BACKGROUND:Nonalcoholic fatty liver disease (NAFLD) is a growing global health concern linked to obesity. METHODS:This study employed a Mendelian randomization approach to explore the causal influence of BMI on metabolic biomarkers and the subsequent risk of NAFLD. We analyzed data from multiple sources, including 249 metabolic traits, to establish direct and mediating relationships among BMI, metabolic factors, and NAFLD risk. RESULTS:Our findings revealed a significant positive correlation between BMI and NAFLD across various datasets. We identified 176 metabolites associated with BMI, of which 106 were also linked to NAFLD. Importantly, 86 metabolites were found to mediate the relationship between BMI and NAFLD risk. Specifically, elevated levels of branched-chain amino acids, triglycerides, and certain cholesterol esters were notably associated with increased NAFLD risk, whereas changes in free cholesterol and phospholipid levels also played critical roles. CONCLUSION:This study highlights the complex interactions between BMI, metabolic biomarkers, and NAFLD risk. By elucidating these relationships, we highlight potential targets for interventions aimed at reducing NAFLD incidence in populations with elevated BMI, ultimately contributing to improved metabolic health.

Gastric cancer, primarily manifested as gastric adenocarcinoma (STAD), remains one of the leading causes of cancer-related deaths worldwide. Ammonia-induced cell death (AID), a newly discovered form of cell death, has gained attention due to its unique mechanisms, including lysosomal alkalinization and mitochondrial dysfunction. However, the role of AID in STAD, particularly its impact on immune regulation and patient prognosis, remains unclear. This study integrated bulk RNA-seq and single-cell RNA-seq (scRNA-seq) data. The Seurat package was used for single-cell gene expression analysis, clustering different cell types, and AID scoring. CellChat software analyzed the ligand-receptor interactions between immune cell subtypes. Differential expression analysis (DEGs) and GO/KEGG enrichment identified key genes associated with AID. We constructed a prognostic risk model based on these findings. Immune cell infiltration was analyzed using the “GSVA” and “xCell” packages. In vitro experiments were conducted on HGC-27 and MKN45 gastric cancer cell lines, where GLS1 was knocked down using siRNA to assess the effects on cell proliferation, migration, invasion, and lysosomal function. Additionally, drug sensitivity tests were used to evaluate the impact of GLS1 overexpression on resistance to various anticancer drugs. Single-cell clustering analysis revealed that T cells had significantly higher ammonia-induced cell death (AID) scores, leading to the classification of cells into high-AID and low-AID groups. Gene expression analysis and pathway enrichment showed significant enrichment in lysosomal and mitochondrial pathways, consistent with known AID mechanisms. A prognostic risk model based on five key genes (C1QA, MARCKSL1, GLS1, N4BP2L2, and CD68) effectively classified patients into high-risk and low-risk groups, with the high-risk group showing stronger immune cell infiltration, including CD4 + and CD8 + T cells, dendritic cells, and macrophages. Among the model genes, GLS1 was identified as the most significant prognostic factor, the strongest risk factor, and the most central gene in the interaction network. In vitro experiments showed that GLS1 inhibition led to increased ammonia levels, elevated lysosomal pH, and reduced lysosomal function, thereby enhancing AID expression. Furthermore, GLS1 overexpression significantly promoted cell proliferation, migration, and tumor growth. Finally, gastric cancer cells with GLS1 overexpression exhibited resistance to multiple anticancer drugs, highlighting the potential value of GLS1 as a therapeutic target. The AID model is a promising biomarker for accurately determining survival and predicting the effectiveness of immunotherapy in STAD patients. GLS1 plays a crucial role in driving tumor proliferation and migration and may act as a potential tumor biomarker of STAD.

Background: Cholangiocarcinoma is a malignant invasive biliary tract carcinoma with a poor prognosis. Anoikis-related genes are prognostic features of a variety of cancers. However, the value of prognostication and therapeutic effect of anoikis-related genes in cholangiocarcinoma have not been reported. The aim of this research was developing an ARGs signature associated with cholangiocarcinoma patients. Methods: We introduced transcriptome data to discover genes that were differentially expressed in cholangiocarcinoma. Subsequently, WGCNA was utilized to screen critical module genes in reference to anoikis. The univariate Cox, Lasso regression and Kaplan-Meier survival were executed to build a prognostic signature. We further performed gene functional enrichment, immune microenvironment and immunotherapy analysis between two risk subgroups. Finally, the pRRophetic algorithm was applied to compare the half inhibitory concentration value of several drugs. Results: A grand total of 1844 genes with differential expression related to the cholangiocarcinoma patients were identified. Furthermore, we obtained 2678 key module genes related to anoikis. Then, a prognostic signature was developed using the 6 prognostic genes (FXYD2, PCBD1, C1RL, GMNN, LAMA4 and HACL1). Independent prognostic analysis showed that risk score and alcohol could function as separate prognostic variables. We found cetain distinction in the immune microenvironment between the two risk subgroups. Moreover, immunotherapy evaluation showed that the anoikis-related gene signature could be applied as a therapy predictor. Finally, Chemotherapeutic drug sensitivity results showed that the low-risk group responded better to bosutinib, gefitinib, gemcitabine, and paclitaxel, while the high-risk group responded better to axitinib, cisplatin, and imatinib. Conclusion: The prognostic signature comprised of FXYD2, PCBD1, C1RL, GMNN, LAMA4 and HACL1 based on anoikis-related genes was established, which provided theoretical basis and reference value for the research and treatment of cholangiocarcinoma.

The study is designed to explore the regulatory network that MALAT1 competitively binds with miR-188-5p to up-regulate PSMD10 to facilitate cholangiocarcinoma cell migration and invasion and suppress apoptosis. qRT-PCR and fluorescence in situ hybridization (FISH) were used to examine the expression and positive signal of MALAT1 and miR-188-5p in cholangiocarcinoma tissues and HIBEC, HCCC-9810, RBE, and QBC939 cells. Western blot, qRT-PCR, and immunohistochemistry were selected to detect PSMD10 expression in cholangiocarcinoma tissues and cell lines. Dual luciferase reporter gene assay was adopted to verify that miR-188-5p targeted MALAT1 and PSMD10. qRT-PCR, pull down, and western blot were used to examine the regulation of MALAT1-miR-188-5p-PSMD10 axis. Transwell, wound healing assay, and Tunel cell apoptosis were adopted to respectively detect the regulatory abilities of MALAT1-miR-188-5p-PSMD10 axis on cell invasion, migration, and apoptosis. Western blot was used to detect the regulation mechanism of MALAT1 on Bax, Bcl-2, and caspase-3 proteins. Nude mice subcutaneous xenograft model of cholangiocarcinoma was established to examine the impacts of MALAT1 on subcutaneous tumor growth. Immunohistochemistry was adopted to examine the positive indicator of Ki67 antibodies and SMD10 antibodies in each group. MALAT1 and PSMD10 were highly expressed in cholangiocarcinoma tissues and cell lines, while miR-188-5p was lowly expressed. MALAT1 could competitively bind to miR-188-5p, and miR-188-5p could negatively regulate PSMD10. MALAT1, In-miR-188-5p, and PSMD10 could facilitate cell invasion and migration and inhibit apoptosis, while siMALAT1, miR-188-5p, and siPSMD10 produced an opposite result. MALAT1-miR-188-5p-PSMD10 axis could promote RBE cell invasion and migration and inhibit apoptosis, whereas siMALAT1-In-miR-188-5p-siPSMD10 axis showed an opposite result. On the other hand, it was verified that up-regulation/down-regulation of MALAT1 can inhibit/promote Bax and caspase-3 proteins and promote/inhibit the expression of Bcl-2 protein. MALAT1 could facilitate subcutaneous tumor growth and enhance cell proliferation and positive signal of PSMD10, while miR-188-5p worked in an opposite direction. MALAT1 competitively binds to miR-188-5p to up-regulate mRNA translation and protein expression of PSMD10, thereby facilitating cholangiocarcinoma cell invasion and migration and inhibiting its apoptosis. However, interfering MALAT1-miR-188-5p-PSMD10 axis could inhibit the occurrence and development of cholangiocarcinoma.

Background:Cholangiocarcinoma is a primary malignant tumor, and its progression involves oncogene activation, the absence of tumor suppressor gene, abnormal signaling pathways and miRNA expression. MiRNAs are abnormally ex pressed in many types of tumors. Objective:This study aims to observe the effects of miR-582 on cholangiocarcinoma cell proliferation, S-phase arrest, migration and invasion and to analyze the regulation of miR-582 on LIS1 to clarify the real role of miR-582 in cholangiocarcinoma development. Materials and Methods: TCGA database of cholangiocarcinoma samples was analyzed. Dual fluorescence reporter and TargetScan were conducted to confirm whether LIS1 was the target gene of miR-582. Effects of miR-582 and LIS1 on HCC-9810 cell proliferation, S-phase cell ratio, migration and invasion were determined by CCK-8, Flow cytometry and Transvvell, respectively, whereas the function of iniR-582 on MMP-2 and P-Akt expression was identified by Western blotting. Nude mice xenograft model of cholangiocarcinoma was established to detect what miR-582 did for tumor growth. Results:TCGA showed that miR-582 was lowly expressed and LIS1 was highly expressed in donor tissues compared with adjacent tissues. MiR-582 targeted LIS1 to inhibit MMP-2 and p-AKT expression. Transfection of miR-582 mimics could suppress HCC-9810 cell proliferation, S-stage arrest, migration and invasion, while LIS1 worked oppositely. MiR-582 inhibitors promoted cell biological behavior, whereas LIS1 siRNA was opposite. In nude mice xenograft model, miR-582 overexpression inhibited tumor growth. Conclusions:It implies that miR-582 could negatively regulate LIS1 to inhibit MMP-2 and P-Akt expression, thus suppressing cell invasion and proliferation in cholangiocarcinoma.

BackgroundLncRNAs have proven to be involved in the initiation and progression of cholangiocarcinoma (CCA), although the mechanism by which this occurs remains unknown.MethodsThe current study reveals that RHPN1-AS1 was overexpressed in CCA patient samples, which predicted poor outcome of CCA patients. RHPN1-AS1 increased in vitro pancreatic carcinoma cell proliferation as well as promoted xenograft growth in vivo. Mechanistically, DANCR upregulated expression of YAP1 by competitively binding to miR-345-5p. Importantly, RHPN1-AS1 level was positively correlated with YAP1 expression level in CCA tissues. Moreover, YAP1 overexpression could predicted a poor outcome of CCA patients.ResultsTaken together, our results suggested that RHPN1-AS1 might be a remarkable biomarker to evaluate prognosis in CCA.ConclusionThe RHPN1-AS1/YAP1 axis may provide new strategies for CCA clinical practice.

Objective:To analyze the relationship between the expression of microRNA (miRNA, miR)-150 and epithelial-mesenchymal transition (EMT) in gallbladder cancer and its mechanism.Methods:The expression of miR-150 in 30 cases of gallbladder cancer was detected by real-time quantitative polymerase chain reaction (qPCR), and 30 cases of chronic cholecystitis tissues was used as control. Simultaneously, the expression of proliferating cell nuclear antigen (PCNA), E-cadherin, N-cadherin and matrix metalloproteinase (MMP)-9 was detected, and the relationship of miR-150 and these genes was analyzed. Human gallbladder cancer cell line GBC-SD was transfected with miR-150 mimics. Then scratch experiments and Transwell chamber experiments were used to detect changes in cell migration and invasion; qPCR and Western blotting were used to detect changes in mRNA and protein of each gene.Results:The expressions of miR-150 and E-cadherin in gallbladder cancer tissues were lower than those in chronic cholecystitis tissues ( t=-7.035, -14.146, P<0.01). The expressions of PCNA, N-cadherin and MMP-9 were significantly higher compared with chronic cholecystitis tissues ( t=3.813, 9.339, 5.616, P<0.01). The expression of miR-150 was related to the degree of tumor differentiation, clinical stage, and lymph node metastasis ( t=3.228, 2.396, -2.604, P<0.05). Pearson correlation analysis showed that miR-150 was positively correlated with E-cadherin ( r=0.630, P<0.05), and negatively correlated with PCNA, N-cadherin, and MMP-9 ( r=-0.769, -0.628, -0.616, P<0.05). After transfection, the migration and invasion ability of GBC-SD cells decreased ( F=1 965.860, 114.940, P<0.01). The expressions of PCNA, N-cadherin, and MMP-9 were decreased, while the expression of E-cadherin was increased ( P<0.05). Conclusion:Decreased miR-150 expression in gallbladder cancer plays a key role in tumor progression, and the mechanism may be that miR-150 participates in tumor EMT process by regulating some genes.

Background Outcomes of gastroesophageal reflux disease (GERD) using Toupet fundoplication (TF) and Stretta radiofrequency (SRF) have not been compared and this study was conducted to compare therapeutic efficacy of the two methods. Methods This retrospective study analyzed a total of 230 patients undergoing TF or SRF at our hospital. Baseline data, reflux symptoms, the DeMeester scores, lower esophageal sphincter (LES) pressure and adverse events were compared over 1 year period. Results A total of 226 patients were included in the study. The time and frequency of reflux and percentage of reflux time before and 12 months after therapy were not significantly different. There were significantly interactions between the therapy method and follow-up time on the DeMeester score and LES pressure. Twelve months post therapy, the DeMeester score was significantly higher in SRF than in TF group, while the LES pressure was lower. At 12 months after therapy, multivariate Cox proportional regression analysis showed that reflux frequency, the DeMeester score and LES pressure were risk factors for poor prognosis in TF group, while reflux frequency and the DeMeester score, and LES pressure were risk factors for poor prognosis in SFR group. Conclusions Compared with TF, SFR can significantly improve the esophageal pH and pressure in GERD patients without increasing the risk of poor prognosis.

LncRNAs have been reported to be involved in the initiation and progression of cholangiocarcinoma (CCA), although the mechanisms by which this occurs remains unknown. The current study aimed to reveal the clinico-pathological relationship of RHPN1-AS1 expressed with CCA, and also investigate the functions of RHPN1-AS1 both in vivo and in vitro. RHPN1-AS1 was overexpressed in CCA cell proliferation as well as promoted xenograft growth in vivo. Mechanistically, DANCR upregulated the expression of YAP1 by competitively binding to miR-345-5p. Importantly, RHPN1-AS1 level was positively correlated with YAP1 expression level in CCA tissues (P<0.05). Moreover, Kaplan-Meier curves showed that YAP1 overexpression predicted a poor outcome of CCA patients (P=0.004). Taken together, our results suggested that RHPN1-AS1 might be a remarkable biomarker to evaluate prognosis in CCA. The RHPN1-AS1/YAP1 axis may provide new strategies for CCA clinical practice.