MetaboLights

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Aligned LC-MS features were therefore represented by unique Feature_POS or Feature_NEG identifiers together with their measured mass-to-charge ratio and retention time. Optional compound annotation fields were left empty; no compound names, formulas, structures, or database identifiers were fabricated.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseChromatographic separation was performed using an ultra-high-performance liquid chromatography system equipped with an ACQUITY HSS T3 column (100 mm x 2.1 mm, 1.8 µm; Waters) maintained at 40 °C. The flow rate was 0.40 mL/min. The supplied method describes formic-acid-containing aqueous/acetonitrile and acetonitrile mobile phases. The injection volume recorded in the RAW-file metadata was 3 µL.A dual-site oral nanovaccine programs anti-colorectal tumor immunity from small intestinal priming to large intestinal boosting.kun yangsouthwest universitycolonsmall intestinemass spectrometry assayApproximately 100 mg of intestinal content was transferred to a 2 mL tube containing a 6 mm grinding bead and 800 uL methanol/water (4:1, v/v) extraction solution containing internal standards, including 0.02 mg/mL L-2-chlorophenylalanine. Samples were homogenized for 6 min at -10 degrees C and 50 Hz, sonicated for 30 min at 5 degrees C and 40 kHz, held at -20 degrees C for 30 min, and centrifuged at 13,000 x g for 15 min at 4 degrees C. Supernatants were collected for LC-MS analysis.Mus musculushttps://www.ebi.ac.uk/metabolights/MTBLS15453kun yang. Southwestern University. Chongqing, General Delivery, kun yang. 1355227608@qq.com.kun yang. southwest university. 1355227608@qq.com.Thermo RAW files were converted to centroided MS1 mzML with ThermoRawFileParser 1.4.5. Features were detected separately for each polarity with pyOpenMS 3.5.0 FeatureFindingMetabo using a 5 ppm mass tolerance, an intensity threshold of 10,000, fixed elution-peak-width filtering, no isotope-filtering model, and removal of single mass traces. Retention times were aligned with MapAlignmentAlgorithmPoseClustering and features were linked across samples with FeatureGroupingAlgorithmKD. Retention time was reported in minutes. Features were retained when detected in all four samples of at least one experimental group, corresponding to at least 80% detection within a group of n=4. Missing abundance values were left blank. Group1355227608@qq.comSmall-intestinal and colonic contents were collected from mice assigned to water control or DPPL@OVA-CpG+NIR treatment groups. Samples were coded JM1 (water, small-intestinal content), JM4 (DPPL@OVA-CpG+NIR, small-intestinal content), JC1 (water, colonic content), and JC4 (DPPL@OVA-CpG+NIR, colonic content). Individual samples were collected for LC-MS-based untargeted metabolomics analysis.Metabolomicsultra-performance liquid chromatography-mass spectrometryThermo Scientific Vanquish UHPLC SystemMetabolomicsMus musculusuntargeted analysisThermo Scientific Accela 1250 Pumpcolonsmall intestineThermo Scientific DSQ IIThermo Scientific Q Exactive HF-X mass spectrometerexperimental sampleuntargeted metabolite profilingultra-performance liquid chromatography-mass spectrometryThermo Scientific Vanquish UHPLC SystemMetabolomicsMus musculusuntargeted analysisThermo Scientific Accela 1250 PumpcolonThermo Scientific DSQ IIsmall intestineThermo Scientific Q Exactive HF-X mass spectrometerexperimental sampleuntargeted metabolite profilingMass spectra were acquired with a Thermo Scientific Orbitrap Exploris 480 mass spectrometer using electrospray ionization in separate positive- and negative-ion assays. The ion-source temperature was 400 °C, sheath gas was 40 arbitrary units, auxiliary gas was 10 arbitrary units, and spray voltages were 3500 V in positive mode and -2800 V in negative mode. Data-dependent acquisition was used with stepped normalized collision energies of 20, 40, and 60. The acquisition range was m/z 70-1050 and the RAW-file run duration was approximately 8 min. falseMetabolites interact with microorganisms and vaccineswe engineered a β-glucan (Dex)-stabilized mulberry leaf lipid-based nanovaccine (DPML@OVA-CpG) that protected vaccine components during GI transit and facilitated their specific delivery to the GALTs. Following oral administration, the DPML@OVA-CpG underwent M cell-mediated transcytosis in the small intestine, activated immune responses within PPs and MLNs, and initiated mucosal and systemic anti-tumor immunity. In parallel, a fraction of the structurally intact nanovaccine persisted along the GI tract and reaches colorectal tumor regions, where near-infrared (NIR) irradiation leverages the carrier-enabled photothermal immunomodulation to induce immunogenic cell death (ICD). The released endogenous tumor-associated antigens (TAAs), together with the locally delivered CpG adjuvant, converted the tumor into an in situ vaccine, thereby establishing a secondary immune activation site within the colorectal TME (Scheme 1). This coordinated small intestinal priming-large intestinal boosting strategy overcomes tumor heterogeneity and synergizes with ICBs to achieve effective control of advanced CRC.2026-08-252026-08-25MTBLS15453