Title: A new model of portal vein thrombosis in rats with cirrhosis induced by partial portal
vein ligation plus carbon tetrachloride and intervened with rivaroxaban
Version: 2 Date: 04 Mar 2024
Reviewer:  Dinesh Mani Tripathi
Reviewer's report:
The present study entitled “Model establishment and pathological mechanism of portal vein
thrombosis in rats with cirrhosis” by Yanan Guo, et.al. Authors Developed and characterized
and animal model of PVT and explained the possible mechanism, based on result they claimed
this model is suitable further translational research in field of vascular complication of cirrhosis.
The research area is very interesting and need scientific attention for better understanding of
many unexplained mechanism and some extend with limited therapeutic options.
There are a number of concerns related to this manuscript namely:
Major Comments:
1. In the context of this PPVL-CCL4 model, the overall survival analysis was not done hence
the mortality due PPVL alone upto 10 weeks remains unaddressed, as partial portal vein ligation
constitutes a model of portal vein obstruction and pre-hepatic portal hypertension, leading to
mortality in animals.
2. Notably, the second insult introduction of further CCL4, a model of cirrhosis and intrahepatic
portal hypertension, to animals already afflicted with portal hypertension can cause mortality.
Emphasizing the mortality rate in this scenario would have been crucial, author may provide
some evidence or can explain in manuscript.
3. Neither sham animal in comparison to PPVL was included in the study Nor any vehicle group
for CCl4 administration was included in entire study.
4. Surprisingly, all the data was compared with only control animals there is no PPVL animals
of different time points i.e 4,6,8,10 weeks. Can author explain how they will address the
pathological changes due to PPVL alone in such a long period of time? Seems they have
postulated there will be no changes due to partial ligation of portal vein after 7 days.
5. Author(s) have also not disclosed how they compared PBFV and Portal Trunk diameter
without any CCL4 Cirrhotic group alone?
6. Cirrhosis itself can lead to intrahepatic portal vein thrombosis, contrasting with the models
of partial portal vein ligation (PPVL) combined with CCl4 induction. Strong recommendation
to add per se group for each time point.
7. What was the reason behind increasing the standard dose of carbon tetrachloride for creating
a model of chronic liver disease from 1ml/kg to 2ml/kg, along with administering 40% of its
total volume?
8. The peak velocity graph suggests a decrease in velocity with the progression of cirrhosis.
This trend would typically imply a reduction in the diameter of the portal vein. However, the
accompanying graph depicting portal vein diameter shows no such change, thus presenting a
contradiction between the two sets of data. Can author(s) explain ?
9. In ,Table 1 8weeks Data, Author(s) please specify why there is no noticeable changes were
found in the levels of d-dimer and FIB/AT III between the control group and the cirrhotic animal
group at the eighth week. How its possible? Please explain in discussion on the basis of strong
evidence. Same for D-Dimer and (ug/ml) AT III (%) values.
10. Number of animal per group in varies at different time points, why?
11. Western blot showing Fibrin and PAI-1 protein levels representation should be n=2 or 3
per group.
12. Rats model with rivaroxaban for 6 weeks from the fifth week to the tenth week. Author(s)
can show the results of rivaroxaban treatment at  6th and 8th weeks also.
Title: A new model of portal vein thrombosis in rats with cirrhosis induced by partial portal
vein ligation plus carbon tetrachloride and intervened with rivaroxaban
Version: 2 Date: 04 Mar 2024
Reviewer:  Dinesh Mani Tripathi
Reviewer's report:
The present study entitled “Model establishment and pathological mechanism of portal vein
thrombosis in rats with cirrhosis” by Yanan Guo, et.al. Authors Developed and characterized
and animal model of PVT and explained the possible mechanism, based on result they claimed
this model is suitable further translational research in field of vascular complication of cirrhosis.
The research area is very interesting and need scientific attention for better understanding of
many unexplained mechanism and some extend with limited therapeutic options.
There are a number of concerns related to this manuscript namely:
Major Comments:
1. In the context of this PPVL-CCL4 model, the overall survival analysis was not done hence
the mortality due PPVL alone upto 10 weeks remains unaddressed, as partial portal vein ligation
constitutes a model of portal vein obstruction and pre-hepatic portal hypertension, leading to
mortality in animals.
2. Notably, the second insult introduction of further CCL4, a model of cirrhosis and intrahepatic
portal hypertension, to animals already afflicted with portal hypertension can cause mortality.
Emphasizing the mortality rate in this scenario would have been crucial, author may provide
some evidence or can explain in manuscript.
3. Neither sham animal in comparison to PPVL was included in the study Nor any vehicle group
for CCl4 administration was included in entire study.
4. Surprisingly, all the data was compared with only control animals there is no PPVL animals
of different time points i.e 4,6,8,10 weeks. Can author explain how they will address the
pathological changes due to PPVL alone in such a long period of time? Seems they have
postulated there will be no changes due to partial ligation of portal vein after 7 days.
5. Author(s) have also not disclosed how they compared PBFV and Portal Trunk diameter
without any CCL4 Cirrhotic group alone?
6. Cirrhosis itself can lead to intrahepatic portal vein thrombosis, contrasting with the models
of partial portal vein ligation (PPVL) combined with CCl4 induction. Strong recommendation
to add per se group for each time point.
7. What was the reason behind increasing the standard dose of carbon tetrachloride for creating
a model of chronic liver disease from 1ml/kg to 2ml/kg, along with administering 40% of its
total volume?
8. The peak velocity graph suggests a decrease in velocity with the progression of cirrhosis.
This trend would typically imply a reduction in the diameter of the portal vein. However, the
accompanying graph depicting portal vein diameter shows no such change, thus presenting a
contradiction between the two sets of data. Can author(s) explain ?
9. In ,Table 1 8weeks Data, Author(s) please specify why there is no noticeable changes were
found in the levels of d-dimer and FIB/AT III between the control group and the cirrhotic animal
group at the eighth week. How its possible? Please explain in discussion on the basis of strong
evidence. Same for D-Dimer and (ug/ml) AT III (%) values.
10. Number of animal per group in varies at different time points, why?
11. Western blot showing Fibrin and PAI-1 protein levels representation should be n=2 or 3
per group.
12. Rats model with rivaroxaban for 6 weeks from the fifth week to the tenth week. Author(s)
can show the results of rivaroxaban treatment at  6th and 8th weeks also.
Please review this paper.
user
Title: A new model of portal vein thrombosis in rats with cirrhosis induced by partial portal
vein ligation plus carbon tetrachloride and intervened with rivaroxaban
Version: 2 Date: 04 Mar 2024
Reviewer:  Dinesh Mani Tripathi
Reviewer's report:
The present study entitled “Model establishment and pathological mechanism of portal vein
thrombosis in rats with cirrhosis” by Yanan Guo, et.al. Authors Developed and characterized
and animal model of PVT and explained the possible mechanism, based on result they claimed
this model is suitable further translational research in field of vascular complication of cirrhosis.
The research area is very interesting and need scientific attention for better understanding of
many unexplained mechanism and some extend with limited therapeutic options.
There are a number of concerns related to this manuscript namely:
Major Comments:
1. In the context of this PPVL-CCL4 model, the overall survival analysis was not done hence
the mortality due PPVL alone upto 10 weeks remains unaddressed, as partial portal vein ligation
constitutes a model of portal vein obstruction and pre-hepatic portal hypertension, leading to
mortality in animals.
2. Notably, the second insult introduction of further CCL4, a model of cirrhosis and intrahepatic
portal hypertension, to animals already afflicted with portal hypertension can cause mortality.
Emphasizing the mortality rate in this scenario would have been crucial, author may provide
some evidence or can explain in manuscript.
3. Neither sham animal in comparison to PPVL was included in the study Nor any vehicle group
for CCl4 administration was included in entire study.
4. Surprisingly, all the data was compared with only control animals there is no PPVL animals
of different time points i.e 4,6,8,10 weeks. Can author explain how they will address the
pathological changes due to PPVL alone in such a long period of time? Seems they have
postulated there will be no changes due to partial ligation of portal vein after 7 days.
5. Author(s) have also not disclosed how they compared PBFV and Portal Trunk diameter
without any CCL4 Cirrhotic group alone?
6. Cirrhosis itself can lead to intrahepatic portal vein thrombosis, contrasting with the models
of partial portal vein ligation (PPVL) combined with CCl4 induction. Strong recommendation
to add per se group for each time point.
7. What was the reason behind increasing the standard dose of carbon tetrachloride for creating
a model of chronic liver disease from 1ml/kg to 2ml/kg, along with administering 40% of its
total volume?
8. The peak velocity graph suggests a decrease in velocity with the progression of cirrhosis.
This trend would typically imply a reduction in the diameter of the portal vein. However, the
accompanying graph depicting portal vein diameter shows no such change, thus presenting a
contradiction between the two sets of data. Can author(s) explain ?
9. In ,Table 1 8weeks Data, Author(s) please specify why there is no noticeable changes were
found in the levels of d-dimer and FIB/AT III between the control group and the cirrhotic animal
group at the eighth week. How its possible? Please explain in discussion on the basis of strong
evidence. Same for D-Dimer and (ug/ml) AT III (%) values.
10. Number of animal per group in varies at different time points, why?
11. Western blot showing Fibrin and PAI-1 protein levels representation should be n=2 or 3
per group.
12. Rats model with rivaroxaban for 6 weeks from the fifth week to the tenth week. Author(s)
can show the results of rivaroxaban treatment at  6th and 8th weeks also.
assistant