BackgroundColon adenocarcinoma (COAD) shows marked clinical heterogeneity, and robust biomarkers with mechanistic relevance are still needed for prognostic stratification. RBMS3 has been implicated in tumor biology, but its pan-cancer significance, prognostic value in COAD, and regulatory mechanisms remain insufficiently defined.MethodsWe performed integrated analyses of The Cancer Genome Atlas pan-cancer and COAD cohorts, followed by external validation in GEO datasets. Prognostic associations were evaluated by Cox and Kaplan-Meier analyses, and clinicopathological relevance was assessed in TCGA-COAD. A prognostic nomogram combining RBMS3 and clinical variables was constructed. Single-cell RNA-seq analysis was used to characterize RBMS3-associated functional programs. In vitro experiments in HCT116 cell and THP-1-derived M2 macrophage co-culture systems were conducted to validate ferroptosis-related effects and upstream STAT3-dependent regulation.ResultsRBMS3 was significantly dysregulated across multiple tumor types and showed prognostic relevance in pan-cancer analysis, with a prominent adverse survival association in COAD. GEO cohorts confirmed decreased RBMS3 expression in tumor samples relative to normal controls, and low RBMS3 expression was associated with unfavorable prognosis, supporting its tumor-suppressive prognostic role. In TCGA-COAD, RBMS3 expression correlated with T stage and pathological stage, and integration of RBMS3 with staging factors improved prognostic stratification in a nomogram model. Single-cell analysis showed enrichment of ferroptosis-related pathways in RBMS3-positive tumor cells. Functionally, RBMS3 overexpression in HCT116 cells reduced viability, decreased GPX4, increased ACSL4, and elevated intracellular Fe ion levels, indicating molecular features consistent with enhanced ferroptotic sensitivity. Mechanistically, M2 macrophages suppressed RBMS3 expression in tumor cells with concomitant STAT3 activation, while pharmacological STAT3 inhibition partially restored RBMS3 expression.ConclusionRBMS3 is a clinically relevant prognostic biomarker in COAD and is linked to ferroptosis-associated tumor-cell programs. The M2 macrophage-STAT3-RBMS3 axis provides a mechanistic connection between the immune microenvironment and tumor intrinsic regulation, highlighting RBMS3 as a potential target for prognostic modeling and translational intervention in COAD.

Colorectal cancer (CRC) is the third most common malignant tumor and the second leading cause of cancer‑related deaths worldwide. Identifying driver genes in CRC development may provide clinical benefits for patients. Zinc finger protein 695 (ZNF695) is a nuclear protein with transcriptional regulatory activity, which has been implicated in tumor progression; however, the role of ZNF695 in CRC is unclear. The clinical relevance of ZNF695 and NIMA‑related kinase 2 (NEK2) in patients with CRC was analyzed based on The Cancer Genome Atlas database. Knockdown was performed by transfecting the cells with small interfering RNAs, whereas overexpression was induced by infecting the cells with a lentivirus. In addition, cell Counting Kit‑8, colony formation, cell cycle and apoptosis assays were carried out to assess the role of ZNF695 and NEK2 in CRC. Chromatin immunoprecipitation‑quantitative PCR (qPCR) and dual luciferase reporter assays were used to examine the transcriptional regulation of ZNF695 on the NEK2 gene. Reverse transcription‑qPCR and western blotting were applied to assess mRNA and protein abundance, respectively. The present study aimed to investigate the clinical relevance, contribution and downstream effects of ZNF695 in CRC. The results revealed that ZNF695 was upregulated in CRC tissues compared with that in non‑cancer tissues. CRC cells also expressed higher ZNF695 expression than normal cells. In vitro, knockdown of ZNF695 suppressed the proliferation of HCT‑8 cells; conversely, overexpression of ZNF695 promoted the malignancy of HT‑29 cells. Moreover, ZNF695 accelerated cell cycle progression and inhibited apoptosis in CRC cells. Mechanistically, it was revealed that ZNF695 upregulated the expression of NEK2 at both the mRNA and protein levels. Luciferase reporter assay demonstrated that ZNF695 enhanced the transcriptional activity of the NEK2 promoter. Furthermore, knockdown of NEK2 reversed the oncogenic function of ZNF695. Additionally, ZNF695 activated the PI3K/Akt/mTOR signaling pathway in CRC cells. Inhibition of this pathway with rapamycin resulted in higher cytotoxicity to CRC cells with ZNF695 overexpression, suggesting that elevated ZNF695 levels may increase the sensitivity of CRC cells to rapamycin. In summary, the current study identified ZNF695 as a tumor‑promoting protein in CRC through activation of the NEK2 and PI3K/Akt/mTOR signaling pathways. Targeting the NEK2 and PI3K/Akt/mTOR signaling pathways may therefore be a promising strategy for the treatment of patients with CRC and high ZNF695 expression.

Langerhans cell histiocytosis (LCH) of the stomach is rare. Moreover, it is usually found in pediatric patients with systemic diseases and may be associated with a poor prognosis. Solitary gastric LCH in adults is extremely rare and is often misdiagnosed or missed. The aim of our study was to review cases of gastric LCH and explore the characteristics of the disease further. A retrospective study of all patients admitted with solitary gastric LCH was conducted between 2013 and 2023. Clinical manifestations, endoscopic and pathological features, immunophenotypes, and molecular changes were collected from medical records. We examined four cases (one female, three males) of gastric LCH. The affected patients were between 33 and 70 years of age. Endoscopically, three patients presented with a solitary polyp or elevated lesions, whereas one patient showed no abnormalities. Under a microscope, all cases showed abnormal proliferation of histiocytoid cells infiltrating in a nested or sheet-like fashion. The tumor cells were medium-sized, with a slightly eosinophilic cytoplasm, irregular or renal-shaped nuclei, folded nuclear membranes, visible nuclear grooves, and the infiltration of inflammatory cells in the background. Immunohistochemically, all lesions expressed CD1a, S-100, langerin, and cyclinD1. One case showed diffuse BRAF V600E positivity. Follow-up data were available for all patients from 4 to 36 months, and all patients were alive without recurrence or progress at the time of manuscript preparation. Combined with previously reported data, solitary adult gastric LCH is more common in male patients, most of whom are asymptomatic or exhibit only mild gastrointestinal symptoms, with a good prognosis. Endoscopy often reveals solitary polyps or protruding lesions; rare cases may progress to multifocal/multisystem lesions, necessitating long-term close follow-up.

Gastric cancer typically originates from the abnormal proliferation of normal cells within the gastric mucosa, eventually forming tumors. The roles of sperm-associated antigen 5 (SPAG5) and abnormal spindle-like microcephaly (ASPM) associated genes in gastric cancer are not yet clear. Gastric cancer datasets GSE51575 and GSE36076 profiles were downloaded from the GPL13607 and GPL570-generated gene expression omnibus database. The analysis included filtering for differentially expressed genes, weighted gene co-expression network analysis, functional enrichment analysis, gene set enrichment analysis, immune infiltration analysis, construction and analysis of the protein-protein interaction network, survival analysis, and Comparative Toxicogenomics Database analysis. Heatmaps of gene expression were also created. A total of 1457 differentially expressed genes were identified. According to gene ontology analysis, they are primarily enriched in the metabolic processes of organic acids, condensed chromosome centromere regions, and oxidoreductase activity. Kyoto Encyclopedia of Gene and Genome analysis showed they are mainly involved in metabolic pathways, P53 signaling pathway, and PPAR signaling pathway. The soft threshold power for weighted gene co-expression network analysis was set to 8. Three core genes (CENPE, SPAG5, and ASPM) were identified. Heatmaps of core gene expression revealed that SPAG5 and ASPM are highly expressed in gastric cancer samples and low in normal samples. Comparative Toxicogenomics Database analysis indicated that the core genes (CENPE, SPAG5, and ASPM) are associated with gastric tumors, gastric diseases, gastritis, gastric ulcers, tumors, inflammation, and necrosis. The SPAG5 and ASPM genes are overexpressed in gastric cancer tissues, and higher expression levels are associated with worse prognosis, may serve as potential prognostic markers.

Colorectal cancer (CRC) is one of the most common malignancies worldwide. The 5‑year survival rate of patients diagnosed with the early stages of the disease is markedly higher than that of patients in the advanced stages. Therefore, identifying novel biomarkers and drug targets for CRC is critical for clinical practice. Zinc finger protein 169 (ZNF169) is a crucial transcription factor, and its role in CRC remains to be explored. The present study aimed to investigate the clinical relevance, function and underlying mechanisms of ZNF169 in CRC growth and proliferation. The Cancer Genome Atlas (TCGA) database was utilized to analyze the clinical relevance of ZNF169 in patients with CRC. Immunohistochemical staining was performed on tissue samples from patients with CRC to detect the expression of ZNF169. The HCT‑116, HT‑29 and RKO cell lines were employed for in vitro experiments. The overexpression and knockdown of ZNF169 were achieved by transfecting the cells with lentivirus and small interfering RNAs, respectively. Cell Counting Kit‑8, colony formation and EdU staining assays were applied to investigate the function of ZNF169 in CRC cells. Dual luciferase activity and chromatin immunoprecipitation (ChIP)‑quantitative PCR (qPCR) assays were performed to identify the regulatory effects of ZNF169 on the ankyrin repeat and zinc‑finger domain‑containing 1 (ANKZF1; also known as ZNF744) gene. Reverse transcription‑quantitative PCR and western blot analysis were performed to measure mRNA and protein expression, respectively. The analysis of TCGA data revealed a positive correlation between ZNF169 and ANKZF1, with the overexpression of ANKZF1 being associated with a poor prognosis of patients with CRC. The experimental results demonstrated that ZNF169 was expression upregulated in CRC tissue compared with that in normal colon tissue. Gain‑of‑function and loss‑of‑function experiments revealed that ZNF169 enhanced the intensity of EdU staining, promoting the growth and proliferation of CRC cells. Furthermore, the overexpression of ZNF169 potentiated the transcriptional activity of the ANKZF1 gene, while the knockdown of ZNF169 produced the opposite results. ChIP‑qPCR confirmed the interaction between ZNF169 and the promoter sequence of ANKZF1. Rescue experiments revealed that ZNF169 accelerated CRC cell growth and proliferation through the upregulation of ANKZF1. Furthermore, there was a positive correlation identified between ZNF169 and ANKZF1, and upregulation of ANKZF1 expression was associated with the poor prognosis of patients with CRC. On the whole, the present study demonstrates that ZNF169 contributes to CRC malignancy by potentiating the expression of ANKZF1. Thus, the regulation of ZNF169 and/or ANKZF1 expression may represent a viable strategy for the treatment patients with CRC with a high expression of ZNF169.

The alteration of metabolism is essential for the initiation and progression of numerous types of cancer, including colorectal cancer (CRC). Metabolomics has been used to study CRC. At present, the reprogramming of the metabolism in CRC remains to be fully elucidated. In the present study, comprehensive untargeted metabolomics analysis was performed on the paired CRC tissues and adjacent normal tissues from patients with CRC (n=35) using ultra-high-performance liquid chromatography-mass spectrometry. Subsequently, bioinformatic analysis was performed on the differentially expressed metabolites. The changes in these differential metabolites were compared among groups of patients based on sex, anatomical tumor location, grade of tumor differentiation and stage of disease. A total of 927 metabolites were detected in the tissue samples, and 24 metabolites in the CRC tissue were significantly different compared with the adjacent normal tissue. The present study revealed that the levels of three amino acid metabolites were increased in the CRC tissue, specifically, N-α-acetyl-ε-(2-propenal)-Lys, cyclo(Glu-Glu) and cyclo(Phe-Glu). The metabolites with decreased levels in the CRC tissue included quinaldic acid (also referred to as quinoline-2-carboxilic acid), 17α- and 17β-estradiol, which are associated with tumor suppression activities, as well as other metabolites such as, anhydro-β-glucose, Asp-Arg, lysophosphatidylcholine, lysophosphatidylethanolamine (lysoPE), lysophosphatidylinositol, carnitine, 5′-deoxy-5′-(methylthio) adenosine, 2′-deoxyinosine-5′-monophosphate and thiamine monophosphate. There was no difference in the levels of the differential metabolites between male and female patients. The differentiation of CRC also showed no impact on the levels of the differential metabolites. The levels of lysoPE were increased in the right side of the colon compared with the left side of the colon and rectum. Analysis of the different tumor stages indicated that 2-aminobenzenesulfonic acid, P-sulfanilic acid and quinoline-4-carboxylic acid were decreased in stage I CRC tissue compared with stage II, III and IV CRC tissue. The levels of N-α-acetyl-ε-(2-propenal)-Lys, methylcysteine and 5′-deoxy-5′-(methylthio) adenosine varied at different stages of tumorigenesis. These differential metabolites were implicated in multiple metabolism pathways, including carbohydrate, amino acid, lipid, nucleotide and hormone. In conclusion, the present study demonstrated that CRC tumors had altered metabolites compared with normal tissue. The data from the metabolic profile of CRC tissues in the present study provided supportive evidence to understand tumorigenesis.

To determine whether oxidative stress and inflammation are associated with constipation by examining the expression of the main producers of reactive oxygen species, nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, and pro-inflammatory cytokines in the colon of patients with chronic functional constipation. The colonic biopsies were collected from 32 patients with chronic functional constipation and 30 healthy subjects who underwent colonoscopy. Colonic mucosal histology was observed. Interleukin (IL)-1β, IL-6, IL-8 messenger RNA (mRNA), and 4 members of NADPH oxidase (NOX1, NOX2, DUOX2, and NOX4) protein and mRNA were assessed by immunohistochemistry, western blotting, and reverse transcription polymerase chain reaction. The tissues from both patients and healthy subjects showed normal histological structure without increase of inflammatory cells. NOX1 protein and mRNA levels were significantly increased compared to controls (P < .05). DUOX2 protein, but not mRNA, was increased by 2-fold compared to controls (P < .05). The levels of NOX2 and NOX4 protein and mRNA demonstrated no significant difference between patients and control subjects. The levels of IL-1β and IL-6 mRNA were significantly higher in constipation patients (P < .05), while IL-8 mRNA level was no different between the 2 groups. NADPH oxidase and pro-inflammatory cytokine might be involved in the pathogeneses of chronic functional constipation.

Background Colorectal cancer (CRC) is the third most frequent cancer worldwide. The AHCYL1 gene is required for CNV and has a close association with the tumor immune microenvironment. However, the predictive value of the AHCYL1 gene in patients with CRC remains unknown. Methods AHCYL1 gene with prognostic potential was comprehensively analyzed. Next, using LASSO Cox regression, we fully examined and integrated the AHCYL1 and AHCYL1-related genes from TCGA database. Meanwhile, TCGA database was used to study the connection between AHCYL1 and the tumor immune microenvironment and tumor mutation burden (TMB) in CRC. The influence of AHCYL1 in tumor growth and the recruiting ability of CD8+ T cells were verified, respectively, in vivo and in tissues. To ascertain the connection between AHCYL1 and AHCYL1-related genes and the prognosis of CRC, a prognostic model was created and validated. Result We demonstrated that AHCYL1 has a differential expression and patients with AHCYL1 deletion get shorter survival in CRC. Additionally, the tissues without AHCYL1 have a weaker ability to recruit the natural killer (NK) cell, CD8+ T cells, and tumor-infiltrating lymphocytes (TILs) and response to immunotherapy. Additionally, knockdown of AHCYL1 promoted tumor growth in the CRC mouse model and recruited lower CD8+ T cells in CRC tissues. TCGA database was used to classify patients into low- and high-risk categories based on the expression of four genes. Meanwhile, we discovered an association between the low-risk group and a lower TMB and a higher response to immunotherapy. Finally, a predictive nomogram based on these genes was developed and verified, yielding a C-index of 0.74. Conclusion For CRC patients, the prognostic model based on AHCYL1 and AHCYL1-related genes showed a high predictive performance in terms of prognosis and immunotherapy response.

目的探讨白介素（IL）-17A和IL-18在慢性功能性便秘患者结肠黏膜内表达情况,分析这两种白介素在慢性功能性便秘发病机制中的作用。方法选择2 0 1 8年10月至2 0 1 9年6月在河南科技大学第一附属医院消化内镜中心进行肠镜检查的32例慢性功能性便秘患者作为试验组,并选取同期30例健康体检者作为对照组;两组研究对象在结肠镜检查过程中取乙状结肠黏膜活检,进行HE染色观察组织形态学、实时定量聚合酶链式反应（PCR）检测IL-17A和IL-18 mRNA表达情况,并应用酶联免疫吸附测定（ELISA）方法测定IL-17A和IL-18蛋白水平,比较这两种炎症因子mRNA和蛋白在两组结肠黏膜的表达差异。结果试验组与对照组结肠黏膜组织形态学均无异常改变,试验组患者结肠黏膜内未发现慢性炎症细胞（如淋巴细胞、浆细胞等）浸润增加。试验组结肠黏膜组织中IL-17A mRNA、IL-17A蛋白表达水平高于对照组,差异有统计学意义（P＜0.05）;而IL-18 mRNA和蛋白表达两组间比较,差异无统计学意义（P＞0.05）。结论 IL-17A在慢性功能性便秘结肠黏膜组织中表达升高,IL-17A可能参与功能性便秘的发病机制或病理过程。

BACKGROUND: Esophageal squamous cell carcinoma (ESCC) in some cases can be diagnosed as esophageal varices (EV). DLG1-AS1 promotes cervical cancer, while its function is other malignancies remains unknown. Our aim for this study is to study the role of DLG1-AS1 in esophageal squamous cell carcinoma. METHODS: Plasma levels of DLG1-AS1 in 66 early stage ESCC patients, 60 EV patients and 60 healthy controls were measured by RT-qPCR. Receiver operating characteristic (ROC) curve was applied to analyze the diagnostic value of DLG1-AS1 for early stage ESCC. Relationship between miR-145 and DLG1-AS1 was analyzed by overexpression experiments. Proliferation of cells was determined by CCK-8 assay. RESULTS: DLG1-AS1 was upregulated in ESCC, but not in EV patients compared with healthy control. DLG1-AS1 overexpression distinguished ESCC patients from healthy controls and EV patients. Plasma miR-145 was inversely correlated with DLG1-AS1 in ESCC patients. Moreover, DLG1-AS1 overexpression resulted in the downregulation of miR-145, while miR-145 mimic transfection did not significantly alter DLG1-AS1. Overexpression of DLG1-AS1 mediated the promoted, while overexpression of miR-145 resulted in inhibited proliferation of ESCC cells. The role of DLG1-AS1 overexpression was inhibited by miR-145 mimic transfection. CONCLUSION: Therefore, DLG1-AS1 may promote ESCC under the repression of miR-145.