Reviewer ’s report  
 
Title: Role of serotonin on the intestinal mucosal immune response to stress -induced diarrhea in  
weaning mice  
 
Version: 0 Date: 28 Apr 2016  
 
Reviewer: Syeda Afroze  
 
Reviewer's report:  
 
The manuscript entitled "Role of serotonin on the intestinal mucosal immune response to stress -
induced diarrhea in weaning mice" by Dong et al, the authors aimed to demonstrate the effect of 
serotonin (which is also known as 5 -hydroxytryptamine (5 -HT)) on the development of diarrhea. 
To explore this study, the authors performed stress induced diarrh ea in weaning mice model. 
Three groups of weaning mice were used to perform the experiments in this study. Those mice 
were (a) control group, (b)), stress induced diarrhea group (by performing intragastric 
administration of folium sennae) and (c) the group  pretreated with para -chlorophenylalanine 
(PCPA) following stress -induced diarrhea group. To determine the role of 5 -HT in stress induced 
diarrhea, the authors performed measurement of different biochemical markers in plasma such as 
blood glucose level, co rtisol level and and 5 -HT level as well in three groups of weaning mice. 
The authors also measured the levels of IEL, B lymphocytes and slgA in different part of small 
intestine and ileum of those three groups of mice. The authors also measured T cell mark ers (T 
lymphocytes proliferation, CD3, CD4 etc) as well as cytokines (such as TNF -α, IL -2, IL -4, and 
IL-10) in the small intestine and ileum of those three different groups of mice. This work is very 
interesting and well done. This resubmitted manuscript i s well described and well written. The 
authors nicely addressed each concerns made by the reviewer. Just the following minor concerns 
are needed to be addressed.  
 
 
 
 
 
Minor Correction:  
 
 
1. In the methods section, there is a sentence," The first group of mice w as subjected to stress -
induced diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration at 2:00 PM after intraperitoneal injection of sterile saline at 
10:00AM.The second group was the PCPA+stress -induced diarrhea group, which was treated by 
intraperitoneal injection of sterile saline containing PCPA (30mg/mL, 300mg/kg body weight) at 
10:00AM followed by the intragastric administration of folium sennae (0.4g/mL, 15 mL/Kg 
body weight) at 2:00 PM"  in he re no need to mention the time.  
 
 
It is better to write the sentence like as "The first group of mice was subjected to stress -induced 
diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration 4 hours af ter intraperitoneal injection of sterilesaline. Similarly, the 
second group was the PCPA+stress -induced diarrhea group, which was treated by intraperitoneal 
injection of sterile saline containing PCPA (30mg/mL,300mg/kg body weight) followed by the 
intragas tric administration of folium sennae (0.4g/mL, 15 mL/Kg body weight)."  
 
 
 
 
 
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.  
 
Yes 
 
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 t he 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 share s 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 inter ests 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  
 
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 au thors'  
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 incl uded 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: Role of serotonin on the intestinal mucosal immune response to stress -induced diarrhea in  
weaning mice  
 
Version: 0 Date: 28 Apr 2016  
 
Reviewer: Syeda Afroze  
 
Reviewer's report:  
 
The manuscript entitled "Role of serotonin on the intestinal mucosal immune response to stress -
induced diarrhea in weaning mice" by Dong et al, the authors aimed to demonstrate the effect of 
serotonin (which is also known as 5 -hydroxytryptamine (5 -HT)) on the development of diarrhea. 
To explore this study, the authors performed stress induced diarrh ea in weaning mice model. 
Three groups of weaning mice were used to perform the experiments in this study. Those mice 
were (a) control group, (b)), stress induced diarrhea group (by performing intragastric 
administration of folium sennae) and (c) the group  pretreated with para -chlorophenylalanine 
(PCPA) following stress -induced diarrhea group. To determine the role of 5 -HT in stress induced 
diarrhea, the authors performed measurement of different biochemical markers in plasma such as 
blood glucose level, co rtisol level and and 5 -HT level as well in three groups of weaning mice. 
The authors also measured the levels of IEL, B lymphocytes and slgA in different part of small 
intestine and ileum of those three groups of mice. The authors also measured T cell mark ers (T 
lymphocytes proliferation, CD3, CD4 etc) as well as cytokines (such as TNF -α, IL -2, IL -4, and 
IL-10) in the small intestine and ileum of those three different groups of mice. This work is very 
interesting and well done. This resubmitted manuscript i s well described and well written. The 
authors nicely addressed each concerns made by the reviewer. Just the following minor concerns 
are needed to be addressed.  
 
 
 
 
 
Minor Correction:  
 
 
1. In the methods section, there is a sentence," The first group of mice w as subjected to stress -
induced diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration at 2:00 PM after intraperitoneal injection of sterile saline at 
10:00AM.The second group was the PCPA+stress -induced diarrhea group, which was treated by 
intraperitoneal injection of sterile saline containing PCPA (30mg/mL, 300mg/kg body weight) at 
10:00AM followed by the intragastric administration of folium sennae (0.4g/mL, 15 mL/Kg 
body weight) at 2:00 PM"  in he re no need to mention the time.  
 
 
It is better to write the sentence like as "The first group of mice was subjected to stress -induced 
diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration 4 hours af ter intraperitoneal injection of sterilesaline. Similarly, the 
second group was the PCPA+stress -induced diarrhea group, which was treated by intraperitoneal 
injection of sterile saline containing PCPA (30mg/mL,300mg/kg body weight) followed by the 
intragas tric administration of folium sennae (0.4g/mL, 15 mL/Kg body weight)."  
 
 
 
 
 
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.  
 
Yes 
 
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 t he 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 share s 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 inter ests 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  
 
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 au thors'  
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 incl uded 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: Role of serotonin on the intestinal mucosal immune response to stress -induced diarrhea in  
weaning mice  
 
Version: 0 Date: 28 Apr 2016  
 
Reviewer: Syeda Afroze  
 
Reviewer's report:  
 
The manuscript entitled "Role of serotonin on the intestinal mucosal immune response to stress -
induced diarrhea in weaning mice" by Dong et al, the authors aimed to demonstrate the effect of 
serotonin (which is also known as 5 -hydroxytryptamine (5 -HT)) on the development of diarrhea. 
To explore this study, the authors performed stress induced diarrh ea in weaning mice model. 
Three groups of weaning mice were used to perform the experiments in this study. Those mice 
were (a) control group, (b)), stress induced diarrhea group (by performing intragastric 
administration of folium sennae) and (c) the group  pretreated with para -chlorophenylalanine 
(PCPA) following stress -induced diarrhea group. To determine the role of 5 -HT in stress induced 
diarrhea, the authors performed measurement of different biochemical markers in plasma such as 
blood glucose level, co rtisol level and and 5 -HT level as well in three groups of weaning mice. 
The authors also measured the levels of IEL, B lymphocytes and slgA in different part of small 
intestine and ileum of those three groups of mice. The authors also measured T cell mark ers (T 
lymphocytes proliferation, CD3, CD4 etc) as well as cytokines (such as TNF -α, IL -2, IL -4, and 
IL-10) in the small intestine and ileum of those three different groups of mice. This work is very 
interesting and well done. This resubmitted manuscript i s well described and well written. The 
authors nicely addressed each concerns made by the reviewer. Just the following minor concerns 
are needed to be addressed.  
 
 
 
 
 
Minor Correction:  
 
 
1. In the methods section, there is a sentence," The first group of mice w as subjected to stress -
induced diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration at 2:00 PM after intraperitoneal injection of sterile saline at 
10:00AM.The second group was the PCPA+stress -induced diarrhea group, which was treated by 
intraperitoneal injection of sterile saline containing PCPA (30mg/mL, 300mg/kg body weight) at 
10:00AM followed by the intragastric administration of folium sennae (0.4g/mL, 15 mL/Kg 
body weight) at 2:00 PM"  in he re no need to mention the time.  
 
 
It is better to write the sentence like as "The first group of mice was subjected to stress -induced 
diarrhea by treating the mice with folium sennae (0.4g/mL, 15 mL/Kg body weight) via 
intragastric administration 4 hours af ter intraperitoneal injection of sterilesaline. Similarly, the 
second group was the PCPA+stress -induced diarrhea group, which was treated by intraperitoneal 
injection of sterile saline containing PCPA (30mg/mL,300mg/kg body weight) followed by the 
intragas tric administration of folium sennae (0.4g/mL, 15 mL/Kg body weight)."  
 
 
 
 
 
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.  
 
Yes 
 
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 t he 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 share s 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 inter ests 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  
 
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 au thors'  
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 incl uded as confidential comments  
to the editors, which will not be published.  
 
I agree to the open peer review policy of the journal  
assistant