Background: Acetaminophen (APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available. Liensinine stands out as a key alkaloid known for its pharmaceutical activities. However, the role of liensinine in mitigating APAP-induced liver injury remains unclear. Objective: The aim of the study was to explore the protective effects of liensinine against APAP-induced liver injury. Methods: C57BL/6 male mice were treated with a dose of 200 mg/kg N-acetylcysteine or varying doses of liensinine (10 or 20 mg/kg) for seven consecutive days. APAP (400 mg/kg, i.g.) was then administered to induce liver damage for 12 hours. Blood samples and hepatic tissues were collected for further analysis. Liver enzyme levels and histopathological analysis were employed to assess liver injury. RNA-seq was conducted to evaluate the dynamic changes in gene expression. Biochemical assays were used to measure oxidative stress and inflammation, while the TUNEL assay was performed to assess hepatocyte apoptosis. Results: The results demonstrated that the administration of liensinine mitigated serum liver enzyme levels and tissue damage resulting from APAP overdose. Transcriptome analysis revealed significant and coordinated changes in genes related to the peroxisome proliferator-activated receptor signaling pathway, mitogen-activated protein kinase signaling pathway, and apoptosis pathway in response to APAP-induced hepatotoxicity. The expression alterations of key genes within these three pathways, associated with inflammation, oxidative stress, and cell apoptosis, were reversed by liensinine, indicating its potential in alleviating APAP-induced liver damage through multiple signaling pathways. This suggests the diverse therapeutic effects of liensinine, including inflammation suppression, oxidative stress reduction, and cell apoptosis inhibition. Indeed, pretreatment with liensinine effectively reduced inflammatory cytokines, oxidative stress indicators, and apoptotic cells induced by APAP. Conclusions: Liensinine mitigates APAP-induced hepatotoxicity in mice through multifaceted pathways, providing anti-inflammatory, antioxidant, and anti-apoptotic benefits.

Background: Acetaminophen (APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available. Liensinine stands out as a key alkaloid known for its pharmaceutical activities. However, the role of liensinine in mitigating APAP-induced liver injury remains unclear. Objective: The aim of the study was to explore the protective effects of liensinine against APAP-induced liver injury. Methods: C57BL/6 male mice were treated with a dose of 200 mg/kg N-acetylcysteine or varying doses of liensinine (10 or 20 mg/kg) for seven consecutive days. APAP (400 mg/kg, i.g.) was then administered to induce liver damage for 12 hours. Blood samples and hepatic tissues were collected for further analysis. Liver enzyme levels and histopathological analysis were employed to assess liver injury. RNA-seq was conducted to evaluate the dynamic changes in gene expression. Biochemical assays were used to measure oxidative stress and inflammation, while the TUNEL assay was performed to assess hepatocyte apoptosis. Results: The results demonstrated that the administration of liensinine mitigated serum liver enzyme levels and tissue damage resulting from APAP overdose. Transcriptome analysis revealed significant and coordinated changes in genes related to the peroxisome proliferator-activated receptor signaling pathway, mitogen-activated protein kinase signaling pathway, and apoptosis pathway in response to APAP-induced hepatotoxicity. The expression alterations of key genes within these three pathways, associated with inflammation, oxidative stress, and cell apoptosis, were reversed by liensinine, indicating its potential in alleviating APAP-induced liver damage through multiple signaling pathways. This suggests the diverse therapeutic effects of liensinine, including inflammation suppression, oxidative stress reduction, and cell apoptosis inhibition. Indeed, pretreatment with liensinine effectively reduced inflammatory cytokines, oxidative stress indicators, and apoptotic cells induced by APAP. Conclusions: Liensinine mitigates APAP-induced hepatotoxicity in mice through multifaceted pathways, providing anti-inflammatory, antioxidant, and anti-apoptotic benefits.

Background: Acetaminophen (APAP)-induced hepatotoxicity has attracted considerable attention in clinical settings due to the limited treatment options available. Liensinine stands out as a key alkaloid known for its pharmaceutical activities. However, the role of liensinine in mitigating APAP-induced liver injury remains unclear. Objective: The aim of the study was to explore the protective effects of liensinine against APAP-induced liver injury. Methods: C57BL/6 male mice were treated with a dose of 200 mg/kg N-acetylcysteine or varying doses of liensinine (10 or 20 mg/kg) for seven consecutive days. APAP (400 mg/kg, i.g.) was then administered to induce liver damage for 12 hours. Blood samples and hepatic tissues were collected for further analysis. Liver enzyme levels and histopathological analysis were employed to assess liver injury. RNA-seq was conducted to evaluate the dynamic changes in gene expression. Biochemical assays were used to measure oxidative stress and inflammation, while the TUNEL assay was performed to assess hepatocyte apoptosis. Results: The results demonstrated that the administration of liensinine mitigated serum liver enzyme levels and tissue damage resulting from APAP overdose. Transcriptome analysis revealed significant and coordinated changes in genes related to the peroxisome proliferator-activated receptor signaling pathway, mitogen-activated protein kinase signaling pathway, and apoptosis pathway in response to APAP-induced hepatotoxicity. The expression alterations of key genes within these three pathways, associated with inflammation, oxidative stress, and cell apoptosis, were reversed by liensinine, indicating its potential in alleviating APAP-induced liver damage through multiple signaling pathways. This suggests the diverse therapeutic effects of liensinine, including inflammation suppression, oxidative stress reduction, and cell apoptosis inhibition. Indeed, pretreatment with liensinine effectively reduced inflammatory cytokines, oxidative stress indicators, and apoptotic cells induced by APAP. Conclusions: Liensinine mitigates APAP-induced hepatotoxicity in mice through multifaceted pathways, providing anti-inflammatory, antioxidant, and anti-apoptotic benefits.

Background::Histological healing is closely associated with improved long-term clinical outcomes and lowered relapses in patients with ulcerative colitis (UC). Here, we developed a novel diagnostic criterion for assessing histological healing in UC patients.Methods::We conducted a retrospective cohort study in UC patients, whose treatment was iteratively optimized to achieve mucosal healing at Shanghai Tenth People’s Hospital of Tongji University from January 2017 to May 2022. We identified an inflammatory cell enumeration index (ICEI) for assessing histological healing based on the proportions of eosinophils, CD177 + neutrophils, and CD40L + T cells in the colonic lamina propria under high power field (HPF), and the outcomes (risks of symptomatic relapses) of achieving histological remission vs. persistent histological inflammation using Kaplan-Meier curves. Intrareader reliability and inter-reader reliability were evaluated by each reader. The relationships to the changes in the Nancy index and the Geboes score were also assessed for responsiveness. The ICEI was further validated in a new cohort of UC patients from other nine university hospitals. Results::We developed an ICEI for clinical diagnosis of histological healing, i.e., Y = 1.701X 1 + 0.758X 2 + 1.347X 3 - 7.745 (X 1, X 2, and X 3 represent the proportions of CD177 + neutrophils, eosinophils, and CD40L + T cells, respectively, in the colonic lamina propria under HPF). The receiver operating characteristics curve (ROC) analysis revealed that Y <-0.391 was the cutoff value for the diagnosis of histological healing and that an area under the curve (AUC) was 0.942 (95% confidence interval [CI]: 0.905-0.979) with a sensitivity of 92.5% and a specificity of 83.6% ( P <0.001). The intraclass correlation coefficient (ICC) for the intrareader reliability was 0.855 (95% CI: 0.781-0.909), and ICEI had good inter-reader reliability of 0.832 (95% CI: 0.748-0.894). During an 18-month follow-up, patients with histological healing had a substantially better outcome compared with those with unachieved histological healing ( P <0.001) using ICEI. During a 12-month follow-up from other nine hospitals, patients with histological healing also had a lower risk of relapse than patients with unachieved histological healing. Conclusions::ICEI can be used to predict histological healing and identify patients with a risk of relapse 12 months and 18 months after clinical therapy. Therefore, ICEI provides a promising, simplified approach to monitor histological healing and to predict the prognosis of UC.Registration::Chinese Clinical Trial Registry, No. ChiCTR2300077792.

Objective:To explore the effect of hyperbaric oxygen（HBO）combined with paclitaxel（PTX）on the proliferation，level of reactive oxygen species（ROS），and cell cycle of human gastric cancer AGS cell line cultured in vitro. Methods:After the completion of in vitro culture，the AGS cells were divided into four groups，including control group，PTX group，HBO group，and HBO+PTX group. Cell Counting Kit-8（CCK-8）assay was used to detect the inhibitory effect of HBO combined with PTX on the proliferation of AGS cells. Flow cytometry was adopted to detect the intracellular ROS level and cell cycle arrest. Western blotting was used to assess the expression levels of cell cycle-related proteins. Results:The survival rate of AGS cells in the HBO+PTX group was significantly lower than those in the control group and the HBO group（ P<0.01），and also lower than that in the PTX group（ P<0.05）. The ROS level of AGS cells in the HBO+PTX group was significantly higher than those in the control group and the HBO group（ P<0.01），and also higher than that in the PTX group（ P<0.05）. PTX blocked the AGS cell cycle in S phase（ P<0.05），and the percentage of cells at S phase in the HBO+PTX group was found slightly higher than that in the PTX group（ P>0.05）. The expression level of Cyclin A1 in the HBO+PTX group was significantly lower than those in the other three groups（ P<0.05 or P<0.01）；the expression level of CDK2 in the HBO+PTX group was lower than those in the control group and the HBO group（ P<0.05），and also slightly lower than that in the PTX group（ P>0.05）. Conclusion:HBO can effectively enhance the inhibitory effect of PTX on the proliferation of AGS cells，increase the intracellular ROS level in AGS cells，and block the cell cycle，thereby inhibiting the differentiation and proliferation of AGS cells.

Objective:To investigate the relationship between the expression level of lymphocyte enhancer-binding factor 1(LEF1) and CTNNB1 and the cycle arrest, apoptosis and radiation resistance of esophageal cancer cells and unravel the related mechanisms.Methods:Recombinantplasmids and empty plasmids expressing LEF1 and CTNNB1were constructed and transfected into esophageal cancer cells. RT-PCR assay was used to detect the transfection efficiency of the plasmids. Clone formation assay, CCK8 assay, cell cycle test by flow cytometry, apoptosis test by flow cytometry and Western blot were performed to detect the differences in theradioresistance, proliferation, cell cycle and apoptosis of esophageal cancer cells before and after transfection.Results:The survival rate of clonal colony cells in the pGEX-LEF1+ pCMV6-CTNNB1 group was significantly better than those in other groups ( P<0.05). The proliferation of clonal colony cellsat 72 h, 96 h and 120 h in the pGEX-LEF1+ pCMV6-CTNNB1 group was significantly better than those in the pGEX+ pCMV6, pGEX-LEF1+ pCMV6 and pCMV6-CTNNB1+ pGEX groups (all P<0.05). The percentage of G 2 phase arrest cells in the pGEX-LEF1+ pCMV6-CTNNB1 group was significantly higher than those in the other groups (all P<0.05). The apoptosis rate of esophageal cancer cells in the pGEX-LEF1+ pCMV6-CTNNB1 group was significantly lower compared with those in the pGEX+ pCMV6, pGEX-LEF1+ pCMV6 and pCMV6-CTNNB1+ pGEX groups (all P<0.05). The expression levels of Bax and Caspase 3 proteins in the pGEX-LEF1+ pCMV6-CTNNB1 group were significantly lower than those in the pGEX+ pCMV6, pGEX-LEF1+ pCMV6 and pCMV6-CTNNB1+ pGEX groups (all P<0.05). The expression level of Bcl-2 protein in the pGEX-LEF1+ pCMV6-CTNNB1 group was significantly higher compared with those in the other groups (all P<0.05). Conclusion:LEF1 and CTNNB1 can regulate the proliferation and G 2 phase arrest of esophageal cancer cells after radiation intervention by mediating the Wnt signaling pathway, and improve the radiation resistance of esophageal cancer cells by inhibiting cell apoptosis.

Objective:To investigate the effect of hyperbaric oxygen（HBO）on the resistance of gastric cancer AGS cells to irinotecan（ITC）and its related regulatory mechanisms.Methods:Gastric cancer AGS cells were cultured in vitro，and the cells were divided into four groups according to the experimental design：control group，ITC group，HBO group，and HBO + ITC group. The effect of ITC combined with HBO on the proliferation rate of gastric cancer cells was detected by Cell Counting Kit-8（CCK-8）；the changes of apoptosis rate after ITC combined with HBO treatment of AGS cells were detected by flow cytometry；and the changes in the expression of apoptosis-related and drug resistance-related proteins in AGS cells were detected by Western blotting assay. Results:Compared with control group and HBO group，the proliferation of AGS cells in the HBO + ITC group was further inhibited，and its proliferation rate was significantly lower than those of the control group and HBO group（ P<0.01），and also lower than that of the ITC group（ P<0.05）. Compared with those in the control group and the HBO group，the apoptosis rate of AGS cells in the HBO+ITC group was significantly higher than those in the control group and the HBO group（ P<0.01），and also higher than that in the ITC group（ P < 0.05）. In the ITC group and HBO+ITC group，the expression of apoptosis-related and drug resistance-related proteins，including Bcl-2，caspase-9 and P-gp，were significantly decreased，while the expression of Bad，caspase-3 and PARP were significantly increased，and the differences were statistically significant（ P<0.05 or P<0.01）. Compared with the control group and HBO group，p-JNK expression was increased and STAT3 expression was decreased in the ITC group（ P<0.01）；compared with the ITC group，p-JNK expression was increased and STAT3 expression was decreased in the HBO+ITC group（ P<0.05 or P<0.01）. Conclusion:HBO combined with ITC can effectively inhibit the proliferation of gastric cancer AGS cells，affect the apoptosis of AGS cells by regulating the JNK/STAT3 signaling pathway，and improve the cells’ drug resistance to a certain extent.

Objective:To investigate the effect of hyperbaric oxygen（HBO）on the resistance of gastric cancer AGS cells to irinotecan（ITC）and its related regulatory mechanisms.Methods:Gastric cancer AGS cells were cultured in vitro，and the cells were divided into four groups according to the experimental design：control group，ITC group，HBO group，and HBO + ITC group. The effect of ITC combined with HBO on the proliferation rate of gastric cancer cells was detected by Cell Counting Kit-8（CCK-8）；the changes of apoptosis rate after ITC combined with HBO treatment of AGS cells were detected by flow cytometry；and the changes in the expression of apoptosis-related and drug resistance-related proteins in AGS cells were detected by Western blotting assay. Results:Compared with control group and HBO group，the proliferation of AGS cells in the HBO + ITC group was further inhibited，and its proliferation rate was significantly lower than those of the control group and HBO group（ P<0.01），and also lower than that of the ITC group（ P<0.05）. Compared with those in the control group and the HBO group，the apoptosis rate of AGS cells in the HBO+ITC group was significantly higher than those in the control group and the HBO group（ P<0.01），and also higher than that in the ITC group（ P < 0.05）. In the ITC group and HBO+ITC group，the expression of apoptosis-related and drug resistance-related proteins，including Bcl-2，caspase-9 and P-gp，were significantly decreased，while the expression of Bad，caspase-3 and PARP were significantly increased，and the differences were statistically significant（ P<0.05 or P<0.01）. Compared with the control group and HBO group，p-JNK expression was increased and STAT3 expression was decreased in the ITC group（ P<0.01）；compared with the ITC group，p-JNK expression was increased and STAT3 expression was decreased in the HBO+ITC group（ P<0.05 or P<0.01）. Conclusion:HBO combined with ITC can effectively inhibit the proliferation of gastric cancer AGS cells，affect the apoptosis of AGS cells by regulating the JNK/STAT3 signaling pathway，and improve the cells’ drug resistance to a certain extent.

Objective:To explore the effect of hyperbaric oxygen（HBO）combined with paclitaxel（PTX）on the proliferation，level of reactive oxygen species（ROS），and cell cycle of human gastric cancer AGS cell line cultured in vitro. Methods:After the completion of in vitro culture，the AGS cells were divided into four groups，including control group，PTX group，HBO group，and HBO+PTX group. Cell Counting Kit-8（CCK-8）assay was used to detect the inhibitory effect of HBO combined with PTX on the proliferation of AGS cells. Flow cytometry was adopted to detect the intracellular ROS level and cell cycle arrest. Western blotting was used to assess the expression levels of cell cycle-related proteins. Results:The survival rate of AGS cells in the HBO+PTX group was significantly lower than those in the control group and the HBO group（ P<0.01），and also lower than that in the PTX group（ P<0.05）. The ROS level of AGS cells in the HBO+PTX group was significantly higher than those in the control group and the HBO group（ P<0.01），and also higher than that in the PTX group（ P<0.05）. PTX blocked the AGS cell cycle in S phase（ P<0.05），and the percentage of cells at S phase in the HBO+PTX group was found slightly higher than that in the PTX group（ P>0.05）. The expression level of Cyclin A1 in the HBO+PTX group was significantly lower than those in the other three groups（ P<0.05 or P<0.01）；the expression level of CDK2 in the HBO+PTX group was lower than those in the control group and the HBO group（ P<0.05），and also slightly lower than that in the PTX group（ P>0.05）. Conclusion:HBO can effectively enhance the inhibitory effect of PTX on the proliferation of AGS cells，increase the intracellular ROS level in AGS cells，and block the cell cycle，thereby inhibiting the differentiation and proliferation of AGS cells.

Objective:To explore the effects of miR-181a-5p on the occurrence and development of gastrointestinal stromal tumors (GIST) through targeting CTDSPL mediating TGF-β signaling pathway.Methods:Surgical treatment of GIST patients in the First People’s Hospital of Shangqiu City from Jan. 2016 to Dec. 2019 were selected as research objects, and tumor tissue and adjacent normal tissue were collected intraoperatively. The clinicopathological data of the patients were analyzed. Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the mRNA expression of CTDSPL gene and miR-181a-5p expression. Western blot was used to detect the protein level of CTDSPL and TGF-β signaling pathway related factors. Human gastrointestinal stromal tumor cell lines (GIST-T1) were transfected with miR-181a-5p mimic, miR-181a-5p inhibitor, or CTDSPL overexpression vector. MTT was used to detect cell proliferation activity, Transwell assay was utilized to detect cell invasion, flow cytometry was used to determine cell apoptosis in each group.Results:Compared with adjacent tissues, expression of miR-181a-5p and TGF-β signaling pathway related factors was activated while CTDSPL expression was inhibited. Tumor size, invasion depth and modified NIH grading were related to the mRNA expression level of CTDSPL gene in GIST tumor tissues (All P<0.05) . Compared with Blank group, inhibition of miR-181a-5p or CTDSPL overexpression had the ability to inhibit the cell viability and invasion, induce apoptosis. The effects of miR-181a-5p mimic on GIST-T1 can be saved by CTDSPL overexpression. Conclusion:miR-181a-5p can promote the occurrence and development of GIST by down-regulating the CTDSPL gene level and activating TGF-β signaling pathway.