Background Colorectal cancer (CRC) is a highly prevalent and lethal digestive - tract malignancy. Its global incidence and mortality are on the rise. Understanding the underlying mechanisms of CRC development is crucial for improving clinical diagnosis and treatment. Methods The study employed Mendelian Randomization (MR) analysis, considering plasma proteins, metabolites, and tissue genes as exposure factors and CRC risk as the outcome. Key molecules were identified, and mediation MR explored their relationships. Bioinformatics analyses, using CRC Bulk RNA-Seq and scRNA-Seq data, validated the expression and functions of key molecules. Finally, the expression and regulatory mechanisms of the identified proteins and metabolites were validated using clinical tissue specimens and cell-mouse experiments. Results MR analysis showed 16 plasma proteins and 87 metabolites had causal relationships with CRC. FABP2, a plasma protein, was negatively correlated with CRC risk, while sphingomyelin was positively associated. Mediation MR indicated sphingomyelin mediated FABP2's protective effect. Bulk RNA-seq data showed that FABP2 expression was significantly lower in CRC tissues compared to normal tissues. GSEA enrichment and PPI analyses revealed that FABP2 is closely associated with lipid metabolic processes. scRNA-seq further revealed that FABP2 is primarily expressed in epithelial cells, and its expression was markedly reduced in epithelial cells from tumor tissues relative to adjacent normal tissues. Clinical validation confirmed that FABP2 expression was significantly downregulated in tumor tissues and CRC patient plasma, whereas sphingomyelin levels were significantly elevated. Finally, vitro and vivo experiments combined with RNA-Seq demonstrated that FABP2 may regulate sphingomyelin metabolism and modulate CRC progression via the PPAR signaling pathway. Conclusion FABP2 and sphingomyelin are closely related to CRC development. FABP2 may inhibit CRC progression by regulating sphingomyelin metabolism through the PPAR signaling pathway, suggesting its potential as a candidate biomarker for CRC diagnosis and a potential target for therapeutic intervention.

Necroptosis, driven by RIP3, is critical for intestinal homeostasis but requires stringent regulation to prevent inflammatory pathology. While inhibitory checkpoints of RIP3 are known, mechanisms enabling its physiological activation remain unclear. We identify PRMT5-mediated symmetric di-methylation of RIP3 at arginine 479 (R479) serves as a gatekeeper specifically suppresses RIP1- but not ZBP1- driven necroptosis in intestinal epithelial cells (IECs). Notably, even though STORM super-resolution imaging and molecular dynamics simulations revealed that R479 methylation adjacent to the RHIM domain sterically blocks RIP3 oligomerization both with RIP1 and ZBP1. Mice harboring IEC-specific Rip3R479K knockin mutation developed spontaneous necroptosis-driven colitis in vivo, directly linking this PTM to pathogenesis. Leveraging these insights, we engineered an SDMA-mimetic peptide that competitively inhibit necroptosis, rescuing Prmt5-deficient mice from lethal colitis without disrupting global signaling homeostasis. Unlike conventional kinase inhibitors, this “methylation mimicry” strategy selectively targets pathogenic cell death, offering a therapeutically actionable advance for IBD and related disorders.

The burden of postoperative liver metastasis (LM) in colorectal cancer remains substantial. Consequently, there is a pressing demand for highly effective predictive biomarkers to expedite early diagnosis and treatment. Using a retrospective design, 273 patients were recruited from Xijing Hospital between October 2015 and July 2021. The cohort comprised 91 patients with LM and 182 without LM. The regions of interest in the liver, spleen, and tumor were delineated using portal venous phase CT scans, followed by extraction of radiomic features. Subsequently, radiomics score (rad-score) was developed based on the optimal features identified in each respective region. Finally, a combined predictive model was developed incorporating rad-score and clinicopathological characteristics. Multivariable analysis confirmed that the liver, spleen, and tumor radiomics scores were each independent predictors of LM. The combined radiomic–clinical model demonstrated strong discriminative ability, with AUCs of 0.866 in the training set and 0.814 in the validation set, and a nomogram was subsequently developed. The calibration curves indicated a strong degree of agreement between the predicted values and the actual event probabilities. Decision curve analysis demonstrated that the nomogram could provide a substantial net benefit, indicating that its use would result in a considerable improvement in clinical outcomes across a broad range of situations. The combined model exhibited satisfactory predictive performance, thereby contributing to the enhancement of clinical diagnostic accuracy and prognostic predictions for LM in CRC patients.

OBJECTIVE:To develop interpretable habitat imaging based on multi-b-value diffusion-weighted imaging (DWI) for characterizing tumor microenvironment features and predicting progression-free survival (PFS) in rectal cancer (RC). METHODS:This study enrolled pathologically confirmed patients with RC (n = 153) who underwent multi-b-value DWI before therapy. Intratumoral and peritumoral habitat subregions were coded using K-means clustering based on three maps (DDC_map, α_map and f_map). The associations between habitat features (fraction and Rad_score) and clinicopathological characteristics, as well as tumor microenvironment metrics (Glasgow Microenvironment Score (GMS)) were analysed using appropriate statistical methods. Univariable and multivariable Cox proportional hazards analysis were performed to assess the prognostic factors for RC. Four models (Clinical, Habitat, Habitat_Rad, and Nomogram) were developed for predicting 3-year PFS in RC. The performance of the four models was evaluated using Harrell's concordance index (C-index) and receiver operating characteristic (ROC) curves. The risk stratification of the Nomogram was investigated using Kaplan-Meier survival analysis and log-rank test. RESULTS:Overall, 153 patients (n = 153; 59.25 ± 11.95 years) with RC were randomly divided into training (n = 107) and validation sets (n = 46). Intratumoral and peritumoral tissues were partitioned into three habitat subregions, respectively. Intratumoral habitat 3 (Intra-H3) fraction demonstrated a significant positive correlation with lymph node metastasis (p = 0.010), mrT-stage (p = 0.041) and GMS grade (p = 0.013), and served as an independent risk factor for PFS (HR = 1.957, 95%CI (1.209---3.167), p = 0.006). Peritumoral (0-3 mm) habitat 2 (Peri3-H2) fraction exhibited significant differences across Klintrup-Makinen (KM) grades (p=0.014). The Nomogram integrating habitat features demonstrated superior performance compared with conventional clinical model in the training (AUC = 0.900, 95% CI (0.835-0.966) VS 0.737, 95% CI (0.625-0.850), p = 0.002) and validation (AUC = 0.826, 95% CI (0.707-0.945) VS 0.697, 95% CI (0.524-0.871), p = 0.101) sets, respectively. The Kaplan-Meier curves of the Nomogram score showed good discriminative ability for 3-year PFS in RC. CONCLUSIONS:Intra-H3 features reflect the tumor microenvironment and enhance predictive performance of 3-year PFS in RC. Integrating habitat features from multi-b-value DWI offers superior predictive performance, facilitating precise identification of high-risk patients.

For patients with mismatch repair-proficient / microsatellite stable (pMMR/MSS) locally advanced rectal cancer (LARC), the NECTAR (NEoadjuvant Chemoradiotherapy plus immunoTherapy for locally Advanced Rectal cancer) trial reported an encouraging pathological complete response (pCR) rate of 40.0%. Given that pMMR/MSS tumors are generally resistant to immunotherapy, this pCR rate highlights the potential of combination therapy to overcome primary resistance. Here, we report the 3-year survival outcomes of the NECTAR trial, using neoadjuvant chemoradiotherapy (NCRT) plus tislelizumab in pMMR/MSS LARC. In this phase 2, multi-center, single-arm trial, eligible pMMR/MSS LARC patients received three cycles of neoadjuvant programmed cell death protein 1 (PD-1) blockade, three cycles of chemotherapy, and long-course radiotherapy followed by total mesorectal excision (TME). This 3-year analysis evaluates disease-free survival (DFS) and safety. From June 2021 to November 2022, 50 patients were enrolled and 46 patients completed neoadjuvant therapy and TME. Of the 46 patients, 20 patients reached pCR. With a median follow-up of 44.2 months, the median DFS was not reached. The 3-year DFS rate was 89.1%. All patients who achieved pCR (20/46) remained DFS at 3 years. Patients with a low neoadjuvant rectal (NAR) score (<8) had the 3-year DFS rate of 100%, suggesting that overcoming therapeutic resistance in immunotherapy may translate into sustained survival benefits. No new grade ≥III treatment-related adverse events (trAEs) occurred during the 3-year follow-up. The 3-year survival outcomes demonstrate promising efficacy and safety of this NECTAR trial, which provides a promising therapeutic avenue for a patient population traditionally resistant to single-agent immunotherapy.

Circadian disruption is linked to colorectal cancer (CRC) progression. We investigated the role of the tumor suppressor N-myc downstream-regulated gene 2 (NDRG2) in this context. Intestinal-specific Ndrg2 knockout exacerbated colorectal tumorigenesis in mice under circadian disruption. We identified NDRG2 as a novel stabilizer of the core clock protein Circadian Locomotor Output Cycles Kaput (CLOCK). Mechanistically, NDRG2 bidirectionally regulates CLOCK ubiquitination, inhibiting its degradation by the E3 ligase STIP1 Homology and U-Box Containing Protein 1 (STUB1) while promoting stabilization by the deubiquitinase Ubiquitin Specific Peptidase 8 (USP8). This NDRG2-CLOCK axis was crucial for oxaliplatin sensitivity. Integrated RNA sequencing and functional studies revealed that CLOCK transcriptionally represses Fibroblast Growth Factor 2 (FGF2), a chemoresistance driver. Thus, NDRG2 enhances CLOCK activity to suppress FGF2 and promote oxaliplatin efficacy. Clinically, oxaliplatin-sensitive CRC tissues showed higher NDRG2/CLOCK co-expression and lower FGF2 levels. Our work defines a pathway wherein NDRG2 inhibits tumorigenesis and chemoresistance by stabilizing CLOCK to transcriptionally represses FGF2, linking circadian regulation to therapy response, and nominating the NDRG2-STUB1/USP8-CLOCK-FGF2 axis as a therapeutic target.

Background: Constipation is frequently accompanied by anxiety and depressive symptoms, suggesting coordinated disturbances in the gut microbiota, intestinal metabolism, and gut-brain axis. Fecal microbiota transplantation (FMT), as a microbiome-targeted intervention, has shown potential in functional gastrointestinal disorders. However, its clinical response and multi-omics mechanisms in constipation complicated by anxiety remain unclear. Methods: Eighteen patients with constipation complicated by anxiety received two cycles of oral FMT capsules, and 10 healthy individuals were enrolled as controls. Phenotypes were assessed before and after treatment using Wexner, PAC-QOL, ODS, SDS, and HAMA scores. Fecal samples from patients before and after FMT and from healthy controls were subjected to shotgun metagenomic sequencing and untargeted metabolomics. Microbial composition, MetaCyc pathways, fecal metabolites, efficacy-stratified features, and microbiota-metabolite-clinical phenotype correlation networks were analyzed. Results: After FMT, Wexner, PAC-QOL, ODS, SDS, and HAMA scores were reduced. Metagenomic analysis showed that baseline gut microbial structure in patients was distinct from healthy controls and shifted toward a healthy-like profile after FMT. Differential species analysis suggested partial microbial recovery, with reversible species mainly involving Clostridium, Blautia, and Lachnospira. Functional pathway analysis showed recovery of selected amino acid metabolism and anaerobic fermentation pathways, whereas pathways related to bacterial surface structures and polysaccharide biosynthesis remained persistently abnormal. Untargeted metabolomics revealed broad fecal metabolic remodeling, characterized by a shift from a relatively suppressed state toward a more active profile. Efficacy-stratified analyses indicated that the Effective and No_Effective groups differed in baseline microbial composition, functional niches, and metabolic background. Integrated correlation network analysis identified associations among differential species, metabolites, and clinical phenotypes. In particular, Butyricicoccus sp. AM29-23AC was negatively correlated with constipation, quality-of-life, anxiety, and depression-related scores and was embedded in a short-chain fatty acid-related microbiota-metabolite network. Conclusions: Oral FMT capsules were associated with improvement in constipation, quality-of-life, anxiety, and depression-related scores in patients with constipation complicated by anxiety, accompanied by coordinated remodeling of gut microbial composition, microbial functional potential, and fecal metabolic profiles. Baseline microbial and metabolic states may influence clinical response to FMT. Butyricicoccus sp. AM29-23AC and short-chain fatty acid-related microbiota-metabolite networks may represent candidate links between FMT-associated ecological remodeling and clinical improvement.

Aims: We aimed to develop Ferroptosis-Related Gene (FRG) signatures to predict overall survival (OS) along with disease-free survival (DFS) in individuals with colorectal cancer (CRC). Background: Prediction of CRC prognosis is challenging. Ferroptosis constitutes a newly reported kind of cell death, and its association with CRC prognosis remains unexplored. Objective: This research endeavored to establish a prognostic risk signature for colorectal cancer by leveraging ferroptosis-related genes (FRGs), with the objective of refining prognostic precision in clinical settings. Methods: The clinical data and mRNA expression profiles were obtained from The Cancer Genome Atlas (TCGA) colorectal cancer cohorts. The Lasso algorithm was employed to develop the overall survival (OS) and disease-free survival (DFS) prediction models. These models were subsequently validated using independent data from GSE38832. Results: Our research unveiled a significant difference in the expression levels of 85% of ferroptosis-related genes (FRGs) between CRC tissues and paracancer tissues. Out of these, 11 prognostic genes were pinpointed through univariate Cox analysis. By employing two models, patients were stratified into low- and high-risk groups based on predicted risk scores, which were subsequently validated as independent prognostic factors via multivariate Cox analysis. The robustness of these models was further confirmed through Receiver Operating Characteristic (ROC) curve analysis. Functional enrichment analysis indicated a predominance of cancer-associated pathways in the high-risk group, including WNT signaling, along with variations in immune status between the two risk categories. Leveraging the Connectivity Map (CMap) database, a total of sixteen potential therapeutic drugs were identified. Additionally, in vitro experiments corroborated that Farnesyl-Diphosphate Farnesyltransferase 1 (FDFT1) was underexpressed in CRC and exhibited tumor suppressive properties. More specifically, FDFT1 may augment ferroptosis in CRC by modulating the expression of the Iron-Sulfur Cluster Assembly Enzyme (ISCU). Conclusion: Our study highlighted the significance of ferroptosis-related genes in the pathogenesis of CRC and underscored the potential of ferroptosis-related gene-based risk signatures as valuable tools for improving prognostic accuracy and tailoring therapeutic strategies. However, the validity of these predictive models required further validation through real- world studies to ensure their reliability and applicability.

BackgroundGastric cancer patients with metachronous liver metastasis (MLM) generally pose a significant clinical challenge, with 5-year survival below 10%. Macrophages are key components of a tumoral microenvironment, but the predictive value of their morphological features for MLM remains unexplored. This study aimed to construct a risk predictive model for MLM based on macrophage morphology in the tumor microenvironment.MethodsTo reduce major baseline imbalances between patients with and without MLM, propensity score matching (PSM) was performed. After matching, a retrospective analysis of 233 gastric cancer patients who underwent radical surgery between 2016 and 2020 was conducted. Macrophage morphological parameters in different tissues were quantified by Qupath. Patients were randomly divided into training (70%) and validation (30%) datasets. The optimal cutoff values for the continuous variables were determined using the Youden index. Univariate and multivariate logistic regression analyses in the training set were used to identify risk factors for MLM. A nomogram was constructed for clinical applications. The model’s value was validated through receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA).ResultsThe incidence of MLM was similar in the training (38.7%, 63/163) and validation cohorts (32.9%, 23/70). The optimal cut off value of macrophage areas and perimeters in the tumor invasive front (IF) region, tumor region, peritumoral stroma (PS) region were 76.75 μm2, 35.77 μm, 71.35 μm2, 32.00 μm, 74.13 μm2, 31.32 μm. Multivariate analysis revealed that only the area and perimeter of macrophages in the IF region, the area of macrophages in the tumor region were independent predictive factors for MLM. These factors were incorporated into a predictive nomogram. The model demonstrated good discrimination, with an area under the curve (AUC) of 0.835 in the training set and 0.798 in the validation set. Calibration and decision curve analyses confirmed its clinical utility.ConclusionMacrophage morphology parameters, particularly in the IF and tumor regions, are significantly associated with MLM in gastric cancer. The morphology-based nomogram developed in this study provides an exploratory tool for risk stratification and may help inform individualized postoperative surveillance.

Proliferation and metastasis are the core malignant characteristics in esophageal squamous cell carcinoma (ESCC) that contribute to poor prognosis. However, the mechanisms underlying cell proliferation and metastasis remain elusive. We explored the function of high-mobility group box (HMGB3) in promoting ESCC progression. HMGB3 expression in ESCC tissues and cell lines was quantified using quantitative PCR, Western blotting, and immunohistochemistry. The proliferative and migratory characteristics of ESCC cells were assessed using in vitro and in vivo assays, respectively. An RNA sequencing analysis was conducted to identify the downstream signaling pathways of HMGB3. Co-immunoprecipitation was performed to identify HMGB3-interacting proteins. HMGB3 transcriptional regulation was investigated using luciferase reporter and chromatin immunoprecipitation assays. Elevated levels of HMGB3 were observed in both patient-derived ESCC tissues and ESCC cell lines and were correlated with poor patient prognosis. HMGB3 up-regulation promoted ESCC proliferation and metastasis, whereas HMGB3 down-regulation inhibited these processes. Mechanistically, homeodomain protein transforming growth factor beta (TGF-u03B2)-induced factor homeobox 2 (TGIF2) transcriptionally up-regulates HMGB3. HMGB3 subsequently activates TGF-u03B2 signaling through its regulation of and interaction with toll-like receptor 3 (TLR3), ultimately promoting ESCC proliferation and metastasis. Clinically, HMGB3 expression was positively correlated with TGIF2 and TGF-u03B2, and patients with ESCC who positively co-expressed TGIF2/HMGB3, HMGB3/TGF-u03B2, or TGIF2/TGF-u03B2 exhibited poor prognosis. The functional role of HMGB3 in ESCC proliferation and metastasis was illustrated in our research. Targeting the TGIF2/HMGB3/TLR3/TGF-u03B2 axis has the potential to serve as a promising therapeutic approach.

The National Comprehensive Cancer Network (NCCN) recommends that at least 12 lymph nodes (LNs) should be harvested during radical surgery of colorectal cancer (CRC). This study aims to clarify the optimal number of LNs that should be examined during the radical surgery for CRC patients with each N stage. The SEER database (n = 48,331) served as the primary cohort, while the Xijing cohort (n = 4447) was used for external validation. A Cox proportional hazards regression model was employed to evaluate risk factors associated with overall survival (OS) in CRC. Moreover, the restricted cubic splines (RCS) method was used to detect inflection points within the risk function and to investigate the correlation between the number of examined lymph nodes (ELNs) and OS. Ultimately, the optimal number of ELNs for patients at different N stages was determined and verified. The analyses from both databases consistently indicated that a higher number of ELNs is an independent prognostic factor for the OS of patients with CRC. 18 and 19 are identified as the optimal numbers of ELNs for N0, N1a, and N1b, N2a stages, respectively. The Kaplan–Meier analysis revealed that there was a statistical difference in the OS between patients in the limited group and those in the adequate group among the neighboring N subgroups. For CRC patients with different N stages, the number of LNs that needs to be cleared varies. Number of ELNs exceeded 18, 18, 19 and 19 could significantly improve the prognosis of patients with N0, N1a, N1b and N2a stages, respectively.

Gastric cancer remains a formidable global health challenge, with drug resistance being a primary contributor to its high mortality rates. Therefore, understanding the mechanisms driving drug resistance is crucial for developing more effective therapeutic interventions. Utilizing high content screening technology, we identified let-7e-5p and miR-125a-5p as the key regulators of drug resistance in GC, both being members of the miR-99b cluster. Findings from GC cells, organoids and PDX models demonstrated that overexpression of the miR-99b cluster sensitized GC to cisplatin, likely through its inhibitory effects on mitochondrial respiratory function, particularly OXPHOS. We further demonstrated that multiple components, especially SDHC subunit of respiratory complex II, were regulated by miR-99b cluster and played critical roles in GC chemoresistance. Then, through a comprehensive screen of epigenetic compounds, we identified KMT5A as a key upstream repressor of the miR-99b cluster in GC, which reinforces the H4K20me1 modification within the miR-99b cluster region. Moreover, elevated KMT5A expression and decreased miR-125a-5p expression indicated both poorer prognosis and chemo-resistance in patients with GC. This study highlights the multifaceted roles of the miR-99b cluster in GC and offers novel perspectives for the development of innovative therapeutics aimed at overcoming chemoresistance and enhancing treatment efficacy for GC patients.

OBJECTIVE/BACKGROUND:The RNPLS-01 trial aimed to evaluate the efficacy and safety of ramucirumab (a VEGFR2 antagonist) combined with nab-paclitaxel, lobaplatin, and S-1 (RNPLS group) versus nab-paclitaxel, lobaplatin, and S-1 alone (NPLS group) in neoadjuvant therapy for advanced gastric cancer (GC), as part of the RNPLS-01 prospective randomized controlled trial. METHODS AND ANALYSIS:RNPLS-01 trial enrolled 140 patients with advanced GC across two cohorts (neoadjuvant therapy: n = 70; conversion therapy: n = 70). This report focuses on the neoadjuvant cohort where patients were randomized 1:1 to receive either RNPLS (n = 35) or NPLS (n = 35). Primary endpoint was pathological complete response (pCR). Secondary endpoints included R0 resection rate, objective response rate (ORR), disease control rate (DCR), and adverse events (AEs). RESULTS:The RNPLS group demonstrated significant improvements in post-operative T stage ( P = 0.008), N stage ( P = 0.001), and AJCC (American Joint Committee on Cancer) clinical stage ( P = 0.003) compared to controls. The RNPLS group demonstrated significantly higher pCR rates compared to the NPLS group (22.9% vs. 2.9%, P <0.0001). The R0 resection rate was 85.7% in the RNPLS group versus 82.9% in the NPLS group. The RNPLS group achieved a significantly higher ORR compared to the NPLS group (68.6% vs. 25.7%, P = 0.001), while DCRs were comparable between the two groups (94.3% vs. 77.1%, P = 0.055). Treatment-related adverse events (TRAEs) were confined to Grade 1-2 severity, with no Grade 3-4 events reported in either group. The RNPLS group showed a TRAE incidence of 80.0%, versus 88.6% in the NPLS group ( P = 0.324). Neither group exhibited surgical complications during the study period. CONCLUSION:Ramucirumab combined with nab-paclitaxel, lobaplatin, and S-1 significantly enhances the pCR rate and ORR in neoadjuvant therapy of locally advanced GC patients, while maintaining an acceptable safety profile.

BackgroundPatients with metachronous liver metastasis (MLM) in gastric cancer generally have a poor prognosis. Early detection and accurate prediction of MLM are crucial for improving clinical outcomes. This study aims to identify the risk factors for MLM through clinical pathological parameters and develop a predictive model for MLM in gastric cancer.MethodsA retrospective analysis of 1248 gastric cancer patients who underwent radical surgery between December 2016 and December 2020 was conducted. Patients were randomly divided into training (70%, n=873) and validation (30%, n=375) datasets. The optimal cutoff values for the continuous variables were determined using the Youden index. Univariate and multivariate logistic regression analyses were used to identify risk factors for MLM. A nomogram was developed based on the results of multivariate analysis. The model’s value was validated through receiver operating characteristic (ROC) curves, calibration curves, and decision curve analysis (DCA).ResultsThe incidence of MLM was comparable between the training (10.3%, 90/873) and validation set (9.9%, 37/375). The optimal cutoff value was 3.315ng/ml for preoperative alpha-fetoprotein (AFP) level, 16.275U/ml for preoperative cancer antigen 125 (CA125) level, 0.280×109/L for monocyte count and 1.430×109/L for lymphocyte count, respectively. Univariate analysis showed that age, tumor size, pathological type, surgical method, T stage, N stage, TNM stage, neural invasion, lymphatic vascular invasion, number of lymph nodes harvested (LNH), preoperative total protein (TP), hemoglobin (HB), albumin (ALB), preoperative carcinoembryonic antigen (CEA), preoperative cancer antigen 19-9 (CA19-9), CA125, AFP levels, monocyte count, lymphocyte count, red blood cell (RBC) count and platelet count were considered as potential variables. Multivariate logistic regression analysis indicated that T stage, N stage, monocyte count, lymphocyte count, preoperative AFP and CA125 levels were independent predictive factors for MLM. The identified risk factors were further used to develop a predictive nomogram for MLM. The nomogram exhibited robust discriminatory performance, with an area under the curve (AUC) of 0.859 in the training set and 0.803 in the validation set. Moreover, the nomogram demonstrated excellent calibration and significant clinical utility.ConclusionThis study successfully developed a predictive nomogram for MLM in gastric cancer. Besides conventional parameters, we identified and incorporated peripheral blood monocyte and lymphocyte counts as novel predictors, demonstrating their independent predictive value. Integrating these factors into nomogram could enhance predictive accuracy of MLM.

SPP1+ macrophages have been identified as key players in the colorectal cancer (CRC) tumor microenvironment, but their function remains unclear. This study integrated single-cell and spatial transcriptomics with bulk sequencing to investigate the roles and mechanisms of SPP1+ macrophages in CRC. Our findings revealed a pronounced elevation of SPP1+ macrophages in CRC, especially within tumor territories. These macrophages served as markers for CRC initiation, progression, metastasis, and potential prognosis. Furthermore, they showed heightened transcriptional activity in genes linked to angiogenesis, epithelial-mesenchymal transition, glycolysis, hypoxia, and immunosuppression. SPP1 protein amplified CRC cell migration and invasion, potentially mediating cellular crosstalk via the SPP1-CD44, SPP1-PTGER4, and SPP1-a4b1 complex axes. Patients with a high proportion of SPP1+ macrophages could benefit more from immune checkpoint blockade therapy. Interestingly, CSF1R expression was significantly enriched in C1QC+ macrophages versus SPP1+ macrophages, possibly explaining limited anti-CSF1R monotherapy effects. In conclusion, we propose an SPP1+ macrophage model in CRC, highlighting such macrophages as a promising therapeutic target due to their malignancy markers.

Proliferation and metastasis are the core malignant characteristics in esophageal squamous cell carcinoma (ESCC) that contribute to poor prognosis. However, the mechanisms underlying cell proliferation and metastasis remain elusive. We explored the function of high-mobility group box (HMGB3) in promoting ESCC progression. HMGB3 expression in ESCC tissues and cell lines was quantified using quantitative PCR, Western blotting, and immunohistochemistry. The proliferative and migratory characteristics of ESCC cells were assessed using in vitro and in vivo assays, respectively. An RNA sequencing analysis was conducted to identify the downstream signaling pathways of HMGB3. Co-immunoprecipitation was performed to identify HMGB3-interacting proteins. HMGB3 transcriptional regulation was investigated using luciferase reporter and chromatin immunoprecipitation assays. Elevated levels of HMGB3 were observed in both patient-derived ESCC tissues and ESCC cell lines and were correlated with poor patient prognosis. HMGB3 up-regulation promoted ESCC proliferation and metastasis, whereas HMGB3 down-regulation inhibited these processes. Mechanistically, homeodomain protein transforming growth factor beta (TGF-β)-induced factor homeobox 2 (TGIF2) transcriptionally up-regulates HMGB3. HMGB3 subsequently activates TGF-β signaling through its regulation of and interaction with toll-like receptor 3 (TLR3), ultimately promoting ESCC proliferation and metastasis. Clinically, HMGB3 expression was positively correlated with TGIF2 and TGF-β, and patients with ESCC who positively co-expressed TGIF2/HMGB3, HMGB3/TGF-β, or TGIF2/TGF-β exhibited poor prognosis. The functional role of HMGB3 in ESCC proliferation and metastasis was illustrated in our research. Targeting the TGIF2/HMGB3/TLR3/TGF-β axis has the potential to serve as a promising therapeutic approach.

Drug resistance in colorectal cancer (CRC) poses a major challenge for cancer therapy and stands as the primary cause of cancer-related mortality. The N6-methyladenosine (m6A) modification has emerged as a pivotal regulator in cancer biology, yet the precise m6A regulators that propel CRC progression and chemoresistance remain elusive. Our study established a significant correlation between m6A regulatory gene expression profiles and CRC severity. Notably, based on the knockout cellular and mouse model created by CRISPR/Cas9-mediated genome engineering, we identified m6A demethylase FTO emerged as a pivotal orchestrator of CRC chemoresistance through the regulation of NUPR1, a critical transcription factor involved in iron homeostasis via LCN2 and FTH1. Mechanistic study revealed that FTO stabilized NUPR1 mRNA by specifically targeting the +451 m6A site, thereby preventing YTHDF2-mediated degradation of NUPR1 mRNA. Moreover, the simultaneous targeting of FTO and NUPR1 dramatically enhanced the efficacy of chemotherapy in CRC cells. Our findings underscore the potential of modulating the m6A methylome to overcome chemoresistance and highlight the FTO-NUPR1 axis as a critical determinant in CRC pathobiology.

INTRODUCTION:Anastomotic leakage (AL) following low anterior resection remains a serious complication related to mortality and recurrence. We propose a novel method for treating AL after radical resection of low rectal cancer. CASE PRESENTATION:The patient was a 36-year-old male who was referred to the department of digestive surgery of our hospital due to intermittent hematochezia for 2 years. Physical examination yielded the following: 9-12 o'clock in the knee-chest (KC) position, about 5 cm from the anal verge, with soft, mobile, unclear edges, and mild tenderness with no blood staining. The patient was pathologically diagnosed as rectal adenocarcinoma before admission and staged as cT3N2M0 by Computed tomography (CT) and magnetic resonance imaging (MRI), long-term radiotherapy (25x45Gy) with chemotherapy(capecitabine), was performed after 4 cycles of CapeOX protocol, and the patient then underwent transanal total mesorectal excision(TaTME). 15 days after the surgery, the patient was readmitted due to hematochezia which caused by AL. We applied modified Bacon operation on this patient, and successfully performed a second operation two weeks later. CLINICAL DISCUSSION:Given its simple operation, few complications and effective treatment, Modified Bacon can be used for remedy anastomotic leakage after TaTME. CONCLUSIONS:We propose modified Bacon operation to deal with anastomotic leakage which provides a new strategy to treat AL after low anterior resection (LAR).

Background Colorectal cancer (CRC) prognosis assessment is vital for personalized treatment plans. This study investigates the prognostic value of dynamic changes of tumor markers CEA, CA19-9, CA125, and AFP before and after surgery and constructs prediction models based on these indicators. Methods A retrospective clinical study of 2599 CRC patients who underwent radical surgery was conducted. Patients were randomly divided into training (70%) and validation (30%) datasets. Univariate and multivariate Cox regression analyses identified independent prognostic factors, and nomograms were constructed. Results A total of 2599 CRC patients were included in the study. Patients were divided into training (70%, n = 1819) and validation (30%, n = 780) sets. Univariate and multivariate Cox regression analyses identified age, total number of resected lymph nodes, T stage, N stage, the preoperative and postoperative changes in the levels of CEA, CA19-9, and CA125 as independent prognostic factors. When their postoperative levels are normal, patients with elevated preoperative levels have significantly worse overall survival. However, when the postoperative levels of CEA/CA19-9/CA125 are elevated, whether their preoperative levels are elevated or not has no significance for prognosis. Two nomogram models were developed, and Model I, which included CEA, CA19-9, and CA125 groups, demonstrated the best performance in both training and validation sets. Conclusion This study highlights the significant predictive value of dynamic changes in tumor markers CEA, CA19-9, and CA125 before and after CRC surgery. Incorporating these markers into a nomogram prediction model improves prognostic accuracy, enabling clinicians to better assess patients’ conditions and develop personalized treatment plans.

Adding PD-1 blockade in the neoadjuvant regimens for locally advanced rectal cancer (LARC) patients with microsatellite stable (MSS) / mismatch repair-proficient (pMMR) tumors is an attractive, but debatable strategy. This phase 2, multicenter, prospective, single-arm study enrolled patients from 6 centers from June 2021 to November 2022. Locally advanced rectal cancer (LARC, cT 3-4a N 0 M 0 and cT 1-4a N 1-2 M 0 ) patients aged ≥18 years with the distance from distal border of tumor to anal verge ≤10 cm (identified by Magnetic Resonance Imaging) were qualified for inclusion. The patients received long-course radiotherapy (50 Gy/25 fractions, 2 Gy/fraction, 5 days/week) and three 21-day cycles capecitabine (850–1000 mg/m2, bid, po, day1–14) and three 21-day cycles tislelizumab (200 mg, iv.gtt, day8) as neoadjuvant. Total mesorectal excision (TME) was 6–12 weeks after the end of radiotherapy to achieve radical resection. A total of 50 patients were enrolled in this study. The pathological complete response rate was 40.0% [20/50, 95% confidence interval (CI): 27.61–53.82%], while 15 (30.0%, 95% CI: 19.1–43.75%), 9 (18.0%, 95% CI: 9.77–30.8%), 2 (4.0%, 95% CI: 1.10–13.46%) patients respectively achieved grade 1, 2, and 3 tumor regression. Treatment-related adverse events (TRAEs) occurred in 28 (56.0%) LARC patients, including 26(52.0%) with grade I-II and 2 (4.0%) with grade III (1 with grade 3 immune-related colitis and 1 with grade 3 rash). PD-1 blockade plus long-course chemoradiotherapy (CRT) showed promising therapeutic effects according to pathological complete response rate and is well-tolerated in LARC patients. A larger randomized controlled study is desired to further validate the above findings.