Acetaminophen (APAP) is the primary cause of drug-induced acute liver failure. Ovarian tumor deubiquitinase 6A (OTUD6A), a recently discovered deubiquitinase of the OTU family, has been primarily studied in tumor contexts. However, its role in APAP-induced liver injury (AILI) remains unclear. Therefore, this study aimed to investigate the involvement of OTUD6A in the pathogenesis of AILI. Our findings demonstrated a substantial upregulation of OTUD6A in both the liver tissue and isolated hepatocytes of mice following APAP stimulation. OTUD6A knockout exacerbated APAP-induced inflammation, hepatocyte necrosis, and liver injury, whereas OTUD6A overexpression alleviated these pathologies. Mechanistically, OTUD6A directly interacted with the enhancer of zeste homolog 2 (EZH2) and selectively removed K48-linked polyubiquitin chains from EZH2, enhancing its stability. This resulted in increased protein levels of EZH2 and H3K27me3, as well as reduced endoplasmic reticulum (ER) stress and cell death in hepatocytes. Collectively, our research uncovers a novel role for OTUD6A in mitigating APAP-induced liver injury by promoting EZH2 stabilization.

Purpose To investigate how IL-6-mediated monocytes/macrophages suppress the effector function of natural killer(NK)cells through the PD-L1/PD-1 axis in acute-on-chronic liver failure(ACLF).Methods HE stai-ning and Masson's trichrome staining were used to assess hepatic inflammatory infiltration and fibrosis.Immunofluores-cence staining was performed to detect IL-6 expression in hepatic macrophages.Flow cytometry was applied to evaluate the phenotypes and functions of monocytes/macrophages and NK cells.Recombinant IL-6 was used to treat peripheral blood mononuclear cells,and PD-L1 expression on monocytes was examined.Monocytes and NK cells were co-cul-tured,and after PD-1 blockade,NK cell effector function was evaluated.Results The ACLF patients,the degree of hepatic fibrosis increased with higher serum IL-6 level.Hepatic macrophages secreted more IL-6,and the circulating monocytes showed an increased percentage[(9.89±0.65)％],absolute count[(0.58±0.04)× 106/mL],prolif-erative capacity[(36.65±6.810)％],and IL-6 secretion[(16.97±2.387)％].In ACLF mice,hepatic inflamma-tory infiltration and fibrosis were aggravated,and IL-6+macrophages exhibited enhanced pro-inflammatory activity.IL-6 had mediated the upregulation of PD-L1 expression in monocytes,while NK cells showed increased PD-1 expression and impaired effector function.When monocytes and NK cells were co-cultured,PD-1 blockade restored,NK cell ef-fector functions.Conclusion In ACLF,elevated IL-6 levels were associated with hepatic fibrosis and disease severi-ty.IL-6 mediated monocytes-induced suppression of NK cell effector functions via the PD-L1/PD-1 axis,suggesting that IL-6 plays a critical role in ACLF-related immune dysregulation.

Objective:To investigate the molecular mechanism by which 2',4'-dihydroxychalcone(D2)inhibits proliferation,migration,and epithelial-mesenchymal transition(EMT)in colorectal cancer cells through miR-7-5p-mediated autophagy.Methods:Human colorectal cancer cell lines HCT-15 and SW620 were treated with D2 at concentrations of 0,12.5,25,and 50 μmol/L.Cell proliferation and clonogenic capacity were evaluated using MTT and colony formation assays.Cell migration was assessed by wound healing and Transwell assays.WB assay was used to detect the expression of EMT-related proteins,autophagy-related proteins,and key components of the PI3K/AKT/mTOR pathway.Autophagosome formation was visualized by immunofluorescence staining.TCGA database and KEGG pathway analyses were performed to evaluate miR-7-5p expression and its association with colorectal cancer.RT-qPCR was used to quantify miR-7-5p expression,and lentiviral transduction was employed to establish stable miR-7-5p knockdown or overexpression cell lines.Results:D2 significantly inhibited colorectal cancer cell proliferation,migration,and EMT(P<0.05 or P<0.01).TCGA and KEGG analyses revealed that miR-7-5p expression was downregulated in colorectal cancer and closely associated with disease progression.D2 treatment(12.5,25,and 50 μmol/L)significantly upregulated miR-7-5p expression in HCT-15 and SW620 cells(P<0.01).At 25 μmol/L,D2 increased the expression of autophagy-related proteins(LC3 and p-ULK1)and inhibited the PI3K/AKT/mTOR signaling pathway(P<0.05),accompanied by increased autophagosome formation(P<0.01).In miR-7-5p-knockdown cells treated with D2,the levels of LC3 and p-ULK1 were significantly reduced compared to D2-only treated cells(P<0.05 or P<0.01).Conclusion:D2 upregulates miR-7-5p to induce autophagy,thereby inhibiting colorectal cancer cell proliferation,migration,and EMT,possibly through suppression of the PI3K/AKT/mTOR signaling pathway.

Purpose To investigate how IL-6-mediated monocytes/macrophages suppress the effector function of natural killer(NK)cells through the PD-L1/PD-1 axis in acute-on-chronic liver failure(ACLF).Methods HE stai-ning and Masson's trichrome staining were used to assess hepatic inflammatory infiltration and fibrosis.Immunofluores-cence staining was performed to detect IL-6 expression in hepatic macrophages.Flow cytometry was applied to evaluate the phenotypes and functions of monocytes/macrophages and NK cells.Recombinant IL-6 was used to treat peripheral blood mononuclear cells,and PD-L1 expression on monocytes was examined.Monocytes and NK cells were co-cul-tured,and after PD-1 blockade,NK cell effector function was evaluated.Results The ACLF patients,the degree of hepatic fibrosis increased with higher serum IL-6 level.Hepatic macrophages secreted more IL-6,and the circulating monocytes showed an increased percentage[(9.89±0.65)％],absolute count[(0.58±0.04)× 106/mL],prolif-erative capacity[(36.65±6.810)％],and IL-6 secretion[(16.97±2.387)％].In ACLF mice,hepatic inflamma-tory infiltration and fibrosis were aggravated,and IL-6+macrophages exhibited enhanced pro-inflammatory activity.IL-6 had mediated the upregulation of PD-L1 expression in monocytes,while NK cells showed increased PD-1 expression and impaired effector function.When monocytes and NK cells were co-cultured,PD-1 blockade restored,NK cell ef-fector functions.Conclusion In ACLF,elevated IL-6 levels were associated with hepatic fibrosis and disease severi-ty.IL-6 mediated monocytes-induced suppression of NK cell effector functions via the PD-L1/PD-1 axis,suggesting that IL-6 plays a critical role in ACLF-related immune dysregulation.

Shaoyao Gancao Decoction(SGD) is one of the prescriptions included in the Catalogue of Ancient Classical Prescriptions(First Batch) published by the National Administration of Traditional Chinese Medicine in 2018. It is composed of Paeoniae Radix Alba and Glycyrrhizae Radix et Rhizoma Praeparata Cum Melle. Experiments and clinical cases have proved that SGD has anti-inflammatory, pain-relieving, antispasmodic, and liver-protecting effects. It is widely used in the clinical treatment of diseases, demonstrating definite effects on digestive system diseases. By reviewing the literature on SGD in recent years, this paper summarizes the applications of SGD in the treatment of digestive system diseases from three aspects of basic prescription and syndrome analysis, clinical application, and mechanism to expounds the research status in this field. Furthermore, this paper put forward the shortcomings and improvement methods of the available studies, aiming to provide reference for the application of SGD in treating digestive system diseases and the research and development of SGD preparations.
Drugs, Chinese Herbal/administration & dosage* ; Humans ; Digestive System Diseases/drug therapy* ; Animals ; Paeonia/chemistry* ; Glycyrrhiza/chemistry*

BACKGROUND: Osteoarthritis (OA), a degenerative joint disorder, is a major reason of disability in adults. Accumulating evidences have proved that bone marrow mesenchymal stem cells (BMSCs)-carried exosomes play a significant therapeutic effect on OA. However, the precise regulatory network remains unknown. METHODS: OA and normal cartilage samples were acquired from patients, and chondrocytes were exposed to IL-1b to conduct a cellular OA model. Exosomes prepared from BMSCs were identified using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Cell viability was determined with CCK-8 assay. Inflammatory injury was assessed by LDH and inflammatory factors (TNF-a and IL-6) using corresponding ELISA kits, respectively. Ferroptosis was evaluated by GSH, MDA and iron levels using corresponding kits, and ROS level with DCFH-DA. The expressions of genes/proteins were determined with RT-qPCR/western bolt. RNA immunoprecipitation and luciferase activity assay were conducted for testing the interactions of small nucleolar RNA host gene 7 (SNHG7)/ferroptosis suppressor protein 1 (FSP1) and miR-485-5p. RESULTS: The expressions of SNHG7 and FSP1 were both reduced in IL-1b-induced chondrocytes and OA cartilage tissues, and there was a positive correlation between them in clinical level. Moreover, SNHG7 was enriched in BMSCsderived exosomes (BMSCs-Exos) and could be internalized by chondrocytes. Functional analysis illustrated that BMSCsExos administration repressed inflammatory injury, oxidative stress and ferroptosis in IL-1b-induced chondrocytes, while these changes were reinforced when SNHG7 was overexpressed in BMSCs-Exos. Notably, FSP1 silencing in chondrocytes abolished the beneficial effects mediated by exosomal SNHG7. CONCLUSIONS Exosomal SNHG7 released from BMSCs inhibited inflammation and ferroptosis in IL-1b-induced chondrocytes through miR-485-5p/FSP1 axis. This work suggested that BMSCs-derived exosomal SNHG7 would be a prospective target for OA treatment.

BACKGROUND: Osteoarthritis (OA), a degenerative joint disorder, is a major reason of disability in adults. Accumulating evidences have proved that bone marrow mesenchymal stem cells (BMSCs)-carried exosomes play a significant therapeutic effect on OA. However, the precise regulatory network remains unknown. METHODS: OA and normal cartilage samples were acquired from patients, and chondrocytes were exposed to IL-1b to conduct a cellular OA model. Exosomes prepared from BMSCs were identified using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Cell viability was determined with CCK-8 assay. Inflammatory injury was assessed by LDH and inflammatory factors (TNF-a and IL-6) using corresponding ELISA kits, respectively. Ferroptosis was evaluated by GSH, MDA and iron levels using corresponding kits, and ROS level with DCFH-DA. The expressions of genes/proteins were determined with RT-qPCR/western bolt. RNA immunoprecipitation and luciferase activity assay were conducted for testing the interactions of small nucleolar RNA host gene 7 (SNHG7)/ferroptosis suppressor protein 1 (FSP1) and miR-485-5p. RESULTS: The expressions of SNHG7 and FSP1 were both reduced in IL-1b-induced chondrocytes and OA cartilage tissues, and there was a positive correlation between them in clinical level. Moreover, SNHG7 was enriched in BMSCsderived exosomes (BMSCs-Exos) and could be internalized by chondrocytes. Functional analysis illustrated that BMSCsExos administration repressed inflammatory injury, oxidative stress and ferroptosis in IL-1b-induced chondrocytes, while these changes were reinforced when SNHG7 was overexpressed in BMSCs-Exos. Notably, FSP1 silencing in chondrocytes abolished the beneficial effects mediated by exosomal SNHG7. CONCLUSIONS Exosomal SNHG7 released from BMSCs inhibited inflammation and ferroptosis in IL-1b-induced chondrocytes through miR-485-5p/FSP1 axis. This work suggested that BMSCs-derived exosomal SNHG7 would be a prospective target for OA treatment.

BACKGROUND: Osteoarthritis (OA), a degenerative joint disorder, is a major reason of disability in adults. Accumulating evidences have proved that bone marrow mesenchymal stem cells (BMSCs)-carried exosomes play a significant therapeutic effect on OA. However, the precise regulatory network remains unknown. METHODS: OA and normal cartilage samples were acquired from patients, and chondrocytes were exposed to IL-1b to conduct a cellular OA model. Exosomes prepared from BMSCs were identified using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Cell viability was determined with CCK-8 assay. Inflammatory injury was assessed by LDH and inflammatory factors (TNF-a and IL-6) using corresponding ELISA kits, respectively. Ferroptosis was evaluated by GSH, MDA and iron levels using corresponding kits, and ROS level with DCFH-DA. The expressions of genes/proteins were determined with RT-qPCR/western bolt. RNA immunoprecipitation and luciferase activity assay were conducted for testing the interactions of small nucleolar RNA host gene 7 (SNHG7)/ferroptosis suppressor protein 1 (FSP1) and miR-485-5p. RESULTS: The expressions of SNHG7 and FSP1 were both reduced in IL-1b-induced chondrocytes and OA cartilage tissues, and there was a positive correlation between them in clinical level. Moreover, SNHG7 was enriched in BMSCsderived exosomes (BMSCs-Exos) and could be internalized by chondrocytes. Functional analysis illustrated that BMSCsExos administration repressed inflammatory injury, oxidative stress and ferroptosis in IL-1b-induced chondrocytes, while these changes were reinforced when SNHG7 was overexpressed in BMSCs-Exos. Notably, FSP1 silencing in chondrocytes abolished the beneficial effects mediated by exosomal SNHG7. CONCLUSIONS Exosomal SNHG7 released from BMSCs inhibited inflammation and ferroptosis in IL-1b-induced chondrocytes through miR-485-5p/FSP1 axis. This work suggested that BMSCs-derived exosomal SNHG7 would be a prospective target for OA treatment.

Objective:To demonstrate the three-dimensional structures of the limbic system and its fiber connections through fiber dissection,and to provide reference for relevant professionals to master the anatomy of the limbic system.Methods:Ten cerebral hemispheres were treated and dissected according to Kelinger method,and the limbic system and its fiber connections were displayed.Results:The limbic system was arranged around the thalamus and corpus cal-losum in a double-layer concentric circle structure.The outer layer structures mainly consisted of the cingulate gyrus and the parahippocampal gyrus,while the inner layer structures included the amygdala,hippocampus and fornix.The main association fiber of the outer layer is the cingulum,whose superior trunk is mainly located in the cingulate gyrus,and the inferior trunk is mainly located in the parahippocampal gyrus.The fiber structures of the inner layer includes the striae terminalis and ansa peduncularis emanating from the amygdala and the fornix of the hippocampus.Conclusion:Limbic system is an important connection structure between telencephalon and diencephalon,and its anatomical struc-ture is complex.Fiber dissection method can effectively demonstrate the complex spatial structure of limbic system,which is of great benefit to relevant professionals to understand its three-dimensional structure.

BACKGROUND: Osteoarthritis (OA), a degenerative joint disorder, is a major reason of disability in adults. Accumulating evidences have proved that bone marrow mesenchymal stem cells (BMSCs)-carried exosomes play a significant therapeutic effect on OA. However, the precise regulatory network remains unknown. METHODS: OA and normal cartilage samples were acquired from patients, and chondrocytes were exposed to IL-1b to conduct a cellular OA model. Exosomes prepared from BMSCs were identified using nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM). Cell viability was determined with CCK-8 assay. Inflammatory injury was assessed by LDH and inflammatory factors (TNF-a and IL-6) using corresponding ELISA kits, respectively. Ferroptosis was evaluated by GSH, MDA and iron levels using corresponding kits, and ROS level with DCFH-DA. The expressions of genes/proteins were determined with RT-qPCR/western bolt. RNA immunoprecipitation and luciferase activity assay were conducted for testing the interactions of small nucleolar RNA host gene 7 (SNHG7)/ferroptosis suppressor protein 1 (FSP1) and miR-485-5p. RESULTS: The expressions of SNHG7 and FSP1 were both reduced in IL-1b-induced chondrocytes and OA cartilage tissues, and there was a positive correlation between them in clinical level. Moreover, SNHG7 was enriched in BMSCsderived exosomes (BMSCs-Exos) and could be internalized by chondrocytes. Functional analysis illustrated that BMSCsExos administration repressed inflammatory injury, oxidative stress and ferroptosis in IL-1b-induced chondrocytes, while these changes were reinforced when SNHG7 was overexpressed in BMSCs-Exos. Notably, FSP1 silencing in chondrocytes abolished the beneficial effects mediated by exosomal SNHG7. CONCLUSIONS Exosomal SNHG7 released from BMSCs inhibited inflammation and ferroptosis in IL-1b-induced chondrocytes through miR-485-5p/FSP1 axis. This work suggested that BMSCs-derived exosomal SNHG7 would be a prospective target for OA treatment.