Review for 10.1186/s12876-019-1041-x (2019)

Reviewer ’s report  
Title: A New Method to Induce Nonalcoholic Steatohepatitis (NASH) in Mice  
Version: 3 Date: 03 May 2019  
 
Reviewer: Reviewer 2  
Reviewer's report:  
PEER REVIEWER ASSESSMENTS:  
OBJECTIVE - Full research articles: is there a clear objective that a ddresses one or several 
testable research questions? (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 minor issues  
 
EXECUTION - Are the experiments and analyses performed with sufficient technical rigor to 
allow confidence in the results?  
Yes - experiments and analyses were performed appropriately  
 
STATISTICS - Is the use of statistics in the  manuscript appropriate?  
Yes - appropriate statistical analyses have been used in the study  
 
INTERPRETATION - Is the current interpretation/discussion of the results reasonable and not 
overstated?  
No - there are minor issues  
 
OVERALL MANUSCRIPT POTENTIAL - Has the author addressed your concerns sufficiently 
for you to now recommend the work as a technically sound contribution? If not, can further 
revisions be made to make the work technically sound?  
Probably - with minor revisions  
 
PEER REVIEWER COMMENTS:  
GENERAL COMMENTS: The authors performed (minimal) changes to the manuscript. Several 
addition/changes requested during R1 revision were not performed, and as authors clearly stated 
will not in the future. The manuscript should be considered as short commu nication since only 
histopathological observations have been made.  
 
REQUESTED REVISIONS:  
Unfortunately R1 version suffers from the same limitation as R0. In addition to histological 
analysis, biomolecular or proteomic measurements have been suggested. How ever, the authors 
clearly stated are not willing to perform any additional measurements/analysis. It has been 
claimed that the authors were not wise enough to save some tissue/sample for further analysis or 
confirmation. The study cannot be improved and sh ould be considered as it it: a brief 
communication with limited evidence or novel observation.  
 
ADDITIONAL REQUESTS/SUGGESTIONS:  
Differently of what stated, the authors did not apply the same amount of digits after the comma 
to all different values in Tabl e 1 and 2.  
Figure 1 is not mentioned anywhere in the text. In Figure 1 several SD bars are missing (with 
n=8 is sounds unlikely that SD is equal to 0). It's reviewer's opinion that bars with cigarette 
smoke exposure should be painted differently and maybe grouped together (since statistical 
comparison is related to C+CS control group). Whiskey plot is probably a better way to show 
results Probably the use of connecting line to better representing the comparison with its own 
relative control group (ctrl and C+CS) is probably better. Similar changes should be applied to 
Figure 3.  
Figure 2: panel labels are not present. In the legend, authors refer as (A), (B), etc…but such 
labelling has not been applied. It is not clear why Fru+CS condition has an additional panel 
(E)(is the magnitude the same as the other images?)  
The evaluation of a critical event in pathogenesis such as inflammatory probably deserves 
additional analysis to one cytokine only (TNFalpha) quantification. It looks like smoking 
condition increase s TNFalpha hepatic concentration of approx.25%. Is such increase consistent 
with the different diets? Please check comparison between smoking and non -smoking diets 
(ctrl/C+CS; Fru/Fru+CS is similar to HFD/HFD+CS or WD/WD+Cs)? Similar comparison to 
Figure 1  is largely suggested.  
Are the methods appropriate and well described?  
If not, please specify what is requi red 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, plea se 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 furth er 
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 competi ng 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 i n 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 competin g 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 
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