Background: Gastric cancer is the third leading cause of cancer-related mortality worldwide. According to The Cancer Genome Atlas (TCGA), it can be classified into four molecular subtypes, including microsatellite instability (MSI) and genomically stable (GS) subtypes, which display distinct clinical and pathological features. However, differences in their tumor microenvironment, particularly metabolic reprogramming, remain poorly understood. Methods: Single-cell RNA sequencing data from gastric cancer patients classified as GS or MSI were enrolled. Cell clusters were identified and annotated to compare cellular landscapes between subtypes. Differential gene expression and pathway analyses were performed among malignant epithelial cells. Key genes related to oxidative phosphorylation were identified using LASSO regression, and their expression was further validated in the TCGA dataset. Patient-derived xenograft models were used to compare tumor growth rates, ATP levels, and expression of oxidative phosphorylation-related genes. Results: Single-cell transcriptomic analysis revealed eight major cell types in MSI tumors. Compared to the GS subtype, MSI samples showed significantly greater infiltration of T cells and a lower proportion of epithelial cells. Malignant cells from MSI samples exhibited increased activity of oxidative phosphorylation pathways. LASSO regression identified five oxidative phosphorylation-related genes that were consistently overexpressed in MSI tumors in both single-cell and TCGA datasets. In Patient-derived xenograft models, MSI tumors grew more rapidly and demonstrated higher ATP levels and elevated expression of the five oxidative phosphorylation-related genes compared to MSS tumors. Conclusion: Our study reveals enhanced oxidative phosphorylation metabolism in MSI gastric cancer at single-cell resolution and identifies five oxidative phosphorylation-related genes that may serve as potential therapeutic targets for this subtype.

Anorectal malignant melanoma (ARMM), originating from melanocytes in the anorectal mucosa, is a rare but highly aggressive tumor characterized by early metastasis and poor prognosis, with a 5-year survival rate of 20.5

BackgroundIschemic-type biliary lesion (ITBL) remains one of the most common complications following liver transplantation. It is imperative to further explore the occurrence and development mechanism of ITBL. Intrahepatic cholangiocyte organoids (ICOs) derived from human liver tissue replicate the structure and function of bile ducts and serve as an innovative experimental tool for in vitro modeling of cholangiopathies.MethodsIn this study, ICOs derived from human liver tissue were cultured under ischemia and hypoxia (IH) conditions to establish an in vitro model of ITBL. Immunofluorescence staining, RT-qPCR, Western blotting, and transcriptomic analysis were performed to investigate the effects of IH on ICOs and evaluate the validity of the ITBL model.ResultsIH significantly reduced the diameter and cell viability of ICOs. After exposure to IH for more than 48 h, the proliferation of ICOs decreased, accompanied by increased inflammatory responses and apoptosis. Transcriptomic analysis revealed a landscape of pathophysiological genetic changes in the ITBL model in response to IH.ConclusionWe have presented a novel ITBL model constructed using expandable human ICOs, which is of great significance for exploring the molecular mechanism and potential therapeutic targets of ITBL.

Diabetic wounds (DWs) are characterized by impaired angiogenesis, which is a key factor contributing to delayed wound healing. However, the specific regulators of angiogenic dysfunction in DWs remain poorly understood. In this study, we identified the abnormal downregulation of tissue inhibitor of metalloproteinases 2 (TIMP2) in skin wounds of diabetic patients through RNA sequencing and histopathological techniques. By upregulating TIMP2 expression via adeno-associated virus (AAV) transfection in vitro and in vivo, we demonstrated that TIMP2 overexpression enhanced vascular endothelial cell proliferation through activation of the PI3K-AKT signaling pathway and promoted wound healing in diabetic mice. Furthermore, using multiple online functional prediction databases and dual luciferase reporter assays, we investigated miR-106a-5p as an upstream regulator of TIMP2, which was upregulated in the wounds of diabetic mice. Inhibition of miR-106a-5p can upregulate TIMP2 mRNA expression in vascular endothelial cells, leading to increased cell proliferation and decreased apoptosis, thereby accelerating wound healing in diabetic mice. Our findings underscore the critical role of the TIMP2 and its upstream regulator miR-106a-5p in angiogenesis during wound healing, providing a promising avenue for diabetic wound repair.

Background: Colorectal cancer is a common malignant tumor of the digestive tract, with a high incidence and mortality rate. Ki67 and vascular endothelial growth factor (VEGF) play important roles in tumor cell proliferation and angiogenesis. Vitamin P is a natural flavonoid compound with various biological activities. Objectives: To explore the effects of vitamin P on the proliferation, angiogenesis, and apoptosis of colorectal cancer cells and its underlying molecular mechanisms. Methods: In vitro cultivation of human colorectal cancer cell lines (HCT116 and SW480) was performed. Vitamin P was added to the cells in varying quantities. Cell proliferation was identified using the CCK-8 technique. The flow cytometry method was used to determine the proportion of apoptotic cells. Real-time fluorescence quantitative PCR and Western blot methods were used to measure the expression levels of the VEGF and Ki67 genes and proteins. Secretion of VEGF was observed via immunofluorescence staining. Results: Vitamin P treatment significantly inhibited the proliferation of colorectal cancer cells in a dose-dependent manner. The half-maximal inhibitory concentrations (ICs50) at 48 hours were 72.3 μM (HCT116) and 85.6 μM (SW480). Moreover, in the vitamin P treatment group (42.7 - 68.9% and 35.4 - 61.2%), the secretion of VEGF decreased by 52.3 - 79.8%. The percentage of apoptotic cells induced by 20 - 80 μM vitamin P increased from 5.3% in the control group to 18.6 - 37.9%, with increased caspase-3 activity. In vivo experiments showed that vitamin P significantly inhibited the growth of colorectal cancer transplanted tumors and reduced the expression of Ki67 and VEGF in the tumors. Meanwhile, vitamin P treatment induced apoptosis of tumor cells, and this effect was closely related to the regulation of the Bax/Bcl-2 ratio and the promotion of caspase-3 activation. Vitamin P may play a regulatory role by inhibiting the phosphorylation of the STAT3 signaling pathway and downregulating the expression of proteins related to the PI3K/AKT pathway. Conclusions: Vitamin P can inhibit the malignant biological behavior of colorectal cancer cells by suppressing Ki67-mediated cell proliferation, blocking VEGF-related angiogenesis pathways, and activating mitochondrial apoptotic pathways.

Background:Hepatocellular carcinoma (HCC) is one of the leading causes of cancer-related death, with limited reliable biomarkers for early diagnosis and prognosis. In this study, we explored the role of DCUN1D5, a potential biomarker, in HCC. Methods:We performed integrated bioinformatics analyses of publicly available HCC datasets, including TCGA, GEO, and ICGC, followed by in vitro experiments on HCC cell lines to validate our findings. Results:DCUN1D5 expression was significantly elevated in HCC tissues compared to normal liver tissues. High DCUN1D5 levels were associated with poor prognosis and advanced tumor stage in HCC patients. Additionally, bioinformatics analysis revealed a correlation between DCUN1D5 expression levels and both immune infiltration and immunotherapy response in hepatocellular carcinoma. Functional assays revealed that DCUN1D5 knockdown inhibited cell proliferation, migration, and invasion in HCC cells. Mechanistically, Enrichment analysis revealed significant correlations between DCUN1D5 and multiple biological pathways; specifically, we observed a negative correlation between the expression level of DCUN1D5 and fatty acid metabolism. Further cellular experiments demonstrated that knockdown of DCUN1D5 can induce accumulation of free fatty acids in hepatocellular carcinoma cells. Conclusion:In summary, our study identified DCUN1D5 as a potential biomarker for poor prognosis in HCC, high expression of DCUN1D5 are closely associated with tumor invasion and proliferation. Knockdown of DCUN1D5 was found to inhibit the proliferation, migration, and invasion of HCC cells and increases its free fatty acid accumulation. These studies suggest new possibilities for treatment strategies for HCC.

BackgroundTumor-associated macrophages (TAMs) are known to facilitate cancer progression. However, the diversity of TAM subsets and their distinct roles in GC remain poorly understood. This study aimed to evaluate the impact of legumain (LGMN)+ macrophages on GC progression and clarify the underlying mechanisms of their roles.MethodsWe used single-cell RNA sequencing (scRNA-seq) and bulk RNA sequencing (bulk RNA-seq) analyses from public databases (GEO and TCGA) to systematically evaluate the clinical prognostic significance of LGMN and to characterize the remodeling of its associated signaling pathways. To investigate the role of LGMN in mouse GC (TAMs), we generated macrophage-specific LGMN conditional knockout mice, LGMNflox/flox; Lyz2-Cre. Utilizing a combination of subcutaneous xenograft tumor models, primary cell isolation and culture, immunofluorescence staining, and tube formation assays, we systematically elucidated the regulatory function and underlying molecular mechanisms of LGMN+ macrophages in GC progression.ResultsWe found that LGMN+ macrophages are significantly enriched in GC tissues, and their high infiltration was significantly associated with poor outcomes. Additionally, scRNA-seq revealed that hypoxia and immune suppression pathways are enriched in LGMN+ macrophages. LGMN+ macrophages infiltration levels showed a significant positive correlation with the infiltration of regulatory T (Treg) cells and endothelial cells. Mechanistically, conditional knockout of LGMN in macrophages inhibits tumor growth by reprogramming TAMs toward an anti-tumor phenotype, reducing Treg cell infiltration, and enhancing the infiltration level of CD8+ T cells. Furthermore, LGMN knockout can inhibit tumor angiogenesis by downregulating VEGF-A expression.ConclusionsLGMN+ macrophages drive GC progression by promoting tumor angiogenesis and establishing an immunosuppressive microenvironment. Therefore, targeting this TAM subset may represent a novel therapeutic strategy for GC.

AIMS:To characterize the progression patterns and analyze the efficacy of immune checkpoint inhibitors (ICIs) in unresectable, recurrent, or metastatic cholangiocarcinoma (CCA). METHODS:A retrospective analysis was conducted in CCA patients treated with ICIs between September 2019 and September 2024. The progression patterns of ICIs were assessed, and survival and prognostic factors were analyzed using Kaplan-Meier and Cox regression models. RESULTS:Among 131 eligible patients, the median overall survival (OS) and progression-free survival (PFS) were 14.1 and 7.9 months, respectively. Multivariable Cox analysis identified lung metastasis, absent abdominal lymph node/retroperitoneal lymph node (LN/RPLN) metastasis, and CA199 ≥ 35 U/ml as independent predictors of shorter PFS, and smoking, no surgery, and CA199 ≥ 35 U/ml as predictors of shorter OS (all p < 0.05). Rising CA199 correlated with poorer PFS (p < 0.05). Progression occurred in 80 patients, 77.5% within 12 months. Oligo-progression was the main pattern (53.8%). Liver (43.8%) and abdominal LN/RPLN (38.8%) were the most common sites of progression. Oligo-progression was associated with superior OS versus systemic progression (median 22.7 vs. 11.7 months, p = 0.003). Among oligo-progression patients, local plus systemic therapy showed a trend toward better OS than systemic therapy alone (28.9 vs. 15.8 months, p = 0.062). CONCLUSIONS:ICIs demonstrate meaningful efficacy in CCA. Oligo-progression represents the predominant failure pattern and is associated with favorable prognosis.

Background Vessels that encapsulate tumor clusters (VETC) is a powerful predictor of aggressive hepatocellular carcinoma (HCC) and associated with poor outcomes of HCC. Imaging surrogates of VETC potentially help predict postsurgical recurrence. Purpose To explore the noninvasive predictive potential of contrast-enhanced computed tomography (CE-CT) for VETC of HCC (VETC-HCC). A web-based prognostic nomogram model including VETC-associated imaging markers was subsequently created to predict postoperative recurrence-free survival (RFS) in HCC patients. Methods A retrospective evaluation was performed on 393 patients with HCC who underwent CE-CT and immunohistochemical staining for CD34 at three different institutions. Patients from institution 1 (n = 241) were split into training (n = 169) and internal test (n = 72) sets. The remaining 152 patients from institutions 2 and 3 were used as the external test set. Univariate logistic regression analyses and six machine learning algorithms were performed on the training set, and the performance of 6 ML models (AUC, DeLong test, etc.) was compared to identify VETC-associated imaging markers (which were calculated as the VETC score), which were validated in the internal and external test sets. An interactive prognostic nomogram model including VETC-associated imaging markers and clinical data was used to predict RFS in the training and internal test sets. The association between the model's stratification and postoperative recurrence after radical resection or liver transplantation was also assessed. Results Nonsmooth margins (P = 0.011), tumor size > 5 cm (P = 0.030), and intratumoral necrosis (P = 0.038) were identified as independent predictors of VETC-HCC, and were combined into 6 machine learning models. Logistic regression (LR) was the final selected model and the VETC score was calculated. In the training, internal test, and external test sets, the VETC score demonstrated effective predictive performance for VETC (AUC: 0.768, 0.742, and 0.724, respectively). The interactive prognostic nomogram model(https://radiology.shinyapps.io/DynNomapp/) including the neutrophil-to-lymphocyte ratio (NLR), serum alpha-foetoprotein (AFP), and VETC score yielded C-index values ranging from 0.805 to 0.783 in the training and internal test sets and produced three prognostically distinct groups. Among patients classified as those associated with medium risk by the model, those who underwent liver transplantation had significantly improved RFS (P < 0.05). In contrast, radical resection had no significant effect on RFS in patients classified as having either low or high risk (both P > 0.05). Conclusion Preoperative CE-CT features can be used to characterize VETC-HCCs. The prognostic nomogram model has prognostic value for the preoperative prediction of RFS can aid in the selection of the appropriate surgical approach.

Hepatocellular carcinoma (HCC) remains a highly aggressive malignancy with limited therapeutic options. The long non-coding RNA (lncRNA) HOTAIR contributes to HCC progression through the regulation of the miR-526b-3p/DHX33 axis. This study investigated the antitumor effects of quercetin (Que) on HCC using both Hep3B and HepG2 cell lines, focusing on the modulation of this molecular pathway. In this regard, the study evaluated Que's effects on HOTAIR, miR-526b-3p, and DHX33 expression using qRT-PCR and ELISA. Cell viability was assessed by MTT assay, while apoptosis was measured through Annexin V-FITC/PI staining and analysis of BCL-2/CASP-3 expression. Additional investigations included cell cycle analysis (cyclin A2 and D1), invasion assays (p53 and PTEN), and assessment of oxidative stress markers (SOD, CAT, MDA). Results demonstrated that Que significantly suppressed HOTAIR and DHX33 while upregulating miR-526b-3p in both cell lines (P-value<0.001). Treatment with Que caused dose-dependent apoptosis (54.3% in Hep3B and 48.9% in HepG2 at 100 mu M) and cell cycle arrest while reducing invasion through TP53 downregulation and PTEN upregulation. Que also ameliorated oxidative stress by enhancing antioxidant enzyme activity (P-value<0.01). These findings suggest Que may exert potent antitumor effects in HCC through modulation of the HOTAIR/miR-526b-3p/DHX33 axis, though further in vivo and clinical investigations are warranted.

The long-term impact of COVID-19 on cancer patients receiving immune checkpoint inhibitors (ICIs) remained unknown. This study aimed to investigate the association between COVID-19 and long-term outcomes in ICIs-treated lung cancer patients. Three hundred eighty-one patients with advanced lung cancer who were treated with ICIs were enrolled and followed for at least 6 months in 10 medical centers in China during Omicron pandemic. The primary endpoints were overall survival (OS) and progression-free survival (PFS). Cox model with time-dependent covariate and landmark analysis were used. The multivariable analysis showed that patients with COVID-19 had significantly worse OS (HR: 2.59 [1.58–4.26], P < 0.001) and PFS (HR: 1.55 [1.02–2.35], P < 0.001). In landmark analyses, COVID-19 that occurred within 3 months after initiation of ICIs was found to be associated with shorter OS (HR: 3.40 [1.70–6.77], P = 0.001) and PFS (HR: 3.40 [1.70–6.77], P = 0.02). In subgroup analysis, both mild and severe COVID-19 were associated with shorter OS (mild, HR: 2.39 [1.33–4.29], P = 0.004; severe, HR 4.46 [2.39–8.33], P < 0.002) and PFS (mild, HR 1.71 [1.05–2.78], P = 0.03; severe, HR 3.32 [1.97–5.60], P < 0.002). Additionally, there were no significant differences in OS or PFS among patients with varying treatment delays. COVID-19 had a negative impact on the long-term outcomes of patients with lung cancer who received ICIs, particularly if the infection occurred during the first 3 months of ICIs treatment. These findings are crucial for addressing the COVID-19 epidemic and other respiratory infectious diseases.

BACKGROUND:Intrahepatic cholangiocarcinoma is a malignant tumor that starts from the epithelium of the bile duct and has a poor prognosis. They are characterized by poor response to chemotherapy and lack of effective targeted therapies; thus, therapeutic options are limited. CASE PRESENTATION:A 59-year-old man was admitted to the hospital for a workup of abnormal CA19-9 levels. He was diagnosed with ICC, underwent surgery and was found to have pT1bNx disease. He developed rapid disease recurrence on adjuvant gemcitabine + capecitabine. Following recurrence, he received first-line systemic pembrolizumab + lenvatinib and second-line pembrolizumab + lenvatinib + chemotherapy and had mild tumor regression followed by progression. Next-generation sequencing was performed on the baseline surgical sample. This revealed a novel RBPMS-MET fusion, and based on the literature, crizotinib 250 mg twice a day was administered. After 3 months of crizotinib treatment, magnetic resonance imaging revealed a significant reduction in liver lesions, and 4 months after initiating treatment, scans demonstrated a partial response. CONCLUSION:Our case report strengthens the evidence that crizotinib may be a viable treatment option for patients with ICC with a c-MET tyrosine kinase fusion, necessitating additional clinical investigation.

Cholangiocarcinoma, a hepatobiliary malignancy, has a poor prognosis and treatment resistance. This study integrated spatial transcriptome and conditional knockdown models to explore the tumor microenvironment (TME) regulatory mechanism. We found that macrophages were located at the boundary between cholangiocarcinoma and normal tissue. Cholangiocarcinoma - secreted SPP1 induced blood monocytes to migrate toward the tumor tissue and polarize into M2 macrophages through the CD44 - PI3K - AKT pathway. In vitro and in vivo experiments confirmed this process. Macrophages polarized by SPP1 promoted tumor proliferation through the TGF - β/P - SMAD2/P - SMAD3 pathway. Targeting SPP1 or TGF - β inhibited tumor progression. This study reveals the bidirectional regulatory mechanism of the SPP1 - TGF - β axis in cholangiocarcinoma, providing new perspectives for developing combined therapies targeting the tumor - immunity interaction network.

Background: A significant portion of primary liver cancer patients in China are diagnosed at intermediate-to-advanced stages, often making them ineligible for curative surgery. Furthermore, high postoperative recurrence rates, reaching up to 70%, pose a major challenge for long-term survival. The emergence of novel systemic treatments, such as immune checkpoint inhibitor combinations, and advancements in locoregional therapies have created new opportunities for conversion and perioperative strategies. This updated consensus aims to standardize the clinical application of these therapies based on the latest evidence, with the objective of improving patient prognosis. Methods: A multidisciplinary committee of 97 experts was convened to revise previous guidelines. The process involved a comprehensive search of medical databases and conference proceedings, with evidence graded according to the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) system. Consensus statements were finalized through a formal electronic voting process, requiring at least 80% agreement for approval, resulting in 18 updated statements. Results: The consensus provides refined definitions for conversion and perioperative therapy. It recommends various strategies for oncological conversion, including systemic therapy with anti-angiogenic drugs plus immunotherapy, and locoregional approaches like precision transarterial chemoembolization (TACE) and hepatic artery infusion chemotherapy (HAIC). The document strongly affirms surgical resection as a crucial step for achieving long-term survival after successful conversion and offers guidance on surgical timing and adjuvant therapy. For resectable patients with high-risk features, neoadjuvant and adjuvant treatments are outlined to mitigate recurrence. The consensus also advocates for using dynamic enhanced magnetic resonance imaging ( MRI) and the modified Response Evaluation Criteria in Solid Tumors (mRECIST) criteria for efficacy assessment and underscores the essential role of a multidisciplinary team in management. Conclusions: This updated consensus offers standardized, evidence-based guidance for clinicians on implementing conversion and perioperative strategies to optimize patient-centered care and highlights the need for continued research to further refine these promising approaches.

This study investigates the antitumor efficacy of cantharidin against cholangiocarcinoma (CCA) using patient-derived organoids (PDOs) that faithfully replicate the histological and genomic features of original tumors. Results demonstrate that cantharidin effectively inhibits CCA PDO growth, showing comparable or superior efficacy to conventional chemotherapeutics such as cisplatin, though slightly less potent than gemcitabine or Adriamycin. Critically, drug sensitivity in PDOs correlated perfectly with clinical responses in five patients, validating the model's predictive relevance. Mechanistic studies revealed that cantharidin suppresses proliferation and induces apoptosis primarily through downregulation of the p-ERK1/2-c-Fos signaling pathway, both in vitro and in patient-derived organoids-based xenografts. These effects were reversible upon treatment with a p-ERK agonist, confirming pathway specificity. The study highlights cantharidin's potential as a targeted therapeutic agent in CCA and underscores the utility of PDOs in personalized drug screening and mechanistic investigation.

Metabolic dysfunction-associated steatotic liver disease (MASLD) remains the most common chronic liver disease worldwide, and appropriate in vitro models are of great significance for investigating pathogenesis and drug screening of MASLD. In this study, human expandable cholangiocyte organoids were derived from adult stem cells of normal liver tissue. After differentiation, liver organoids (LOs) exhibited the functional characteristics and genomic features of mature hepatocytes. To induce steatosis, LOs were incubated with a gradient concentration oleic acid, and it was found that the model could recapitulate the development of lipid accumulation and inflammation. In addition, the drug sensitivity of the hepatic steatosis model was further verified through anti-steatosis drug testing. In summary, LOs have great potential for disease modeling, and the results indicate that the hepatic steatosis model may serve as a useful tool for exploring the molecular mechanisms and drug screening of MASLD.

BACKGROUND:The pursuit of novel non-invasive strategies for cancer therapy, particularly those capable of modulating the immunosuppressive tumor microenvironment (TME), is a significant focus in oncology research. Piezoelectric materials, which can generate electrical signals and reactive oxygen species (ROS) under mechanical stimulation, represent a promising platform for biomedical applications. OBJECTIVE:This study aimed to synthesize and evaluate the antitumor efficacy of piezoelectric Ba(Zr₀.₂₅Ti₀.₇₅)O₃ (BZT) nanocubes activated by ultrasound (US) both in vitro and in vivo, and to elucidate the underlying immunomodulatory mechanisms. METHODS:BZT nanocubes with a pure perovskite structure were synthesized via a hydrothermal route method and characterized by XRD, SEM, TEM, and XPS. The piezoelectric catalytic activity for ROS generation was confirmed by methylene blue degradation. The antitumor effects were assessed on hepatocellular carcinoma (HCC) organoids, cell lines (Huh7, Hepa1-6), and an orthotopic mouse model. Cellular proliferation, apoptosis, migration, and viability were evaluated using Ki-67/PCNA staining, TUNEL, EdU, scratch, and CCK-8 assays. Immune cell infiltration and polarization in the TME were analyzed by immunofluorescence and flow cytometry. The mechanistic role of calcium-NF-κB signaling in macrophage polarization was investigated using RNA sequencing, western blotting, and specific inhibitors. KEY RESULTS:US-activated BZT generated a significant piezoelectric current (7.5 nA) and ROS, leading to apoptosis in HCC cells. In the orthotopic mouse model, US+BZT treatment markedly inhibited tumor growth. Crucially, it reprogrammed the immunosuppressive TME by increasing the infiltration and cytotoxicity of CD8+ T cells and driving the repolarization of macrophages from the M2 to the M1 phenotype. In vitro experiments confirmed that the US+BZT-induced M1 polarization enhanced macrophage phagocytosis and CD8+ T cell activation. Mechanistically, the effect was mediated by US-induced piezoelectric activation of BZT, which triggered calcium influx via TRP channels, subsequently activating the PKC-IKK-NF-κB signaling axis. CONCLUSION:Our findings demonstrate that US-activated piezoelectric BZT nanocubes exert potent antitumor effects through direct piezoelectric catalysis and, more importantly, through the remodeling of the TME via the calcium/NF-κB pathway. This study highlights the great potential of piezoelectric nanomaterials as a multifaceted and immunomodulatory strategy for cancer therapy.