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The authors improved the manuscript after the review and can now be accepted.
[# PeerJ Staff Note - this decision was reviewed and approved by Vladimir Uversky, a PeerJ Section Editor covering this Section #]
Dear Authors
I appreciate your efforts in improving the article. However, some aspects still need to be clarified:
a) how was the sample size calculated? What method was used to determine the sample size?
b) the raw data spreadsheet is a little confusing. My impression is that the sample size varies in the same spreadsheet (see for example the spreadsheet Fig. 3 eNOS "). The raw data from Figure 7 is missing. Please review the whole spreadsheet.
[# PeerJ Staff Note: Please note that this revision has only been reviewed by the Academic Editor at this point. Once you have addressed their comments, it might still need to be sent out for peer review. #]
Your manuscript "Therapeutic effect of and mechanisms underlying the effect of miR-195-5p on subarachnoid hemorrhage-induced vasospasm and brain injury in rats” has been reviewed by the two experts. I append their reviews below. I have raised specifics issues about the description of the materials/methods and statistics.
I, therefore, invite you to revise and resubmit your manuscript, considering the points below. Let me reinforce some of the reviewer’s comments and add some more:
1. Please describe how was the sample size calculated?
2. Statistical analyses: the authors should include the values from the appropriate statistical test (e.g., F(x,x) = xx; n = x; P = x.xxx).
3. Considering that small RNAs are unstable and peripheral delivery requires carriers to prevent degradation before achieving access to the brain, how can the authors claim that these compounds are accessing the central nervous system? It seems a limitation of this study.
4. The article should be reviewed by someone fluent in the English language.
[# PeerJ Staff Note: Please ensure that all review comments are addressed in a rebuttal letter and any edits or clarifications mentioned in the letter are also inserted into the revised manuscript where appropriate. It is a common mistake to address reviewer questions in the rebuttal letter but not in the revised manuscript. If a reviewer raised a question then your readers will probably have the same question so you should ensure that the manuscript can stand alone without the rebuttal letter. Directions on how to prepare a rebuttal letter can be found at: https://peerj.com/benefits/academic-rebuttal-letters/ #]
[# PeerJ Staff Note: The Academic Editor has identified that the English language must be improved. PeerJ can provide language editing services - please contact us at [email protected] for pricing (be sure to provide your manuscript number and title) #]
The manuscript should be edited by a native speaker of English. Additionally, the raw data of immunobolt was missing.
Methods failed to offer sufficient information to be reproducible by another investigator.
The raw data of immunobolt was missing.
In the current study, Tsai et al investigated the role of miR-195-5p in the pathological process following subarachnoid hemorrhage (SAH). And they concluded that administration of miR-195-5p alleviated SAH-induced vasospasm and brain injury. It’s an interesting study. However, several issues must be addressed.
1. Method. How did the author choose the delivey route and dose of miR-195-5p in this study?
2. Method. Had the author determined the concentration of miR-195-5p in brain?
3. Method. Did the authors perfuse rats with formalin before the basilar artery morphometric analysis?
4. Method. How many animals were used for each test, including Basilar Artery Morphometric Analysis, Cell Death Assay, immunoblot, and TNF-α evaluation? They wrote that 96 animals for each test, which was quite complexing.
5. Method. As mentioned in Statistical Analysis, the difference among the groups were analyzed by ANOVA. How were the data analyzed to ensure they were normally distributed, a requirement of ANOVA?
6. Data points should be indicated on all statistical graphs. Providing the bar graph and only statistical parameters can suggest that the data underlying any particular bar are normally distributed and contains no outliers, when this may not be the case.
7. Generally, microRNA selience specific gene through binding to the DNA fragments. How could the miR-195-5p modulate the level of eNOS and iNOS, as well as regulate the phosphorylation p-65 and IκBα?
This study aimed to determine the effects of miR-195-5p on an experimental model of SAH in male rats. It was unclear why authors selected this specific miRNA for studies. No clear rationales or justifications were provided except listing a couple of studies showing beneficial effects of miR-195 mainly in cancers. No prior studies that associate SAH with miR-195 were provided to support the scientific premise of the study.
The miR-195 and control were in suspensions delivered in tail vein injections. It is questionable how this method could ensure BBB penetrability and exhibit efficacy on brain changes as authors assumed. The small RNAs are unstable and peripheral delivery requires carriers to prevent degradation before getting access into the brain. No validation studies were provided about whether miR195 was efficiently getting into the brain and increasing the levels in the brain. With this questionable method for studies, the validity of findings is questionable.
see above. The validity of the findings is questionable.
n/a
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