abstract:
Objective To study the effects and potential mechanisms of the aquaporin 1(AQP1) inhibitor acetazolamide(AZ) on the proliferation, apoptosis, and inflammatory response of rheumatoid arthritis(RA) fibroblast-like synoviocytes(FLS). Methods TNF-α-induced RA-FLS was served as in vitro RA model. MTT assay, IF staining, and EdU incorporation assay were applied to study AZ′s effects on RA-FLS proliferation. Hoechst staining, flow cytometry analysis of Annexin V-FITC/PI-stained cells, and mitochondrial membrane potential detection experiments were used to detect cell apoptosis. ELISA and quantitative real-time PCR methods were used to measure pro-inflammatory cytokine production. Cell autophagy was evaluated using IF staining and mCherry-GFP-LC3B puncta assay. Western blot was performed to detect the levels of autophagy, apoptosis, and proliferation-related proteins. Moreover, the role of autophagy inhibition in AZ′s effects on RA-FLS was examined by co-treating with the autophagy activator rapamycin(RAPA) or the autophagy inhibitor 3-methyladenine(3-MA). Results AZ dose-dependently inhibited cell proliferation, promoted apoptosis, and reduced the production of pro-inflammatory cytokines in RA-FLS. Furthermore, AZ suppressed cytoprotective autophagy in these cells, as evidenced by decreased LC3B levels(P

abstract:
Objective To explore the role and molecular mechanism of parthenolide(PTL) in regulating tumor-associated macrophage(TAM) polarization to inhibit the malignant progression of gallbladder cancer cells. Methods M0 macrophages were co-cultured with human gallbladder cancer cell line GBC-SD and treated with PTL, and the cells were divided into control group, TAM group, TAM+PTL group. Flow cytometry was used to determine the levels of CD86 and CD206 in macrophages. RT-qPCR and Western blot were used to determine the expressions of CD163, interleukin-10(IL-10) and arginase-1(Arg-1). ELISA and Western blot were used to determine the secretion of CC chemokine ligand 2(CCL2) in the supernatant of GBC-SD cells and the expression of CCL2 in the cells. GBC-SD cells were transfected with a CCL2 overexpression vector and divided into TAM, TAM+PTL, TAM+PTL+OE-NC, TAM+PTL+OE-CCL2 groups. Flow cytometry was used to assess CD86 and CD206 levels in macrophages, RT-qPCR and Western blot were used to assess CD163, IL-10 and Arg-1 expression. CCK-8 assay, plate-based colony formation assay and Transwell chamber assay were used to assess GBC-SD cell proliferation, migration and invasion, respectively. Results Compared with the TAM group, the TAM+PTL group showed increased CD86 expression and decreased CD206 expression in macrophages, the mRNA and protein expressions of CD163, IL-10 and Arg-1 were downregulated, the CCL2 content and protein expression in GBC-SD cells decreased, cell proliferation activity, colony formation, migration and invasion were all reduced(P

abstract:
Objective To investigate the effect and mechanism of long non-coding RNA RMRP(LncRNA RMRP) on oxygen-glucose deprivation/reperfusion(OGD/R)-induced ferroptosis in mouse HL-1 cardiomyocytes by regulating miR-766-5p. Methods HL-1 cells were cultured in vitro, and OGD/R models were established. The expression levels of LncRNA RMRP in HL-1 cells at various reperfusion time points were subsequently quantified using qRT-PCR. The LncRNA RMRP small RNA interference fragment(si-RMRP) and its corresponding negative control(si-NC), as well as the miR-766-5p inhibitor and its respective negative control(inhibitor-NC), were transfected into HL-1 cells. Subsequently, the cells were subjected to OGD/R treatment. CCK-8 assay was employed to evaluate cell viability. Assay kits were employed to measure the levels of lactate dehydrogenase(LDH) in the cell supernatant, as well as the intracellular levels of malondialdehyde(MDA), superoxide dismutase(SOD), glutathione(GSH), and ferrous ion(Fe2+). qRT-PCR analysis was conducted to assess the expression levels of LncRNA RMRP and miR-766-5p. Western blot analysis was conducted to assess the expression levels of proteins associated with ferroptosis including GPX4, SLC7A11, and FTH1. Dual-luciferase reporter assays were performed to investigate the sponge adsorption relationship between LncRNA RMRP and miR-766-5p. Results As reperfusion time extended, the expression level of LncRNA RMRP in cells progressively increased(P2+ levels within the cells, and decreased the activities of SOD and GSH in cells(P2+ levels within the cells, and promoting SOD and GSH activities in cells(P

abstract:
Objective To identify ferroptosis-related genes associated with gastric cancer prognosis and investigate their potential molecular functions. Methods Gene expression profiles and clinical information of gastric cancer tissues and adjacent normal tissues were obtained from TCGA database. Differential expression analysis of ferroptosis-related genes was performed using the "DESeq2" package in R software. Key genes were identified and a prognostic model for gastric cancer was constructed through Cox regression analysis based on the LASSO algorithm. Patients were stratified into high-risk and low-risk groups according to the median risk score. The accuracy of the model was evaluated using Kaplan-Meier survival analysis and ROC curve analysis. Immune cell infiltration in gastric cancer patients was assessed with the "CIBERSORT" package. The mRNA expression of differentially expressed genes(DEGs) with prognostic significance was examined in both gastric cancer and adjacent normal tissue samples. In vitro experiments were conducted to validate the impact of hydroxycarboxylic acid receptor 1(HCAR1) on the malignant biological behavior of gastric cancer. Results Based on ferroptosis-related genes from the TCGA database, a novel prognostic model was constructed. It demonstrated robust predictive power for survival in both training and validation cohorts. RT-qPCR analysis of 8 pairs of gastric cancer and normal tissues revealed that the expression patterns of 6 prognostic DEGs in cancer tissues were consistent with those predicted by the model. In vitro experiments confirmed that downregulation of the key gene HCAR1 could inhibit the proliferation, invasion, and metastasis of gastric cancer cells. Conclusion The ferroptosis-related gene based prognostic model exhibits robust predictive capability, allowing for accurate determination of prognosis and survival in individuals with gastric cancer.

abstract:
Objective To explore the active components, key targets, and mechanism of action of turnip in alleviating pulmonary fibrosis(PF) based on network pharmacology and animal experiments. Methods The active components and targets of Brassica rapa L. were screened using the traditional Chinese medicine systems pharmacology database and analysis platform database, and PF-related targets were obtained from disease databases such as online mendelian inheritance of man(OMIM) and DrugBank. The intersection targets were used to construct a protein-protein interaction(PPI) network to identify core targets, followed by gene oncology(GO)/Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis. In the animal experiments, a bleomycin-induced PF mouse model was established. Pathological changes in lung tissue were evaluated using HE and Masson staining. qRT-PCR was used to detect the mRNA expression of tumor necrosis factor-α(TNF-α), phosphatidylinositol 3-kinase(PI3K), and akstrain transforming 1(AKT1), and immunofluorescence staining was used to measure the protein expression of TNF-α, PI3K, and AKT1. Results The 68 active components identified in Brassica rapa L. may regulate PI3K-Akt signaling pathway by acting on 89 potential targets such as TNF-α and AKT1. The results of animal experiments showed that polysaccharide of Brassica rapa L.(BRPs) could significantly reduce the degree of bleomycin induced pulmonary fibrosis in mice; HE and Masson staining of lung tissue showed that compared with the model group, the damage of alveolar structure, the infiltration of inflammatory cells and the deposition of collagen fibers in the BRPs treatment group were significantly reduced. Further mechanism studies showed that BRPs could significantly down-regulate the mRNA and protein expression levels of TNF-α, PI3K and AKT1 in lung tissue of pulmonary fibrosis mice. Conclusion Brassica rapa L. can synergistically alleviate pulmonary fibrosis through “multi-component, multi-target and multi-channel” approach; BRPs is one of the main active components, and plays an anti-fibrosis role by inhibiting TNF-α/PI3K Akt signaling pathway.

abstract:
Objective To determine the expression of melanoma-associated antigens D4(MAGED4) and SIRT7 in human glioma, and to analyze the potential effects of MAGED4 and SIRT7 on glioma cell proliferation. Methods The MAGED4 and SIRT7 expression levels and their correlation were compared by the China glioma genome atlas(CGGA), human protein atlas(HPA), and UALCAN databases. Survival analysis, ROC curve analysis, and Cox regression analysis were used to predict the outcome of MAGED4 and SIRT 7 in glioma patients. Gene ontology(GO) and Kyoto encyclopedia of genes and genomes(KEGG) signaling pathway enrichment analysis were used to explore the biological functions of MAGED4 and SIRT7 in glioma. Western blot experiment was used to investigate whether MAGED4 protein exerted its regulatory effects on the activity of the PI3K/AKT signaling pathway via SIRT7. The effect of MAGED4 on cell proliferation in glioma through SIRT7 was explored by CCK-8. Results The analysis results of CGGA, UALCAN, and HPA databases showed that the expression levels of MAGED4 and SIRT7 in glioma tissues were higher than those in normal brain tissue, and the expression were positively correlated. Results of survival, ROC, and Cox analysis showed that high expression of MAGED4 and SIRT7 mRNA were risk factors for poor prognosis in glioma. Results of KEGG enrichment analysis showed that MAGED4 and SIRT7 were associated with the PI3K/AKT signaling in glioma, and Western blot results showed that MAGED4 activated the PI3K/AKT signaling pathway by regulating SIRT7. The CCK-8 results showed that MAGED4 promotes the proliferation of glioma cells through SIRT7. Conclusion MAGED4 and SIRT7 are highly expressed in glioma and associated with poor prognosis, and MAGED4 promotes glioma cell proliferation through activation of the PI3K/AKT signaling pathway by SIRT7.

abstract:
Objective To investigate the role mechanism of doublecortin(DCX) in glioma and screen DCX overexpression-related differentially expressed genes, so as to provide new targets for glioma targeted therapy. Methods Using rat glioma C6 cells as the target cells, LV-DCX-EGFP, LV-Cas9-Puro, and LV-Ctrl-EGFP lentiviruses were constructed. DCX overexpressing and control C6 cell lines were established through Cas9 dual-vector system transfection and screening. Transcriptome sequencing was used to obtain the differentially expressed gene profiles. iDEP and DESeq2 were applied to screen differentially expressed genes with the threshold of |Log_2FC|>2 and P0.7, indicating good diagnostic value. RT-qPCR validation showed that the mRNA expression levels of some upregulated genes(POU2F3, LPAR6, SREBF1) significantly increased(P

abstract:
Objective To establish a mouse model of lung injury induced by self-antigens and explore its pathological mechanisms to provide a reliable animal model for studying the pathogenesis of rheumatoid arthritis-associated interstitial lung disease(RA-ILD). Methods The mice were injected intradermal with antigenemulsion made of complete freund′s adjuvant(CFA) and lung tissue protein, and the emulsion prepared with incomplete Freund′s adjuvant(IFA) was used to enhance immunity. HE staining was used to observe the histopathological changes in the mice. Masson staining was used to detect pulmonary fibrosis. The mRNA levels of rheumatoid factor(RF), krebs von den lungen-6(KL-6) and surfactant protein D(SP-D) were detected by qPCR. The levels of ACPA, IgG1, IgG2a and IgG3 antibodies were detected by ELISA. The changes of inflammatory cells and α-smooth muscle actin(α-SMA)expression in lung tissue were detected by immunofluorescence staining. The protein expression levels of Collagen I and α-SMA were detected by Western blot. The changes of immune cells were detected by flow cytometry. Results HE staining showed that inflammatory cell infiltration increased and tissue structure changed significantly in lung tissue after the model was established by self-lung tissue antigen. Masson staining showed increased collagen deposition in lung tissue of model mice. qPCR tests revealed elevated mRNA levels of RF, KL-6 and SP-D. ELISA tests revealed elevated levels of ACPA, IgG1 and IgG3 antibodies. Immunofluorescence results showed that monocytes and T cells increased, and α-SMA expression increased in the model group. Western blot results showed increased protein expressions of Collagen Ⅰ and α-SMA. The flow cytometry results showed an increase in T cells and monocytes in the lung tissue. Conclusion The mouse model of lung injury induced by self-antigens is successfully established, and T cells and monocytes may be involved in the occurrence and progression of the disease.