ObjectiveThis paper aims to explore the effects and mechanisms of TAK-242, a specific inhibitor of Toll-like receptor 4 (TLR4), on the intestinal mucosal barrier in mice with ulcerative colitis (UC).MethodsA total of 30 C57BL/6 mice were randomly divided into a normal group (Normal group), a model group (Model group), a low-dose TAK-242 group (TAK-242-L group), a high-dose TAK-242 group (TAK-242-H group), and a 5-aminosalicylic acid group (5-ASA group). The Normal group was fed with normal feed. The Model group was given a 5% sodium dextran sulfate (DSS) solution as drinking water for modeling. The TAK-242-L and TAK-242-H groups were given TAK-242 intervention (3 mg/kg, 10 mg/kg) at the same time of modeling, and intraperitoneally injected once every day. The 5-ASA group was given 5-ASA (500 mg/kg) orally once a day during modeling. Each group was treated continuously for 7 days, and the changes in body mass and disease activity index (DAI) score were observed. After euthanizing the mice on the 8th day, the colon length was measured. H-E staining was used to observe the pathological changes of colon mucosa. ELISA was used to detect the expression levels of TNF-, IL-1, and IL-6 in various colon tissues. Western blotting was used to detect the expression levels of tight junction proteins Claudin-1 and ZO-1 in colon tissue, as well as the expression levels of TLR4, MyD88, and NF-B p65 proteins related to the TLR4 signaling pathway.ResultsCompared with the Normal group, the Model group shows a significantly increase in DAI score, a marked shortening colon length, a significant increase in histological injury score, and significantly elevated expression levels of TNF-, IL-1, and IL-6 in colon tissue. Additionally, the expression levels of Claudin-1 and ZO-1 proteins in colon tissue are significantly decreased, while the expression levels of TLR4, MyD88, and NF-B p65 proteins are significantly increased (P<0.05). Compared with the Model group, the TAK-242-H and 5-ASA groups show significantly reduced DAI scores, increased colon lengths, decreased histological injury scores, and significantly lowered expression levels of TNF-, IL-1, and IL-6 in colon tissue. Moreover, the expression levels of Claudin-1 and ZO-1 proteins in colon tissue are significantly increased, whereas the expression levels of TLR4, MyD88, and NF-B p65 proteins are significantly reduced (P<0.05).ConclusionThe TLR4 inhibitor TAK-242 can alleviate intestinal inflammation and improve intestinal mucosal barrier function in UC by targeting the TLR4/MyD88/NF-B signaling pathway.

ObjectiveThis paper attempts to investigate the expression of transient receptor potential cation channel subfamily M member 8 (TRPM8) during the carcinogenesis process of ulcerative colitis (UC), and to explore its relationship with disease activity, lesion range and prognosis of patients.MethodsA total of 60 UC patients, 60 ulcerative colitis with intraepithelial neoplasia (IN) patients, and 60 colon cancer patients who were hospitalized at the First Hospital of Qinhuangdao from June 2021 to June 2024 were divided into the UC group, the UC-IN group, and the colon cancer group, respectively. Significant colonic mucosal tissue samples and tumor tissue specimens were collected from the UC and UC-IN groups. Another 30 healthy individuals who underwent physical examination at the same hospital, whose normal colon mucosal tissue were obtained (20 cm from the anus) through colonoscopy were selected as the control group. Real time fluorescence quantitative PCR and immunohistochemical staining were used to detect the expression levels of TRPM8 mRNA and protein in colon tissue, and the relationship between TRPM8 and disease activity and lesion range in UC patients was analyzed. After a 6-month follow-up of UC patients after discharge, ROC curve analysis was used to evaluate the predictive value of TRPM8 for poor prognosis in UC patients.ResultsCompared with the control group, the positive expression rates of TRPM8 mRNA and protein are significantly increased in the UC group, UC-IN group, and colon cancer group, with statistically significant differences among the three groups (P<0.05). The positive expression rates of TRPM8 mRNA and protein in the UC-HGIN group are significantly higher than those in the UC-LGIN group (P<0.05). Additionally, the positive expression rates of TRPM8 mRNA and protein in the poorly differentiated cancer group are significantly higher than those in the moderately and well-differentiated cancer groups (P<0.05). In UC patients, the relative expression level and positive protein expression rate of TRPM8 mRNA in the severe active phase group are significantly higher than those in the moderate active, mild active, and clinical remission groups (P<0.05). The relative expression in the moderate activity group is also significantly higher than in the mild activity and clinical remission groups (P<0.05), and the mild active phase group shows significantly higher expression than the clinical remission phase group (P<0.05). There is no statistically significant difference in TRPM8 mRNA expression level or protein positivity across UC patients with different lesion extents (P>0.05). Follow-up results show that the relative expression level and protein positive rate of TRPM8 mRNA in the poor prognosis group are significantly higher than those in the good prognosis group (P<0.05). ROC curve analysis shows that the area under the curve (AUC) for TRPM8 protein, TRPM8 mRNA, and their combined detection in predicting poor prognosis in UC patients are 0.734, 0.762, and 0.823, respectively.ConclusionsTRPM8 expression gradually increases during UC-associated carcinogenesis and is significantly correlated with the pathological differentiation, disease activity, and poor prognosis. It has the potential as an early diagnostic and prognostic biomarker for UC related colon cancer.

ObjectiveThis paper intends to investigate the clinical value of serum long non-coding RNA (lncRNA) OTUD6B-AS1 and lncRNA ITGB8-AS1 in the early diagnosis and prognosis of patients with colorectal cancer (CRC).MethodsA total of 108 CRC patients admitted to the Rehabilitation University Qingdao Central Hospital from March 2020 to March 2021 were selected as the study group, while 108 healthy individuals who underwent physical examinations at the same period served as the control group. Real-time quantitative PCR was used to detect the expression levels of serum lncRNA OTUD6B-AS1 and lncRNA ITGB8-AS1. Multivariate logistic regression analysis was conducted to identify factors affecting CRC patient mortality. ROC curve analysis was used to assess the predictive value of lncRNA OTUD6B-AS1 and lncRNA ITGB8-AS1 for CRC patient mortality.ResultsCompared with the control group, the expression level of lncRNA OTUD6B-AS1 in the study group is significantly reduced, while that of lncRNA ITGB8-AS1 is significantly increased (P<0.05). Follow-up results show that among 108 CRC patients, 33 died (designated as the death group), while 75 survived (designated as the survival group). There are no statistically significant differences between the two groups in terms of age, gender, BMI, tumor diameter, tumor location, tumor histological type, tumor differentiation grade, CRC family history, surgical treatment or not, or chemotherapy or not (P>0.05), while differences in TNM staging are statistically significant (P<0.05). Compared to the survival group, the expression level of serum lncRNA OTUD6B-AS1 is significantly lower, and serum lncRNA ITGB8-AS1 expression level is significantly higher in the death group (P<0.05). The multivariate logistic regression analysis show that TNM stage and serum lncRNA ITGB8-AS1 expression level is independent risk factors for CRC patient mortality (P<0.05), while serum lncRNA OTUD6B-AS1 expression level is an independent protective factor (P<0.05). ROC curve analysis shows that the area under the curve (AUC) for predicting CRC patient mortality using serum lncRNA OTUD6B-AS1, lncRNA ITGB8-AS1, and their combined detection are 0.848, 0.796, and 0.933, respectively, indicating that the predictive efficacy of combined detection is higher than that of individual detection (P<0.05).ConclusionsSerum lncRNA OTUD6B-AS1 expression level is lower in CRC patients, while lncRNA ITGB8-AS1 expression level is higher. Additionally, serum lncRNA ITGB8-AS1 is an independent risk factor and lncRNA OTUD6B-AS1 is an independent protective factor for CRC patient mortality. Both lncRNA OTUD6B-AS1 and lncRNA ITGB8-AS1 show promise as biomarkers for predicting mortality in CRC patients.

ObjectiveThis paper intends to explore the mechanism by which pachymic acid (PA) inhibits the proliferation and migration of colorectal cancer (CRC) cells through the glycogen synthase kinase 3 (GSK3)/-catenin signaling pathway.MethodsHuman colon cancer cell line HCT116 was cultured routinely and divided into four groups: control (NC) group (untreated), PA group (treated with 8.50 mol/L PA for 24 h), 5-fluorouracil (5-FU) group (treated with 0.70 μg/mL 5-FU for 24 h), and PA+5-FU group (treated with 4.25 mol/L PA and 0.35 μg/mL 5-FU for 24 h). The CCK-8 assay and cell colony formation assay were used to assess the effect of PA on the proliferation of HCT116 cells, respectively. A Transwell experiment was conducted to evaluate the effect of PA on the invasion of HCT116 cells. A cell scratch healing experiment was performed to detect the effect of PA on the migration of HCT116 cells. Western blotting was used to detect the effect of PA on the expression of GSK3/-catenin pathway related proteins GSK3, phosphorylated -catenin (p--catenin), -catenin, CyclinD1, and c-Myc in HCT116 cells.ResultsPA has a dose-dependent effect on inhibiting the proliferation of HCT116 cells. Compared with the NC group, the number of cell clones formed in the PA group is significantly reduced (P<0.01), and the migration and invasion abilities of HCT116 cells are significantly weakened (P<0.01). The protein expression levels of GSK3 and p--catenin in HCT116 cells are significantly increased (P<0.01), while the protein expression levels of -catenin, CyclinD1, and c-Myc are significantly decreased (P<0.01).ConclusionPA can inhibit the proliferation and migration of HCT116 cells by activating the GSK3/-catenin signaling pathway.

ObjectiveThis paper intends to explore the influence of BRCA1 on the biological behavior and angiogenesis of colon cancer cells.MethodsSpecimens of colon cancer tissues and adjacent tissues (>5 cm from the tumor margin) were collected from 58 patients who underwent radical resection of colon cancer at Tai'an Cancer Hospital from June 2019 to June 2020 were selected. BRCA1 expression in colon cancer tissues was detected by immunohistochemistry. The relative expression levels of BRCA1, vascular endothelial growth factor (VEGF), and ERCC1 were measured in human normal colonic epithelial cells HCoEpiC and human colon cancer cell lines SW480, SW620, DLD-1, Caco-2, HT-29, and HCT-116 using real-time fluorescence quantitative PCR. SW480 cells were divided into three groups: the colon cancer group (untreated), the NC group (transfected with NC siRNA), and the interference group (transfected with BRCA1 siRNA). Cell survival rate, apoptosis rate, invasion ability and migration ability were detected using the MTT assay, flow cytometry, Transwell chamber assay, and scratch wound assay, respectively. The expression levels of VEGF and ERCC1 in SW480 cells were analyzed by Western blotting.ResultsThe positive expression rate of BRCA1 in colorectal cancer tissues is 44.83% (26/58), significantly higher than that in adjacent non-cancerous tissues (13.79%), with a statistically significant difference (P<0.05). Real-time fluorescence quantitative PCR show that BRCA1 expression in SW480 cells is higher than that in other colon cancer cells. Compared with the colon cancer group and the NC group, the interference group exhibits significantly decreased cell viability and invasion, reduced migration distance, and increased apoptosis, with statistically significant differences (P<0.05). Compared with the colon cancer group and the NC group, both expression levels of VEGF and ERCC1 mRNA and protein in SW480 cells are significantly downregulated in the interference group (P<0.05).ConclusionsDownregulation of BRCA1 expression significantly inhibits the survival, invasion, and migration of colon cancer cells, promotes apoptosis, and suppresses the expression of VEGF and ERCC1, suggesting that BRCA1 may play a key role in colon cancer progression.

ObjectiveThis paper aims to explore the predictive value of serum catechinostatin (CST), lectin-like oxidized low-density lipoprotein receptor-1 (LOX-1), and cholinesterase (ChE) levels for postoperative survival in patients with decompensated cirrhosis undergoing splenectomy.MethodsA total of 115 patients with decompensated liver cirrhosis who underwent splenectomy at Nanchong Central Hospital from April 2016 to January 2021 were selected as the study subjects. Patients were followed up for 2 years after surgery and categorized into a survival group and a death group based on their survival status. ELISA was used to detect and compare the serum levels of CST, LOX-1, and ChE between the two groups. Clinical data were also analyzed. Multivariate Cox regression analysis was performed to explore factors affecting survival. ROC curve analysis was used to analyze the predictive value of serum CST, LOX-1, and ChE for postoperative survival.ResultsStatistically significant differences are observed between the survival and death groups in terms of gender, portal vein diameter, splenic vein diameter, presence of portal vein thrombosis, platelet count (PLT), and D-dimer levels (P<0.05). Compared with the survival group, the death group shows significantly higher serum CST and LOX-1 levels and lower serum ChE levels (P<0.05). Multivariate Cox regression analysis shows that gender, portal vein diameter, splenic vein diameter, postoperative portal vein thrombosis, PLT, D-dimer, CST, LOX-1, and ChE are factors affecting survival of patients with decompensated cirrhosis after splenectomy (P<0.05). The area under the ROC curve (AUC) for CST, LOX-1, and ChE in predicting postoperative survival in patients are 0.878, 0.853, 0.882, and 0.964, respectively. The predictive value of the three combined tests is higher than that of any individual prediction (Ztriple combination-CST=3.238, Ztriple combination-LOX-1=3.601, Ztriple combination-ChE=2.945, P<0.05).ConclusionsIn patients with decompensated cirrhosis who died after splenectomy, serum CST and LOX-1 levels are significantly increased, while serum ChE levels are significantly decreased. Serum CST, LOX-1, and ChE have the potential to be used as valuable biomarkers for predicting postoperative survival of decompensated liver cirrhosis patients, with combined detection offering superior predictive performance.

ObjectiveThis paper attempts to investigate the expression level of protein arginine methyltransferase 3 (PRMT3) in hepatocellular carcinoma (HCC) tissues and analyze its relationship with oxaliplatin resistance and the prognosis of patients.MethodsFrom January 2019 to June 2022, tumor and adjacent normal tissues (>5 cm from tumor margin) were collected from 52 patients with hepatitis B-related HCC (set as the HCC group) hospitalized in Cangzhou People's Hospital. For comparison, liver tissue samples were obtained via ultrasound-guided puncture from 38 patients with chronic hepatitis B (set as the chronic hepatitis B group) and 38 patients with hepatitis B-related cirrhosis (set as the hepatitis B-related cirrhosis group), and from 38 patients undergoing surgical resection for liver hemangioma (set as the control group). PRMT3 expression levels in tissues were assessed via immunohistochemical staining. PRMT3 expression levels in tissues were assessed via immunohistochemical staining. All HCC patients received the FOLFOX4 chemotherapy regimen, with oxaliplatin as the primary agent, and the correlations between PRMT3 expression, chemotherapy efficacy, and patient prognosis were analyzed. SNU-398 cells in logarithmic growth phase were divided into the Lv-shNC group (transfected with negative control lentivirus) and the Lv-shPRMT3 group (transfected with PRMT3 interfering lentivirus). After treatment with different concentrations of oxaliplatin, the effect of PRMT3 on oxaliplatin cytotoxicity was observed. Cells were taken from the Lv-shNC group and the Lv-shPRMT3 group, and added to culture medium containing 20 mol/L oxaliplatin (designated as the Lv-shNC+OXA group and the Lv-shPRMT3+OXA group, respectively). The proliferation, migration, invasion, and apoptosis abilities of each group of cells were analyzed.ResultsThe positive expression rate of PRMT3 in HCC tissue is significantly higher compared to adjacent tissues and to tissues from the chronic hepatitis B group, the hepatitis B-related cirrhosis group, and control group (P<0.05). ROC curve analysis show that the area under the curve (AUC), sensitivity, and specificity of PRMT3 for diagnosing HCC in HCC tissues are 0.831 (95%CI: 0.763 to 0.899), 75.00%, and 91.23%, respectively. Compared with the PRMT3 negative expression group, the total effective rate of treatment in the PRMT3 positive expression group is significantly reduced (2=4.392, P=0.036), and so is the 2-year survival rate (2=5.148, P=0.023). The relative expression level of PRMT3 in the Lv-shPRMT3 group cells is significantly lower than that in the Lv-shNC group (t=10.844, P<0.05). As the concentration of oxaliplatin added increases, the cell inhibition rates of both groups significantly increase. Compared with the Lv-shNC group, the number of monoclonal cells formed and the number of transmembrane cells in the Lv-shPRMT3 group and the Lv-shNC+OXA group are significantly reduced, the apoptosis rate is significantly increased, and the scratch healing rate is significantly decreased (P<0.05). Compared with the Lv-shPRMT3 group and the Lv-shNC+OXA group, the number of monoclonal cells formed and the number of transmembrane cells in the Lv-shPRMT3+OXA group are significantly reduced, the apoptosis rate is significantly increased, and the scratch healing rate is significantly decreased (P<0.05).ConclusionsPRMT3 is highly expressed in HCC tissues, and downregulation of PRMT3 expression can inhibit cell proliferation, migration, and invasion, and can promote cell apoptosis, thereby increasing the sensitivity of HCC cells to oxaliplatin and reducing the rate of drug resistance. PRMT3 is expected to become a diagnostic biomarker and therapeutic target for HCC.

ObjectiveThis paper aims to explore the role and mechanism of the gut microbe metabolite trimethylamine oxide (TMAO) in regulating ferroptosis in colon cells through the reactive oxygen species (ROS)/c-Jun N-terminal kinase (JNK)/glutathione peroxidase 4 (GPX4) signaling pathway in slow transit constipation (STC).MethodsA total of 50 rats were selected and randomly assigned to the blank group, the STC group, the TMAO group, the SP600125 (JNK inhibitor) group, and the TMAO+SP600125 group, with 10 rats in each group. Except for the blank group, all other groups were orally administered 10 mg/kg/d of compound difenofibrate suspension via gavage to establish the STC model. After successful modeling, the TMAO group received daily intraperitoneal injections of TMAO (100 mg/kg); the SP600125 group received intraperitoneal injection of SP600125 (15 mg/kg) twice a week; the TMAO+SP600125 group received daily intraperitoneal injections of TMAO (100 mg/kg) along with SP600125 (15 mg/kg) twice a week. All treatments lasted for 3 weeks. Each group was given a single gavage of activated carbon suspension to observe the time required for the first black feces excretion and the small intestinal propulsion rate. H-E staining was used to observe pathological changes in colon tissues. Colorimetric assays were employed to detect the levels of superoxide dismutase (SOD), glutathione (GSH), malondialdehyde (MDA), and ferrous ions (Fe2+) in colon tissues. Immunofluorescence assay was used to detect the expression of ROC in colon tissue. Immunohistochemical staining was used to detect the expression of JNK protein in colon tissues. Western blotting was used to detect the expression levels of ferroptosis-related proteins.ResultsCompared with the blank group, the STC group has a longer time required for the first black feces excretion, along with significantly increased levels of MDA, Fe2+, ROS, and JNK protein expression (P<0.05), while the small intestine propulsion rate, SOD and GSH levels, as well as the expression levels of GPX4 and FTH1 proteins are significantly reduced (P<0.05). After modeling, colonic mucosal tissue in rats shows bleeding, morphological structural damage, epithelial cell degeneration, and necrosis, and inflammatory cell infiltration. TMAO can cause more significant changes in the above indicators of STC rats, while SP600125 can reverse the effect of TMAO on the above indicators of STC rats.ConclusionTMAO can exacerbate the condition of STC, and its mechanism of action may be to induce ferroptosis in colon cells by regulating the ROS/JNK/GPX4 signaling pathway.