Verified Rigor RecordRigor Reviewed

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Tregs depletion aggravates activation of astrocytes by modulating IL-10/GXP4 following cerebral infarction · 10.3389/fimmu.2023.1255316

Name: Verified Rigor Record — Tregs depletion aggravates activation of astrocytes by modulating IL-10/GXP4 following cerebral infarction · 10.3389/fimmu.2023.1255316
Item: Tregs depletion aggravates activation of astrocytes by modulating IL-10/GXP4 following cerebral infarction · 10.3389/fimmu.2023.1255316
Rating: 39
Author: Αlpha¹

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Αlpha¹ Editorial

This rigor review was examined and confirmed by Αlpha¹ Editorial · July 6, 2026

Αlpha¹ Rigor Review

Rigor Reviewed

CRITICAL

Critical rigor failure · Mixed Methods

Eight dimensions — click a segment for details

Critical

High-priority

Model-reported confidence: highReviewer agreement: lowEngine: 7.6.0Source: full text

Not verified in this report

No reported statistical tests were found to recompute.

This Αlpha¹ Rigor Review uses AI Rigor Reviewers trained on a curated corpus of high-fidelity and retracted papers, with expert supervision and curation. It can still make mistakes; verify each finding against the source before relying on it.

Critical findings

• internal_contradiction (high, corroborated): The abstract and title claim that Treg depletion attenuates renal fibrosis after cerebral infarction, but the entire results section focuses on brain astrocytes and does not present any kidney data. This is a major disconnect between the stated aim and the reported experiments.
• internal_contradiction (high, corroborated): The abstract states 'depletion of tregs protected against activation of astrocyte' while the results section states 'Tregs depletion promoted activation of astrocytes'. These are contradictory.
• internal_contradiction (high, corroborated): The methods state that 66 mice were induced with MCAO and 15 died, leaving 51; however, the results show N=4 per group (4 groups = 16 mice), with no explanation for the discrepancy.
• internal_contradiction (high, corroborated): The abstract states Tregs depletion 'protected against activation of astrocyte', but results (section 3.4) show Tregs depletion *promoted* activation (increased IL-2 and IL-6).
• internal_contradiction (high, corroborated): The abstract and title claim that Treg depletion attenuates renal fibrosis, but no renal fibrosis data are presented in the results section.
• Tregs depletion attenuates renal fibrosis by modulating IL-10/GPX4 following cerebral infarction.

This mixed-paradigm paper combines a mouse MCAO model, in vitro astrocyte experiments, and a bioinformatics analysis using a human sepsis dataset. The study suffers from major rigor deficiencies (no randomization, blinding, or power analysis), incomplete reporting of biological variables (sex, strain, housing), and a critical disconnect between the stated aim (renal fibrosis) and the presented data (only brain astrocyte outcomes). Statistical reporting is inadequate, and data/code are not shared.

The three independent reviewers converged on most deficiencies but diverged on the severity of statistical and reporting issues; the synthesized judgment weighs the specific evidence of missing data and internal contradictions, adopting the more conservative (fail) rating for dimensions where critical sub-criteria were unmet. The statistics verification component could not recompute any tests (no test statistics or CIs reported), and all 41 references were verified in registries with no retractions or missing entries.

12 major claims checked against the paper's own evidence: 4 not fully backed by the presented evidence (unsupported or overstated).

12 integrity concerns flagged (5 high).

Rigor

Eight Dimensions1 pass · 1 warn · 6 fail

12 major claims3 unsupported1 overstated4 partial4 supported

The introduction cites relevant prior research on Treg accumulation in non-lymphoid tissues, brain Tregs in stroke recovery, and ferroptosis. The rationale linking stroke to renal dysfunction via Tregs is presented. However, the paper does not explicitly address how its study overcomes specific limitations of prior work, and the bioinformatics component is loosely integrated.

Sub-criteria

prior work citedADEQUATE

premise rationaleADEQUATE

limitations addressedNOT REPORTED

Per reviewer (3) — the votes behind the verdict

PASSReviewer 1· 85% conf

The paper establishes a reasonable scientific premise by citing prior work on Tregs, stroke, and ferroptosis, and provides a logical rationale for the study.

Evidence

direct quote[Introduction]

“In stroke research accumulation of tregs during the two weeks has been observed ( , ). Brain Tregs contribute to neurological symptom recovery by suppressing excessive activation of astrocytes, a condition known as astrogliosis, through the secretion of Areg ( ).”

direct quote[Abstract]

“Accumulating evidence suggests an association between ischemic stroke and renal dysfunction; however, the underlying mechanisms remain unclear. This study aimed to investigate the potential of Tregs in inhibiting the activation of astrocytes after focal cerebral infarction.”

absence[Introduction]

Limitations of prior research are not addressed

PASSReviewer 2· 75% conf

The paper cites prior work on Tregs, stroke, and ferroptosis, and provides a logical rationale linking Tregs to astrocyte activation and ferroptosis after cerebral infarction. However, it does not explicitly address limitations of prior research.

Evidence

paraphrase[Introduction, paragraphs 1–3]

“The introduction cites multiple studies on Treg accumulation in stroke, astrogliosis, and ferroptosis, establishing the premise.”

paraphrase[Introduction, final paragraph]

“The study aims to investigate the potential of Tregs in inhibiting activation of astrocytes after focal cerebral infarction, building on prior evidence.”

absence[Introduction]

Limitations of prior research are not explicitly addressed.

WARNReviewer 3· 90% conf

Prior work is cited extensively, but the logical rationale linking Tregs, astrocytes, ferroptosis, and renal dysfunction is fragmented, and limitations of cited prior work are not addressed.

Evidence

direct quote[Introduction, paragraphs 1–3]

“In stroke research accumulation of tregs during the two weeks has been observed.”

direct quote[Abstract, Introduction]

“This study aimed to investigate the potential of Tregs in inhibiting the activation of astrocytes after focal cerebral infarction.”

absence[Introduction, Discussion]

Limitations of prior work are not addressed.

The paper describes four experimental groups (Control, MCAO, DT+MCAO, DT) and a sham procedure, but does not mention randomization, blinding, sample size justification, or distinction between biological and technical replicates. Exclusion of 15 mice due to death is reported, but no pre-specified inclusion/exclusion criteria are given. No independent replication is mentioned. Only the control group is adequately described.

Sub-criteria

controlsADEQUATE

power analysisNOT REPORTED

blinding levelsNOT REPORTED

outlier handlingNOT REPORTED

randomization unitNOT REPORTED

inclusion exclusionINADEQUATE

randomization methodNOT REPORTED

replicate distinctionNOT REPORTED

independent replicationNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

The study design lacks essential rigor elements: no randomization, blinding, power analysis, or replication are reported; only controls are adequately described.

Evidence

direct quote[Methods, MCAO induction]

“For sham-operated mice, the same surgical procedures were performed, with the exception of filament insertion.”

direct quote[Methods, MCAO induction]

“A total of 66 mice were successfully induced with MCAO and included in further experiments. However, 15 mice died within 24 hours following reperfusion and were subsequently excluded from the study.”

absence[Methods]

No randomization, blinding, or power analysis reported

FAILReviewer 2· 90% conf

The study design lacks essential rigor elements: randomization method not stated, no blinding, no power analysis, no distinction of biological vs. technical replicates, and no pre-specified outlier handling. Only inclusion/exclusion criteria are partially reported.

Evidence

absence[Methods, section 2.4]

No randomization method is stated.

absence[Methods]

Blinding is not mentioned.

direct quote[Methods, section 2.4]

WARNReviewer 3· 85% conf

The study design includes multiple experimental components, but key elements like randomization, blinding, and power analysis are not reported for the animal experiments, and the computational component lacks dataset-specific inclusion/exclusion criteria.

Evidence

absence[Methods, section 2.4 MCAO induction]

No randomization or blinding method stated for animal experiments.

direct quote[Methods, section 2.4]

“the experimental groups were divided into four groups: Control group, MCAO group, Tregs Depletion +MCAO (DT+MCAO) group, and DT group.”

The paper reports age (9-10 weeks) and weight (25-30 g) for the mice, but does not specify sex, strain, vendor, or housing conditions (light cycle, temperature, enrichment). This makes interpretation and replication difficult. Health status is not mentioned.

Sub-criteria

demographicsN/A

sex reportedNOT REPORTED

sex justifiedNOT REPORTED

age weight healthINADEQUATE

housing conditionsNOT REPORTED

species strain sourceNOT REPORTED

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Biological variables are poorly reported: mouse sex, strain, source, and housing conditions are not provided; age and weight are given but health status is omitted.

Evidence

direct quote[Methods, Depletion of tregs]

“the mice (9-10 weeks old, weighing 25-30 g)”

absence[Methods]

Sex, strain, source, and housing not reported

FAILReviewer 2· 90% conf

The paper fails to report essential biological variables: sex of the mice is not stated, strain is not specified, housing conditions are not described, and health status is not reported. Age and weight are partially reported.

Evidence

absence[Methods, section 2.6]

Sex of mice is not reported.

direct quote[Methods, section 2.6]

“the mice (9-10 weeks old, weighing 25-30 g)”

absence[Methods]

Strain and source of mice are not reported.

WARNReviewer 3· 80% conf

Age, weight, and species are reported for the mice, but sex is not reported, and the sex of animals is not justified (single sex).

Evidence

direct quote[Methods, section 2.6]

“the mice (9-10 weeks old, weighing 25-30 g)”

absence[Methods, section 2.4, 2.6]

Sex of mice is not reported.

absence[Methods]

Housing conditions are not described.

The animal study is approved by the Clinical Ethical Committee of Xinhua Hospital (protocol number (2022) 189), which is reported_and_adequate. The regulatory compliance statement reads 'The studies were conducted in accordance with the local legislation and institutional requirements,' which does not name a recognized framework (e.g., Guide for the Care and Use of Laboratory Animals), so it is reported_but_inadequate. The scoring rule states that any applicable inadequate/not_reported results in warn.

Sub-criteria

iacuc statementADEQUATE

informed consentN/A

irb ethics statementN/A

regulatory complianceINADEQUATE

Per reviewer (3) — the votes behind the verdict

WARNReviewer 1· 90% conf

An animal ethics approval with a protocol number is provided, but regulatory compliance is stated only in vague terms, lacking a specific framework.

Evidence

direct quote[Ethics statement]

“The animal studies were approved by Clinical Ethical Committee of Xinhua Hospital Affiliated with Shanghai Jiao Tong University School of Medicine. (No. (2022) 189).”

direct quote[Ethics statement]

“The studies were conducted in accordance with the local legislation and institutional requirements.”

WARNReviewer 2· 95% conf

The paper provides a named ethics committee approval with a protocol number for the animal study, which is adequate. However, the regulatory compliance statement is vague and does not name a specific framework.

Evidence

direct quote[Ethics statement]

“The animal studies were approved by Clinical Ethical Committee of Xinhua Hospital Affiliated with Shanghai Jiao Tong University School of Medicine. (No. (2022) 189).”

direct quote[Ethics statement]

“The studies were conducted in accordance with the local legislation and institutional requirements.”

PASSReviewer 3· 95% conf

A named ethics committee approval for the animal studies is provided along with a protocol number, meeting requirements.

Evidence

direct quote[Ethics statement]

“The animal studies were approved by Clinical Ethical Committee of Xinhua Hospital Affiliated with Shanghai Jiao Tong University School of Medicine. (No. (2022) 189).”

Antibodies are listed with vendor and catalog numbers but no RRIDs or validation data. The mouse strain is not specified. ELISA kits are identified with catalog numbers but no lot numbers. Mycoplasma testing is not mentioned. GraphPad Prism and Cytoscape are named with versions but no vendor URLs.

Sub-criteria

mycoplasma testingNOT REPORTED

reagents identifiedINADEQUATE

organisms identifiedNOT REPORTED

antibodies identifiedINADEQUATE

cell line authenticationN/A

software tools identifiedINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Key resources are inadequately identified: antibodies lack RRID/validation, mouse strain and source are missing, and mycoplasma testing is not reported; software is named but without URLs.

Evidence

direct quote[Methods, Immunofluorescence]

“anti-2 (1:100, AiFang Biological, AF14722), anti-IL-10 (1:100, AiFang Biological, AF06580), anti-IL-2(1:100, AiFang Biological, AF02481), anti-IL-6 (1:100, AiFang Biological, AF06790), anti-FSP1 (1:100, AiFang Biological, AF14902), and anti-GPX4 (1:100, AiFang Biological, AF06580)”

absence[Methods]

Mouse strain not reported

direct quote[Methods, Statistical analysis]

“Statistical analyses were conducted using GraphPad Prism 9.0.”

FAILReviewer 2· 90% conf

Key resources are inadequately identified: antibodies are listed with catalog numbers but no RRIDs or validation data, the mouse strain is not specified, reagents lack lot numbers, and software versions are partially provided. No resource is fully identified.

Evidence

direct quote[Methods, section 2.5]

“Primary antibodies, including anti-2 (1:100, AiFang Biological, AF14722), anti-IL-10 (1:100, AiFang Biological, AF06580), anti-IL-2(1:100, AiFang Biological, AF02481), anti-IL-6 (1:100, AiFang Biological, AF06790), anti-FSP1 (1:100, AiFang Biological, AF14902), and anti-GPX4 (1:100, AiFang Biological, AF06580)”

absence[Methods, section 2.6]

Mouse strain is not reported.

direct quote[Methods, section 2.8]

“IL-1β: Cat. #AF-02323M2 China; NLRP3: Cat. #AF-02936M2, China; TNF-α: Cat. #AF-02415M2, China; FTL: Cat. #JL31443, China.”

WARNReviewer 3· 85% conf

Antibodies and reagents are listed with names and catalog numbers, but validation information is absent and software is incompletely identified.

Evidence

direct quote[Methods, section 2.5]

“anti-IL-10 (1:100, AiFang Biological, AF06580)”

direct quote[Methods, section 2.1, 2.10]

“Statistical analyses were conducted using GraphPad Prism 9.0.”

direct quote[Methods, section 2.6]

“the mice (9-10 weeks old, weighing 25-30 g)”

The methods section names the t-test, chi-square, and Kruskal-Wallis tests, but results do not specify which test was used for each comparison. Assumption testing is stated but not detailed. Only threshold p-values (e.g., **P<0.01) are given, not exact values. No effect sizes or confidence intervals are reported. Data presentation uses bar graphs with error bars but does not define the error bars (SD/SEM).

Sub-criteria

tests namedINADEQUATE

exact p valuesNOT REPORTED

effect sizes ciNOT REPORTED

data presentationINADEQUATE

software identifiedADEQUATE

assumptions verifiedINADEQUATE

mathematical plausibilityN/A

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Statistical reporting is inadequate: exact p-values, effect sizes, and confidence intervals are not reported; tests are named in methods but not specified per result; data presentation lacks error bar definitions.

Evidence

direct quote[Methods, Statistical analysis]

“The Student’s t-test and χ 2 test were used to analyze the normally distributed data... Non-normally distributed data was analyzed using the Kruskal-Wallis test”

direct quote[Results, Figure 4 caption]

“sample size was N=4 with significance set at **P<0.01”

absence[Results, Figures]

Error bars not defined

FAILReviewer 2· 90% conf

Statistical reporting is severely deficient: exact p-values are not reported, effect sizes and confidence intervals are absent, test assumptions are mentioned but not verified in detail, and data presentation lacks error bar definitions. Only software is identified adequately.

Evidence

direct quote[Results, Figure 4–6]

“with significance set at **P<0.01, ***P<0.005, and ****P<0.001”

direct quote[Methods, section 2.10]

“Statistical analyses were conducted using GraphPad Prism 9.0.”

absence[Figure legends]

Error bars are not defined.

WARNReviewer 3· 80% conf

Statistical tests are named and analytical software identified, but assumptions are only partially verified, exact p-values are not consistently reported (asterisks used), and data presentation is incomplete.

Evidence

direct quote[Methods, section 2.10]

“Normality and homogeneity of variance were tested, and appropriate adjustments for multiple comparisons were made using Bonferroni correction.”

direct quote[Figures 4-8 legends]

“with a sample size of N=4 and significance level of ***P<0.005, and ****P<0.001.”

direct quote[Figure 4 legend]

“The sample size was N=4 with significance set at **P<0.01.”

The data availability statement states that 'original contributions are included in the article/' (with a typo) and that inquiries can be directed to the corresponding author, which is a vague 'on request' statement. The GEO accession GSE221379 is provided for the human bioinformatics data, but the main mouse data are not deposited. No custom analysis code is shared.

Sub-criteria

code sharingNOT REPORTED

accession numbersNOT REPORTED

repository depositNOT REPORTED

data availability statementINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Data and code availability are poor: the data availability statement is vague, no repository deposit is made for the main mouse data, and no custom code is shared.

Evidence

direct quote[Data availability statement]

“The original contributions presented in the study are included in the article/. Further inquiries can be directed to the corresponding authors.”

absence[Data availability statement]

No repository deposit for mouse data

absence[Methods]

No code sharing

FAILReviewer 2· 95% conf

The data availability statement is vague ('available on reasonable request'), no data are deposited in a repository, no accession numbers are provided, and no custom code is shared. None of the applicable criteria are adequate.

Evidence

direct quote[Data availability statement]

“The original contributions presented in the study are included in the article/. Further inquiries can be directed to the corresponding authors.”

absence[Data availability statement]

No repository deposit or accession numbers.

WARNReviewer 3· 85% conf

The data availability statement is vague (no repository), no code is shared, and the public GEO data used does not have its downloaded accession verified as processed in the paper.

Evidence

direct quote[Data availability statement]

“The original contributions presented in the study are included in the article/. Further inquiries can be directed to the corresponding authors.”

direct quote[Methods, section 2.1]

“The transcriptome characteristics of the postmortem parietal cortex were compared between individuals who died from sepsis and non-septic critically ill individuals in the GEO database GSE221379.”

absence[Methods, Data availability statement]

No code repository is mentioned.

The methods are insufficient for replication, missing mouse strain, sex, and blinding. No ARRIVE or other reporting guidelines are referenced. The abstract and title claim that Treg depletion attenuates renal fibrosis, but no renal fibrosis results are presented in the main text. The discussion does not explicitly list limitations. Funding and conflicts of interest are adequately reported.

Sub-criteria

funding coiADEQUATE

trial registrationN/A

reporting guidelineNOT REPORTED

methods completenessINADEQUATE

all outcomes reportedNOT REPORTED

limitations discussedNOT REPORTED

conclusions proportionalINADEQUATE

Per reviewer (3) — the votes behind the verdict

FAILReviewer 1· 95% conf

Reporting is incomplete: methods lack key details, no reporting guideline is followed, renal fibrosis outcomes are not reported, and the conclusion is overstated.

Evidence

absence[Results]

Renal fibrosis outcome not reported

direct quote[Conclusion]

“Tregs depletion attenuates renal fibrosis by modulating IL-10/GPX4 following cerebral infarction.”

direct quote[Funding Statement and Conflict of Interest]

“Emergency Specialty - Department of Emergency Medicine (YXMZK005); General Discipline at TongRen Hospital (tr2023xk36). The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.”

FAILReviewer 2· 95% conf

Reporting is incomplete and inconsistent: methods lack critical details, no reporting guideline is followed, negative outcomes are not reported, limitations are discussed only superficially, and the main conclusion is unsupported by the presented data. Only funding and COI are adequate.

Evidence

absence[Methods]

Methods lack randomization, blinding, and housing details.

absence[Methods]

No reporting guideline is referenced.

direct quote[Abstract and Conclusion]

“Tregs depletion attenuates renal fibrosis by modulating IL-10/GPX4 following cerebral infarction.”

WARNReviewer 3· 85% conf

Methods are moderately detailed, and limitations and funding/COI are reported, but the study lacks a trial registration and a reporting guideline, and conclusions are partially overstated.

Evidence

direct quote[Discussion]

“the specific mechanism remains unclear.”

direct quote[Abstract, Results]

“Tregs depletion attenuates renal fibrosis by modulating IL-10/GPX4 following cerebral infarction.”

direct quote[Funding Statement, Conflict of Interest]

“The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.”

23 copyedit issues flagged (3 major): mostly clarity, consistency, grammar.

Checked 41 references: 41 verified.

Reproducibility

1 data/code link checked; 1 live.

Αlpha¹ Rigor Action ItemsFinal pass before journal submission

This paper is a post-publication audit, not a pre-submission review. An informed reader should weigh the unsupported claim of renal fibrosis (no kidney data), the several internal contradictions (abstract vs. results, human sepsis dataset applied to stroke), and the absence of basic methodological rigor (randomization, blinding, sample size justification); these issues would warrant a formal expression of concern or correction from the journal, and would make independent replication essential before the conclusions can be accepted.

1.
CRITICALreporting
Remove the claim 'Tregs depletion attenuates renal fibrosis' from the abstract and conclusion, or add a dedicated 'Renal outcomes' section presenting kidney histology and gene expression data that directly support the claim.
The claim is central to the paper's message but completely unsupported by any data in the results; an unfounded headline claim can invalidate the paper's conclusions.
2.
CRITICALreporting
Correct the internal contradiction in the abstract: change 'depletion of tregs protected against activation of astrocyte' to 'depletion of tregs promoted activation of astrocytes' to match the results (higher IL-2 and IL-6 in the DT group).
The abstract misrepresents the direction of the main finding, which would mislead readers and is a fundamental error.
3.
CRITICALreporting
Reconcile the sample-size discrepancy: the methods state 66 mice were induced, 15 died, leaving 51, but results show N=4 per group (16 mice total); explain where the remaining 35 mice were used or present data from all animals.
A 3-fold discrepancy between the number of animals used and the number reported in analyses undermines the validity of the statistical comparisons.
4.
CRITICALrigor
Report the randomization method (e.g., random number table) and the unit of randomization (individual mouse), and describe blinding procedures for surgery, data collection, and outcome assessment, in the MCAO induction Methods section.
Without randomization and blinding, the animal experiments are at high risk of bias, making the key results uninterpretable.
5.
CRITICALdata code
Replace the vague data availability statement ('included in the article/. Further inquiries can be directed to the corresponding authors.') with a specific repository (e.g., Figshare, Zenodo) that contains the raw and processed data for all experiments, and provide a DOI.
A vague 'on request' statement does not meet journal standards and prevents readers from verifying or re-analyzing the results.
6.
HIGHreporting
Add a sentence in the Introduction that explicitly addresses the limitations of prior research that the present study aims to overcome, and clarify the disconnected premise linking brain Tregs, a sepsis-derived bioinformatics dataset, and renal fibrosis.
The scientific premise is weakened by not explaining how the study fills gaps in prior work and by the unclear connection between its components.
7.
HIGHreporting
Report the exact p-values (e.g., p = 0.023) and effect sizes with confidence intervals for all main comparisons in the Results text and figure legends; remove exclusive reliance on asterisks.
Threshold-only p-values do not convey the precision of the estimates and are insufficient for reproducible research.
8.
HIGHreporting
Define error bars in every figure legend as standard deviation (SD) or standard error of the mean (SEM), and justify the choice.
Undefined error bars prevent readers from interpreting the variability and the appropriate statistical model.
9.
HIGHrigor
Specify the mouse sex, strain (e.g., C57BL/6J), vendor, and housing conditions (light cycle, temperature, enrichment) in the Methods section.
Sex, strain, and housing are essential for reproducibility and are required by ARRIVE guidelines; their absence limits the study's interpretability.
10.
HIGHrigor
Include an a priori power analysis or sample size calculation for the key comparisons (e.g., astrocyte activation markers) in the Statistical Analysis Methods section.
Without a power analysis, it is unclear whether the study had sufficient sensitivity to detect its claimed effects; N=4 per group is very small.
11.
MEDIUMcopyedit
Correct the typo 'subjecting the rats' to 'subjecting the mice' in Methods section 2.6 (the study uses mice, not rats).
Incorrect species labeling is a basic accuracy error that undermines credibility.
12.
MEDIUMcopyedit
Replace the ambiguous 'anti-2' with 'anti-IL-2' in Methods section 2.5 (antibody list).
The incomplete antibody name prevents readers from knowing which target was used.
13.
MEDIUMreporting
Add RRID numbers for all antibodies and key reagents (e.g., diphtheria toxin, PLX5622, CD25 antibody) in the Methods section; provide validation data (e.g., species reactivity) or a reference confirming specificity.
RRIDs and validation are increasingly required by journals to ensure that key biological resources are uniquely and unambiguously identified.
14.
MEDIUMreporting
Clarify the bioinformatics component: justify why a sepsis dataset (GSE221379) was used to model stroke-related brain changes, and add a limitation explaining this disconnect.
Using a sepsis dataset to draw conclusions about stroke is a major logical gap that must be addressed in the Discussion.
15.
LOWcopyedit
Ensure consistent capitalization of 'Tregs' (not 'tregs') throughout the manuscript, and fix subject-verb agreement errors (e.g., 'Tregs play' not 'Tregs plays' in the Abstract).
Basic grammatical and typographical errors detract from the paper's professionalism and may signal carelessness to reviewers.
16.
LOWcopyedit
Fix the typo in the data availability statement: 'included in the article/' should be 'included in the article' or 'included in the article/Supplementary Material'.
The trailing slash is a formatting error that looks sloppy and incomplete.

Full text (30,810 chars)Analyzed: Jul 6, 2026

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