manuscript. miRNA profile and
analysis required supportive description and motivation for the cut-off and statistical analysis. Once
again, comparison between ctrl and HFD+A group is required and motivated. Transfection experiments
in primary liver cells would be the better choice and implemented to give strength to the study. Cell
line may be used a preliminary approach to set up technique or only when primary cells have been
proved inefficiently in such approach. Long-term expression/effects should also be monitored. Finally,
statistical analysis should be motivated and reconsidered. Histological images need to be uploaded to
higher resolutions to be evaluated
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
No
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
No
Are you able to assess any statistics in the manuscript or would you recommend an
additional statistical review?
If an additional statistical review is recommended, please specify what aspects require further
assessment in your comments to the editors.
I am able to assess the statistics
Quality of written English
Please indicate the quality of language in the manuscript:
Acceptable
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below.
If your reply is yes to any, please give details below.
This reviewer has been recruited by a partner organization, Research Square. Reviewers with declared
or  apparent  competing  interests  are  not  utilized  for  these  reviews.  This  reviewer  has  agreed  to
publication of their comments online under a Creative Commons Attribution License attributed to
Research Square and was paid a small honorarium for completing the review  within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer recommendation.
I agree to the open peer review policy of the journal. I understand that my name will be included
on my report to the authors and, if the manuscript is accepted for publication, my named report
including any attachments I upload will be posted on the website along with the authors'
responses. I agree for my report to be made available under an Open Access Creative Commons
CC-BY license (http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments
which I do not wish to be included in my named report can be included as confidential comments
to the editors, which will not be published.
I agree to the open peer review policy of the journal
Please review this paper.
user
Reviewer’s report
Title: Uric acid induced hepatocytes lipid accumulation through regulation of miR-149-5p/FGF21
axis
Version: 0 Date: 06 Nov 2019
Reviewer: Reviewer 2
Reviewer's report:
PEER REVIEWER ASSESSMENTS: 
OBJECTIVE - Full research articles: is there a clear objective that addresses  a testable research
question(s) (brief or other article types: is there a clear objective)?
Yes - there is a clear objective
DESIGN - Is the current approach (including controls and analysis protocols) appropriate for the
objective?
No - there are major issues
EXECUTION - Are the experiments and analyses performed with technical rigor to allow confidence
in the results?
No - there are major issues
STATISTICS - Is the use of statistics in the manuscript appropriate?
No - there are issues with the statistics in the study
INTERPRETATION  -  Is  the  current  interpretation/discussion  of  the  results  reasonable  and  not
overstated?
No - there are major issues
OVERALL MANUSCRIPT POTENTIAL - Is the current version of this work technically sound? If
not, can revisions be made to make the work technically sound?
Maybe - with major revisions
PEER REVIEWER COMMENTS: 
GENERAL COMMENTS: 
The proposed study provides insight view and novel strategy potential applicable into the clinical
setting for the treatment of NAFLD and hyperuricemia in particular. The study is well-described but
important limitations are related to the sole use of cell lines.
REQUESTED REVISIONS: 
The proposed study is interesting but requiring additional refinements and supportive data to reach
(clinically) relevant conclusions. Both mouse (AML-12) and human (HepG2) cell lines have been
tested, while more accurate and clinically relevant results would be collected by the use of primary
(human and murine) hepatocytes. The technique to isolate primary liver cells is quite well-established,
and cryopreserved primary cells are commercially available. Variability, in particular in human samples
need to be considered during such analysis and it would potentially provide a much better insight view
and mechanistic outcome. Regarding the in vivo experiments: Interestingly, the authors did not find
important to analysis or comment difference between control group (SCD) and the allopurinol-treated
group. Moreover, Introduction section is not covering the allopurinol use and the  reasons to design the
study around such pharmacological treatment only. Large parts  of the Results section is actually
introductive or commentary to the results. Conclusions and comments on the provided results should be
avoided in the Results sections and only address in the final part of the manuscript. miRNA profile and
analysis required supportive description and motivation for the cut-off and statistical analysis. Once
again, comparison between ctrl and HFD+A group is required and motivated. Transfection experiments
in primary liver cells would be the better choice and implemented to give strength to the study. Cell
line may be used a preliminary approach to set up technique or only when primary cells have been
proved inefficiently in such approach. Long-term expression/effects should also be monitored. Finally,
statistical analysis should be motivated and reconsidered. Histological images need to be uploaded to
higher resolutions to be evaluated
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
No
Does the work include the necessary controls?
If not, please specify which controls are required in your comments to the authors.
No
Are the conclusions drawn adequately supported by the data shown?
If not, please explain in your comments to the authors.
No
Are you able to assess any statistics in the manuscript or would you recommend an
additional statistical review?
If an additional statistical review is recommended, please specify what aspects require further
assessment in your comments to the editors.
I am able to assess the statistics
Quality of written English
Please indicate the quality of language in the manuscript:
Acceptable
Declaration of competing interests
Please complete a declaration of competing interests, considering the following questions:
1. Have you in the past five years received reimbursements, fees, funding, or salary from an
organisation that may in any way gain or lose financially from the publication of this
manuscript, either now or in the future?
2. Do you hold any stocks or shares in an organisation that may in any way gain or lose
financially from the publication of this manuscript, either now or in the future?
3. Do you hold or are you currently applying for any patents relating to the content of the
manuscript?
4. Have you received reimbursements, fees, funding, or salary from an organization that holds
or has applied for patents relating to the content of the manuscript?
5. Do you have any other financial competing interests?
6. Do you have any non-financial competing interests in relation to this paper?
If you can answer no to all of the above, write 'I declare that I have no competing interests' below.
If your reply is yes to any, please give details below.
This reviewer has been recruited by a partner organization, Research Square. Reviewers with declared
or  apparent  competing  interests  are  not  utilized  for  these  reviews.  This  reviewer  has  agreed  to
publication of their comments online under a Creative Commons Attribution License attributed to
Research Square and was paid a small honorarium for completing the review  within a specified
timeframe. Honoraria for reviews such as this are paid regardless of the reviewer recommendation.
I agree to the open peer review policy of the journal. I understand that my name will be included
on my report to the authors and, if the manuscript is accepted for publication, my named report
including any attachments I upload will be posted on the website along with the authors'
responses. I agree for my report to be made available under an Open Access Creative Commons
CC-BY license (http://creativecommons.org/licenses/by/4.0/ ). I understand that any comments
which I do not wish to be included in my named report can be included as confidential comments
to the editors, which will not be published.
I agree to the open peer review policy of the journal
assistant