2026-10-14T18:56:16Z https://www.scidb.cn/oai

10.57760/sciencedb.011yh 2026-09-14T15:09:19Z

2026-09-14 Exosomal RNAs in Tuberculous and Lung Adenocarcinoma Pleural Effusions doi:10.57760/sciencedb.011yh en 1. Data Set Generation Process, Processing Methods/Steps, and Equipment and Tools Used(1) Sample Collection and GroupingStudy Subjects: 10 patients with pleural effusion, divided into two groups—the malignant pleural effusion (MPE) group (n=5, all associated with lung adenocarcinoma) and the tuberculous pleural effusion (TPE) group (n=5).Inclusion/Exclusion Criteria: Strictly limited to patients who had not received antituberculosis or anticancer treatment, had no other malignant tumors or autoimmune diseases, and presented with a single type of pleural effusion.Sample Volume: Approximately 100 mL of pleural effusion was collected from each patient via thoracentesis.(2) Exosome Isolation and CharacterizationIsolation method: Exosomes were isolated from pleural effusion using ultracentrifugation.Morphological and particle size characterization: Exosome morphology (typical cup-shaped) was observed using transmission electron microscopy; particle size distribution (40–150 nm) was determined via particle size analysis techniques.Protein Marker Validation: The exosome protein concentration was determined using the BCA method, and the expression of the exosome marker protein CD63 was detected via Western blot.(3) Exosome RNA Extraction and Library ConstructionRNA Extraction: Total exosome RNA was extracted using the miRNeasy Mini Kit.RNA Quality Assessment: RNA concentration and quality were determined using a Quantus fluorometer; the RIN must be > 7.5 and the 28S/18S ratio must be ≈ 2:1.Amplification and Library Construction: Perform micro-amplification of RNA according to the instructions for the RNALib Single Cell WTA Kit; purify cDNA using magnetic beads; take 1 ng of cDNA and construct a DNA library using the Illumina LifeTn Transpose DNA Library Preparation Kit.Library Quality Control: Perform preliminary concentration measurement using the Qubit 3.0 and dilute to 1 ng/μL; verify the effective concentration (must be > 10 nM) using the Bio-RAD CFX96 real-time quantitative PCR system.(4) High-Throughput SequencingSequencing Platform: Cluster generation was performed on the QSeq 100 platform using paired-end sequencing with a read length of 150 bp.Data Output: LncRNA and mRNA expression profiles were obtained for 10 exosome samples.(5) Data AnalysisDifferentially Expressed Gene Screening: Differentially expressed lncRNAs (DELs) and mRNAs (DEMs) were identified using the criteria |log₂ fold change| ≥ 1 and P < 0.05.Functional Annotation: Annotation was performed using databases such as NCBI, UniProt, GO, KEGG, Ensembl, GENCODE, and HGNC.Enrichment Analysis: GO (FDR < 0.05) and KEGG (p < 0.05) enrichment analyses were conducted.Diagnostic Performance Evaluation: ROC curves were plotted, and AUC, sensitivity, and specificity were calculated. exosomal RNAs; pleural effusion; lung adenocarcinoma Han Yijia Zhang Jiaqiang PUBLIC https://creativecommons.org/licenses/by/4.0/ dataset Science Data Bank