好的，根据您的需求，我将以转化医学研究期刊审稿人的身份，从后续正式撰写学术论文的角度，对您这份实验报告的设计和内容进行评估，并指出需要补充的资料或检测指标。 首先，针对您的核心需求——**“从后续文章撰写的角度看，这份实验报告写得好不好，需要补充什么资料或者检测指标”**，我的总体评估如下： 这份研究报告的设计思路清晰，科学假说明确，体现了非常好的转化医学研究潜质。它成功地构建了一个从临床观察（AR患者菌群失调、丁酸水平下降）到动物模型验证（丁酸干预改善AR），再到体外细胞实验揭示分子机制（HDAC抑制-Treg分化）的完整证据链。这是一个非常扎实的研究框架，具备冲击高水平期刊的潜力。 然而，要将这份报告转化为一篇具有高影响力的学术论文，以下几个关键环节需要补充和深化，以增强研究的严谨性和结论的说服力： **需要补充的关键实验和检测指标：** 1. **强化“肠-鼻轴”的直接证据 (Strengthen the Gut-Nose Axis Link):** * **核心问题:** 目前研究证明了系统性（血液）丁酸水平与鼻腔免疫状态相关，但缺乏丁酸直接作用于鼻腔黏膜的证据。 * **补充建议:** 在动物实验中，增加检测**鼻腔灌洗液或鼻黏膜组织匀浆中的丁酸浓度**。这将提供丁酸通过循环系统到达并富集在鼻腔病变部位的直接证据，是连接“肠”和“鼻”的关键一环。 2. **深化Treg细胞的功能与归巢机制 (Deepen Treg Homing and Function Mechanisms):** * **核心问题:** 报告中提到“功能受损的Treg无法有效归巢至鼻黏膜”，但这只是一个推论，缺乏实验支持。 * **补充建议:** * **归巢能力检测:** 采用流式细胞术检测外周血和鼻黏膜Treg细胞表面**趋化因子受体**的表达，例如**CCR4、CCR8**等与黏膜归巢相关的分子。比较AR患者与健康人、模型小鼠与丁酸干预小鼠之间这些受体的表达差异。 * **Treg抑制功能验证:** 从小鼠模型中分选出Treg细胞，进行**体外抑制实验**。即将Treg细胞与效应T细胞（如Th2细胞）共培养，观察Treg对效应T细胞增殖和分泌细胞因子（IL-4, IL-5）的抑制能力。这将直接证明丁酸干预后Treg的“功能”确实得到了增强。 3. **优化动物模型分组与因果关系论证 (Refine Animal Model for Causality):** * **核心问题:** 抗生素处理组和FMT组的预期结果（“无明显改善，甚至加重”）逻辑上不够清晰，未能强有力地支持“菌群失调是AR的原因”。 * **补充建议:** * **重设抗生素组实验:** 建议增加一个**“抗生素+丁酸补充”**的救援组（Rescue group）。如果抗生素处理加重了AR症状，而补充丁酸能够逆转这种恶化，这将更有力地证明肠道菌群是通过丁酸来发挥保护作用的。 * **明确FMT组的假设:** 将AR患者的粪菌移植给无菌小鼠或抗生素处理后的小鼠，预期结果应该是**诱导或加重AR样症状**，而不是“无明显改善”。这才能证明AR患者的失调菌群具有“致病性”。 4. **增强机制研究的特异性 (Enhance Specificity of the Mechanism):** * **核心问题:** 证明了丁酸是HDAC抑制剂，但如何证明丁酸对Treg的调控作用*必须*通过HDAC通路？ * **补充建议:** 在体外T细胞分化实验中，增加一个**HDAC激动剂**（或另一种非丁酸的HDAC抑制剂作为阳性对照）组。如果HDAC激动剂能够阻断丁酸对Treg的促进作用，则可以更牢固地确立丁酸-HDAC-Foxp3这条信号轴的特异性。 总结来说，您目前的研究框架非常出色。如果能补充上述四个方向的实验数据，您的研究将不仅仅是揭示一种现象，而是能够清晰地阐明从肠道菌群到鼻腔炎症的完整因果链条和分子机制，这将极大提升文章的科学价值和影响力。 ---

Reviewer Report

### **Phase 1: Target Journal Analysis** Based on the manuscript's design, which integrates clinical samples, animal models, and in vitro mechanistic studies, the work is positioned as translational research. A suitable target journal would be one that values studies elucidating disease mechanisms with clear clinical relevance, such as the *Journal of Allergy and Clinical Immunology*, *Mucosal Immunology*, or *Science Translational Medicine*. These journals typically prefer: * **Novelty:** The "gut-nose axis" in allergic rhinitis (AR) is a timely and novel concept. While the butyrate-Treg-HDAC link is known in other contexts (e.g., gut immunity), its specific application to the nasal microenvironment in AR provides sufficient novelty. * **Mechanistic Depth:** They require a robust and specific mechanism. The current proposal linking butyrate to Treg differentiation via HDAC inhibition is strong but would be significantly enhanced by experiments confirming causality and specificity (e.g., rescue experiments, validation of Treg homing). * **Translational Significance:** A clear "bench-to-bedside" narrative is essential. The study design strongly supports this, but direct evidence of the metabolite (butyrate) acting at the site of pathology (nasal mucosa) is a critical piece of data that is currently missing. ### **Phase 2: Peer Review Report** **To**: The Editor **From**: Senior Reviewer **Manuscript ID**: [Hypothetical ID: JTM-2025-11-04-001] **Manuscript Title**: Mechanism of the gut microbiota metabolite butyrate in regulating allergic rhinitis by remodeling the nasal immune microenvironment **Date**: 2025-11-04 #### **Part 1: Summary for the Editor & Overall Recommendation** This manuscript investigates the role of the gut microbiota metabolite butyrate in the pathogenesis of allergic rhinitis (AR) via the "gut-nose axis". The authors propose a comprehensive study combining clinical data, a murine AR model, and in vitro experiments. The central hypothesis is that gut dysbiosis in AR patients leads to reduced systemic butyrate, which impairs the differentiation and function of regulatory T cells (Tregs) through HDAC inhibition, ultimately failing to suppress Th2/ILC2-mediated inflammation in the nasal mucosa. The study design is logical, ambitious, and addresses a significant knowledge gap in the field of allergic diseases. The proposed use of modern techniques like 16S rRNA sequencing, metabolomics, and single-cell RNA sequencing is appropriate and powerful. If the expected results are obtained, the findings would provide a novel mechanistic link between gut health and respiratory allergy, offering a strong rationale for microbiota-based therapies. However, the manuscript, in its current form as a research plan with expected results, contains several conceptual gaps and methodological weaknesses that must be addressed to substantiate its main conclusions. The link between systemic butyrate and its local action in the nose is correlational, the mechanism of Treg homing is speculative, and the design of the animal model control groups (antibiotic and FMT) is not robust enough to establish clear causality. Therefore, my recommendation is **Major Revision**. The study has high potential, but requires substantial additional experimental evidence to strengthen its key claims before it can be considered for publication. #### **Part 2: Detailed Review Comments** **1. Title** The title is accurate and informative, clearly stating the subject, intervention, and mechanism. It is appropriate for the scope of the study. **2. Abstract** An abstract was not formally provided, but based on the report, a structured abstract for the final manuscript should concisely summarize the background, methods, key quantitative findings (e.g., fold-change, p-values), and the main conclusion regarding the butyrate-Treg-HDAC axis in AR. **3. Introduction** The introduction provides a clear and well-referenced background on AR, the gut-organ axis, and the preliminary evidence linking butyrate to AR. The scientific question and hypothesis are well-defined. **4. Methods** * **(Major Concern)** The justification for the clinical sample size (n=60/group) is missing. A power analysis should be conducted and reported in the manuscript to ensure the study is adequately powered to detect the expected differences. * **(Major Concern)** The design of the animal model control groups requires significant revision. * The antibiotic treatment group is intended to deplete gut microbiota. The expected result of "no obvious improvement, or even worsened" is vague. A clearer hypothesis would be that microbiota depletion exacerbates AR symptoms. Crucially, a rescue group (antibiotics + butyrate) should be included to demonstrate that the effects of microbiota depletion are mediated specifically by the loss of butyrate. * The Fecal Microbiota Transplantation (FMT) group using feces from AR patients should be designed to test for causality. The expected outcome should be the induction or exacerbation of AR-like symptoms in recipient mice, not simply "no obvious improvement". This would provide evidence that the AR-associated microbiota is pro-inflammatory. * **(Minor Concern)** The methods section should provide more specific details for reproducibility, such as catalog numbers and dilutions for antibodies, specific qPCR primer sequences, and detailed parameters for the bioinformatic analysis of sequencing data. **5. Results** The manuscript presents "expected results," which are logical but require actual data for evaluation. The following points are critical for the final manuscript: * **(Major Concern)** A crucial piece of evidence is missing: the direct measurement of butyrate concentration in the nasal mucosa or nasal lavage fluid of the mice. This is essential to prove that systemically administered or gut-derived butyrate reaches the target organ at a physiologically relevant concentration. Without this data, the "gut-nose axis" remains an indirect association. * **(Major Concern)** The manuscript claims that Tregs "fail to home effectively to the nasal mucosa." This is a key part of the hypothesis but is not supported by any proposed experiment. To validate this, the authors must analyze the expression of homing receptors (e.g., CCR4, CCR8) on Tregs from both peripheral blood and nasal tissue. * **(Minor Concern)** The authors should plan to include in vitro Treg suppression assays. This would involve co-culturing Tregs isolated from control vs. butyrate-treated mice with effector T cells to functionally confirm that butyrate enhances the suppressive capacity of Tregs. **6. Discussion** * The planned discussion points are relevant. However, the interpretation of the results will depend heavily on the additional data requested above. * **(Major Concern)** The limitations section correctly identifies the sample size and animal model fidelity as potential issues. However, it understates the most significant limitation: the lack of direct evidence for butyrate's local presence and action in the nasal microenvironment. This point should be explicitly discussed. **7. References** The use of PMIDs indicates that the literature review is based on published work. The final manuscript should ensure all references are up-to-date and formatted according to the journal's guidelines. **8. Figures & Tables** Not provided. High-quality figures (e.g., flow cytometry plots with gating strategies, histology images with scale bars, scRNA-seq UMAP plots, and summary graphs with clear statistical annotations) will be essential for the final manuscript. **9. Point-by-point comments on key conceptual gaps** * **Section 1.3 (Scientific Hypothesis):** The hypothesis mentions Tregs failing to home. This must be experimentally tested. * **Section 2.3.2 (Animal experiment):** The antibiotic and FMT group designs are not optimal for proving causality. They should be redesigned as suggested in the "Methods" section review. * **Section 3.2 (Animal experiment expected results):** The link between systemic intervention (butyrate gavage) and local effect (nasal immunity) needs to be bridged by measuring local butyrate concentrations. * **Section 3.3 (In vitro mechanism):** To confirm the specificity of the HDAC pathway, an experiment using an HDAC activator to counteract butyrate's effect, or another HDAC inhibitor as a positive control, would greatly strengthen the conclusion. **10. Consistency/Contradictions** The document is internally consistent as a research proposal. No contradictions were noted. #### **Part 3: Recommended journals**

Journal Name	Impact Factor (2024)	CAS Classification	JCR Classification	Review Period	Recommended Reason
Allergy	~12.4	Medicine, Q1 Top	Q1	~4-6 weeks	Top journal in the field of allergy. A good fit if the additional mechanistic data (especially Treg homing and function) is provided.
Mucosal Immunology	~7.3	Immunology, Q1 Top	Q1	~4-8 weeks	Highly relevant as the study focuses on the nasal mucosal immune microenvironment and the gut-organ axis.
Frontiers in Immunology	~6.4	Immunology, Q1	Q1	~8-12 weeks	A well-respected open-access journal with a broad scope in immunology. Suitable for this comprehensive study.
Journal of Leukocyte Biology	~4.5	Immunology, Q2	Q2	~4-6 weeks	Focuses on the biology of immune cells, making it a good venue for the detailed analysis of Tregs, Th2, and ILC2s.
Clinical & Experimental Allergy	~5.0	Medicine, Q2	Q1	~6-8 weeks	Strong focus on translational research in allergy, aligning well with the study's design.
Microbiome	~15.5	Microbiology, Q1 Top	Q1	~6-10 weeks	An ambitious target, but suitable if the microbiota's causal role is robustly proven with the revised FMT and antibiotic experiments.

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