IntroductionMyasthenia gravis (MG) is an autoimmune disorder of neuromuscular transmission. Gut dysbiosis has been implicated in autoimmune pathogenesis, yet integrated microbial and metabolomic profiling in MG remains scarce. To characterize gut microbiota and the fecal metabolome in MG, identify diagnostic biomarkers, and explore associations between microbial taxa, metabolites, and clinical severity.MethodsFecal samples from 29 MG patients and 10 healthy controls underwent 16S rRNA sequencing and UHPLC-Q-TOF MS metabolomics. LEfSe identified differential taxa; metabolites were screened by VIP > 1.0, P < 0.05, FDR q < 0.05. Random Forest and Spearman correlation assessed biomarker performance and microbiota – metabolite – clinical associations.ResultsMG patients showed significantly reduced alpha- and beta-diversity. LEfSe identified 232 discriminative taxa, with depletion of butanoic acid-producing commensals (Faecalibacterium prausnitzii, Ruminococcus bromii, Bifidobacterium bifidum) and enrichment of Klebsiella. Metabolomics revealed 567 altered metabolites (424 downregulated), including reduced short-chain fatty acids and secondary bile acids (lithocholic, isolithocholic, and allolithocholic acid). The Random Forest metabolite model achieved AUC = 1.0. Spearman analysis revealed that lithocholic acid (P < 0.05) and allocholic acid (P < 0.001) showed positive correlations with the QMGS, while Ruminococcus abundance was positively correlated with butanoic acid (P < 0.01). KEGG analysis implicated cholinergic synapse, bile secretion, sphingolipid signaling, and mTOR pathways.ConclusionsMG patients exhibit a distinct profile of gut dysbiosis and metabolic disturbances. The specific microbial and metabolic biomarkers identified in this study may offer novel insights for auxiliary diagnosis of MG and guide future microbiota-targeted intervention strategies.

ABSTRACT Clostridioides difficile (CD) is a leading cause of antibiotic‐associated diarrhoea in both hospitalized and non‐hospitalized patients. This study explores the role of extracellular vesicles (EVs) derived from Clostridioides difficile strain 630 (CD630‐EVs), which released spherical EVs during in vitro culture, in modulating pro‐inflammatory cytokine production in macrophages. Proteomic analysis identified a total of 1064 proteins within CD630‐EVs, including four immune‐related proteins: FliC, TrxA, TrxA2 and HtpG. Notably, FliC exhibited the highest expression intensity within immune‐related pathways. CD630‐EVs significantly stimulated the production of inflammatory factors, such as interleukin (IL)‐6, monocyte chemoattractant protein‐1 (MCP‐1), IL‐1α and TNF‐α, in mouse macrophages. However, the addition of TH1020, a flagellin receptor inhibitor, markedly suppressed cytokine production. Protein docking analysis further demonstrated that TH1020 disrupts the interaction between Toll‐Like Receptor 5 (TLR5) and FliC, thereby attenuating FliC‐mediated inflammatory responses in macrophages. FliC likely plays a pivotal role in mediating the inflammatory response induced by CD630‐EVs in macrophages. This effect is potentially driven by the activation of innate immunity through FliC's interaction with TLR5 in the host, which may subsequently influence adaptive immune responses. In conclusion, this study elucidates the immune‐modulatory effects of CD630‐EVs on host macrophages during Clostridioides difficile infection (CDI). CD630‐EVs contain a diverse array of proteins, with FliC emerging as a key mediator of pro‐inflammatory responses in macrophages. These findings suggest that FliC may play a critical role in intestinal mucosal injury during CDI, highlighting its potential as a therapeutic target for mitigating inflammation in CDI.

Background: Endobronchial hamartoma (EBH) is an exceptionally rare benign neoplasm frequently misdiagnosed as an obstructive malignancy. The therapeutic paradigm is shifting from traditional anatomical resection toward parenchyma-preserving interventional techniques. This study evaluates the efficacy and safety profiles of contemporary bronchoscopic interventions versus surgical management for EBH. Methods: A retrospective analysis was conducted on a clinical cohort of 17 patients treated between 2013 and 2026, alongside a comprehensive systematic review of 31 contemporary studies (2013-2025). The primary endpoint was the treatment success rate at 3 months, while secondary outcomes included perioperative complications, re-intervention rates, and successful lung parenchyma preservation. Results: Within the analyzed cohort (median age, 58 years), lesions exhibited a significant right-sided predilection (70.6%). Preoperative imaging uniformly revealed non-specific masses, with 41.2% displaying secondary obstructive manifestations. Definitive interventions comprised bronchoscopic management (n = 11, 64.7%) and surgical resection (n = 6, 35.3%). The technical success rate was 100%, with zero major perioperative complications and only minimal-to-scant intraoperative bleeding reported. Over a median follow-up of 3 months, local recurrence was observed in three cases (17.6%)-notably spanning both surgical (n = 2) and bronchoscopic (n = 1) modalities. The systematic review corroborated these findings, underscoring the exemplary safety profile and superior lung-sparing capacity of bronchoscopic interventions. Conclusions: Within the limits of this retrospective cohort and literature review, interventional bronchoscopy appears to be a safe and lung-sparing approach. It may be considered as a preferable initial treatment option for anatomically suitable EBHs. Traditional surgical resection remains necessary for anatomically complex lesions or cases with irreversible distal parenchymal destruction. Vigilant longitudinal surveillance is advised across all modalities.

110 Background: Immune checkpoint blockade (ICB) shows limited efficacy in mismatch repair proficient (pMMR) locally advanced rectal cancer (LARC). Recent clinical studies have shown that neoadjuvant chemoradiotherapy (nCRT) plus ICB improve response. However, radiotherapy is associated with toxicities, including radiation proctitis, anastomotic leakage, and anal dysfunction. Moreover, radiotherapy is not feasible for high position LARC. Radiotherapy-free neoadjuvant strategy warrants further exploration. The FIRM trial is the first phase 2, proof-of-concept study evaluating neoadjuvant immuno-chemotherapy (nICT) with mFOLFOX6 plus PD1 blockade for pMMR LARC. Methods: Patients (pts) with T3/4 or N+, pMMR LARC located ≤15 cm from anal verge were eligible. Pts received 6 cycles of mFOLFOX6 and serplulimab followed by surgery. Primary endpoints were pathological complete response (pCR) rate and major pathological response (MPR) rate. Enrollment of 30 pts was planned, with hypothesis of an increased pCR of 24% compared with reported data of 6.6% after mFOLFOX6 (FOWARC trial, JCO 2016). Multi-omics profiling (single cell RNAseq, bulk RNAseq, whole exome sequencing) was performed to elucidate response mechanisms and develop an immuno-response molecular subtype (IRMS) for immunotherapy stratification. Results: 30 pts were enrolled, 28 completed ≥4 cycles of nICT. 2 discontinued due to adverse events (AE). The pCR and MPR rate were 42.9% and 67.9%, respectively. Tumor regression grade (TRG) 0, 1, 2, and 3 were observed in 44.4%, 11.1%, 40.7%, and 3.7% of pts. 6 pts experienced grade 3 AEs, with no grade 4-5 AE or anastomotic leakage observed. Baseline profiling identified two novel subsets predicting nICT response: inflammatory/migratory tumor cells (IMT) and ITGAX⁺ activated B cells (IAB). Post-nICT samples showed increased enrichment of CD8⁺ T, cDC1, pDC, memory B and Tfh cells, together with reduced exhausted CD8⁺ T, Tregs, extracellular matrix deposition and tumor angiogenesis. IRMS stratified pts into immune-sensitive and immune-resistant types and showed strong predictive performance in internal cohort (AUC =0.87, sensitivity =0.74, specificity =1.0, accuracy =0.81), outperforming conventional biomarkers including CPS (AUC = 0.48) and TMB (AUC =0.47). Notably, IRMS was validated across multiple external immunotherapy cohorts (melanoma, colorectal, lung, breast cancers), showing robust predictive value (AUC =0.70-0.86). Conclusions: These findings provide the first evidence supporting immuno-chemotherapy as a promising radiotherapy-free neoadjuvant strategy for pMMR LARC. nICT converts the immune-cold tumors into immune-activated state by orchestrating a dual-pathway immune remodeling involving IMT-cDC1-CD8 and IAB-CD4 axes. The IRMS provides a high-precision, pan-cancer tool to guide patient selection and personalized immunotherapy. Clinical trial information: NCT06688786 .

Low-molecular-weight (LMW) compounds, including both endogenous and exogenous molecules, are critical in characterizing diverse life processes, such as cancer. Specifically, the comprehensive characterization of LMW compounds aids in understanding the pharmacological effects of traditional Chinese medicine (TCM), enabling the identification of active ingredients and assessment of therapeutic efficacy against tumor cells. However, challenges arise in laser desorption/ionization mass spectrometry (LDI MS) due to the limited availability of existing matrices and the detection preference for specific molecules. Here, ZnS/Au (ZA) composites are developed as matrices for LDI MS, achieving a 100% detection coverage of 78 standard analytes in dual-ion modes. The composites are optimized with a loose ZnS substrate densely anchored with large Au nanoparticles, forming a Schottky heterostructure at the interface. This design results in the generation of stable hotspot layers and long-lifetime electron-hole pairs. Consequently, a wide coverage of phytochemicals (12 categories, 50 types) is identified from 11 TCMs. Further, the growth-inhibitory of TCMs on A549 lung cancer cells are validated based on metabolic profiles of endogenous molecules. This study provides an efficient tool to elucidate the active components and action mechanisms of the complex TCM system at the molecular level, thereby advancing the modernization and standardization of TCM.

119 Background: PD-1 inhibitor shows poor efficacy in MSS locally advanced rectal cancer (LARC). Radiotherapy activates tumor immunogenicity, and current evidence have shown that combination of neoadjuvant chemoradiotherapy (nCRT) with PD-1 inhibitor enhances tumor regression and improves pathological complete response (pCR) rate in MSS LARC. However, nCRT induces radiation toxicities including radiation proctitis, anastomotic fistula and anal dysfunction. Moreover, radiotherapy is not feasible for high position LARC. Radiotherapy-free neoadjuvant strategy warrants further exploration. Neoadjuvant mFOLFOX6 circumvents radiation toxicity but achieves only 6.6% pCR rate (FOWARC trial, JCO 2016). Preclinical studies suggested oxaliplatin and 5FU have immune-sensitizing effects. Thus, we conducted the first proof of concept trial exploring a radiotherapy-free neoadjuvant immuno-chemotherapy (nICT) regimen, which employed a multicenter, single arm, phase 2 design to evaluate the efficacy and safety of mFOLFOX6 plus serplulimab in MSS/pMMR LARC. Methods: Patients (pts) with cT3/4 or cN+, MSS and pMMR LARC located ≤15 cm from anal verge were eligible. Pts received 6 cycles of neoadjuvant mFOLFOX6 and serplulimab, followed by surgery and adjuvant mFOLFOX6. The primary endpoints were pCR rate and major pathological response (MPR) rate. Enrollment of 30 pts was planned, with hypothesis of an increased pCR of 24% compared with reported data of 6.6% after conventional mFOLFOX6. The per-protocol set (PPS) was defined as pts who completed ≥4 cycles of nICT for primary endpoint assessment. Results: Among 30 enrolled patients, 28 completed ≥4 cycles of nICT and were included in PPS. 2 discontinued due to adverse events. The pCR was achieved in 42.9% (12/28) of pts, and MPR in 67.9% (19/28). For high LARC (>10 cm from anal verge, ineligible for nCRT), the pCR and MPR rates were 71.4% and 85.7%. Radiologic response of tumor regression showed complete response in 57.1% (16/28) and partial response in 39.3% (11/28) of pts. One pt had stable disease and received nCRT. 27 pts proceeded to TME and were included in surgical set. TRG 0, 1, 2, and 3 were observed in 44.4%, 11.1%, 40.7%, and 3.7% of pts. Downstaging was achieved in 22 pts (81.5%). The Grade 3 TRAEs were lymphocyte decrease (4 pts), neutropenia (3 pts) and bilirubin elevation (1 pts). No Grade 4-5 TRAEs or anastomotic fistula were observed. Conclusions: mFOLFOX6 plus serplulimab showed promising efficacy and manageable safety in MSS LARC. pCR rate unexpectedly reached 42.9%, showing an improvement compared with the reported 6.6% pCR rate of chemotherapy, and were comparable to nCRT plus anti-PD-1 regimens. This is the first proof of concept trial to explore a nICT strategy for MSS LARC, which spares pts from radiation toxicities and shows particularly applicability for high LARC. We are launching a randomized controlled trial (FIRM02) for further validation. Clinical trial information: NCT06688786 .

Thymic epithelial tumors (TETs), rare yet clinically significant malignancies, face diagnostic challenges due to their occult presentation and lack of noninvasive risk-stratification tools, leading to systemic overtreatment and poor prognoses for high-risk subtypes. To address this unmet need, we developed a Fe3O4@Fe metal-organic framework heterojunction-enhanced laser desorption ionization mass spectrometry (LDI MS) platform for the efficient analysis of serum metabolic fingerprints (SMFs). Engineered through gradient pyrolysis, this nanomaterial synergizes ultraviolet absorption and photothermal conversion from its two constituent components with enhanced charge separation, achieving 1,000-fold improvement in sensitivity and thus enabling direct SMF acquisition from 1 μL of serum. Coupled with machine learning, the platform demonstrates robust diagnostic performance, yielding area under the curve (AUC) of 0.960 for distinguishing TETs from benign control and AUC of 0.856 for hierarchical risk stratification, outperforming clinical workflows. Beyond advancing material design for LDI MS, this work establishes a clinically translatable framework for rapid, large-scale screening, addressing critical gaps in TET management through metabolic-driven stratification.

BACKGROUND Portal hypertension is typically associated with cirrhosis, but acute etiologies must be considered in atypical cases. Post-splenectomy arteriovenous fistula (AVF) is a rare vascular complication that can lead to acute portal hypertension with nonspecific symptoms. CASE SUMMARY A 43-year-old man presented with acute abdominal pain, hematemesis, and melena, eight years after open splenectomy. Computed tomography angiography revealed a splenic AVF. Emergency laparotomy with splenic artery ligation was performed. The patient recovered uneventfully and remained asymptomatic at five-year follow-up. CONCLUSION Splenic AVF should be considered in patients with new-onset portal hypertension, a history of splenectomy, and no liver disease. Prompt diagnosis and surgical intervention yield excellent outcomes.

BackgroundUlcerative colitis (UC) is subtype of inflammatory bowel disease that is frequently comorbid with anxiety disorders. However, effective dual-targeting therapies are still lacking. Hyperoside (HYP), a natural flavonoid, exhibits anti-inflammatory and neuroprotective properties, yet its potential therapeutic effects on UC and associated anxiety, as well as the underlying mechanisms, remain largely unexplored.MethodsA murine model of DSS-induced colitis was established and treated with HYP. Disease activity was assessed through body weight, colon length, and histopathology. Anxiety-like behaviors were evaluated using open field and elevated plus maze tests. Neuroinflammation was examined through immunohistochemistry of BDNF expression and microglial activation. Gut microbiota composition was profiled by metagenomic sequencing, and metabolomic profiling was conducted using the Q300 Kit. Network pharmacology and molecular docking were employed to predict signaling pathways, which were further validated by Western blotting. Additionally, antibiotic depletion experiments were conducted to determine microbiota dependency.ResultsHYP administration significantly ameliorated DSS-induced colitis, as evidenced by attenuated weight loss, restored colon length, and improved histopathology. It suppressed pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) and restored intestinal barrier integrity by upregulating Mucin-2 and ZO-1. Furthermore, HYP also alleviated anxiety-like behaviors and mitigated neuroinflammation by increasing BDNF levels and suppressing microglial activation. HYP treatment also restored gut microbial homeostasis, enriching beneficial bacteria such as Enterobacter ludwigii while reducing the abundance of Enterobacter hormaechei, Escherichia coli, and Acinetobacter baumannii. Metabolomic analysis revealed that HYP significantly promoted arginine biosynthesis. Network pharmacology and molecular docking identified the MAPK, PI3K-Akt, and NF-κB pathways as potential targets, with HYP showing strong binding affinity to MAPK3, AKT1, and NFκB1. Importantly, the therapeutic effects of HYP were abolished in microbiota-depleted mice.ConclusionOur findings demonstrate that HYP effectively alleviates DSS-induced colitis and comorbid anxiety-like behaviors. Its efficacy is dependent on the gut microbiota and is associated with the restoration of microbial homeostasis, enhancement of arginine metabolism, and modulation of the MAPK/PI3K-Akt/NF-κB signaling pathways. HYP represents a promising microbiota-targeting therapeutic candidate for UC and its neuropsychiatric comorbidities.

Infectious complications (ICs) in acute pancreatitis (AP) are primarily driven by intestinal bacterial translocation, significantly increasing mortality and hospital stays. Despite this, the role of the gut microenvironment, particularly its metabolic aspects, in AP remains poorly understood. In this study, we investigated a cohort of patients with AP, and conducted supplemental murine studies, to explore the relationship between the gut metabolome and the development of ICs. Metabolomic analysis revealed that disruptions in gut tryptophan metabolism - especially reductions in serotonin and indole pathways - are key features associated with IC occurrence. Additionally, elevated plasma levels of tryptophan metabolites within the kynurenine pathway were identified as valuable predictive biomarkers for ICs. Mechanistic studies in murine models demonstrated that an impaired intestinal Th17 response, modulated by these tryptophan metabolites, plays a critical role in IC development. Serotonin supplementation enhanced Th17 responses, reducing IC incidence, while administration of kynurenic acid, a kynurenine metabolite, exacerbated pancreatic infections, potentially through immunosuppressive effects. These findings highlight the pivotal role of tryptophan metabolites in AP pathogenesis, emphasizing their potential as both predictive markers and therapeutic targets in IC management.

Photodynamic therapy (PDT) has received much attention as a promising modality for tumor treatment. However, the weak targeting ability of conventional photosensitisers and the metastasis of malignant tumors have severely limited the development of PDT. To address this, an esterase-activated prodrug (BPYM) has been developed for imaging-guided photodynamic therapy cascade immunotherapy for the treatment of pancreatic cancer. Upon reaction with esterase, BPYM releases the photosensitiser BPY and exhibits strong red fluorescence emission, which is further enhanced by the aggregation-induced emission (AIE) characteristics of BPY. Interestingly, the activation of the fluorescence signal simultaneously indicates the activation of photosensitivity capabilities. Under white light irradiation, activated BPYM can generate large amounts of reactive oxygen species (ROS) to induce apoptosis in pancreatic cancer cells. More importantly, BPYM-mediated PDT can trigger immunogenic cell death (ICD) and elicit a systemic anti-tumor immune response. Ultimately, this imaging-guided PDT not only precisely ablates the primary pancreatic cancer tumors, but also inhibits the growth of distant tumors through an immune response. In summary, we report a strategy to achieve photodynamic immunotherapy for the treatment of pancreatic cancer through the rational design of an esterase-activated prodrug.

A multicenter cross-sectional study was conducted across six Chinese hospitals between September 2023 and April 2024 to investigate the prevalence, molecular epidemiology, antimicrobial resistance (AMR) and genetic relatedness of C. difficile in pediatric populations. Among 1,442 stool samples collected, 242 C. difficile isolates were recovered (16.8%), including 188 from asymptomatic carriers (13.0%) and 11 from confirmed C. difficile infection (CDI) cases (0.98%). The highest rate of asymptomatic carriage was observed in children aged 1-2 years (29.3%). Multilocus sequence typing (MLST) identified ST3 as the predominant sequence type (30.6%), followed by ST42, ST54, ST2, ST102, and ST110. Core genome phylogenetic analysis revealed nine distinct genetic lineages, with lineage VI being the most prevalent among CDI cases. High resistance to clindamycin (64.5%) was observed, primarily mediated by ermB, while all isolates remained susceptible to metronidazole and vancomycin. Moxifloxacin resistance (10.7%) was associated with gyrA T82I mutations, particularly in toxigenic ST3 strains. Transposon analysis indicated ST-specific carriage of AMR genes, with Tn6218(ermB) prevalent in nontoxigenic ST3 isolates and Tn5801 (tet(M)) predominant in ST54. Genomic relatedness (≤2 SNPs) was detected in 12.0% of asymptomatic carriers, with most links associated with hospital-wide contact, suggesting possible transmission events. This study highlights the importance of asymptomatic colonized children as a reservoir for C. difficile, maintaining resistant lineages and disseminating AMR, thereby underscoring the need for enhanced surveillance and targeted antimicrobial stewardship in Chinese healthcare settings.

Mass spectrometry (MS) is the primary tool to detect small metabolite ions for profiling bio-systems. However, small metabolite profiling in MS still needs to be improved and calls for new rules in ionization. Herein, we studied the Br & oslash;nsted acid-induced multiple alkali metal cation adductions (BAI-mAMCA) in laser desorption/ionization MS, with annotation of BAI-mAMCA in complex bio-samples and improved profiling of small metabolites. Firstly, we engineered the enhanced mAMCA (relative intensities increased from 0.1-4.6 % to 76.1-89.9 %, p < 0.05) of small metabolites with desirable reproducibility via a nanoparticle-enhanced approach. Subsequently, we revealed the rule of BAI-mAMCA according to the proton dissociation ability of small metabolites qualitatively and quantitatively (|r| = 0.824-0.857), with validation using density functional theory calculation. Finally, we integrated a computational workflow of mAMCA-identity networking to annotate BAI-mAMCA in complex bio-samples and demonstrated excellent diagnostic performance (AUCs = 0.963-0.977) when applied to 300 serum samples from lung adenocarcinoma patients and healthy controls. We also achieved improved label-free quantitation (r(2) = 0.989-0.997, p < 0.05) and direct analysis of specific isomers based on the established BAI-mAMCA rules. This work provides both fundamental mechanisms and practical engineering strategies for advancing small metabolite profiling in MS, offering potential for integration into clinical diagnostic platforms.

Clostridioides difficile infection (CDI) is a major cause of healthcare-associated diarrhea, leading to inflammation due to the action of large clostridial toxins. Madecassoside (MA), a pentacyclic triterpene extracted from Centella asiatica, has broad anti-inflammatory effects. However, its therapeutic impact and mechanisms in CDI have not been thoroughly investigated. Here, we initially performed a CDI mouse model to analyze the improvement of symptoms by MA in vivo. Subsequently, we isolated primary bone marrow-derived macrophages (BMDM) in vitro and constructed an inflammation model stimulated by C. difficile isolates or toxins in macrophages, and analyzed the effects of MA. This study found that the administration of low and high concentrations of MA via continuous intragastric administration alleviated the symptoms in CDI mice, including improvements in body weight, colon length, and histopathological damage. ELISA and immunohistochemical staining showed that MA reduced the secretion of IL-1β in a dose-dependent manner in vivo and in vitro. Furthermore, the hyperactive state of inflammasome-related signaling pathway from the stimulated by C. difficile and toxin B was inhibited. Co-immunoprecipitation and confocal microscopy results indicated that MA inhibited the formation of the inflammasome complex and ASC speck. Molecular docking and SPR studies revealed MA's strong binding affinity for ASC. In conclusion, our study suggests that MA can reduce the activation of inflammasomes triggered by C. difficile toxins, potentially through its specific binding to ASC, which may pave the way for new treatment strategies against CDI.

Bloodstream infections (BSIs) annually claim millions of lives, contributing to one‐third of global mortality. Mortality rates predominantly depend on the duration between symptom onset and the administration of appropriate antibiotics. Clinical protocols for identifying pathogens typically take a 2–5 day wait for blood culture, due to the limited pathogen load present in blood at the initial stage. Currently, stratifying infections into high‐risk subpopulations and administering prompt therapy rely on separate platforms. Here, multifunctional bimetallic nanoreactors are designed to develop a rapid platform for integrative analysis of both host and pathogen characteristics directly from blood samples. These nanoreactors enable colorimetric quantitation of host immune responses and mass spectrometric profiling of pathogen metabolic fingerprints. The platform predicts BSI in 100% of suspected cases and achieves 91.67% match in species identification within a 60 min sample‐to‐answer time, holding promise for implementation in clinical practice.

Gallbladder cancer (GBC) frequently mimics gallbladder benign lesions (GBBLs) in radiological images, leading to preoperative misdiagnoses. To address this challenge, we initiated a prospective, multicenter clinical trial (ChicCTR2100049249) and proposed a multimodal, non-invasive diagnostic model to distinguish GBC from GBBLs. A total of 301 patients diagnosed with gallbladder-occupying lesions (GBOLs) from 11 medical centers across 7 provinces in China were enrolled and divided into a discovery cohort and an independent external validation cohort. An artificial intelligence (AI)-based integrated model, GBCseeker, is created using cell-free DNA (cfDNA) genetic signatures, radiomic features, and clinical information. It achieves high accuracy in distinguishing GBC from GBBL patients (93.33% in the discovery cohort and 87.76% in the external validation cohort), reduces surgeons' diagnostic errors by 56.24%, and reclassifies GBOL patients into three categories to guide surgical options. Overall, our study establishes a tool for the preoperative diagnosis of GBC, facilitating surgical decision-making.

BACKGROUND:This study aimed to evaluate the potential of 68 Ga-NODAGA-SNA006 PET/CT imaging as a noninvasive method for assessing immune cell infiltration and predicting treatment response in lung cancer patients undergoing immunotherapy. PATIENTS AND METHODS:A prospective study enrolled 8 patients with histologically confirmed lung cancer who received first-line chemotherapy combined with anti-PD-1/PD-L1 immunotherapy. 68 Ga-SNA006 PET/CT imaging was performed before treatment. The primary endpoints analyzed the correlation between tumor SUVmax and CD8 + T-cell infiltration, as well as associations of multiparametric imaging parameters (SUVmax, TBR, TLR, TSR, and TMR) with treatment outcomes. RESULTS:The SUVmax significantly correlated with stromal CD8 + T-cell infiltration (R 2 = 0.8218, P = 0.0338) but not with intratumoral infiltration (R 2 = 0.5178, P > 0.05), indicating its stromal-specific association with immune activity. Among 8 patients, 6 achieved partial response and 2 stable diseases after 2 treatment cycles. Baseline blood CD8 + T cells, Ki67%, and PD-L1% showed no prognostic significance, nor did SUVmax correlate with posttreatment lesion reduction. In 24 lesions analyzed, TBR demonstrated the strongest correlation with lesion reduction (R 2 = 0.3178, P < 0.01). ROC analysis further revealed that TBR had the highest diagnostic efficacy (AUC = 0.87) for predicting treatment response, with a sensitivity of 62.5% and specificity of 93.3% at a cutoff value of TBR >2.120, and a sensitivity of 87.5% and specificity of 66.7% at TBR >5.535. CONCLUSIONS:68 Ga-NODAGA-SNA006 PET/CT provides valuable insights into immune cell infiltration and treatment response in lung cancer patients. The study highlights the potential of TBR as a prognostic biomarker and underscores the importance of integrating multiparametric imaging into clinical decision-making.