Abstract Gut microbiota dysbiosis has been closely associated with COVID‐19 progression, and probiotic supplementation has been regarded as a potential adjuvant therapy for COVID‐19. However, large‐scale clinical evidence regarding the efficacy of probiotics against the currently predominant Omicron variant remains scarce. This multicenter retrospective cohort study employed a 1:2 propensity score‐matched analysis of hospitalized Omicron‐infected patients from 10 hospitals in Henan Province (probiotics group: 4205 cases vs. control group: 8410 cases). The primary outcome was all‐cause mortality, and the secondary outcome was the incidence of composite disease progression. The results demonstrated that probiotic supplementation significantly reduced all‐cause mortality by 27% (hazard ratio [HR] = 0.73, 95% confidence interval: 0.654‒0.811), with more pronounced benefits observed in patients not receiving antiviral therapy (HR = 0.67) and those with severe disease (HR = 0.69). However, no significant difference was found in the composite disease progression outcome (HR = 1.05, p = .312). Safety assessment indicated that probiotic supplementation was generally well‐tolerated, although transient fluctuations in serum creatinine levels warranted attention. In conclusion, based on the real‐world evidence from a multicenter Omicron‐infected patients’ cohort, the probiotic supplementation could reduce the all‐cause mortality rate of hospitalized patients infected with Omicron, especially in severe cases and without antiviral treatments cases. This study provides the first large‐scale real‐world data and new microbiome‐modulating strategies against future respiratory viral infections.

Introduction:Patients with mental disorders are at increased risk of adverse outcomes from COVID-19, but prognostic evidence specific to this population remains limited. This study aimed to develop and validate machine-learning models for predicting 31-day mortality among hospitalized patients with mental disorders and laboratory-confirmed COVID-19. Methods:Data were retrospectively collected from 439 hospitalized patients across 10 hospitals in Henan Province, China. Patients were randomly divided into a training cohort (n = 308) and an independent test cohort (n = 131). Oversampling was applied during model development to address class imbalance. Candidate predictors were selected using LASSO, Boruta, and random forest methods, and eight machine-learning algorithms were trained. SHAP analysis was used for model interpretation, and Kaplan-Meier analysis compared survival between model-defined risk groups. Results:Patients were generally older, 63.1% were female, and comorbidities were common. Several models showed good discrimination in the training cohort, although some showed overfitting. In the test cohort, the neural network model with LASSO-selected features performed best, with an AUC of 0.911 (95% CI: 0.832-0.990). SHAP analysis identified concomitant hormone therapy, alkaline phosphatase, and lymphocyte count as the leading predictors. The high-risk group had significantly higher cumulative mortality than the low-risk group (log-rank P < 0.0001). Discussion:A machine-learning model based on routine clinical and laboratory variables may support short-term mortality risk stratification in this regional multicenter cohort.

Ulcerative colitis (UC) is an incurable inflammatory bowel disease characterized by chronic mucosal inflammation, with a continuously increasing global prevalence. Although infrared (IR) therapy has demonstrated anti-inflammatory potential, conventional devices that can only emit single-wavelength IR often exhibit limited tissue penetration and poor suboptimal spectral overlap with mammalian absorption. Herein, we develop a broad-spectrum infrared (BSIR) device enabled with an almost defect-free graphene based radiator to deliver high output IR aligned with mammalian IR absorption spectra. In a mouse model of UC, BSIR treatment significantly alleviated disease symptoms and promoted mucosal recovery. Crucially, this study shows that BSIR radiation induces the relocation of T lymphocytes to the spleen, leading to reduced immune cell infiltration and inflammation in the colon. Gene expression analysis further reveals enhanced innate immune activity and cell regeneration in colonic tissue. Collectively, these findings demonstrate that BSIR represents a safe, noninvasive therapeutic strategy for UC. More broadly, this study highlights spectrum-matched IR irradiation as a novel modality for immune modulation, with potential translational relevance for other deep-tissue inflammatory diseases.

IntroductionThe anticancer drug camptothecin (CPT) has limited clinical applications due to severe toxic reactions.MethodsWe combined CPT with PHBVHHx (PHA) nanoparticles by a modified emulsion method for the first time construct a novel nanomedical drug (CPT-PHA-NPs, CPNs).ResultsIn vitro experiments verified the drug loading level (89%), sustained-release properties (40% release within 48 h; near-complete release over 21 days), and inhibition ability of the compound on HT-29 cell activity (IC50 = 0.44 μM). In vivo, CPN-treated mice showed significantly less body weight reduction (P < 0.05 from day 7) and markedly improved liver and kidney function markers compared to controls. Histological analysis confirmed that CPN effectively prevented hepatocyte necrosis and renal inflammation observed with free CPT, demonstrating higher biosafety and lower toxicity. Crucially, 16S rRNA sequencing revealed that CPT severely depleted probiotics (Akkermansia, Lactobacillus, Candidatus_Arthromitus, and Bacilli_unclassified) while promoting pathogenic taxa (Lachnospiraceae_NK4A136_group, [Eubacterium]_xylanophilum_group, and Faecalibaculum), whereas CPNs attenuated these microbial disruptions. Metabolomics further showed CPNs' milder effects on phenylalanine and essential amino acid metabolism vs. CPT.DiscussionIn conclusion, this novel type of nanomaterial not only possesses excellent performance but also can reduce the impact of CPT on tissues, intestinal flora and serum metabolism, providing a new strategy for anti-tumor treatment that takes into account both microbial homeostasis and metabolic safety.

Background Patients with initially unresectable intrahepatic cholangiocarcinoma (iCCA) have poor prognoses and the current first-line treatments remain unsatisfactory. Our study aimed to evaluate gemcitabine-based chemotherapy plus PD-1/PD-L1 inhibitors and tyrosine kinase inhibitors (TKIs) for iCCA. Methods In this multicenter retrospective cohort study, 392 patients were included in the full cohort analysis. For the primary analysis, 88 patients received gemcitabine-based chemotherapy (GEMCIS or GEMOX), and 177 patients received the same chemotherapy backbones combined with PD-1/PD-L1 inhibitors and TKIs. The primary outcome was to evaluate overall survival (OS). The propensity score matching (PSM) method was utilized to reduce potential confounders. Results Overall, 392 patients were included from 12 hospitals across China, with data from January 2016 to December 2024. In the full cohort analysis, the combination of GEMCIS/GEMOX with PD‑1/PD‑L1 inhibitors and TKIs significantly prolonged median OS compared to GEMCIS/GEMOX alone (26.7 vs. 15.3 months; hazard ratio [HR] 0.47, 95% CI: 0.35–0.63, P < 0.001). In the primary analysis, the combination strategy was associated with a median OS of 24.8 months and median progression-free survival (PFS) of 11.0 months, with HRs of 0.56 (95% CI: 0.38–0.83, P = 0.004) for OS and 0.46 (95% CI: 0.32–0.66, P < 0.001) for PFS after PSM. Importantly, our treatment strategy increased the rate of conversion surgery to 42% for locally advanced disease, ultimately resulting in significantly improved overall outcomes (HR for OS: 0.17 [95% CI: 0.05–0.61]). Conclusion Combining gemcitabine-based chemotherapy with PD-1/PD-L1 inhibitors and TKIs may provide a promising new first-line treatment option for patients with initially unresectable iCCA.

Post-COVID-19 sequelae have become an emerging global health issue, but the mechanisms for the sustained susceptibility of convalescents to the sequelae remain poorly understood. Here we report the use of a restricted open-search approach to explore the molecular imprints of SARS-CoV-2 infection left on the proteome of 412 COVID-19 patients and convalescences. A total of 827 non-standard amino acid variations, chemically modified residues as well as post-translational modifications, termed non-coded amino acids (ncAAs), are found spreading over 29,814 sites in patient's serum proteins. Markedly, widespread ncAAs are induced and sustainedly imprinted on the serum proteome predominately perturbing the immunoglobulin-mediated immune response, complement activation and coagulation regulation even 12 months after recovery. Sustained amino acid variations and chemical modifications are found in the complementary‑determining regions (CDRs) of the variable region of immunoglobulin contributing to the interactions between the emerging antibody and antigens; durable chemical amino acid modifications found in the hyper ncAA-modified regions of the constant region of immunoglobulin important for the interaction with the complement and regulatory receptors. In the complement system, inducible ncAAs are memorized in the components essential for the complement activation, amplification cascades and membrane attack processes. Thus, the workflow described in this study can be used to identify the molecular imprints of viral infection at the proteomic scale, particularly the specific antibodies and the immune targets left in COVID-19 patients and convalescents.

Esophageal cancer (EC) is a leading cause of cancer-related mortality worldwide and early detection strategies and precise postoperative interventions must be developed. However, the identification of noninvasive biomarkers for the diagnosis and prognosis remains limited. We performed 16S rRNA gene sequencing on tongue-coating samples from 440 participants, including 157 EC patients, 167 healthy controls (HCs) and 120 EC patients who received radiotherapy. We characterized the oral microbiome and constructed microbial diagnostic and prognostic classifiers. Furthermore, the oral microbiome of EC who received radiotherapy (n = 120) was characterized. The oral microbial diversity of EC patients was increased, with differences in the microbial community between EC patients and HCs. In EC, the genera Veillonella, Streptococcus and Actinomyces were enriched, whereas Porphyromonas and Rothia were depleted. The classifier based on six optimal microbial markers was constructed using random forest algorithm and achieved area under the curves (AUCs) of 93.69

Acute myeloid leukemia (AML) is driven by complex cellular interactions within the bone marrow microenvironment, in which tumor-associated macrophages (TAMs) and leukemia stem cells (LSCs) play essential regulatory roles. This study investigated how the N6-methyladenosine (m6A) reader protein heterogeneous nuclear ribonucleoprotein C (HNRNPC) modulates macrophage function and influences AML progression. Single-cell RNA sequencing (scRNA-seq) identified sixteen distinct cell types and revealed enhanced macrophage communication with other cell populations, accompanied by reduced interactions involving M1 macrophages in AML. Integrated transcriptomic and metabolomic analyses identified HNRNPC as a key m6A-related regulator of macrophage activity through the modulation of metabolic pathways. In vitro, silencing of HNRNPC increased tricarboxylic acid (TCA) cycle activity and adenosine triphosphate (ATP) production, thereby promoting M1 polarization, enhancing tumor cell recognition and phagocytosis, and strengthening antitumor immune responses. In vivo, macrophage-specific deletion of HNRNPC significantly suppressed tumor growth and extended survival in AML-bearing mice. Collectively, the findings identify HNRNPC as a central mediator linking m6A modification to macrophage metabolic reprogramming and immune activation, offering a promising potential for therapeutic intervention in AML.

Severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2) infection induces perturbations in the human oral microbiome. However, alterations of the oral microbiome in patients reinfected with SARS‐CoV‐2 omicron variant (ORs) remain uncharacterized. We enrolled 2046 participants providing tongue‐coating specimens, with 1637 samples meeting inclusion criteria for sequencing and analysis, including 702 ORs, 184 OR primary‐infected patients, 50 wild‐type strain primary‐infected patients (WTPs), 154 common cold patients, and 547 heathy controls (HCs). While α‐diversity showed no significant differences between OR and HC groups, compositional analysis revealed notable taxonomic shifts: ORs exhibited elevated abundance of 13 genera (including Veillonella and Prevotella ) alongside depletion of 12 genera (e.g., Actinomyces and Neisseria ) relative to HCs. Across SARS‐CoV‐2 evolutionary progression and host infection stages, maximal microbial dysbiosis was observed in WTPs, contrasting with minimal perturbation in ORs. The abundance of Fusobacterium and Porphyromonas gradually increased during this process, while Parascardovia and Pyramidobacter gradually decreased. Furthermore, a diagnostic model for OR was constructed. High AUC values were obtained in discovery phase (97.25%, 97.07%) and independent validation phase (89.61%). Importantly, two cross‐regional validation phases (70.90%, 73.97%) also showed high efficiency, and three cross‐disease validation phases (92.34, 95.46, and 85.10%) also revealed good disease specificity. This study is the first to characterize the oral microbiome of ORs. Our findings indicate that dynamic alterations of oral microbiome were closely associated with the severity of COVID‐19. Moreover, oral microbiome analysis contributes to diagnosis for ORs.

Background Hepatic fibrosis represents a major global health burden with no approved first-line therapeutic agents. The urgent need for innovative targeted small molecule interventions has prompted investigation of natural compounds with anti-fibrotic potential. While α-amyrin demonstrates established anti-inflammatory properties, its therapeutic efficacy against hepatic fibrosis remains unexplored. Purpose The present study was designed to assess the anti-fibrotic efficacy of α-amyrin and determine its molecular mechanisms of action. Study design Firstly, the efficacy of α-Amyrin was evaluated through the liver fibrosis model. Further, the mechanism of α-Amyrin was analyzed using multi-omics and molecular biology experiments, and was verified using the target knockdown model. Finally, the effective components of α-Amyrin were analyzed through bioinformatics analysis and experimental verification. Methods In this study, we use an in vitro model of fibrosis in human hepatic stellate LX-2 cells induced with TGF-β1 and an in vivo mouse model with CCl4 to evaluate the anti-fibrosis potential of α-Amyrin using multiple approaches. Employing multi-omics and molecular biology techniques to investigate the mechanism of α-Amyrin counteracting hepatic fibrosis. The interaction targets of α-Amyrin were examined through molecular docking, molecular dynamics simulations, and surface plasmon resonance. The anti-hepatic fibrosis mechanism of α-Amyrin was further validated using both in vitro and in vivo PPARα knockout or inhibition models. Results In vivo, α-Amyrin significantly inhibited the formation of hepatic pseudolobules and suppressed hepatic inflammation compared with the model control group. In vitro, α-Amyrin significantly down-regulated the expression levels of well-established markers of liver fibrosis. The results of transcriptomics and metabolomics provided clues to the mechanisms of the PPARα pathway and fatty acid metabolism. Further experimental validation of these mechanisms was conducted. In vitro and in vivo, α-Amyrin significantly promoted fatty acid oxidation levels which is consistent with up-regulation of PPARα pathway involved in this process, including Sirt1, PPARα, PGC-1α, CPT1A, ACOX1, and its downstream targets. Moreover, α-Amyrin markedly enhanced PPARα nuclear translocation and facilitated the Sirt1-PGC-1α interaction. Molecular docking, dynamics simulations, and surface plasmon resonance confirmed direct binding of α-Amyrin to PPARα. Critically, the anti-hepatic fibrosis effect of α-Amyrin was significantly compromised in both the in vivo PPARα inhibitor model and the in vitro PPARα knockout model. Conclusions The possible mechanism of α-Amyrin's anti-fibrosis effect is to target the PPARα axis to promote the reprogramming of fatty acid oxidation metabolism, which in turn exerts an anti-inflammatory effect and reverses the inflammation-fibrosis pathological process. The above results are expected to provide an innovative strategy for the development of small molecule drugs targeting liver fibrosis.

BACKGROUND Aging is the primary risk factor for numerous chronic diseases, cognitive deterioration, and mortality. The gut microbiota is increasingly implicated in the aging process, yet a comprehensive understanding of its dynamic compositional and functional shifts throughout the human lifespan and its mechanistic contributions to aging remains unclear. AIM To study the gut microbiota profiles of age-related changes during aging in both human and rat cohorts, with particular focus on microbiota involved in lipid metabolism. Subsequently, to intervene fatty acid oxidation inhibitor trimetazidine (TMZ) was used in the aging process. METHODS The metagenomic sequencing and 16S rRNA sequencing were performed to detect gut microbiota in 300 individuals of different age groups and rats of different weeks, mapping the evolutionary profiles of microbiota during aging across all age groups and focusing on analyzing significantly changed gut microbiota and their biological functions. Based on clues obtained from the above analysis that microbiota with fatty acid oxidation function was closely related to aging, and validation was conducted using fatty acid oxidation inhibitor TMZ in aging animal models. RESULTS Multicohort analysis revealed that gut microbial diversity follows a nonlinear trajectory, initially increasing until age 10, remaining relatively stable until approximately 70 years and declining thereafter. Functional enrichment analysis demonstrated a significant, age-associated increase in lipid metabolism pathways in both human and animal models, which was consistent with marked changes in abundance in the gut microbiota involved in lipid metabolism such as Luteipulveratus (P < 0.05). TMZ, a fatty acid oxidation inhibitor, reshaped the gut microbiota structure and suppressed the abundance of lipid metabolism-associated gut microbiota in aging rats. Further molecular validation confirmed that the TMZ inhibited fatty acid beta-oxidation and significantly downregulated the expression levels of key senescence marker proteins and genes. CONCLUSION In conclusion, gut microbiota undergoes age-dependent remodeling, with significant enrichment of the fatty acid oxidation-related microbiome. The fatty acid oxidation inhibitor TMZ may attenuate aging phenotype through the dual modulation of gut microbial composition and lipid metabolism and may provide an antiaging strategy.

The immunosuppressive tumor microenvironment (TME) undermines the efficacy of many cancer therapies. This study investigated the immunomodulatory and anti-tumor activity of Azvudine (FNC), alone or in combination with anti-PD-1 blockade. We established syngeneic tumor models in immunocompetent mice. Single-cell RNA sequencing, flow cytometry, and immunological assays were employed to analyze immune cell reconstitution and functional changes following FNC administration. FNC demonstrated dose- and time-dependent tumor inhibition. It significantly expanded memory T cells, natural killer (NK) cells, and CD8+ cytotoxic T lymphocytes, while reducing the abundance of myeloid-derived suppressor cells (MDSCs). Flow cytometry confirmed these immunological shifts, showing enhanced infiltration of effector immune cells within the TME. Moreover, FNC induced hallmark features of immunogenic cell death (ICD), including the release of damage-associated molecular patterns such as high-mobility group box 1 (HMGB1) and calreticulin. When combined with anti-PD-1 therapy, FNC produced a synergistic anti-tumor effect, leading to durable tumor remission in all treated mice. FNC remodels the TME by mitigating immunosuppression and amplifying anti-tumor immunity, offering a promising strategy to augment existing immunotherapies. Further clinical evaluation is warranted to ascertain the translational potential of FNC in diverse oncologic settings.

Cancer patients are at an elevated risk for SARS-CoV-2 infection and require vigilant monitoring. The efficacy and safety of Azvudine in treating COVID-19 among this vulnerable group remain under-researched. We conducted a multicenter, retrospective cohort study of nine hospitals involving cancer patients with COVID-19 hospitalized from December 2022 to January 2023. To minimize immortal time bias, patients who died or were discharged within 24 h of drug administration were excluded. A 2:1 propensity score matching (PSM) was performed to balance baseline characteristics, explicitly adjusting for tumor metastasis, tumor classification, and COVID-19 vaccination status. The primary outcome was 30-day all-cause death, and the secondary outcome was composite disease progression. Robustness was verified using Probit regression and double robust estimation with Augmented Inverse Probability Weighting (AIPW). A total of 1,829 patients were included in the final matched cohort (1,200 in the control group and 629 in the Azvudine group). Azvudine treatment was significantly associated with a reduced risk of all-cause death (adjusted hazard ratio [HR]: 0.65, 95

Growing evidence suggests a role for the gut microbiome in progression of cholangiocarcinoma (CCA), however, its diagnostic and therapeutic potential remains incompletely characterized. Here, metagenomic sequencing was performed on fecal samples (n = 785) from individuals across East, Central, and Northwestern China. Gut microbial dysbiosis in CCA was characterized by depletion of short-chain fatty acids-producing species and enrichment of potential pathobionts (Klebsiella aerogenes, Clostridium symbiosum). Diagnostic models built using species-level markers demonstrated superior performance, compared to pathway-based models, achieving area under the curve (AUC) values of 98.63% and 99.42% in the discovery cohort, with robust cross-regional validation (AUC = 80.89% and 80.43%). The model effectively distinguished CCA from hepatocellular carcinoma (AUC = 97.86%) and liver fibrosis (AUC = 98.73%) and nonalcoholic fatty liver disease (mean AUC = 96.86%). Analysis of public datasets encompassing 6847 samples across 31 studies and 11 disease states revealed moderate disease specificity influenced by biomarker overlap across conditions. Mechanistically, depleted Bifidobacterium pseudocatenulatum suppressed CCA progression, associated with inhibition of the PI3K-AKT-mTOR pathway. Collectively, this study supports the potential of fecal metagenomic signatures as a complementary noninvasive aid for CCA detection, and provides functional evidence for a candidate protective microbe.

Oral microbiota is related to the severity and recovery of SARS-CoV-2 infection. This study aims to predict clinical classification after SARS-CoV-2 infection using oral microbiota before infection. Herein, we collected tongue-coating samples before infection and then monitored clinical information after infection. Oral microbiota was detected by MiSeq sequencing. We randomly assigned participants from Zhengzhou into discovery and validation cohorts to develop a predictive model and conducted cross-region verification using Xinyang and Hangzhou cohorts. Sixteen asymptomatic patients (AP), 257 mild patients (MP), 106 common patients (CP), and 7 severe patients (SP) were enrolled. Oral microbiota diversity was decreased in CP versus MP. At genus level, 11 microorganisms, including Rothia and Gemella, were increased, while 5 microorganisms, including Selenomonas and Lachnoanaerobaculum, were decreased in CP versus MP. Moreover, the classifier based on 15 optimal markers showed high prediction efficiency in discovery cohort (area under the curve [AUC]: 98.35%), validation cohort (AUC: 81.91%), Xinyang cohort (AUC: 74.34%), and Hangzhou cohort (AUC: 94.44%). Interestingly, a higher abundance of Selenomonas was associated with milder clinical symptoms. In conclusion, our study established a good model to predict clinical classification after SARS-CoV-2 infection using oral microbiota before infection, providing a novel strategy for precise prevention and treatment.

Hypertension is widely acknowledged as a major risk factor for disease severity and death in patients with coronavirus disease 2019 (COVID-19). Azvudine is recommended for COVID-19 patients in China. However, its clinical efficacy and safety for individuals with hypertension remain unclear. This nine-center retrospective cohort study included 32,864 hospitalized COVID-19 patients in Henan Province, China, from December 2022 to January 2023. Among these patients, those with hypertension were identified and divided into the Azvudine and control groups (standard treatment without antiviral medication) after propensity score matching (PSM) at a 1:1 ratio. The primary outcomes measured were all-cause mortality and composite disease progression. Subgroup analyses and sensitivity tests were conducted to verify the robustness of the results. Safety was assessed based on adverse events (AEs). After PSM to balance baseline characteristics, the analysis included 2434 Azvudine recipients and 2434 controls, forming a final matched cohort. Azvudine was associated with a lower risk of all-cause mortality (HR: 0.64, 95% CI 0.519-0.780; P < 0.001) and composite disease progression (HR: 0.84, 95% CI 0.719-0.985; P = 0.032) in hypertensive patients with COVID-19. In five sensitivity analyses, Azvudine showed a highly robust effect in reducing all-cause mortality, while the evidence for a reduction in the progression of composite disease progression was less consistent. No significant difference in severe AEs (≥ Grade 3) was observed between groups. These real-world findings suggest Azvudine may be a promising antiviral option for hypertensive COVID-19 patients, but further prospective trials are necessary to confirm these results.