Ursolic acid (UA) and Rosuvastatin (RST) improves diabetic nephropathy (DN). However, the combination therapy of UA and RST on DN and its possible mechanism requires exploration. Streptozotocin (STZ)-induced DN model cells were treated with UA and RST alone or in combination. Cell viability and apoptosis were assessed using Cell Counting Kit-8 (CCK-8), flow cytometry, and TUNEL staining. The inflammation-related factors' release was examined by enzyme linked immunosorbent assay (ELISA). Potential mechanism was analyzed through network pharmacology. The reactive oxygen species (ROS) accumulation was detected using flow cytometry. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), and catalase (CAT) were measured using commercial reagent kits. UA and RST increased viability while reducing apoptosis of STZ-induced cells. UA and RST suppressed the release of inflammation-related factors in STZ-induced cells. Through network pharmacology analysis, the inflammation and oxidative stress were thought to be implicated in the treatment of UA and RST in DN. UA and RST downregulated ROS and MDA while upregulating SOD and CAT in STZ-induced cells. Compared with RST and UA used individually, their combination showed a more potent effect in increasing viability, mitigating apoptosis, and suppressing the inflammation and oxidative stress in STZ-induced cells. Combined UA and RST therapy ameliorated the inflammation and oxidative stress in DN model cells, providing the theoretical basis for pharmacological research of DN.

DNA Damage-Inducible Transcript 4 (DDIT4), a conserved stress-responsive protein with dual nuclear and cytoplasmic localization, has recently emerged as a critical regulator in multiple cancer types. However, its functional role and molecular mechanisms in hepatocellular carcinoma (HCC) remain poorly understood. In this study, we systematically investigated DDIT4's biological significance through comprehensive in vitro analyses. Clinical specimen analysis revealed significant downregulation of DDIT4 protein in HCC tumor tissues compared to adjacent non-tumor controls. Functional studies demonstrated that DDIT4 knockdown markedly enhanced HCC cell proliferation, whereas its overexpression exerted potent anti-proliferative effects. Mechanistically, our in vitro experiments revealed two key regulatory pathways: (1) DDIT4 overexpression suppressed AKT/mTOR signaling activation, and (2) DDIT4 underwent ubiquitin-mediated proteasomal degradation via specific interaction with the E3 ligase Seven in Absentia Homologue 2 (SIAH2). These findings establish DDIT4 as a tumor-suppressive protein in HCC pathogenesis, whose oncogenic downregulation is mediated through SIAH2-dependent ubiquitination. The resultant decrease in DDIT4 protein levels creates a permissive microenvironment for tumor growth by releasing AKT/mTOR pathway inhibition. Our results nominate DDIT4 as a promising therapeutic target for HCC intervention. Further preclinical and clinical investigations are warranted to validate DDIT4's translational potential and explore targeted strategies to stabilize its tumor-suppressive functions.

Membranous nephropathy (MN) is an autoimmune kidney disease, and its pathogenesis is related to inflammation and podocyte injury. Both disulfiram and baicalin can relieve the symptoms of MN through anti-inflammatory effects. However, it is still unclear whether disulfiram and baicalin can enhance the therapeutic effect of MN by combination therapy. We screened the optimal drug concentration of disulfiram and baicalin for incubation of AB8/13 cells by CCK-8 assay. Angiotensin II was used to stimulate AB8/13 cells to construct a MN cell model. Subsequently, cell proliferation assays, network pharmacology analysis, ELISA assay, and pyroptosis detection were employed to investigate the effects of disulfiram combined with baicalin in MN. Firstly, 250 nM disulfiram and 5 μmol/L baicalin were selected as the optimal dose for the treatment of MN. Cell proliferation assay showed that the combined administration of disulfiram and baicalin could reduce Angiotensin II-induced podocyte injury. Subsequently, a total of 129 genes were identified as potential therapeutic targets of the combination therapy involving disulfiram and baicalin for MN. These targets mainly focused on regulating inflammatory-related functions and pathways. Disulfiram combined with baicalin inhibited the inflammation of MN by suppressing the levels of TNF-α, IL-8 and C-reactive protein and promoting the levels of TGF-β and IL-10. Disulfiram combined with baicalin significantly attenuated Angiotensin II-induced podocyte pyroptosis, as indicated by reduction of Caspase-1 and GSDMD-N, lactate dehydrogenase release and the percentage of podocytes stained with propidium iodide. Compared with monotherapy using either disulfiram or baicalin alone for MN, the combination has a protective role in podocyte injury by inhibiting inflammation and pyroptosis. This discovery provides a new perspective for clarifying the pharmacological effects of disulfiram and baicalin to podocyte injury in MN.

The relationship between human papillomavirus (HPV) and immune cells is vital for understanding the pathophysiology of infection and its role in neoplastic progression. High-risk human papillomavirus (HR-HPV) is the main cause of cervical cancer (CC). Thus, the association between immune response cells, the virus, and its behavior according to cervical disease development could provide new ways for understanding the entire process. Since the role and the presence of the immune response cells in the uterus cervix considering HPV infection has not been elucidated so far, this study aimed to identify the immune cells involved in high-grade intraepithelial lesions (HSIL) and CC development related to uterine cervical infection caused by HR-HPV. The study population included women who had positive molecular tests for HPV. Through the databases MEDLINE, EMBASE, LILACS, Cochrane, Scopus, Web of Science, CINAHL, Science Direct, and Google Scholar we identified 6,698 studies at the beginning. After the systematic review steps, the final number of included studies was 22. Cervical lesions were distributed according to the severity of lesions in HSIL, low-grade squamous intraepithelial lesions (LSIL), and negative for intraepithelial lesions or malignancy (NILM). The cellular phenotypes presented in these publications were T lymphocytes (LT), regulatory T lymphocytes (Tregs), macrophages (MØ), natural killer cells (NK), natural killer T cells (NKT), Langerhans cells (LC), and dendritic cells (DC). Among the observed associations with cervical lesions and HR-HPV, we highlight the DC/LC and MØ being 36.4% of the cell types, followed by Tregs (31.8%) and LT CD4 / CD8 with 27.3%. The increased findings in innate and adaptive immunological response may imply both are acting together, with the innate response cells and Tregs being the most prominent. Since these cells have great importance in the maintenance and balance of the immunological system, the present study highlights the essential role of MØ and Treg cells in the process of cervical lesion severity associated with HPV, suggesting that they may be focused as prognostic markers and immunotherapeutic targets.

Epithelial ovarian carcinoma (EOC) is a highly lethal gynecological malignancy with limited treatment options. This study aimed to explore the regulatory roles of IRE1α and YAP1 in EOC progression and identify potential therapeutic targets. Blood and tissue samples were collected from 26 EOC patients and 10 patients with ovarian cysts. The expression of inflammatory factors in the blood was measured using ELISA. The proliferation, migration, invasion, and cell cycle of human ovarian cancer cell lines SK-OV-3, SW626, and Anglene were evaluated using MTT assays, scratch tests, Transwell assays, and flow cytometry. The effects of IRE1α inhibition on EOC cell proliferation, migration, and apoptosis were investigated using pharmacological inhibitors and shRNA knockdown. IRE1α was highly expressed in EOC patients and was negatively correlated with patient survival rates. Additionally, IRE1α scores in EOC patients were positively correlated with serum levels of TNF-α and VEGF-a. Compared to normal controls, significantly higher expressions of IRE1α and XBP1 were observed in ovarian cancer tissues and cells. Knockdown of IRE1α in ovarian cancer cells led to a significant reduction in the expression of IRE1α and XBP1s, as well as inhibited cell proliferation and survival. The IRE1α inhibitors STF-083100 and 4μ8C suppressed the proliferation, invasion, and migration of SK-OV-3 cells and reduced the expression levels of related factors. 4μ8C inhibited the degradation of YAP within SK-OV-3 cells while downregulating the expression of Cyclin D1 protein. Compared to the group treated with 4μ8C alone, the combined intervention of 4μ8C and a YAP inhibitor showed a more pronounced inhibitory effect on the proliferation of SK-OV-3 cells.This study first reveals that the IRE1α/YAP signal drives the malignant progression of EOC through the regulation of cell proliferation, migration, and invasion. The dual-targeted synergistic inhibition of IRE1α/YAP1 offers an innovative therapeutic paradigm for the treatment of EOC.

Primary Sjögren's syndrome (pSS) is characterized by molecular heterogeneity and immune-metabolic dysregulation. This study aimed to identify diagnostic biomarkers and characterize monocyte-macrophage differentiation trajectories in pSS. Bulk RNA-seq data were first batch-corrected and subjected to differential expression analysis, immune infiltration profiling, and weighted gene co-expression network analysis (WGCNA) to identify key gene modules. For single-cell RNA-seq, 42,157 salivary gland cells were clustered and annotated; Monocle3 pseudotime analysis reconstructed the monocyte-macrophage differentiation trajectory, and CellChat quantified intercellular signaling networks. Candidate immune-metabolism biomarkers were then screened by integrating WGCNA results with three machine-learning algorithms, LASSO regression, SVM-RFE, and random forest, and validated in the external GSE84844 cohort. Finally, functional relevance was tested in PMA-induced THP-1 macrophages by knocking down ISG15 and measuring effects on NF-κB activation and secretion of TNF-α and IL-1β. Pseudotime analysis delineated a monocyte-to-macrophage differentiation axis and pinpointed 182 dynamic marker genes. Integrated machine-learning and network analysis highlighted four immune-metabolism genesISG15, OAS1, LAP3, and PARP9each achieving AUC > 0.80 in the GSE84844 validation set. In vitro, ISG15 knockdown markedly attenuated NF-κB activation and reduced TNF-α/IL-1β release, confirming its pivotal role in pSS-associated inflammation. This integrated transcriptomic analysis highlights key monocyte-macrophage trajectories and identifies four robust immune-metabolism biomarker candidates for pSS diagnosis. Functional validation of ISG15 suggests it plays a pivotal role in driving macrophage-mediated inflammatory responses in pSS.

In this paper, we compared the ability to expand/supercharge NK cells (sNK) by either processed osteoclasts (pOCs) or live osteoclasts (OCs). pOCs induced similar levels of cell expansion in NK cells compared to live OCs. pOC-generated sNK cells exhibited lower lysis of oral squamous carcinoma stem-like cells (OSCSCs) tumors in comparison to live OC-generated sNK cells. Slightly lower levels of IFN-γ secretion were also observed in pOC-generated sNK cells when compared to live OC-generated sNK cells. Cytotoxic function and secretion levels of IFN-γ remained low in pOC-sNK cultures compared to OC-sNK cultures even after adding the supernatants harvested from live OCs to pOC-sNK cultures. pOCs were equally capable of selecting CD8+ T cells in sNK cell cultures when compared to live OCs. Overall, even though live OCs are capable of activating slightly better than the processed osteoclasts, the use of pOCs is preferable for the expansion of sNK cells due to shorter NK expansion period, higher cost-effectiveness, and faster availability for patient infusion.