Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: FLATFOOT IN AFRICA, THE CIRRIPEDE CHTHAMALUS IN THE WEST INDIAN OCEAN
Review round: 1
Reviewer: 2

Output:
Basic reporting: In general, this is a well-written manuscript. The figures are appropriate, especially the phylogenetic trees (Figs. 2 and 3) are well-designed. I suggest you add a map with sampling locations and the distribution of all chthamalid species for me and anybody else not familiar with the geographic area.
The english is sufficient. I suggest the authors check spelling before final submission.
However, I find references are missing in several instances, which I marked by “REF” in annotated PDF. I also suggest you include the following relevant literature:
	Wares, J. P. (2020). Small, flat, and gray: Cryptic diversity in chthamalid barnacles in the 	global context of marine coastal biogeography (Cirripedia: Balanomorpha: 	Chthamalidae). The Journal of Crustacean Biology, 40(1), 1-16.
Wares (2020) points out the gaps in chthamalid phylogeography, which you are filling with this MS

	Wares, J. P., Pankey, M. S., Pitombo, F., Daglio, L. G., & Achituv, Y. (2009). A “Shallow 	Phylogeny” of Shallow Barnacles (Chthamalus). PLOS ONE, 4(5), e5567.

	Chan, B. K. K., Corbari, L., Rodriguez Moreno, P. A., & Tsang, L. M. (2017). Molecular 	phylogeny of the lower acorn barnacle families (Bathylasmatidae, Chionelasmatidae, 	Pachylasmatidae and Waikalasmatidae) (Cirripedia: Balanomorpha) with evidence for 	revisions in family classification. Zoological Journal of the Linnean Society, 180(3), 542–	555. https://doi.org/10.1093/zoolinnean/zlw005

Wares et al. (2009) and Chan et al. (2017) reconstruct chthamaid phylogenies, and their results should be included into the phylogenetic discussion.

I have made some additional remarks directly into the pdf of the MS, which you should consider.
Experimental design: The experimental design is good. Sample sizes are adequate. The morphological analysis is excellent. The use of two genetic markers, one mitochondrial and one nuclear, is also recommendable.
Validity of the findings: The findings are overall valid. The phylogenetic discussion is, in my opinion, the weakest point. On the one hand, large scale phylogenetic reconstructions exist (Wares et al. 2009, Chan et al. 201). On the other hand, this discussion point seems out of place for this manuscript. I would focus on biogeographic or phylogeographic inferences that can be drawn from your data.
Additional comments: NA


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION OF A CIRCADIAN GENE SIGNATURE THAT PREDICTS OVERALL SURVIVAL IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: A few minor points mentioned in the 'General comments for the author'.
Experimental design: A few points mentioned in the 'General comments for the author' for improvement.
Validity of the findings: A few points mentioned in the 'General comments for the author' for improvement especially from 9 to 11.
Additional comments: In this manuscript, Gao et al. established a 5-circadian clock genes signature as a prognostic and predictive marker in TCGA and GSE72094 Lung adenocarcinoma (LUAD) datasets. Though the study is interesting and promising, the following points are recommended to be addressed to make this work more informative, confident, and reliable for the researchers and clinicians.

1. A couple of more lines in the background of the abstract should be mentioned. e.g., about the notoriousness of LUAD, what is circadian clock, and most importantly, the study's purpose.

 2. Elaborate line 89, "Count data were normalized" adding more details, how and why.

3. Line 120, one-tailed or two-tailed Student's t-test?

4. Line 139, ‘of the’ is redundant.

5. Why did the authors do PCA, t-SNE analysis, and LASSO Cox regression analysis? Details will help the researchers who do not have the bioinformatics and statistics background e.g., the authors can frame the sentence like this: in order to find….we used PCA and showed…

6. Usually, in these types of studies, more than one validation cohort are used. Here only GSE72094 as the external validation cohort is used. Why specifically this cohort, and why only one being used? The signature of just 5 genes sometimes can lead to false-positive results physiologically; hence validation in more than one cohort will make this signature more reliable for the prognosis.

7. The results about the risk factors from table 1 and figure 4 are not discussed. These results need the discussion and what did the authors conclude for them.

8. Figure legends are not very informative. Please expand them and add more information to make them self-explanatory.

9. For Fig. 1B, the representation of the KM plot for all the five genes separately will be more informative for TCGA and GSE72094.

10. The heat map in Fig. 1C and its legend lacks the proper label and description. Is it mRNA expression?

11. For Fig. 1C, the representation of the differential expression of these genes between the tumor and the normal tissues, along with the heat map, will be more intuitive if the authors could show them as the box-plot or the violin-plot with individual points. At the moment, we do not know if the difference is significant or not because there are no p-values.

12. If possible, please look into other cancer types if this signature is impacting survival. This analysis is not necessary but will be valuable.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION OF A CIRCADIAN GENE SIGNATURE THAT PREDICTS OVERALL SURVIVAL IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
Experimental design: The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical. Statistical methods are chosen correctly.
Validity of the findings: All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.
Additional comments: This paper investigates the function of expression of circadian genes in  overall survival(OS) of lung adenocarcinoma(LUAD) by analyzing TCGA datasets. The authors identified a novel circadian gene signature by LASSO-penalized Cox regression analysis. Moreover, the authors validate their findings in a GEO dataset. To explore further about the related pathways, the authors demonstrate that circadian genes are functionally related to cell proliferation, gene damage repair, proteasomes, and immune and autoimmune diseases utilizing the GSEA method.

Editorial Criteria
BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical. Statistical methods are chosen correctly.
VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.

Overall, I think this paper is novel and will be of interest to the community of lung cancer genetics, especially LUAD research. The statistical part is valid and makes sense. The authors make it comprehensive by integrating analysis of multiple sources including GEO and TCGA. The main strengths of this paper is that it addresses an interesting and unexplored question, finds a novel discovery based on a carefully selected set of bioinformatic procedures. As such this article represents an excellent and elegant bioinformatics study which will almost certainly influence our thinking about the function of circadian genes in LUAD. Some of the weaknesses are the lack of in vitro or in vivo validation experiments. In general, the work is convincing except some major and minor comments below:

Major Comments:

I do see multiple platforms and normalization methods for LUAD samples in GDC, which includes HTSeq - Counts (sample size: 594), HTSeq - FPKM(sample size: 594) and HTSeq - FPKM-UQ(sample size: 594). Please explain why the sample size in the manuscript is 515 rather than 594. Please also explain why “HTSeq - FPKM” was chosen for analysis, rather than the other two?

I’m wondering if there are any ongoing clinical trials focusing on circadian clock genes in LUAD or other types of lung cancer? It will be very strong evidence for the significance of the current study if so.

I’m just wondering if the differences in sequencing platforms affects the results, since the discovery dataset(TCGA) is RNA-seq, and the validation dataset(GEO) is microarray. I’m wondering if a RNA-seq LUAD dataset as a validation dataset will produce more significant results.

Minor Comments:
Line 74: please replace “were extracted from the publicly available TCGA database” with “were retrieved from the NCI Genomic Data Commons”.

Line 63:  please replace “using data extracted from The Cancer Genome Atlas (TCGA) database” with “using The Cancer Genome Atlas (TCGA) data obtained from the NCI Genomic Data Commons[add reference: https://www.nature.com/articles/s41588-021-00791-5 ]” .

It is great that a session of abbreviations was there to list all the abbreviations for the database names. I would recommend to also include abbreviations like LUAD, TCGA, TNM, GEO, DEG, LASSO, KEGG etc in that list.

All the gene names should be italic for all the gene names.

Figure 2E and 3E: the P value is not correctly formatted.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION OF A CIRCADIAN GENE SIGNATURE THAT PREDICTS OVERALL SURVIVAL IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: The authors addressed most of my points satisfactorily hence making this manuscript far better in every way. It can now be considered for publication!


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: IDENTIFICATION OF A CIRCADIAN GENE SIGNATURE THAT PREDICTS OVERALL SURVIVAL IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 2

Output:
Basic reporting: The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
Experimental design: The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical. Statistical methods are chosen correctly.
Validity of the findings: All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.
Additional comments: This paper investigates the function of expression of ferroptosis-related genes in  overall survival(OS) and prognosis of lung adenocarcinoma(LUAD) by analyzing TCGA datasets. The authors identified a ferroptosis-related gene signature by univariate Cox regression analysis. Moreover, the authors validate their findings in a GEO dataset. To explore further about the related pathways, the authors demonstrate that risk score was prominently enriched in ferroptosis processes. In short, this study identified the ferroptosis-related gene signature as a potential predictor for the prognosis of LUAD.

Overall, I think the corrections look good to me. All my comments have been answered in a proper manner. Please find the original comments and feedbacks below:

Editorial Criteria
BASIC REPORTING
The title, abstract, introduction, methods, results and discussion are appropriate for the content of the text. Furthermore, the article is well constructed, the experiments are well conducted, and analysis is well performed. The figures are relevant, high quality, well labelled and described.
EXPERIMENTAL DESIGN
The experimental design is original and the research is within the scope of the journal.. Research question is well defined, relevant and meaningful. The methods are highly technical, ethical and logistical. Statistical methods are chosen correctly.
VALIDITY OF THE FINDINGS
All underlying data have been provided in detail. The findings are meaningful. The conclusions are well stated and relevant to the research questions.

Overall, I think this paper is novel and will be of interest to the community of lung cancer genetics, especially LUAD research. The statistical part is valid and makes sense. The authors make it comprehensive by integrating analysis of multiple sources including GEO and TCGA. The main strengths of this paper is that it addresses an interesting and unexplored question, and finds a novel discovery based on a carefully selected set of bioinformatic procedures. As such this article represents an excellent and elegant bioinformatics study which will almost certainly influence our thinking about the function of ferroptosis-related genes in LUAD. Some of the weaknesses are the lack of in vitro or in vivo validation experiments. In general, the work is convincing except some major and minor comments below:

Major Comments:

The Methods of the Abstract are too simple, I would recommend expanding it and adding details like what datasets were used, what statistical methods were used to construct the prediction model, what pathway analysis methods were used. Some contents in the Results could be moved to the Methods.

Feedback: I did see the dataset and statistical methods were added in the method of the abstract. It looks good to me.

I do see multiple platforms and normalization methods for LUAD samples in GDC, which includes HTSeq - Counts (sample size: 594), HTSeq - FPKM(sample size: 594) and HTSeq - FPKM-UQ(sample size: 594). Please explain why the sample size in the manuscript is 528 rather than 594. Please also explain why “HTSeq - FPKM” was chosen for analysis, rather than the other two platforms?

Response:
There are 535 tumor samples (including 20 repetitive samples) and 59 normal tissue samples from 515 LUAD cases in TCGA. So there are only 515 cases with clinical data and 500 patients with OS data. We have described it in Line 79-83 and Line 136-145.

Feedback:
Thanks for adding the details. They look good.

I’m wondering if there are any ongoing clinical trials focusing on ferroptosis-related genes identified in this study in LUAD or other types of lung cancer? It will be very strong evidence for the significance of the current study if so.

Response:
We searched the website clinicaltrial.gov using “circadian” and “lung cancer”. There are only four ongoing or completed clinical trials. But all of them are about circadian rhythms and lung cancer. There is no clinical trial focusing on circadian clock genes in lung cancer.

Feedback:
Thanks for checking the clinical trials information. It’s good to know.

I’m just wondering if the differences in sequencing platforms affects the results, since the discovery dataset(TCGA) is RNA-seq, and the validation dataset(GEO) is microarray (GPL96 Affymetrix GeneChip Human Genome). I’m wondering if a RNA-seq LUAD dataset as a validation dataset will produce more significant results.

Response:
In this article, we use two independent data sets as the test set and the validation set. It is true that RNA-seq and microarray data are different from experimental methods, data acquisition, data normalization, and data standardization. That is why we did not combine the two datasets for analysis. Analyzing the two datasets separately avoids the bias caused by different experimental methods. And for LUAD, RNA-seq data with a large sample size cannot be found for verification in other databases.

Feedback:
Thanks for the explanation. It makes sense to me.

Minor Comments:
Line 63: : please replace “using data extracted from The Cancer Genome Atlas (TCGA) database” with “using The Cancer Genome Atlas (TCGA) data obtained from the NCI Genomic Data Commons” .https://www.nature.com/articles/s41588-021-00791-5
Answer: Agreed. Line 68.
Feedback: looks good now.

It is great that a session of abbreviations was there to list all the abbreviations for the database names. I would also recommend including abbreviations like LUAD, TCGA, GEO, DEG, GO, KEGG etc in that list.
Answer: It’s a good idea. If the editor needs it, we will list them.
Feedback: sounds good. Thanks!

All the gene names should be italic for all the gene names.
Answer: Revised.
Feedback: looks good. Thanks!

Figure 2E and 3E: the P value is not correctly formatted.
Answer: Revised.
Feedback: looks good. Thanks!


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A REVIEW OF CORAL BLEACHING SPECIMEN COLLECTION, PRESERVATION, AND LABORATORY PROCESSING METHODS
Review round: 1
Reviewer: 1

Output:
Basic reporting: This manuscript provides simple and elegant insight into how methodology varies in the field of coral bleaching research. I have limited criticism, mostly consisting of grammatical or typographical errors. The introduction is adequate, the justification for the study clearly identified.
Experimental design: The main point thatI think needs addressing is why the limited time frame was chosen, as the full dataset from 1981 surely would provide interesting temporal insight. Further, I recognise that this manuscript was likely developed at the end of 2020 and for that reason only a partial year is included, however since we are now in May 2021 it would be best if the authors could complete this final year’s dataset to increase comparability between years. On that note, it may be interesting to comment briefly on the temporal patterns of sample collection – i.e. is there evidence for increased sample collection coinciding with/following bleaching events?
Validity of the findings: Conclusions are well stated and well supported.
Additional comments: -	Line 153 – why was this particular 6.5 year window selected? Some justification is needed.
-	Line 155 – how did you define “shallow-water” species?
-	Line 176 – “included” should be “including”
-	Line 195 – I believe this should be Fig S1
-	Line 303 – “out” should be “our”
-	Line 341 – “who” should be “that”
-	Line 344 – “causes” should be “cause”
-	Line 357 – what specifically do you mean by tissue parameters?
-	Line 424 – I don’t think the snippet in brackets is necessary
-	Line 430 – missing second closed bracket after reference
-	Lines 433-434- colony size can even influence bleaching susceptibility https://link.springer.com/article/10.1007%2Fs00338-018-1677-y
-	This may be a personal preference, but references to figures / tables in the discussion is distracting to me. Perhaps these can be reduced.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: A REVIEW OF CORAL BLEACHING SPECIMEN COLLECTION, PRESERVATION, AND LABORATORY PROCESSING METHODS
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In ‘a review of coral bleaching specimen collection, preservation, and laboratory processing methods’ McLachlan review and discuss the common practices for coral collection and processing in coral bleaching research over the last 6.5 years. In general, the manuscript is well written and will appeal to the broad readership of Peer J. The authors have done well to synthesize the large volume of research in this field and present a nice and clear manuscript, well done. However, I feel what was ultimately lacking was a ‘best practices’ section to direct future research. Based on what we know about sample degradation (from other models if not corals) etc, what is the best approach for preservation and processing for multiple downstream analyses? Obviously, this depends on the questions and methods, but perhaps a diagram or flow chart that could walk researchers through best practices depending on the desired data? I think such a section would be extremely valuable to other researchers when planning field work and research.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANTI-INFLAMMATORY AND ANTIAGING PROPERTIES OF CHLOROGENIC ACID ON UV-INDUCED FIBROBLAST CELL
Review round: 1
Reviewer: 1

Output:
Basic reporting: This study is very innovative, and hypothesis directed. I feel they did most of the experiments to prove their hypothesis. I feel the following may be necessary to be included in the paper,
1.	UV could change the property of fibroblast, but how it is correlating with aging skin fibroblasts.
2.	Was the supernatant collected on the last day or everyday during CA treatment?
Experimental design: I think IL-1β and TNFα results should be in one figure instead of two.
Fig 3. and Fig. 5 should be in the supplementary section.
After removing the CA treatment, how long did the fibroblast show less production of IL-1β, TNFα and ROS?
Validity of the findings: I think this study is unique and all the points they covered in this study are crucial to show the effect of CA in a aging skin.
Additional comments: This study is well designed and shows the effect of natural product on anti-inflammatory responses in skin. I have suggested few concerns. The paper seems to shed light in the area of skin immunology.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANTI-INFLAMMATORY AND ANTIAGING PROPERTIES OF CHLOROGENIC ACID ON UV-INDUCED FIBROBLAST CELL
Review round: 1
Reviewer: 2

Output:
Basic reporting: Chlorogenic Acid is an interesting compound to study. It appears a lot of work has already been done on it. The perspective of using UV as an insult is interesting. However multiple concerns need to be addressed. The authors must address the language in the paper and make grammatical corrections all throughout the paper.
Experimental design: Only 1 cell line has been tested which is inadequate
Validity of the findings: This study is done in 1 cell line therefore replication of findings in at least 2 more cell lines is required to increase the impact of the outcome and validity of this study.
Additional comments: Overall, it is an interesting area. However, multiple issues need to be addressed to ensure better quality of the paper. My concerns are as follows

In introduction - Mention other external factors that affect skin aging. Add more background to the pathways being addressed

Line 36 – Why especially in women? Does skin aging happen only/more in women? Or is there is a reason why it is more of a problem in women than men?

Lines 50-51, add references for this statement- The inhibition of oxidative stress now becomes an important approach for treating skin damage due to aging.

Materials – Lines 65-69 – Why are cells treated with multiple antibacterial and antimycotic agents? What were the controls and were they in a separate incubator?
How was the 75-minute UV exposure decided? What kind of UV?
It is important to make sure the study is repeated in at least 2 more cell lines for the study to be acceptable.

Line 80 – What is meant by a normal control? Also, 1% DMSO is quite high, DMSO should be in 1:1000 ratio, any reason for 1% DMSO? Also, there should be another control group of DMSO with UV.

Line 85- What is mg protein?

Bar graphs are presented in a complicated manner and the significance is difficult to understand. Also, why are 2 methods of representation needed –pg/ml and pg/mgprotein?

Line 148 and 161- State what the significant difference to controls are.

Note: DMSO affects number of apoptotic cells, reinstating the importance of reducing amount of DMSO and using that as a control


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANTI-INFLAMMATORY AND ANTIAGING PROPERTIES OF CHLOROGENIC ACID ON UV-INDUCED FIBROBLAST CELL
Review round: 2
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No comment


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 1
Reviewer: 1

Output:
Basic reporting: The paper seems to contains interesting material.
The state-of-the-art discussion must be improved.
More explanations are needed in several parts of the work.
Experimental design: The paper seems technical sound but more explanation of the goal and methods are required.
Validity of the findings: The degree of novelty is not high however the paper seems technical sound and the results are interesting.
Additional comments: The paper seems to contains interesting material.  More explanations are needed in several parts of the work. See my comments  below.

- Please, write mathematically and properly the used ARIMA model, defining all the variables, distributions, constants etc. Clarifying what is known and what is unknown. Please, clarify the meaning of each variable which variable represents which physical amount etc.

- Please, explain better the contents of the Tables in the text and in their caption.

- Please, clarify the degree of novelty and the your contribution in the introduction in a better way.

- The state-of-the-art discussion must be improved. Please, clarify which method you use to make the inference as MCMC or Adaptive Importance Sampling (AIS) methods. Even if you do not use AIS or MCMC it is important to clarify that in a general Bayesian inference  they are needed. For instance, consider the important and relevant review papers

M. Bugallo et al, "Adaptive Importance Sampling in Signal Processing", Digital Signal Processing, Volume 47, Pages: 36-49, 2015,

L. Martino, "A Review of Multiple Try MCMC algorithms for Signal Processing", Digital Signal Processing, Volume 75, Pages: 134-152, 2018,

W.J. Fitzgerald, "Markov chain Monte Carlo methods with applications to signal processing", Signal Processing, Volume 81, Issue 1, Pages 3-18, 2001.

This discussion can improve the number of interested readers.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The paper was easy to read and I appreciated the brevity.

Code and data were available, I was easily able to reproduce the findings and figures. Good job.

## Major issues

Currently the paper is a bit hard to understand on its own as I don’t think BSTS are common enough model to be known to readers (I never heard of it myself before). Methods should include a brief description of the structure and basic assumptions of BSTS models and the specifics of the model you actually used (the components of the models, the number of seasons, the fact that you modelled each time series individually, …). A short argument why you expect the assumptions of the model to not be severely violated by the data could also be helpful.

Similarly, an overview of the assumptions and mechanism of CausalImpact should be mentioned.

You should probably also cite and discuss another modelling effort for Pakistan by Khan et al 2020: Modelling and forecasting of new cases, deaths and recover cases of COVID-19 by using Vector Autoregressive model in Pakistan https://doi.org/10.1016/j.chaos.2020.110189

## Minor issues

I find the Figure 1 too crowded - pure line plot would in my view be preferable.

The color palettes are in my view not very nice and also not accessible to people with impaired color perception - red-green contrasts are particularly hard to perceive for a lot of people. Consider using palettes from R packages viridis, colorbrewer or scico which have been assessed for accessibility (and in my view look better). Also use line type, not only colour to distinguish the individual series.

I would expect to have figures and tables embedded in text for submission version, I understand some journals still require this way of submitting, but I don’t think PeerJ does and it makes my job as a reviewer unnecessarily harder.

Minor linguistic suggestion can be found in the attached PDF.
Experimental design: The paper tackles an important problem and I believe Pakistan deserves the best predictions it can get. I am also a big supporter of using Bayesian statistics and I am glad authors chose to use it.

Generally, I think the authors need to decide whether the aim is to provide high-quality predictions for Pakistan (and thus multiple models with very different assumptions should be fit to the Pakistan data to assess uncertainty from model choice), or whether the aim is to evaluate to what extent BSTS are a sensible tool for Covid/disease prediction in general (and thus datasets from more countries should be used).

One possible model to use/compare against would be the epidemia package (https://imperialcollegelondon.github.io/epidemia/) or other models mentioned covid19forecasthub.org

The biggest risk with the current design is in my view that the model doesn’t use the structure of the data (e.g. that all cases eventually become death or recoveries, all deaths/recoveries were once cases, the number of cases is limited by the total population, …). The epidemia package uses this structure and would thus be an interesting point of comparison (though I in no way want to force the authors to use this specific package).

Similarly, most successful models I know of do not focus on cumulative cases, but on new cases, as the cumulative data can be “too smooth” and thus underestimate the uncertainty. Comparing to/pairing with a method that focuses on new cases (or generally the time derivative of the time series) would be beneficial (but I don’t think it is required).

Evaluating predictive ability for just one time point is quite a weak check. Testing prediction should be done starting from multiple historical time points (preferably even a “sliding window” approach where say each data point is predicted from data say 5, 10 and 30 days prior, but that’s not completely necessary).

I also do take issue with the metrics used:  while RMSE and others can be useful, in a Bayesian and decision making setting, it is important to also check if the uncertainty is correctly quantified. The calibration of the model is important (i.e. across multiple starting points, do X% of actual data fit within the X% prediction interval for various values of X?).

On the other hand I do NOT think BSTS need to be shown to be a great method for the manuscript to be publishable. If a revised version of the paper provides a set of carefully checked predictions for Pakistan and/or a good evaluation of BSTS across multiple Covid datasets, I don’t care if the conclusions are not favorable to BSTS and would happily recommend the publication for acceptance.
Validity of the findings: The two following issues are in my view major and need to be addressed.

1) Using any MCMC algorithm, it is important to check that the MCMC chain converged. Multiple chains should be compared to assess if the whole posterior is explored.  Unfortunately this has not been done. Some basic tools for convergence checks for a Gibbs sampler would be traceplots, R-hat, and Effective sample size - see e.g. Vehtari et al. 2020, http://doi.org/10.1214/20-BA1221

And actually some of the parameters, especially “sigma.trend.level” seems to not have converged. Here’s my code I used to check convergence for some of the parameters, using RStan’s implementation of R-hat and ESS. Note that R-hat should roughly be < 1.01 and bulk ESS of at least 50 and usually much more is needed to accurately estimate a quantity (ess_tail should also be used and be high enough if tail-quantities, like the 95% interval are of interest).

fits <- list()
n_chains <- 4
for(i in 1:n_chains) {
  fits[[i]] <- bsts(ts, state.specification = ss, niter = 500, ping=0, seed=2016 + (i - 1) * 157555)
}

for(par_name in c("sigma.obs", "sigma.trend.level", "sigma.trend.slope", "sigma.seasonal.2" ) ) {
  draws <- matrix(nrow = length(fits[[1]][[par_name]]) - burn, ncol = n_chains)
  for(i in 1:n_chains) {
    draws[, i] <- fits[][[par_name]][(burn + 1) : length(fits[][[par_name]])]
  }
  cat(par_name, ", Rhat: ", rstan::Rhat(draws), ", ESS: ", rstan::ess_bulk(draws), "\n")
}

This should be done for all parameters (and preferably a bit neater, the code is a bit hacky, sorry).

The traceplots also do show that the chains actually explore a slightly different part of the posterior.

Thus more iterations, longer burnin, and possibly thinning need to be introduced to make the computation trustworthy.

2) I also find it very weird that different number of seasons is used for different pandemic counts. Seasons are of length 1 (days), but why are 4 or 2 day trends chosen? I would expect the pandemic to have a weekly pattern, but why should there be a 2-day or 4-day pattern? How were these parameters chosen?

## Additional note

Comparing to actual numbers for their target date (August 10th) the reality is on the lower end of the confidence intervals and the number of active cases (17 799) falls quite outside the 95% prediction interval [22,568;102,717]. I suspect this is because the model seems to fit a continued exponential growth, which seems to have not happened. I don’t think it is reasonable to fail the authors for not predicting accurately, as this is very hard. But - if I understand the model correctly, I think the structure of the model may have been unable to model a plateau for the number of total cases, which is in my opinion an important oversight.
Additional comments: I wish you best of luck with modelling and with the overall pandemic situation in general. I hope you are safe.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 1
Reviewer: 3

Output:
Basic reporting: The authors proposed a computational model for forecasting the COVID-19 trends in Pakistan, thus I think the prediction here is not "pattern" like the authors mentioned in the title. It should be "trends" or "outbreak"?

There are a lot of grammatical errors and typos in this manuscript. The authors should re-check and revise carefully. The use of English language should be also improved significantly. For example, some errors are as follows:
- the first sentence of abstract: Though Pakistan is among the countries where the COVID-19 entered quite later, but currently it is on full flow ...
-  To tackle with challenges in the coming months ...
- And timeline of these forecasts have passed now.
- due to which the active number of cases are expected to decrease during ...
- These are quite efficient measure of forecast accuracy of a model.
- ...

There are missing literature review on COVID-19 outbreak prediction in other countries.
Experimental design: Methodology was poorly written. Now it contains two short paragraphs with few information. With these information, the readers could not replicate the methods.

Forecast/prediction model is a well-known problem and there are a lot of models have been developed on this purpose, from machine learning (i.e., PMID: 31055655, PMID: 27475771), or deep learning (PMID: 28643394, PMID: 31921391). Therefore, the authors should show more detail on their model information. Also, it needs more references to attract broader readership in this part.

How did the authors select the optimal hyperparameters of their model?
Validity of the findings: The application is narrow. The authors only focused on COVID-19 outbreak in a specific country (Pakistan). Even in Pakistan, the authors have not validated the model on an unseen data.

I'm wondering whether the model could be efficient in predicting the COVID outbreak in another dataset from the other countries. The authors should implement and discuss this point. It strongly support the hypothesis to provide a useful method in this problem.

The authors should compare the performance results with the previous works on the same dataset.
Additional comments: No comment.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The paper has been improved, however there are still some points that are clear.

For instance, the model ARIMA in Eq. (1)  is instead an ARMA model? where is "d" in Eq. (1)? I believe that you are using and ARMA.
Experimental design: The paper seems technically sound and experiments are good enough.
Validity of the findings: The paper seems technically sound and experiments are good enough.
Additional comments: The paper has been improved, however there are still some points that are clear.

For instance, the model ARIMA in Eq. (1)  is instead an ARMA model? where is "d" in Eq. (1)? I believe that you are using and ARMA.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 2
Reviewer: 2

Output:
Basic reporting: The reporting has been substantially improved from previous version, I have no further bigger requirements.

Minor points:
The authors have not improved the palettes used in their plots and I still find the plots to be not very accessible to people with impaired vision, neither pretty.
Experimental design: Authors have expanded their evaluation and I have no bigger remaining issues.

Minor points:
Authors have not reported calibration, as I asked to in my previous review. I don't think it is strictly necessary, but it would still be useful. (my older suggestion: "The calibration of the model is important (i.e. across multiple starting points, do X% of actual data fit within the X% prediction interval for various values of X?)."). Note that low prediction error does not imply good calibration (or vice versa).
Validity of the findings: Only important problem I noticed:

"From these results, it can be seen that P-values for the Ljung Box test(Q-statistic) are greater than 0.05 for all variables at different lags. Therefore, the residuals from the proposed models are white noise."

This is not a correct interpretation of p-values. The test indicates that the residuals are consistent with white noise (as far as the test is able to discern), but they still could be structure that just is not discernible with the current data.
Additional comments: The article was substantially improved. Not all of my concerns were addressed, but I don't find any value in further gatekeeping the publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 2
Reviewer: 3

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: My previous comments have been addressed.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 3
Reviewer: 1

Output:
Basic reporting: The paper has been improved following my suggestions.
I believe that is ready for publication.
Experimental design: The paper has been improved following my suggestions.
I believe that is ready for publication.
Validity of the findings: ok
Additional comments: The paper has been improved following my suggestions.
I believe that is ready for publication in its current form.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: ANALYSIS AND FORECASTS FOR TRENDS OF COVID-19 IN PAKISTAN USING BAYESIAN MODELS
Review round: 3
Reviewer: 2

Output:
Basic reporting: The manuscript was OK in the previous version, it is also OK after the minor changes.
Experimental design: The manuscript was OK in the previous version, it is also OK after the minor changes.
Validity of the findings: The manuscript was OK in the previous version, it is also OK after the minor changes.
Additional comments: The manuscript was OK in the previous version, it is also OK after the minor changes.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EFFECTS OF EXERCISE TRAINING WITH BLOOD FLOW RESTRICTION ON VASCULAR FUNCTION IN ADULTS: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 1
Reviewer: 1

Output:
Basic reporting: No comment
Experimental design: No comment
Validity of the findings: No comment
Additional comments: The authors set out to write a systematic review and meta-analysis over the effects of blood flow restricted exercise on vascular function in humans.

The authors have done very well in following all of the best-practice guidelines as they relate to conducting such reviews and analyses. I have read the manuscript several times over, and I cannot find anything material that I feel absolutely must be added to the manuscript.

I am in agreement with the articles that have been included in this review and analysis, and all included articles meet the authors' stated criteria for inclusion. The meta-analyses appear to be performed correctly. The writing is clear and concise.

I appreciate the authors' inclusion of careful wording with regard to the wide variety of BFR protocols, the limited number of studies in this particular research area, and that all results of these analyses should be viewed cautiously as a result. Ideally, this review will allow other laboratories to focus their expertise on some of the gaps in the literature that the authors noted.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EFFECTS OF EXERCISE TRAINING WITH BLOOD FLOW RESTRICTION ON VASCULAR FUNCTION IN ADULTS: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The authors of this manuscript did a very nice job on reporting the information retrieved from their hypotheses and presenting the information in numerous figures/tables that make it easy for the reader to understand the results. Their raw data was reported and appears to reflect the analyses performed in the MA.
Experimental design: The authors did a commendable job in providing context for the experimental question and appropriately narrowed their inclusion criteria to reflect the BFR literature. They identified current gaps in the literature and where this manuscript would seek to address these gaps. Methods are appropriately described and depicted in figure 1 to allow for reproduction.
Validity of the findings: The authors provided comprehensive depictions of the data retrieved in Tables 1-5. They provide appropriate conclusions as to what can be determined based on the current body of evidence and provide future investigators some context/guidance to promote future homogeneity in the newer research being conducted on BFRE and its effects on vascular structure/function. They also accurately describe the caution to the reader on applying their results outside of the major populations studied (ie limited generalizability to clinical populations but more relevant for healthy populations).
Additional comments: The authors of this manuscript performed a very comprehensive systematic review and meta-analysis on the effects of BFRE on vascular structure and function along with relevant secondary outcomes (adherence/negative side effects). The authors should be commended at the level of data synthesis performed in this review and the clear reporting in the tables/figures.
Minor edits:
Ln 162-3 – Smart et al needs to be referenced appropriately

Ln 164 – “Studies were not excluded based on score for methodological quality.” One sentence to elaborate on why this was the case could help further clarify for the reader. Was this to increase study inclusion to be able to perform a MA?

Ln 196-7 – I2 needs to be superscripted x 2

Ln 216 – extra space between “reported” and “on”

Ln 238 – What does (%) mean? Is there a number that should precede the %?

Ln 269 – extra % in (46%%) needs removing

Ln 357 – Figure 3 appears to be referenced incorrectly and should be Figure 2.

Ln 372 – Figure 2 appears to be referenced incorrectly and should be Figure 3

Ln 387 – “meta regression” should be “meta-regression” or some consistent term

Ln 427-429- “A single study reported an initial sensation of leg numbness when one participant commenced BFRE, which did not persist beyond the first minute of exercise [39] (2 out of 16 hypertensive older women).” – would suggest changing wording to two participants since the study reported two adverse events in 16 participants, not just one participant.

Ln 452 – extra space present between “favored” and “dynamic”

Ln 466 – remove “the” before both

Ln 485 – needs period at the end

Ln 504 – need space between “40] and”

Ln 506 – Wilvebrand spelled wrong

Ln 514-524 – I appreciated this section in addressing the significant limitations in the literature and how that reflected in your analysis.

Ln 534 – Principals -> principles

Ln 545 – The article (Patterson, 2019) is now not referred to as a position stand, so this should be reflected in the manuscript

Ln 546 – space needs removing between recommending and BFR

Ln 555 – The article (Patterson, 2019) is now not referred to as a position stand, so this should be reflected in the manuscript

Ln 557 – needs a number before the percentage to inform reader of the exact % not using LOP as a standardization approach (also – you have used AOP prior – thoughts on sticking to one term throughout the manuscript?)

Ln 575-76 – additional spacing needs to be deleted, along with the comma between review and excluded

Ln 578 – stick with one variation of the word – comorbidities or co-morbidities and keep consistent. This has comorbidities but on 575 its co-morbidities.

May be good to mention https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5374335/ even though there was no exercise control, it was RCO design and illustrates some of what you are talking about with respect to the at-risk populations (and was excluded from analysis based on your inclusion criteria) and shows some potentially negative changes in both the trained and untrained limb following 3 weeks of training in overweight individuals.

Ln 583 – “side” should have another word attached – ie “side events” if removing the preposition still doesn’t make sense.

Ln 610 – The article (Patterson, 2019) is now not referred to as a position stand, so this should be reflected in the manuscript; along with spacing between “tolerability” and “and”


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: EFFECTS OF EXERCISE TRAINING WITH BLOOD FLOW RESTRICTION ON VASCULAR FUNCTION IN ADULTS: A SYSTEMATIC REVIEW AND META-ANALYSIS
Review round: 2
Reviewer: 1

Output:
Basic reporting: The authors made the recommended adjustments to the manuscript and is suitable for publication.
Experimental design: The authors made the recommended adjustments to the manuscript and is suitable for publication.
Validity of the findings: The authors made the recommended adjustments to the manuscript and is suitable for publication.
Additional comments: The authors made the recommended adjustments to the manuscript and is suitable for publication.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NMDAR IN BLADDER SMOOTH MUSCLE IS NOT A PHARMACOTHERAPY TARGET FOR OVERACTIVE BLADDER IN MICE
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: In this study, Xie and co-authors used a smooth muscle-specific NMDAR knockout mouse to investigate the potential role of NMDAR in OAB, they found that NMDAR is not a pharmacotherapy target  in BSM.   The overall experimental design is excellent, results from this investigation are robust and well presented.   The speculations of the results are reasonable and the conclusions based on theses results are convincing.
I only have one suggestion, which is related to the contractility results, mostly presented in Fig 3.  Are the thickness of the muscle well controlled ? It can be very hard to match the width and thickness of the muscle strip during dissection.   Based on the method section and it seem, in some experiments, the width of the muscle strip is not very well controlled.  In that case,  the authors should consider using stress instead of force to present contractility in Fig 3.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NMDAR IN BLADDER SMOOTH MUSCLE IS NOT A PHARMACOTHERAPY TARGET FOR OVERACTIVE BLADDER IN MICE
Review round: 1
Reviewer: 2

Output:
Basic reporting: no comments
Experimental design: no comments
Validity of the findings: no comments
Additional comments: General comments;
This study was comprehensively performed to confirm that there is no role of NMDAR in bladder smooth muscle in organization of normal bladder morphology and function as well as in the development of CPY-induced abnormal bladder morphology and bladder dysfunction. Although this study is a completely negative study, the experimental methods are reasonable and the results are clearly stated. This study would be a valuable literature in the field of investigation on the control of the lower urinary tract.

Specific comments;
   Wordings should be revised at several sentences.
Line 52:   nycturia  →　nocturia
Line 105:  euthanasia was performed  →  Euthanasia was performed
Line 137:  isolated strips  →  Isolated strips
Line 144:  function. CMG  →  function, CMG
Line 340:  a recent report that revealing →  that revealed


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NMDAR IN BLADDER SMOOTH MUSCLE IS NOT A PHARMACOTHERAPY TARGET FOR OVERACTIVE BLADDER IN MICE
Review round: 1
Reviewer: 3

Output:
Basic reporting: In this manuscript, the authors use clear and professional English throughout. All the sections are well explained, and the results seem to address the claims made by the authors. Literature references are provided and seem to be in order. Article structure is acceptable, and figures are understandable.
Experimental design: The experimental design seems valid. Research question is well defined and relevant to the audience. There are technical discrepancies in the model which are discussed in greater detail in the relevant section of the review. Methods description is adequate.
Validity of the findings: Some of the major concerns in the manuscript are summarized below-

1. The authors discuss generation of SMNRKO mice. However, Is this F1 generation of mice that are used for this study? In this case the mice are still heterozygous. A tissue specific KO, which is needed for this manuscript as the authors plan to knockout SMRKO only in the bladder smooth muscle (BSM). For this, two rounds of breeding is generally necessary. The authors must state if this was so in the methods section.
2. The efficacy of SMNRKO is not validated by the authors. This is most important, especially because the authors find no difference in WT vs KO. It could be simply because there was no KO. The authors must present stainings of NMDAR in bladder smooth muscles to prove complete KO in BSM and as a control perform staining on other tissues from the same mice to show presence of NMDAR in other tissues. This will validate the KO model.
3. Authors must also present genotyping data of the KO mice vs wild type mice to analyze the KO site. Again, BSM as well as other control tissues must be used to isolate DNA.

Other minor concerns are as follows-
1. Statistical softwares that were used must be mentioned in the methods section.
2. In the experimental design section, full names of methods must be written instead of abbreviations. This will enhance readers' experience.
3. Many figures have missing statistical presentation on graphs. This has to be rectified.
4. It is generally a good idea to include figure legends in all the panels of figures. The authors have skipped this which may confuse the readers.
5. Box and whisker plots in Figure 5 have two sets of statistics. The authors must explain why?
6. In the same figure some plots have p values written on them while others have asterisks. Consistency must be maintained. If not significant ns should be written.
7. In the abstract line 37- it should be induced 'by' CYP.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NMDAR IN BLADDER SMOOTH MUSCLE IS NOT A PHARMACOTHERAPY TARGET FOR OVERACTIVE BLADDER IN MICE
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The new data from authors suggests that the muscle thickness is indeed well control. My concerns have been addressed.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: NMDAR IN BLADDER SMOOTH MUSCLE IS NOT A PHARMACOTHERAPY TARGET FOR OVERACTIVE BLADDER IN MICE
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: The authors have addressed all the major concerns in their re-submission. I would recommend the authors to present their genotyping data, mRNA expression and western blot (comment 2 response) as a supplementary figure for the benefit of readers.
I congratulate the authors on their excellent work.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 1

Output:
Basic reporting: A few minor points mentioned in the 'General comments for the author'.
Experimental design: A few minor points mentioned in the 'General comments for the author'.
Validity of the findings: This part is robust!
Additional comments: In this study, Tu et al. established a 15-genes ferroptosis signature. Ferroptosis is a relatively new cell death phenomenon that is dependent on iron and lip peroxides. From the TCGA patients database and a validation dataset, they concluded significantly shorter overall survival of the lung adenocarcinoma patients in the high-risk gene signature group based on the expression of this signature. These patients encountered a more potent immune suppression, especially in the antigen presentation process and a higher level of PD-L1 expression. This could serve as a biomarker for predicting the response to immunotherapy in lung adenocarcinoma. It is a promising study for a better-personalized treatment opportunity in lung adenocarcinoma patients. Though the in-vitro and in-vivo experiments need to be performed for validation, they are out of the scope of this in-silico study paper and are destined for future research. I would suggest the following recommendations to the authors in order to make this manuscript better:

1.	A couple of lines about ferroptosis should be mentioned in the abstract as well. The authors described it in the introduction, but it is relatively a less studied phenomenon, so a brief description in the abstract would attract the readers.
2.	If a new 'R' script is written for this manuscript, please consider it depositing in the public database like Github and give its reference in the manuscript.
3.	As a general note, there is no space between the first brace of the references and the preceding word. A space should be incorporated. e.g. in line 51...cancer(Wei et al. 2018) should be cancer (Wei et al. 2018).
4.	In line 107, elaborate on the criteria for discarding the unrelated genes.
5.	Please replace volcano with volcano plots wherever mentioned e.g. in line 126.
6.	Usually, in these types of studies, more than one validation cohorts are used. Here only GSE72094 as the external validation cohort is used. Why specifically this cohort, and why only one being used?
7.	Figure legends are not very informative. Please expand them and add more information to make them self-explanatory.
8.	It seems like the legends for the supplementary figures are missing. Could authors double-check it?
9.	Fig. 1B is not very informative due to its small size. I would recommend the authors to keep it as it is in this figure as a representation and included the full-page zoomed version in the supplementary so that the names of differentially expressed genes are visible.
10.	For the volcano plot in Fig. 1C, label the most significant top hits (4 to 7).
11.	For the survival plots in Fig. 3 and elsewhere, mention the number of patients.
12.	Please include a good discussion about the enriched common mutations based on Fig. 7. It is missing now!
13.	From Fig. 5, some immune-related cells/functions are enriched in high-risk groups and some in low-risk groups. What biological significance could be drawn from this? This should be discussed.
14.	A conclusion figure (graphical abstract) will be very useful for the readers.
15.	In the discussion, from line 303 onwards about the description of the 15 genes, it would be better and more useful if the authors could present this description in a table format, including the references.
16.	In line 53, ‘in’ is missing from Recent years.
17.	Line 79- typo, a proximately.
18.	Line 105- typo, iron iron
19.	Reference for line 267 missing.
20.	Line 286 is incomplete.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 2

Output:
Basic reporting: Tu et al, report a ferroptosis -related signature in patients with lung adenocarcinoma. To begin with, the authors state an inadequate claim starting from the title. The response to immunotherapy is not supported at all from their data.

In the introduction section, the authors do not include important and highly impactful reports regarding the role of the SLC7A11 in lung cancer progression, stating that not many reports are available (line 72) (The Journal of clinical investigation, 130(4)., Oncogene, 37(36), pp.5007-5019., Redox biology, 38, p.101801., Cancer discovery, 9(12), pp.1673-1685.)

In several areas of the manuscript (line 51, 82, 263-269) the writing needs significant editing to improve comprehension. In many cases, the references used by the authors do not exist, displayed, properly interpreted (Tang et al., 2020, Huang et al 2019). IN other cases, teh authors use vague expressions to justify their findings and conceptual flow without citing literature accordingly  (line 263).

The figures are not adequately prepared with poorly labelled graphs with limited flow between the different topics.
Experimental design: There are several concerns regarding the overall experimental design. The ‘validation’ cohort used in this study (GSE72094) contains mutated patients specifically, and many of them harbor KRas mutations. A recent report showed that inhibition of SLC7A11 in KRAS mutated lung adenocarcinoma causes synthetic lethality in these patients. Based on these, the conclusions of the analysis of this dataset is biased and may be affected by the dependency of the ferroptosis pathway in the biology of KRAS mutated lung adenocarcinoma. The authors are strongly advised to expand their ‘validation’ in several publicly available lung adenocarcinoma datasets in GEO.

In the methods section it is unclear how the authors performed the GSEA for the different cell types that they propose. They should also provide the GO IDs that they used for the latter analysis.
Validity of the findings: The proposed role of the gene signature in immunotherapy response is correlative. The fact that CD274 mRNA expression is differentially expressed between the groups does not mean that it affects the response to immunotherapy. The authors are advised to remove these claims.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 1
Reviewer: 3

Output:
Basic reporting: In the present manuscript the authors used publicly available data bases in order to provide a novel 15-gene ferroptosis-related gene signature depicting the overall survival outcome as well as the possible response to immunotherapy in lung adenocarcinoma patients.

This is an interesting paper which highlights the role of ferroptosis in cancer progression and immunotherapy. Based on a 15 ferroptosis-related gene signature the patients of the two data bases used in this study were stratified into two risk groups with the ones in the high risk showing reduced OS accompanied by strong immunosuppression compared to the patients in the low risk group.

However, there are a few comments that should be addressed to further improve the manuscript.
Experimental design: 1. The authors should consider providing the PPI network from the STRING database as well as the correlation network of the ferroptosis related genes identified in the present study.
2. Survival analyses of each of the 15 genes should be included in the second section of the results.
3. In the last section of the functional analysis, the DEGs between the two groups should be used for the Gene Ontology analysis in addition to the KEGG pathway.
Validity of the findings: The critical role of ferroptosis in cancer treatment is well established from numerous recent studies in different cancer types. However, this specific 15-gene signature is novel and this is the reason why the authors should provide additional information regarding the correlation network of these genes, the pathways they participate or their biological function, in order to give a possible explanation about the molecular mechanisms that might be responsible for the phenotypes presented in this manuscript.
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 1

Output:
Basic reporting: -
Experimental design: -
Validity of the findings: -
Additional comments: I am satisfied with the revision of the authors. The manuscript can now be considered for publication!


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 2

Output:
Basic reporting: Tu et al., return a significantly revised and improved manuscript of their work on the ferroptosis signature in patients with lung adenocarcinoma.
Although the manuscript has been improved significantly, eliminating many mistakes and correlative claims, still there are some important points that should be revised.
The main concern is that in several sections of the manuscript, the effect that the authors trying to present is purely correlative.
Experimental design: The new proposed experiments are well designed. All the raw data are nicely organized and provided in the manuscript.
The authors have done a solid work on providing  detailed and transparent supportive data. In the previous version, the experimental designed was poorly biased, a phenomenon much improved in this version of the manuscript.

My main concern is still focusing on the ssGSEA. I appreciate the fact that the new method is well explained and provided in Github.
Nevertheless, the interpretation of the data is inaccurate and does not support the clinical claim that the authors wish to state.
Validity of the findings: In line 283 the authors identify in an unbiased way, that the high expression of the ferroptosis signature correlates with immune response and immune processes.

Furthermore, in figure 9, the authors present the expression of several gene-signature that are markers of immune cell functions in the high and lo-w groups.

Nevertheless, in line 295  the authors interpret these data as different infiltration of the patient tumors by these immune cell lines. The authors have no data whatsoever to support the claim of immune infiltration. This is just a correlation, not a causation.

The fact that the ferroptosis signature is oncogenic in patients with lung adenocarcinoma, could clearly affect the clinical status of these patients, leading to the differential immune system function. Furthermore, the fact that the proposed signature is oncogenic, could also correlate with more aggressive therapeutic regiment, know to affect the immune cell population, function and distribution in these patients.
This is a very important point that is inaccurate and needs to be corrected.
The authors need to remove these claims for the discussion section as well.

The authors should only interpret these finding as they really are. A correlation of the ferroptosis signature with a proposed immune cellular function. Still a very important finding, that needs a crisp and precise description.
Additional comments: The manuscript is significantly improved and more impactful. If the authors edit some important data interpretation , I would support the publication fo this interesting concept.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors have addressed all my concerns and therefore I support the publication of this manuscript.
Experimental design: No comment
Validity of the findings: No comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: CONSTRUCTION AND VALIDATION OF A 15-GENE FERROPTOSIS SIGNATURE IN LUNG ADENOCARCINOMA
Review round: 3
Reviewer: 1

Output:
Basic reporting: No comments.
Experimental design: No comments.
Validity of the findings: No comments.
Additional comments: The authors return a revised manuscript with a very careful and accurate interpretation of their data.
I recommend the publication of this interesting concept.
Congratulation to the authors.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 1
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I am so appreciated the author presenting this clinical useful research article emphasizing that  high prevalence of spinal pain in physiotherapists. However, the results were well known. My comments are as follows
1.Please clarify the workplace of the patients were enrolled, in clinic of local hospital , or medical center.
2.The health professionals could self-treat pain using drug. Any healthy effects in your series.
3. Table 1, 2 : Any difference among the  Kinesitherapy, physical therapy, Massage group?, please clarify
4. Please provide the number of daily treated patients in each groups?
5.The conclusion is nice


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 1
Reviewer: 2

Output:
Basic reporting: The introduction section is too lengthy, it will be better if it's made more concise and the total number of words is reduced.
Experimental design: 1. How sample size was calculated?
2. The results section is also very extensive. If information is given in tables and figures then there is no need to write it again in the results section. If some information is not given in tables and figures then they can be written in the results section. Thus it would be better if the results section is also made more concise and the total number of words is reduced.
3. Instead of Chi2, Chi-square can be written.
4. Was any particular inclusion and exclusion criteria chosen? The participants studied had the age of a much wider range i.e. 19-63 years. Would it not be appropriate if a smaller range was chosen for study e.g. 20-40 years or 40-60 years so that the effects of old age and PT profession can be separated? Were PTs having traumatic spine injury or history of spinal surgery excluded from study?
Validity of the findings: 1. Conclusion section is not written separately from discussion section.
Additional comments: 1. The topic is relevant for PTs. But the overall manuscript should be made more concise and precise so that readers can get the best of your research without spending much time on reading or without spending much time searching for the best finding among all the findings.
2. What does W mean in line no. 127?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 1
Reviewer: 3

Output:
Basic reporting: 1. Revise the title and make it more specific to the regional population (country name) as so many researches on prevalence and risk have been published in different countries.
Consider writing country name at the last of this topic as “Prevalence and risk of spinal pain among physiotherapists in Poland!”
2. There is a need for Professional English editing by either an English native language editor/reviewer Or English language editing services agency as there is a lot of grammatical mistakes, repeated, spelling mistakes, the informal language used, and chaotic sentences found throughout the manuscript. English editing is strongly recommended.
3. What was the study design? Consider t write the study design in the methodology section of the Abstract and in the text of the manuscript as well.
4.Line 28: The first line should start mainly with addressing spinal pain as the most common problem among working professionals worldwide and are the common source of work-related disabilities and economic burden in the societies and so on… Then its prevalence and incidences, in general, indicating neck pain LBP.
5.Line 42: Avoid using non-formal words. Consider elaborating the term industrialized countries in a general term as “developing or developed countries”
6. Line 46: Consider writing “irregular or abnormal posture” instead of uncomfortable postures.
7. Line 51-55 & 56-59: Since occupation spinal neck pain and back pain are having the same root of origin, therefore, Both paragraphs should be merged into one paragraph and combine all risk factors and cause into 1-2 lines to describe the spinal pain including neck and back pain. Similarly, write 1-2 lines to explain the symptoms of NP & LBP. No need to describe in separate for NP and LBP.
8. Line 64-69: The work-related challenging factors including working environment and settings of equipment, the height of couches, warm-up exercises and muscle strength & conditioning for the physiotherapist, working hours, and rest-period between two consecutive patients, number of patients per day, etc. should be mentioned here. The mechanism of action of these conditions/postures/spinal malalignments, muscle weakness, etc., how badly affect the spinal alignment and make a reason for spinal neck and back pain.
9. Line 79: Write here the justification of the study that how this study would be new from other published studies that are similar so far? If it is specific to the country where research conducted, then add the name of that country in the title and here to justify this study as it was not done before here?
Experimental design: 10. What was the study design?
Consider writing the study design in the methodology section of the Abstract and the text of the manuscript as well.
11. Line 94: Was it an actual or greater or lower sample size?
Explain in detail, How the sample size estimated? explain parameters, method of power analysis, and effect size and source of data to calculate the sample size!
12.Line 97: What was the inclusion and exclusion criteria for the study? Consider explaining the method and procedures for the sample collection and recruitment here in detail! How many samples meet the criteria and denied participation in the study? Add a paragraph:
13. Line 98-99:  Was the questionnaire designed by the authors of this study tested for its accuracy, reliability & validity, if yes, then explain the procedure in brief?
14. Line 101-104: Was all sections of the questionnaire designed by the authors reviewed thoroughly and approved by the team of medical professionals such as senior consultant/physiotherapist, head/dean, or registered body of physiotherapy practice before being utilized? Did you find any weakness or drawback or extra strength of this questionnaire designed by the authors, if yes, consider explaining here?
15. Line 101-104: Consider writing the scoring method and psychometric analysis of the questionnaire designed by the authors here!
16. Line 110-112:Consider including the scoring method in detail and psychometric properties of the NDI questionnaire here!
17. Line 113-115: Consider including the psychometric properties of the ODI questionnaire here!
18. Line 118-112: Consider to explain in sentences rather than point-wise!
19. Line: 130: Here, all tests such as ANOVA, Tukey’s post hoc analysis, and others should be enumerated with their role to determine in the analysis!
Validity of the findings: 20. Results: The authors have mentioned all values of quantitative data in the results section although they are already mentioned in the tables. Therefore, It is strongly recommended to revise the result writing and consider to write only those values which are not mentioned in the tables. No repetitions of data rather than refer to the tables will be encouraged!
21. Line 1323-136: Consider these lines to write as “there were 158 PTs specialized in kinesitherapy (65.8%), 37 physical therapists (15.4%), and 45 masseurs (18.8%) as presented in Table 1.”
22. Line:138: Consider to include standard deviation in the bracket along with mean as “the mean (SD) age of women …. And so on till the end of the paragraph.
Don’t explain the demographic details here if it is already present in tables. Explain those details which are not mentioned in the tables.
Don’t repeat every time like “mean age of women … mean height of women… mean weight of women… so on.” Consider writing in one sentence including all details.
23. Overall, the results section needs to be revised and limited to enumerate the data that are not mentioned in the table rather than the repetition of the same data. Authors should not write the result in a conclusive-form, they are required to just enumerate the findings in the result section.
Additional comments: Discussion:
24. Line:282: Give here an updated reference!
25. Line 291: A separate paragraph is required to write the mechanism of spinal pain (neck and back pain) due to lifting, sitting, and other mechanical/postural factors. How these factors create spinal pain and affect the activities of physiotherapists or other professionals?
26. Line 311-316 & 317-323: These paragraphs should be written in accordance with the purpose of the study. In actuality, It is the answer to your research hypothesis. Therefore, it should be written in a way to answer your hypothesis or fulfill the purpose and objective of your study. Consider to revise the whole paragraph and answer your hypothesis/purpose of the study.
27. Line:325: Give the reference to justify this statement?
28. Line 326: Taking the history of injuries is the basic part of the assessment. Your study was on prevalence and risk of spinal pain, then how you didn’t ask about the previous musculoskeletal injuries that might be imitating the occupational pain? Did you mention this point in the exclusion criteria of the study?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 2
Reviewer: 1

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: I appreciate the author presenting  this research article to emphasize the high prevalence and risk of spinal pain among physiotherapists in Poland. My comments are as follows
1.  This manuscript is well designed but the presentation of results were not easy to read, for example (1) in Table 3 no painkiller used data but the authors mentioned in the manuscript (2). Figure 1, 2, 3 didn't show the p value.
2. Two questions need to be clarified
    a. Previous study showed that pregnancy induces changes in spinal posture and weakens the joint
        structure, which increases the risk for back pain. Any pregnancy in your cases?
    b. Previous studies showed younger physiotherapists have a greater chance of developing back pain
       because of they did not protect themselves in daily practice. Do you agree this mention?


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 2
Reviewer: 2

Output:
Basic reporting: no comment
Experimental design: no comment
Validity of the findings: no comment
Additional comments: No additional comments


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PREVALENCE AND RISK OF SPINAL PAIN AMONG PHYSIOTHERAPISTS IN POLAND
Review round: 2
Reviewer: 3

Output:
Basic reporting: The authors did not answer satisfactorily the following Line 61 and 79/questions no. 13-15, which was important to answer in the revised manuscript. [Line numbers are the same as in the original submitted manuscript ].

Line 61: The authors did not cover the suggested work-related challenging factors that contribute to spinal pain among physiotherapists. They must mention the complete risk factors as the reviewer suggested before.

Line 79: The authors did not add a paragraph for the justification of the study that how this study is novel from other previous studies as the reviewer suggested before. Without justification of the study, how the author can write their purpose. This comment must be addressed.

13. Line 98-99: Was the questionnaire designed by the authors of this study tested for its accuracy, reliability & validity, if yes, then explain the procedure in brief?
14. Line 101-104: Was all sections of the questionnaire designed by the authors reviewed thoroughly and approved by the team of medical professionals such as senior consultant/physiotherapist, head/dean, or registered body of physiotherapy practice before being utilized? Did you find any weakness or drawback or extra strength of this questionnaire designed by the authors, if yes, consider explaining here?
15. Line 101-104: Consider writing the scoring method and psychometric analysis of the questionnaire designed by the authors here!

Line 133-134. Why so many tests used for the normality of distribution. Either consider writing the use of each test Or just mention the one test which data you used in the table or study.
Experimental design: N/A
Validity of the findings: This question was not answered by the authors although it is a must answer.

In discussion, question and line numbers are the same as in the originally submitted manuscript.

26. Line 311-316 & 317-323: These paragraphs should be written in accordance with the purpose of the study. In actuality, It is the answer to your research hypothesis. Therefore, it should be written in a way to answer your hypothesis or fulfill the purpose and objective of your study. Consider revising the whole paragraph and answer your hypothesis/purpose of the study.
Additional comments: The authors of this study replied to most of the questions/comments raised by the reviewer however, there are few questions that are still required to be answered and they are very important for the suitability and acceptance of the manuscript. These questions are indicated with the previous question number and line numbers. Unless these questions will not be answered, the reviewer is not ready to accept this manuscript to be published.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PROGNOSTIC FACTORS AND TREATMENT OF NEUROENDOCRINE TUMORS OF THE UTERINE CERVIX BASED ON THE FIGO 2018 STAGING SYSTEM: A SINGLE-INSTITUTION STUDY OF 172 PATIENTS
Review round: 1
Reviewer: 1

Output:
Basic reporting: I could not evaluate the article because there were discrepancies between the data described in the text and the Figures/Tables.
On line 182-190, authors described the multivariate analyses in the entire series using Figure 4 ant Table 2.
Figure 4 was comparison of survival curves and was not the result of multivariate analysis.
The title of Table 2 was prognostic factors (n¬¬=172). But in the Table 2, the total number of patients was not 172 and there was not the data of HR about FIGO stage, age and chemotherapy regimen that were described as the prognostic factors in the multivariate analyses.
The discrepancy was also seen on line 191 to 196.
Authors described about the multivariate analysis in the patients who underwent surgery using Figure 5 and Table 3.
But Figure 5 was not the data of multivariate analysis (just comparison of survival curves).
Table 3 was not the data of 108 patients because total number of patients was not 108.
Authors should carefully check the manuscript before submitting.
Experimental design: Authors described the efficacy of chemotherapy regimen using multivariate analyses. But the patients who did not receive chemotherapy should be included in the analysis.
Validity of the findings: Although the disease is rare, many observational studies have been conducted so far. No new discoveries are found in this paper.
Additional comments: Authors should carefully check the manuscript before submitting.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PROGNOSTIC FACTORS AND TREATMENT OF NEUROENDOCRINE TUMORS OF THE UTERINE CERVIX BASED ON THE FIGO 2018 STAGING SYSTEM: A SINGLE-INSTITUTION STUDY OF 172 PATIENTS
Review round: 1
Reviewer: 2

Output:
Basic reporting: The number of cases is sometimes inconsistent throughout the manuscript.  In line 137 in Results, the number of total patients is 195, but in line 88 in Methods, the number is 192.

Table 2 and Table 3 should be exchanged.

In Figure 1, the images are not small cell carcinoma but large cell neuroendocrine carcinoma.  Please confirm the pathological diagnosis.
Experimental design: In Method, the authors classified neuroendocrine cervical tumors into carcinoid tumor, atypical carcinoid tumor (ACT), small cell carcinoma (SCNEC), and large cell neuroendocrine carcinoma (LCNEC). However, thereafter, they did not mention about that classification in text and in Tables.  They should present clinical significance of the histological classification, i.e., ACT, SCNEC, and LCNEC, in Results and Tables 2 and 3.

In Table 2 and 3, the choice of parameters for multivariable analyses appears somewhat arbitrary.  In Table 2, in overall survival analysis, they do not include parametrial involvement/depth of stromal invasion, and in progression-free survival analysis, they do not include tumor size, para-aortic lymph node metastasis, and depth of stromal invasion.  In Table 3, in overall survival analysis, they do not include pelvic lymph node metastasis, para-aortic lymph node metastasis, and in progression-free survival analysis, they do not include age, pelvic lymph node metastasis.
Validity of the findings: In the section of 3.4 Efficacy of treatment, they mention that in patients with stage IV disease, primary treatment containing at least six cycles of chemotherapy was associated with significantly better 2-year OS (83.3% vs. 9.1%, P < 0.001) and 2-year PFS (57.1% vs. 0%, P = 0.01) rates than those primary treatment containing less than six cycles of chemotherapy.  However, 5-year OS and 5-year PFS rates of all these groups were 0%.  Such numbers appear unnatural.  The reasons why the patients who received primary treatment containing <6 cycles of chemotherapy died within 2 years but a majority of patients who received primary treatment containing ≧6 cycles of chemotherapy did not suffer progression of cancers or death should be explained and discussed in detail in Discussion.  For that purpose, for stage IV patients, they should also examine the data of therapeutic effect by RECIST and patients’ performance status.

In Discussion, line 228, the authors mention that the TP/TC and EP regimens were the most effective, with similar efficacies and toxicities.  However, it is unclear with which regimen the authors compared the TP/TC and EP regimens.  From Tables 2 and 3, chemotherapy regimens were classified into TP, EP, and others. However, the group of “others” is footnoted only as that including vincristine, bleomycin, Lipitor, idosfamide, temozolomide, and so on, and does not appear to be a single regimen. Some of them are not indication for cervical cancer or not anti-cancer agent. Therefore, it does not appear appropriate to compare TP and EP with the group of “others”.
Additional comments: Chen J. et al. studied prognostic factors and treatment of neuroendocrine tumors of the uterine cervix based on the FIGO 2018 staging system. They analyzed all stage patients together and drawn conclusion about prognostic factors and chemotherapeutic regimens to be recommended.  However, the rationale of these recommendations is not sufficient from the present data.  Especially, the conclusion that they recommend TP/TC and EP is not supported because the control against TP/TC and EP is the mixture of miscellaneous regimens to which detail is not described.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PROGNOSTIC FACTORS AND TREATMENT OF NEUROENDOCRINE TUMORS OF THE UTERINE CERVIX BASED ON THE FIGO 2018 STAGING SYSTEM: A SINGLE-INSTITUTION STUDY OF 172 PATIENTS
Review round: 2
Reviewer: 1

Output:
Basic reporting: .
Experimental design: .
Validity of the findings: .
Additional comments: Chen J et al. retrospectively examined prognostic factors and treatment in 172 patients with cervical neuroendocrine tumors (NETs) in a single institute.  The findings that they described in this study were not of very high novelty and were not conclusive in a sense with regard to the effectiveness of therapies.  Nonetheless, the analyses of a relatively large number of cases in a single institute appear to be worth reporting. The following comments are addressed for the improvement of contents as follows:

In Results 3.1, the authors compared survival between the 108 patients who received surgery as their primary treatment and 53 patients who received radiotherapy as their primary treatment.  In Table 4, the number of patients who received primary surgery is 107 and that of patients who received primary radiotherapy was 47.  Why did the authors exclude other 7 patients from the survival analyses?

Likewise, in Results 3.4 and Table 4, they compared OS and PFS between the patients who received treatment containing at least 6 cycles of chemotherapy and those who received treatment containing less than 6 cycles of chemotherapy in FIGO stage IV cases only.  Why did not they compare the same parameters for Stage IIB-III patients and, if corresponding presents are present, for stage I-IIA2 patients?

The authors should discuss the reason why the prognosis of FIGO stage IV patients who received treatment containing less than 6 cycles of chemotherapy was extremely poor.  The poor prognosis does not appear to have caused only by the insufficient amount of chemotherapy.  There might be other major reasons that repelled the introduction or continuation of chemotherapy, for example, severe adverse effect, underlying systemic diseases, high performance status.

In Conclusion from line 390-391, they mention that adjuvant chemotherapy, adjuvant radiotherapy and neoadjuvant chemotherapy did not improve the outcomes of patients with NETs.  However, they did not present data that support the statement/conclusion in this manuscript. They did not compare each treatment group with the control group.

In Figure 3, yellow curves are very hard to see.  They should use other color or another pattern of black line.

Typographic errors, e.g., analysdis in line 132, dirrerent in legends for Figures 4 and 5.

In the legend of Figure 1, they did not present the image of chromogranin A (CgA).  They should delete “chromogranin A (CgA)” from the legend.


Instruction: You are one of the reviewers, your task is to write a review for the article. You will be given the title of the article, the number of the round in which the article is located, and your order among the reviewers.

Input:
Title: PROGNOSTIC FACTORS AND TREATMENT OF NEUROENDOCRINE TUMORS OF THE UTERINE CERVIX BASED ON THE FIGO 2018 STAGING SYSTEM: A SINGLE-INSTITUTION STUDY OF 172 PATIENTS
Review round: 3
Reviewer: 1

Output:
Basic reporting: The questions and comments have been answered appropriately.
Experimental design: The questions and comments have been answered appropriately.
Validity of the findings: Although there are some concern about the expression of the sentences in Abstract, they may be within a tolerable range.  Examples of concern are: the 2nd line "to determine appropriate treatment guidelines"; and the last two lines "standardized and complete multimodality treatment may improve the survival of patients with NETs", which is somewhat inconsistent with the former sentence "adjuvant chemotherapy, adjuvant radiotherapy and neoadjuvant chemotherapy did not improve the outcomes of patients with NETs".
Additional comments: No comments.