IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-06-06 DOI: 10.1016/j.ymeth.2025.06.002

Satyajit Ghosh , Surojit Ghosh , Aniket Jana , Rajsekhar Roy , Surajit Ghosh

Exosomes, small extracellular vesicles originating from endocytic processes, have garnered increasing attention due to their roles in both physiological functions and pathological conditions. Initially identified in the 1980 s, exosomes are formed within multivesicular bodies (MVBs) through the invagination of the endosomal membrane, leading to the creation of intraluminal vesicles (ILVs). These ILVs can either be degraded by lysosomes or released into the extracellular space as exosomes, facilitating intercellular communication. In the nervous system, exosomes are implicated in various functions, including neural development and the progression of neurodegenerative diseases such as Alzheimer’s and Parkinson’s disease. This study presents a novel protocol for the isolation and proteomic analysis of exosomes derived from the substantia nigra (SN) of rat brains. By employing a combination of differential centrifugation and immunocapture techniques, we achieved a purer exosome fraction and higher exosome yield compared to traditional ultracentrifugation methods. Our proteomics analysis identified 51, 48, and 70 proteins from three distinct exosome samples (SN-EV-1, SN-EV-2, and SN-EV-4), with Gene Ontology annotation revealing their involvement in diverse biological functions. This research not only establishes a reliable method for isolating brain-derived exosomes but also sets the stage for comparative studies between healthy and neurodegenerative conditions. Ultimately, our findings aim to enhance the understanding of exosomal roles in disease mechanisms and contribute to the identification of potential biomarkers and therapeutic targets for neurodegenerative disorders.

外泌体是起源于内吞过程的细胞外小泡，由于其在生理功能和病理条件中的作用而受到越来越多的关注。外泌体最初是在20世纪80年代被发现的，它通过内陷内体膜在多泡体（MVBs）内形成，导致腔内囊泡（ILVs）的产生。这些ilv可以被溶酶体降解或作为外泌体释放到细胞外空间，促进细胞间的通讯。在神经系统中，外泌体参与多种功能，包括神经发育和神经退行性疾病（如阿尔茨海默病和帕金森病）的进展。本研究提出了大鼠脑黑质外泌体的分离和蛋白质组学分析的新方案。通过结合差速离心和免疫捕获技术，与传统的超离心方法相比，我们获得了更纯净的外泌体部分和更高的外泌体产量。我们的蛋白质组学分析从三个不同的外泌体样本（SN-EV-1、SN-EV-2和SN-EV-4）中鉴定了51、48和70个蛋白质，并通过基因本体注释揭示了它们参与多种生物学功能。这项研究不仅建立了一种可靠的分离脑源性外泌体的方法，而且为健康和神经退行性疾病之间的比较研究奠定了基础。最终，我们的研究结果旨在增强对外泌体在疾病机制中的作用的理解，并有助于识别神经退行性疾病的潜在生物标志物和治疗靶点。

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引用次数: 0

cfDNAFE: Comprehensively extracting multi-omics features of cell-free DNA for noninvasive diagnosis cfDNAFE：综合提取游离DNA的多组学特征，用于无创诊断。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-06-03 DOI: 10.1016/j.ymeth.2025.05.013

Wanxin Cui , Junjie You , Wenlong Jie , Zihao Li , Xiaoqing Peng

The tissues-of-origin of circulating cell-free DNA (cfDNA) holds great promise for non-invasive diagnosing cancers, monitoring allograft rejection, and prenatal testing. Many features for inferring the tissues-of-origin of cfDNAs are being revealed from different angles, including genetics, epigenetics, and fragmentomics, with whole-genome sequencing (WGS) and whole-genome bisulfite sequencing (WGBS) data of cfDNA. However, it lacks integrative toolkits for automatically extracting the revealed features from the WGS and WGBS data of cfDNA samples. Here, we propose cfDNAFE, a comprehensive and easy-to-use python package for extracting multi-omics features from the aligned cfDNA sequencing data. It covers three aspects: cfDNA genetic features, cfDNA methylation features, and cfDNA fragmentation features, including 13 types of feature profiles. The genetic features include substitution mutations, mutation signatures and copy number variations. The methylation features are the proportions of methylated fragments, unmethylated fragments, and mixed methylated fragments on cell-type-specific markers. The fragmentation features related to the fragment sizes, end/breakpoint motifs, and nucleosome positions are also integrated. To verify the functions of cfDNAFE, we perform analysis on the WGS/WGBS data of cfDNA samples based on the feature profiles extracted by cfDNAFE. The comparison between the cfDNA samples of hepatocellular carcinoma (HCC) patients and normal controls suggests HCC cfDNA samples exhibit significant difference in fragment size related features and breakpoint/end motif patterns, and obtain significant higher OCF values in the liver-specific open regions than the health controls. Conclusively, cfDNAFE is a most comprehensive toolkit which covers the most features for inferring the tissues-of-origin of cfDNAs in existing studies up to date. It will facilitate researchers to build machine learning models for auxiliary diagnosis based on these features. Availability and implementation: https://github.com/Cuiwanxin1998/cfDNAFE.

循环无细胞DNA （cfDNA）的起源组织在非侵入性诊断癌症、监测同种异体移植排斥反应和产前检测方面具有很大的前景。利用cfDNA的全基因组测序（WGS）和亚硫酸盐全基因组测序（WGBS）数据，从遗传学、表观遗传学和片段组学等不同角度揭示了推断cfDNA起源组织的许多特征。然而，目前还缺乏从cfDNA样本的WGS和WGBS数据中自动提取揭示特征的集成工具。在这里，我们提出cfDNAFE，一个全面且易于使用的python包，用于从校准的cfDNA测序数据中提取多组学特征。涵盖cfDNA遗传特征、cfDNA甲基化特征和cfDNA片段化特征三个方面，包括13种类型的特征谱。遗传特征包括替代突变、突变特征和拷贝数变异。甲基化特征是甲基化片段、非甲基化片段和混合甲基化片段在细胞类型特异性标记上的比例。还集成了与片段大小、末端/断点基序和核小体位置相关的片段特征。为了验证cfDNAFE的功能，我们基于cfDNAFE提取的特征轮廓对cfDNA样本的WGS/WGBS数据进行了分析。肝细胞癌（HCC）患者cfDNA样本与正常对照的比较表明，HCC cfDNA样本在片段大小相关特征和断点/末端基序模式上存在显著差异，并且在肝脏特异性开放区域获得显著高于健康对照的OCF值。最后，cfDNAFE是一个最全面的工具包，涵盖了迄今为止现有研究中推断cfdna起源组织的大多数特征。这将有助于研究人员基于这些特征构建辅助诊断的机器学习模型。可用性和实现：https://github.com/Cuiwanxin1998/cfDNAFE。

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引用次数: 0

Label-free spectral confocal reflectance microscopy for ex vivo neuroimaging and neural structure visualization 无标记光谱共聚焦反射显微镜用于离体神经成像和神经结构可视化。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-06-02 DOI: 10.1016/j.ymeth.2025.06.001

Reinher Pimentel-Domínguez , Rainald Pablo Ordaz , Abraham J. Cisneros-Mejorado , Rogelio O. Arellano , Remy Avila

Confocal microscopy is an essential technique in the field of life sciences, commonly used to study molecules in a variety of preparations, ranging from cells and tissues to entire organisms. In the field of neuroscience, it is a widely utilized tool for both anatomical-structural and functional studies. However, a lesser-known application of this microscopy method is confocal reflectance microscopy, which involves image acquisition based on reflected light from the sample. In this study, we present the use of spectral confocal reflectance microscopy (SCoRe) to explore multiple structures in the rat brain, without the use of any dyes or immunolabeling. Our results demonstrate that this technique allows for the distinction between different brain structures with high spatial resolution, enabling the observation of fibers in the cerebral cortex, corpus callosum, hippocampus, cerebellum, and other regions. These findings highlight the ability of SCoRe to provide detailed anatomical insights that are comparable to those obtained through conventional methods. Additionally, we have shown that SCoRe is compatible with samples prepared using traditional techniques, such as histochemical staining and immunofluorescence. This research emphasizes the value of SCoRe as a cost-effective and label-free method for high-resolution brain imaging, which can improve neuroscience studies and reduce long-term expenses.

共聚焦显微镜是生命科学领域的一项重要技术，通常用于研究从细胞和组织到整个生物体的各种制剂中的分子。在神经科学领域，它是一种广泛应用于解剖结构和功能研究的工具。然而，这种显微镜方法的一个鲜为人知的应用是共聚焦反射显微镜，它涉及基于样品反射光的图像采集。在这项研究中，我们提出使用光谱共聚焦反射显微镜（SCoRe）来探索大鼠脑中的多种结构，而不使用任何染料或免疫标记。我们的研究结果表明，该技术能够以高空间分辨率区分不同的大脑结构，从而能够观察到大脑皮层、胼胝体、海马、小脑和其他区域的纤维。这些发现突出了SCoRe提供与传统方法相当的详细解剖信息的能力。此外，我们已经证明SCoRe与使用传统技术（如组织化学染色和免疫荧光）制备的样品兼容。本研究强调SCoRe作为高分辨率脑成像的成本效益和无标签方法的价值，它可以改善神经科学研究并减少长期费用。

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引用次数: 0

Development and interpretation of a pathomics-based model for the prediction of immune therapy response in colorectal cancer 基于病理的结直肠癌免疫治疗反应预测模型的发展和解释。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-31 DOI: 10.1016/j.ymeth.2025.05.012

Yan Luo , Qiang Tian , Lei Xu , Dongmei Zeng , Hua Zhang , Tianyu Zeng , Hua Tang , Chao Wang , Yihua Chen

Colorectal cancer (CRC) is the third most common malignancy and the second leading cause of cancer-related deaths worldwide, with a 5-year survival rate below 20 %. Immunotherapy, particularly immune checkpoint blockade (ICB)-based therapies, has become an important approach for CRC treatment. However, only specific patient subsets demonstrate significant clinical benefits. Although the TIDE algorithm can predict immunotherapy responses, the reliance on transcriptome sequencing data limits its clinical applicability. Recent advances in artificial intelligence and computational pathology provide new avenues for medical image analysis. In this study, we classified TCGA-CRC samples into immunotherapy responder and non-responder groups using the TIDE algorithm. Further, a pathomics model based on convolutional neural networks was constructed to directly predict immunotherapy responses from histopathological images. Single-cell analysis revealed that fibroblasts may induce immunotherapy resistance in CRC through collagen-CD44 and ITGA1 + ITGB1 signaling axes. The developed pathomics model demonstrated excellent classification performance in the test set, with an AUC of 0.88 at the patch level and 0.85 at the patient level. Moreover, key pathomics features were identified through SHAP analysis. This innovative predictive tool provides a novel method for clinical decision-making in CRC immunotherapy, with potential to optimize treatment strategies and advance precision medicine.

结直肠癌（CRC）是全球第三大最常见的恶性肿瘤，也是癌症相关死亡的第二大原因，其5年生存率低于20% %。免疫治疗，特别是基于免疫检查点阻断（ICB）的治疗，已成为结直肠癌治疗的重要途径。然而，只有特定的患者亚群显示出显著的临床益处。尽管TIDE算法可以预测免疫治疗反应，但对转录组测序数据的依赖限制了其临床适用性。人工智能和计算病理学的最新进展为医学图像分析提供了新的途径。在本研究中，我们使用TIDE算法将TCGA-CRC样本分为免疫治疗应答组和无应答组。此外，构建了基于卷积神经网络的病理模型，直接预测组织病理图像的免疫治疗反应。单细胞分析显示成纤维细胞可能通过胶原- cd44和ITGA1 + ITGB1信号轴诱导结直肠癌免疫治疗耐药。所建立的病理模型在测试集中表现出优异的分类性能，在贴片水平上的AUC为0.88，在患者水平上的AUC为0.85。此外，通过SHAP分析确定了关键的病理特征。这一创新的预测工具为CRC免疫治疗的临床决策提供了一种新的方法，具有优化治疗策略和推进精准医学的潜力。

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引用次数: 0

Metabolic impacts of long-chain fatty acids on cardiomyocyte maturation in neonatal mammalian hearts 长链脂肪酸对新生哺乳动物心脏心肌细胞成熟的代谢影响

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-29 DOI: 10.1016/j.ymeth.2025.05.010

Seong-Eung Cha , Mi Nam Lee , Eung-Sam Kim

Cardiomyocytes are essential models for cardiac disease modeling, drug development, and regenerative therapies. Specifically, human-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) have emerged as widely used cellular models with high reproducibility. However, cardiomyocytes generated in vitro tend to remain immature and insufficient in replicating the electrophysiological and mechanical functions of adult cardiomyocytes, limiting the clinical and experimental applications of these models. Thus, various biochemical and biophysical strategies have been explored to promote the maturation of cardiomyocytes, to address these limitations, and more accurately mimic the characteristics of mature cardiomyocytes. This review summarizes recent studies on multiple methodologies employed to induce cardiomyocyte maturation, with a particular emphasis on the role of long-chain fatty acids (LCFAs). The evidence summarized in this review is derived from studies utilizing cardiomyocytes from neonatal mice or rats and hiPSC-CMs. Meanwhile, immature cardiomyocytes have been demonstrated to predominantly rely on glycolysis, transitioning to oxidative phosphorylation through maturation, which enhances electrical stability, contractility, and structural organization. LCFAs play a key role in the cardiomyocyte maturation process by serving as key metabolic factors that generate ATP through mitochondrial β-oxidation, thereby improving metabolic efficiency. Additionally, LCFAs are involved in activating cytoskeletal components and signaling pathways integral to cardiomyocyte contractility. Importantly, studies suggest that when multiple biochemical and biophysical stimuli are simultaneously applied, various aspects of cardiomyocyte maturation are synergistically accelerated. Therefore, future studies focusing on the coordinated application of these regulatory factors are expected to enhance the maturation process, ultimately contributing to the generation of mature cardiomyocytes suitable for regenerative medicine and other advanced applications.

心肌细胞是心脏病建模、药物开发和再生治疗的重要模型。具体来说，人类诱导的多能干细胞衍生的心肌细胞（hiPSC-CMs）已经成为广泛使用的具有高重复性的细胞模型。然而，体外生成的心肌细胞在复制成人心肌细胞的电生理和机械功能方面往往仍然不成熟，不足，限制了这些模型的临床和实验应用。因此，人们探索了各种生物化学和生物物理策略来促进心肌细胞的成熟，以解决这些限制，并更准确地模拟成熟心肌细胞的特征。本文综述了近年来用于诱导心肌细胞成熟的多种方法的研究，特别强调了长链脂肪酸（LCFAs）的作用。本综述总结的证据来自利用新生小鼠或大鼠心肌细胞和hiPSC-CMs的研究。同时，未成熟心肌细胞已被证明主要依赖于糖酵解，通过成熟过渡到氧化磷酸化，从而增强电稳定性、收缩性和结构组织。LCFAs作为关键代谢因子，通过线粒体β-氧化产生ATP，从而提高代谢效率，在心肌细胞成熟过程中发挥关键作用。此外，LCFAs还参与激活细胞骨架成分和心肌细胞收缩性不可或缺的信号通路。重要的是，研究表明，当多种生化和生物物理刺激同时应用时，心肌细胞成熟的各个方面协同加速。因此，未来的研究重点是这些调节因子的协同应用，有望促进成熟过程，最终有助于生成适合再生医学和其他高级应用的成熟心肌细胞。

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引用次数: 0

Development and validation of the Azu-Eos as a fluorescence-free staining method for enhanced bright-field microscopy in the comet assay Azu-Eos作为一种无荧光染色方法的开发和验证，用于增强的彗星分析中的亮场显微镜。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-27 DOI: 10.1016/j.ymeth.2025.05.011

Penyo Ivanov, Dessislava Staneva, Milena Georgieva, George Miloshev

The comet assay is a valuable technique for assessing cellular DNA damage due to intrinsic and environmental factors, particularly in studies of genotoxicity and mutagenesis. However, its broader application is limited by the need for fluorescent staining, the requirement for expensive fluorescent microscopes, and the inability to preserve and re-examine slides over time. We developed the Azure B-Eosin Y (Azu-Eos) staining method to visualise the comets under a standard bright-field microscope to overcome these limitations. The newly developed DNA-staining technique eliminates the need for fluorescence and allows unlimited slide storage and re-examination without quality deterioration. We optimised various factors to ensure the reliability and quality of bright-field stained comets, achieving results comparable to and even better than those obtained with traditional fluorescent dyes. This cost-effective, sensitive, and fluorescence-free method has broad potential applications in medicine, ecology, and environmental monitoring, enabling fast and on-site genotoxicity testing.

彗星分析是一种有价值的技术，用于评估由于内在和环境因素引起的细胞DNA损伤，特别是在遗传毒性和诱变研究中。然而，由于需要荧光染色，需要昂贵的荧光显微镜，并且无法保存和重新检查载玻片，因此其更广泛的应用受到限制。为了克服这些限制，我们开发了Azure B-Eosin Y （Azu-Eos）染色方法，在标准亮场显微镜下观察彗星。新开发的dna染色技术消除了对荧光的需要，允许无限的切片储存和重新检查，而不会导致质量下降。我们优化了各种因素，以确保亮场染色彗星的可靠性和质量，取得了与传统荧光染料相当甚至更好的结果。这种低成本、灵敏、无荧光的方法在医学、生态和环境监测方面具有广泛的潜在应用，可以实现快速和现场的遗传毒性检测。

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引用次数: 0

Surface-enhanced Raman scattering (SERS) technology: Emerging applications in cancer imaging and precision medicine 表面增强拉曼散射（SERS）技术：在癌症成像和精准医学中的新兴应用。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-21 DOI: 10.1016/j.ymeth.2025.05.009

Biqing Chen, Jiayin Gao, Haizhu Sun, Zhi Chen, Xiaohong Qiu

Recent advancements in Surface-enhanced Raman Scattering (SERS) bioprobes have substantially enhanced their bioimaging capabilities for disease theranostics. This review systematically analyzes three categories of engineered SERS probes: noble metal nanostructures, metal oxide hybrids, and multifunctional composite materials, emphasizing their optimized designs for targeted tumor detection and image-guided surgery. Key developments include improved in vitro biosensing platforms for rapid tumor screening and advanced in vivo probes enabling real-time intraoperative imaging with molecular specificity. The integration of SERS with complementary modalities (fluorescence, photoacoustic, MRI) is critically examined as a strategy to overcome individual technical limitations and achieve multiscale tissue characterization. Technical progress in spatial resolution enhancement, multiplex biomarker detection, and biocompatibility optimization is quantitatively highlighted. Current challenges in signal consistency across biological systems and scalable probe manufacturing are discussed, proposing standardized evaluation frameworks as essential for clinical translation. This work establishes SERS as a multimodal imaging cornerstone for precision oncology, particularly in tumor margin delineation and metastatic lesion identification.

表面增强拉曼散射（SERS）生物探针的最新进展大大增强了其在疾病治疗中的生物成像能力。本文系统分析了三种工程SERS探针：贵金属纳米结构、金属氧化物杂化物和多功能复合材料，重点介绍了它们在靶向肿瘤检测和图像引导手术中的优化设计。主要发展包括改进的体外生物传感平台，用于快速肿瘤筛查和先进的体内探针，实现具有分子特异性的实时术中成像。SERS与互补模式（荧光，光声，MRI）的整合被严格检查为克服个人技术限制和实现多尺度组织表征的策略。在空间分辨率增强、多重生物标志物检测和生物相容性优化等方面的技术进步得到了定量的强调。讨论了生物系统和可扩展探针制造中信号一致性的当前挑战，提出了标准化评估框架，作为临床翻译的必要条件。这项工作建立了SERS作为精确肿瘤学的多模态成像基石，特别是在肿瘤边缘划定和转移灶识别方面。

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引用次数: 0

Enhancing RNA Delivery: Practical insights into NeoLNPTM transfection reagent 增强RNA传递：对NeoLNPTM转染试剂的实际见解

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-21 DOI: 10.1016/j.ymeth.2025.05.007

Xiaobao Chen , Yan Wu , Li Liu , Wei Wang

The rapid advancement of RNA-based therapeutics, particularly in the wake of COVID-19 vaccine success, has prompted significant research into optimizing RNA delivery mechanisms. This study evaluates the NeoLNP^TM RNA Transfection Kit developed by Scindy Pharmaceutical, which utilizes lipid nanoparticles (LNPs) for efficient RNA encapsulation and delivery. We systematically investigate various parameters affecting transfection efficiency, including RNA concentration, RNA/LNP volume ratios, mixing techniques, LNP stability, and culture media. Our results demonstrate that the optimal RNA concentration for transfection efficiency is around 40–60 ng/µL, with a 1:0.75–1:1 RNA-to-LNP ratio yielding the highest protein expression. Additionally, we find that gentle mixing techniques outperform harsher methods, and the stability of LNP-RNA complexes significantly influences transfection outcomes. This research provides practical guidelines for enhancing RNA transfection efficiency, paving the way for more effective RNA therapeutics.

基于RNA的治疗方法的快速发展，特别是在COVID-19疫苗成功之后，促进了对优化RNA递送机制的重要研究。本研究评估了由Scindy制药公司开发的NeoLNPTM RNA转染试剂盒，该试剂盒利用脂质纳米颗粒（LNPs）进行有效的RNA封装和传递。我们系统地研究了影响转染效率的各种参数，包括RNA浓度、RNA/LNP体积比、混合技术、LNP稳定性和培养基。我们的研究结果表明，转染效率的最佳RNA浓度约为40-60 ng/µL， RNA与lnp的比例为1:0.75-1:1，蛋白表达量最高。此外，我们发现温和混合技术优于苛刻的方法，LNP-RNA复合物的稳定性显著影响转染结果。本研究为提高RNA转染效率提供了实用指南，为开发更有效的RNA治疗方法铺平了道路。

引用次数: 0

Quorum sensing: An essential factor in culturing the human promyelocytic leukemia cell line HL-60 and its neutrophil-related functions 群体感应：人早幼粒细胞白血病HL-60细胞培养及其中性粒细胞相关功能的重要因素。

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-20 DOI: 10.1016/j.ymeth.2025.05.008

Danfeng Li , Tian Yang , Siyuan Ma , Xinwei Lyu , Cheng Hu , Jiayin Yan , Lihong Guo , Jiali Tan

HL-60 cells are frequently employed as a standard in vitro model for neutrophil research and extensively utilized. However, the cultivation of HL-60 cells presents a recurring challenge. Historically, cell culture density has been ignored in the consistency of culture conditions. Here, we optimized the culture protocol and explored the impact of culture density on HL-60 cells. Additionally, we investigated the differentiated rate and antibacterial potential of differentiated HL-60 (dHL-60) neutrophils across varying cell density cultures. The findings revealed a positive correlation between cell proliferation activity and cell density, suggesting that increased density facilitates enhanced cell proliferation. Furthermore, as the density of the cell culture increased, there was a concomitant rise in the differentiation rate of HL-60 cells into neutrophils upon stimulation. Importantly, this elevated density also led to significantly higher levels of mitochondrial reactive oxygen species (ROS) production and bacterial phagocytosis. Further investigation revealed that small extracellular vesicles (sEVs) are crucial communicator in quorum sensing within HL-60 cells. Supplementation of HL60-derived sEVs (hEVs) in low-density cell populations resulted in a restoration of cell proliferation, in dose-dependent tendency. Conversely, the inhibition of EV-secretion in HL-60 cells restrains cell growth and proliferation. Overall, our study not only optimized the HL-60 cell culture protocol but also elucidated the critical role of culture density in enhancing HL-60 cell proliferation and antibacterial activity. This finding offers a noteworthy consideration for in vitro experiments of HL-60 cells and suggests the involvement of a quorum sensing mechanism within the neutrophil microenvironment.

HL-60细胞经常被用作中性粒细胞研究的标准体外模型，并被广泛应用。然而，HL-60细胞的培养提出了一个反复出现的挑战。从历史上看，细胞培养密度在培养条件的一致性中被忽略了。本实验优化培养方案，探讨培养密度对HL-60细胞的影响。此外，我们研究了HL-60 （dHL-60）中性粒细胞在不同细胞密度培养中的分化率和抗菌潜力。研究结果显示细胞增殖活性与细胞密度呈正相关，表明细胞密度增加有利于细胞增殖。此外，随着细胞培养密度的增加，HL-60细胞在刺激下向中性粒细胞的分化率也随之上升。重要的是，这种升高的密度也导致线粒体活性氧（ROS）产生和细菌吞噬水平显著提高。进一步的研究表明，小的细胞外囊泡（sev）是HL-60细胞群体感应的重要通讯载体。在低密度细胞群中补充hl60衍生的sEVs （hEVs）导致细胞增殖恢复，并呈剂量依赖趋势。相反，抑制HL-60细胞中ev的分泌会抑制细胞的生长和增殖。总的来说，我们的研究不仅优化了HL-60细胞培养方案，而且阐明了培养密度在促进HL-60细胞增殖和抗菌活性方面的关键作用。这一发现为HL-60细胞的体外实验提供了值得注意的考虑，并表明中性粒细胞微环境中参与了群体感应机制。

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引用次数: 0

The influence of structured light scanning probe configuration on the 3D scanning accuracy of the maxillofacial region in a smiling state: An in vitro study 结构光扫描探针配置对微笑状态下颌面部区域三维扫描精度的影响：体外研究

IF 4.2 3区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

Methods

Pub Date : 2025-05-16 DOI: 10.1016/j.ymeth.2025.05.006

Miaosheng Guan , Xiaoyu Wang , Xue Li , Yaqi Wang , Kun Yan , Ran Huo , Tao Song , Lin Liu , Hongbo Li

Currently, single-unit, dual-unit, and triple-unit structured light 3D scanning technologies have become the predominant facial scanning methods. However, the impact of different unit strategies on facial scanning accuracy remains unclear. A standardized 3D facial model in a smiling state was established. Key point reference coordinates and 3D data were obtained using a coordinate measurement instrument and an industrial-grade laser 3D scanner. Three structured light scanning techniques (single-, dual-, triple-unit) were utilized to capture the 3D information of the model. Linear distance deviations and 3D surface deviations (trueness and precision) of the three scanning strategies were compared. The triple-unit scanning strategy exhibited the lowest deviation among 20 trueness indicators and 22 precision indicators for linear distance measurements (P < 0.05). Furthermore, the accuracy of the triple-unit strategy (trueness: 0.1607 ± 0.0201 mm, precision: 0.0161 ± 0.0112 mm) for overall facial scanning was significantly lower than that of the single-unit and dual-unit strategies, particularly in critical regions for oral and maxillofacial aesthetic analysis, such as the orbital, nasal, and perioral regions. The triple-unit structured light scanning strategy significantly enhances the accuracy of facial 3D scanning, particularly when acquiring 3D facial information from the midline and perioral regions. This in vitro study demonstrates that the triple-unit structured light 3D scanning strategy effectively improves the accuracy of facial scanning, especially in the oral-maxillofacial aesthetic regions. This approach provides a foundation and support for both preoperative planning and postoperative evaluation of aesthetic restoration.

目前，单单元、双单元和三单元结构光三维扫描技术已成为主要的面部扫描方法。然而，不同单元策略对面部扫描精度的影响尚不清楚。建立了微笑状态下的标准化三维人脸模型。利用坐标测量仪和工业级激光三维扫描仪获得关键点参考坐标和三维数据。利用三种结构光扫描技术（单、双、三单元）捕获模型的三维信息。比较了三种扫描策略的线性距离偏差和三维表面偏差（真实度和精度）。在线性距离测量的20个真实度指标和22个精度指标中，三单元扫描策略的偏差最小(P <；0.05)。此外，对于整个面部扫描，三单元策略的准确性（真实度：0.1607±0.0201 mm，精度：0.0161±0.0112 mm）显著低于单单元和双单元策略，特别是在口腔和颌面美学分析的关键区域，如眶、鼻和口周区域。三单元结构光扫描策略显著提高了面部3D扫描的准确性，特别是在从中线和口周区域获取3D面部信息时。体外实验表明，三单元结构光三维扫描策略有效提高了面部扫描的准确性，特别是在口腔颌面美学区域。该方法为美学修复的术前规划和术后评价提供了基础和支持。

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