Reviewer’s report
Title: 
Genistein attenuated gastric inflammation and apoptosis in Helicobacter
pylori-induced gastropathy in rats
Version: 
2 
Date: 
01 May 2020
Reviewer's report:
This is an animal study aimed to investigate whether the effect of Genistein on
the gastropathy of H. pylori-infected rats. 
They found the NF-
κ
B expression and
apoptotic cells in the rats' gastric mucosa were significantly attenuated after
Genistein gavage. In addition, the degree of inflammatory cell infiltration and H.
pylori colonization were also attenuated by Genistein gavage. 
Please see below for my comments:
Major comments:
1. Although the author discuss the probable anti-bacterial effect of Genistein?
They could not rule out the possibility that "the gavage procedure eradicated H.
pylori". It is possible that the gavage procedure led to poor colonization of H.
pylori in the experimental group, not the Genistein. To clarify the issue, the
author may perform 0.1% DMSO gavage with same volume and same duration
as Genistein in the H. pylor-infected group. 
2. No symbols for statistical significance were denoted in Fig. 1 & Fig. 2. Please
add. 
3. In Fig. 4b & 4c, the TUNNEL stain overlapped with the nuclear stain, making
it difficult to differentiate which cells were stained positive. I suggest the author
to perform TUNNEL stain on another serial section of samples that are not
stained for nucleus. In other word, one section for nuclear stain, and another
serial section for TUNNEL stain. This can improve the calculation of apoptotic
cell numbers and percentages. Moreover, the readers will not be confused about
which cells are stained positive for TUNNEL.
4. Fig. 2 used bar-chart to show the percentages of NF-
κ
B positive and apoptotic
cells. While Fig. 3 & Fig. 4 showed the representative images of NF-
κ
B positive
and apoptotic cells. I suggest these 3 figures (Fig.2, 3 & 4) can be merged into
one Figure to enhance comparison between them, and improve understanding of
the data. 
5. In Fig. 5, the H. pylori are small and difficult to identify in the gastric mucosa.
I suggest the author can do anti-H pylori antibody immunostain or H pylori silver
staining to improve recognition of H. pylori. 
6. In Fig. 5, the gastric inflammation intensities were similar among the 3
groups. The authors should provide typical images to show both H. pylori
colonization and gastric inflammation intensities. 
Minor comments:
1. In result, paragraph 2, the sentence "The average percentages of gastric
epithelial cells in all the groups are shown in Figure 2a" is not clear. Should it be
"The average percentages of NF-
κ
B positive gastric epithelial cells in all the
groups are shown in Figure 2a"?
2. H. Pylori should be H. pylori in page 5, paragraph 3.
3. In many parts of the article, H.pylori should be H. pylori. 
4. In discussion, page 11, αβ1- integrins should be β1-integrin.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
Yes
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.
Yes
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.
I declare that I have no competing interests
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
Reviewer’s report
Title: 
Genistein attenuated gastric inflammation and apoptosis in Helicobacter
pylori-induced gastropathy in rats
Version: 
2 
Date: 
01 May 2020
Reviewer's report:
This is an animal study aimed to investigate whether the effect of Genistein on
the gastropathy of H. pylori-infected rats. 
They found the NF-
κ
B expression and
apoptotic cells in the rats' gastric mucosa were significantly attenuated after
Genistein gavage. In addition, the degree of inflammatory cell infiltration and H.
pylori colonization were also attenuated by Genistein gavage. 
Please see below for my comments:
Major comments:
1. Although the author discuss the probable anti-bacterial effect of Genistein?
They could not rule out the possibility that "the gavage procedure eradicated H.
pylori". It is possible that the gavage procedure led to poor colonization of H.
pylori in the experimental group, not the Genistein. To clarify the issue, the
author may perform 0.1% DMSO gavage with same volume and same duration
as Genistein in the H. pylor-infected group. 
2. No symbols for statistical significance were denoted in Fig. 1 & Fig. 2. Please
add. 
3. In Fig. 4b & 4c, the TUNNEL stain overlapped with the nuclear stain, making
it difficult to differentiate which cells were stained positive. I suggest the author
to perform TUNNEL stain on another serial section of samples that are not
stained for nucleus. In other word, one section for nuclear stain, and another
serial section for TUNNEL stain. This can improve the calculation of apoptotic
cell numbers and percentages. Moreover, the readers will not be confused about
which cells are stained positive for TUNNEL.
4. Fig. 2 used bar-chart to show the percentages of NF-
κ
B positive and apoptotic
cells. While Fig. 3 & Fig. 4 showed the representative images of NF-
κ
B positive
and apoptotic cells. I suggest these 3 figures (Fig.2, 3 & 4) can be merged into
one Figure to enhance comparison between them, and improve understanding of
the data. 
5. In Fig. 5, the H. pylori are small and difficult to identify in the gastric mucosa.
I suggest the author can do anti-H pylori antibody immunostain or H pylori silver
staining to improve recognition of H. pylori. 
6. In Fig. 5, the gastric inflammation intensities were similar among the 3
groups. The authors should provide typical images to show both H. pylori
colonization and gastric inflammation intensities. 
Minor comments:
1. In result, paragraph 2, the sentence "The average percentages of gastric
epithelial cells in all the groups are shown in Figure 2a" is not clear. Should it be
"The average percentages of NF-
κ
B positive gastric epithelial cells in all the
groups are shown in Figure 2a"?
2. H. Pylori should be H. pylori in page 5, paragraph 3.
3. In many parts of the article, H.pylori should be H. pylori. 
4. In discussion, page 11, αβ1- integrins should be β1-integrin.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
Yes
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.
Yes
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.
I declare that I have no competing interests
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
Please review this paper.
user
Reviewer’s report
Title: 
Genistein attenuated gastric inflammation and apoptosis in Helicobacter
pylori-induced gastropathy in rats
Version: 
2 
Date: 
01 May 2020
Reviewer's report:
This is an animal study aimed to investigate whether the effect of Genistein on
the gastropathy of H. pylori-infected rats. 
They found the NF-
κ
B expression and
apoptotic cells in the rats' gastric mucosa were significantly attenuated after
Genistein gavage. In addition, the degree of inflammatory cell infiltration and H.
pylori colonization were also attenuated by Genistein gavage. 
Please see below for my comments:
Major comments:
1. Although the author discuss the probable anti-bacterial effect of Genistein?
They could not rule out the possibility that "the gavage procedure eradicated H.
pylori". It is possible that the gavage procedure led to poor colonization of H.
pylori in the experimental group, not the Genistein. To clarify the issue, the
author may perform 0.1% DMSO gavage with same volume and same duration
as Genistein in the H. pylor-infected group. 
2. No symbols for statistical significance were denoted in Fig. 1 & Fig. 2. Please
add. 
3. In Fig. 4b & 4c, the TUNNEL stain overlapped with the nuclear stain, making
it difficult to differentiate which cells were stained positive. I suggest the author
to perform TUNNEL stain on another serial section of samples that are not
stained for nucleus. In other word, one section for nuclear stain, and another
serial section for TUNNEL stain. This can improve the calculation of apoptotic
cell numbers and percentages. Moreover, the readers will not be confused about
which cells are stained positive for TUNNEL.
4. Fig. 2 used bar-chart to show the percentages of NF-
κ
B positive and apoptotic
cells. While Fig. 3 & Fig. 4 showed the representative images of NF-
κ
B positive
and apoptotic cells. I suggest these 3 figures (Fig.2, 3 & 4) can be merged into
one Figure to enhance comparison between them, and improve understanding of
the data. 
5. In Fig. 5, the H. pylori are small and difficult to identify in the gastric mucosa.
I suggest the author can do anti-H pylori antibody immunostain or H pylori silver
staining to improve recognition of H. pylori. 
6. In Fig. 5, the gastric inflammation intensities were similar among the 3
groups. The authors should provide typical images to show both H. pylori
colonization and gastric inflammation intensities. 
Minor comments:
1. In result, paragraph 2, the sentence "The average percentages of gastric
epithelial cells in all the groups are shown in Figure 2a" is not clear. Should it be
"The average percentages of NF-
κ
B positive gastric epithelial cells in all the
groups are shown in Figure 2a"?
2. H. Pylori should be H. pylori in page 5, paragraph 3.
3. In many parts of the article, H.pylori should be H. pylori. 
4. In discussion, page 11, αβ1- integrins should be β1-integrin.
Are the methods appropriate and well described?
If not, please specify what is required in your comments to the authors.
Yes
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.
Yes
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.
I declare that I have no competing interests
Reviewer Publication Consent.
 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
) if this manuscript is accepted for
publication. Any comments that I do not wish to be included in the published
report have been included as confidential comments to the editor, which will not
be published.
If you are not happy for us to publish this report, please contact the editorial
office before completing the review.
If you wish, you can include your name in your published report. Please note
you must decide whether to include your name at the start of the process and
confirm this decision whenever prompted. If you change your decision later, we
will not be able to publish your name.
I agree to the terms of the CC-BY 4.0 license; please 
do not
 publish my name
with my report. (default)
assistant