Lappaconitine (LA) is a diterpene alkaloid isolated from Aconitum species and has been used in traditional Chinese medicine as an analgesic. Previous studies have also suggested that LA exerts anti-inflammatory and immunomodulatory effects. Neuroinflammation and oxidative stress, both of which are associated with mitochondrial dysfunction and apoptosis, contribute to the development and maintenance of neuropathic pain. We hypothesized that LA may protect SH-SY5Y cells against inflammation- and apoptosis-associated injury. To investigate whether LA attenuates TNF-α-induced inflammatory injury and apoptosis in SH-SY5Y cells and to explore the potential mechanisms involved. SH-SY5Y cells were pretreated with LA (0.1 or 1 μM) for 24 h and subsequently exposed to TNF-α (10 ng/mL) for 12 h. Apoptosis, mitochondrial function, inflammatory cytokine production, and the expression of mitophagy-related proteins were assessed in LA-pretreated and non-pretreated cells. LA pretreatment (0.1 or 1 μM) reduced apoptosis in TNF-α-exposed SH-SY5Y cells, increased Bcl-2 expression, and decreased the expression of Bax, Caspase-3, and Caspase-9. LA pretreatment also suppressed NF-κB activation and reduced IL-1β and IL-6 production. In addition, LA preserved mitochondrial integrity, as indicated by maintenance of mitochondrial membrane potential, reduced cytochrome c release, and decreased ROS accumulation. The expression of the mitophagy-related proteins PINK1 and Parkin was also increased following LA pretreatment. LA attenuated TNF-α-induced inflammatory injury and apoptosis in SH-SY5Y cells. These protective effects were associated with modulation of apoptosis-related signaling, suppression of inflammatory responses, preservation of mitochondrial function, and increased expression of PINK1/Parkin-related proteins. These findings suggest that LA may exert neuroprotective effects under inflammatory conditions.

The orphan nuclear receptor Nur77 is a multifunctional regulator involved in diverse cellular processes, including proliferation, survival, and apoptosis, through both transcription-dependent and -independent mechanisms. This regulatory complexity underscores the need for mechanistic studies in defined biological contexts. Here, we uncover a previously unrecognized non-genomic function of Nur77 at the centrosome that promotes mitotic progression in cancer cells. We show that Nur77 is phosphorylated at threonine 143 by cyclin-dependent kinase 1 (Cdk1), leading to its accumulation at the centrosome, where it binds the scaffold protein Cep192. This interaction is critical for maintaining centrosome integrity in tumor cells and facilitating the recruitment of Polo-like kinase 1 (PLK1), a key driver of centrosome maturation. Notably, Cdk1-mediated phosphorylation of Nur77 is aberrantly elevated in tumors, contributing to malignant proliferation through its mitotic role. Depletion of Nur77 or treatment with NMA39, a novel small-molecule Nur77 modulator that disrupts the Nur77-Cep192 interaction, results in mitotic arrest and cell death in tumor cells. These findings reveal a tumor-selective mitotic function of Nur77 and establish a mechanistic rationale for targeting phospho-Nur77 signaling as a cancer vulnerability.

Atherosclerosis (AS) and its complications are the leading causes of death worldwide. Endothelial cell (EC) senescence plays a crucial role in the development of AS by aggravating endothelial erosion and plaque instability. Neuronal pentraxin 1 (NPTX1) is a secreted glycoprotein that plays a key role in various neurological functions, such as synaptic function, plasticity at excitatory synapses, and neurite damage. Although NPTX1 is upregulated in senescent ECs, its role in EC senescence and AS remains unknown. Here, we investigated the role of NPTX1 in ischemic stroke and its potential mechanism in regulating EC senescence and AS. Using a middle cerebral artery occlusion (MCAO) mouse model, we observed dynamic NPTX1 expression: levels were minimal at 1 h post-MCAO but surged by 12 h and persisted for 28 days. Moreover, exogenous NPTX1 administration worsened post-stroke brain injury and blood-brain barrier disruption. Notably, MCAO accelerated both AS progression and EC senescence in vivo, while neuron-derived NPTX1-enriched conditioned medium induced EC senescence in vitro. Mechanistically, RNA sequencing and pharmacological inhibition revealed that NPTX1 promotes human umbilical vein EC senescence via an AKT-related pathway, as confirmed by senescence-associated β-galactosidase staining, impaired tube formation, and altered senescence markers. Critically, adeno-associated virus-mediated NPTX1 knockdown attenuated post-stroke AS and EC senescence in mice. Finally, the knockdown of NPTX1 through a short-hairpin RNA adeno-associated virus ameliorates AS and EC senescence in post-stroke mice. These findings establish NPTX1 as a key regulator of stroke-induced EC senescence and AS pathogenesis, highlighting its therapeutic potential for AS treatment in stroke patients.

Nur77 is an orphan nuclear receptor for which no endogenous ligand has yet been identified. It has been demonstrated that there is aberrant expression or dysfunction of nur77 in breast cancer (BC), however, its role in different types of breast cancer remains contentious. Despite mounting evidence that Nur77 exerts influence over mitochondrial dynamics, including fission, fusion and mitophagy of mitochondria in diverse systems, the role and mechanism of mitochondrial dynamics regulated by Nur77 in tumor cells remain opaque. In the present study, significant differences in Nur77 levels were observed in various BC cell types, particularly in the Luminal A-type cell lines MCF-7 and T47D. Nur77 was more highly expressed in T47D cells with the p53 L194F mutation and significantly promoted the growth of T47D cells. In T47D cells, the knockout of Nur77 unequivocally disrupted mitochondrial function, inducing excessive mitochondrial fragmentation and inactivating mitophagy. Further mechanistic studies demonstrated that only the mutant p53 L194F protein in T47D regulated p-Drp1-S616 in comparison to the wild-type p53 protein in MCF-7 cells. Nur77 up-regulated p53 L194F expression at the transcriptional level and exerted a stronger effect on the interaction of Drp1 with mutant p53 L194F. These results suggest that the Nur77/p53 L194F/ mitofission axis may be involved in the mitochondrial homeostasis of specific types of BC cells to maintain BC cell growth. The discovery of this axis provides an important experimental basis for the fine classification of Luminal A BC and the identification of new therapeutic targets.

[This corrects the article on p. 458 in vol. 11, PMID: 33575081.].

The role of extracellular vesicles (EVs) derived from inflammatory chondrocytes in EV-based therapy for osteoarthritis (OA) has received little attention. We examined the effects of EVs derived from both normal rat chondrocytes (nEVs) and IL-1β-treated rat chondrocytes (iEVs) on IL-1β-treated rat chondrocytes, macrophages, and osteoblasts, alongside mRNA-seq and miRNA-seq analyses of both them. Additionally, nEVs and iEVs were administered intra-articularly in the joints of rat models subjected to anterior cruciate ligament transection (ACLT), and the morphological alterations across the joints were assessed. These findings indicated that iEVs, compared with nEVs, significantly enhanced collagen II synthesis in IL-1β-treated chondrocytes, accompanied by marked increases in ER stress and autophagy. In comparison to nEVs, iEVs exhibited a greater effect on facilitating M2-type macrophage polarization while simultaneously diminishing M1-type polarization, a process likely mediated by the downregulation of chemotactic cytokines such as Cxcl10, Ccl5, Cxcl9, Cxcl1, and Cxcl11. iEVs exerted a more pronounced influence on the phenotypic characteristics of IL-1β-treated osteoblasts than nEVs. In the ACLT-rat model, iEVs, akin to nEVs, effectively mitigated articular cartilage degradation. However, there was no significant difference in OARSI Scores between the two groups, despite iEVs exerting a greater effect on increasing hyaline cartilage thickness and proteoglycan content. iEVs were superior to nEVs in attenuating synovium inflammation and promoting trabecula formation in the femur subchondral bone. Consequently, iEVs, akin to nEVs, significantly alleviated OA-induced damage. Moreover, iEVs outperformed nEVs in certain aspects, notably in augmenting hyaline cartilage, reducing synovium inflammation, and promoting trabecular formation in the subchondral bone during the early stage of OA.

Pancreatic cancer is an aggressive malignancy with a poor prognosis. It is characterized by low surgical resection rates, frequent development of chemoresistance, unsatisfactory treatment outcomes, and a high potential for recurrence and metastasis. Compound F41, a naturally occurring indole-diterpenoid secondary metabolite, was isolated from the entomopathogenic fungus Penicillium sp. Its anti-pancreatic cancer activity has not been previously reported. Our study demonstrates that F41 significantly inhibits DNA replication, invasion, and proliferation in pancreatic cancer cells. By elevating intracellular reactive oxygen species (ROS) levels in pancreatic cancer cells, F41 induces endoplasmic reticulum stress, ultimately leading to apoptosis. These findings suggest that F41 could effectively overcome gemcitabine resistance in a clinical setting, indicating its promise as a potential therapeutic agent for pancreatic cancer.

The synergistic approach of combining photodynamic immunotherapy with endogenous clearance of PD-L1 immune checkpoint blockade therapy holds promise for enhancing survival outcomes in glioblastoma (GBM) patients. The observed upregulation of O-GlcNAc glycolysis in tumors may contribute to the stabilization of endogenous PD-L1 protein, facilitating tumor immune evasion. This study presents a pH-adapted excited state intramolecular proton transfer (ESIPT)-isomerized β-ketoamide-based covalent organic framework (COF) nanoplatform (denoted as OT@COF-RVG). Temozolomide (TMZ) and OSMI-4 (O-GlcNAc transferase inhibitor) were integrated into COF cavities, then modified on the surface with polyethylene glycol and the rabies virus peptide RVG-29, showing potential for sensitizing TMZ chemotherapy and initiating photodynamic therapy (PDT). By inhibiting O-GlcNAc and promoting lysosomal degradation of PD-L1, OT@COF-RVG enhanced the effectiveness of immune checkpoint blockade (ICB) therapy. Additionally, treatment with OT@COF-RVG led to a notable elevation in reactive oxygen species (ROS) levels, thereby re-establishing an immunostimulatory state, inducing immunogenic cell death (ICD). In summary, our research unveiled a correlation between O-GlcNAc in GBM and the evasion of immune responses by tumors, while showcasing the potential of OT@COF-RVG in reshaping the immunosuppressive microenvironment of GBM and offering a more effective approach to immunotherapy in clinical settings.

Glioblastoma multiforme (GBM) is the deadliest brain tumour with an extremely poor prognosis. Tryptophan catabolism could enhance an array of protumour-genic signals and promoted tumour progression in GBM. However, the mechanisms of oncogenic signalling under tryptophan catabolism and potential therapy targeting this pathway have not been completely understood. Interleukin 4-induced 1 (IL4I1) is newly defined as a tryptophan metabolic enzyme and the potential function in GBM cells still remains unclear. In our study, we found IL4I1 was upregulated in GBM patients and predicted poor prognosis. Upregulation of IL4I1 inhibited GBM ferroptosis in vitro and in vivo. Further, we found that indole-3-pyruvic acid (I3P) from tryptophan mediated by IL4I1 could scavenge free radical and had an impressive role in inhibiting ferroptosis. To clarify the potential mechanism of I3P in GBM ferroptosis, we performed transcriptomic analyses of GBM cells treated with I3P and found that Nrf2 related genes was upregulated. Further, we found that the ubiquitination of Nrf2 could be attenuate by I3P binding with Nrf2 directly. Knockdown of Nrf2 attenuated the induction of anti-ferroptosis by IL4I1, pointing to Nrf2 as a key mediator of this process. In vivo, overexpression of IL4I1 with ML385 in GBM xenografts promoted ferroptosis. Collectively, this study emphasises the crucial roles of IL4I1 in anti-ferroptosis through Nrf2 signalling pathway but not AHR pathway by catabolism tryptophan, suggesting IL4I1 and tryptophan reprogramming as potential therapeutic targets for GBM.

Aim: This study explored the prognostic value of N-glycan biosynthesis (NGB) in lower-grade glioma (LGG) and aimed to develop a machine learning model for enhanced prognostic accuracy. Method: LGG patient transcriptome data were analyzed to identify NGB-related genes. Consensus clustering identified subgroups based on NGB expression. A prognostic NGB signature (pNGB) was developed using machine learning. The pNGB score's association with cell proliferation, inflammation, treatment response, tumor recurrence, and the immune microenvironment was also explored. Results: A 22-gene pNGB signature was identified, with MGAT1 and TUSC3 having the highest and lowest hazard ratios, respectively. Two distinct clusters (C1 and C2) with differential pNGB expression and survival outcomes were revealed. NGB pathway analysis indicated an overall poor prognosis, except for MGAT4C and TUSC3. The Enet-based survival model showed superior discriminatory power and reliability. The NGB risk score correlated with increased cell proliferation, inflammation, and altered immune landscape. Additionally, the score is linked to treatment response and tumor recurrence. Conclusion: This study highlights the critical role of NGB in LGG progression and proposes a pNGB-based model for prognosis. The NGB risk score shows promise as a prognostic biomarker and potential therapeutic target in LGG.

Glioblastoma multiforme (GBM) is the most prevalent and lethal primary intracranial neoplasm in the adult population, with treatments of limited efficacy. Recently, bufotalin has been shown to have anti-cancer activity in a variety of cancers. This investigation aims to investigate the effect of bufotalin on GBM and elucidate its potential underlying mechanism. Our results show that bufotalin not only inhibits the proliferation and epithelial-mesenchymal transition (EMT) but also triggers apoptosis in GBM cells. The result of RNA-seq indicated that bufotalin could induce mitochondrial dysfunction. Moreover, our observations indicate that bufotalin induces an excessive accumulation of intracellular reactive oxygen species (ROS) in GBM cells, leading to mitochondrial dysfunction and the dephosphorylation of AKT. Moreover, bufotalin improved TMZ sensitivity of GBM cells in vitro and in vivo. In conclusion, bufotalin enhances apoptosis and TMZ chemosensitivity of glioblastoma cells by promoting mitochondrial dysfunction via AKT signaling pathway.

Scabrol B and Scabrol C, two newly identified iridoid derivatives (1 and 2) and six known compounds (3–8), were extracted from the roots of Patrinia scabra. The structures of these derivatives, including their absolute configurations, were elucidated via comprehensive NMR analysis, chemical derivatization, and quantum chemical ECD calculations. All isolated compounds were evaluated for their anti-renal fibrosis activity. The results demonstrate that compounds 1 and 2 showed dose-dependent protective effects against renal fibrosis in vitro by reducing the expression of fibronectin, collagen I, and alpha-smooth muscle actin (α-SMA) in NRK-49f cells mediated by TGF-β1.

Jacaranone derived from Senecio scandens, a traditional Chinese medicine used for centuries, has been documented to exhibit anti-inflammatory and antiproliferative properties in various tumor cell lines. However, the mechanism of action and relationship between inflammation and apoptosis induced by jacaranone remain inadequately elucidated. In this study, the targets of jacaranone and cancer were identified from various databases, while potential targets and pathways were predicted through the analysis of the protein–protein interactions (PPI) network and pathway enrichment. Through a comprehensive network pharmacology analysis and corroborating experimental findings, we revealed that jacaranone induces tumor cell death by fine-tuning the tumor necrosis factor receptor 1 (TNFR1) downstream signaling pathway. TNFR1 serves as a key node that assembles into complexes I and II, regulating pathways including the nuclear factor (NF)-κB signaling pathway and the cell apoptosis pathway, which play crucial roles in cellular life activities. Jacaranone successfully guides survival signaling pathways to apoptotic mechanisms by inhibiting the assembly of complex I and promoting the formation of complex II. In particular, the main action mechanism of jacaranone lies in inducing the degradation of the inhibitor of apoptosis protein (cIAP)-2. cIAP-2 serves as an E3 ubiquitin ligase that ubiquitinates receptor-interacting serine/threonine-protein kinase 1 (RIPK1), thereby hindering the formation of complex I and effectively reducing the phosphorylation of Inhibitor of κB kinase (IKK) β. When the deubiquitylation process of RIPK1 is triggered, it may promote the formation of complex II, which ultimately leads to cell apoptosis. This fully demonstrates the key role of jacaranone in regulating TNFR1 complexes, especially through the degradation of cIAP-2. Taken together, jacaranone hinders the assembly of TNFR1 complex I and promotes the formation of complex II to induce apoptosis of cancer cells. Our findings unveil a novel mechanism underlying jacaranone, while also presenting a fresh approach for the development of new pharmaceuticals.

Lines of evidence have indicated that type 2 diabetes mellitus (T2DM) is an independent risk factor for osteoarthritis (OA) progression. However, the study focused on the relationship between T2DM and OA at the transcriptional level remains empty. We downloaded OA- and T2DM-related bulk RNA-sequencing and single-cell RNA sequencing data from the Gene Expression Omnibus (GEO) dataset. Differential expression analysis and weighted gene co-expression network analysis (WGCNA) were performed to screen out hub genes between OA and T2DM, and functional enrichment was done. Single-cell sequencing analysis was further used to screen key genes on OA and T2DM datasets. Rat chondrocytes and human articular cartilage were used to validate biomarkers among OA and T2DM. Sixty-eight hub genes were obtained, which were mainly enriched in the inflammatory response. We found that the hub gene TNFAIP6 is not only closely related to OA and T2DM but also a marker of prehypertrophic chondrocytes, which are closely related to the progression of OA. TNFAIP6 was found to be significantly elevated in CD14 + monocytes in T2DM patients, and this group of cells can promote inflammation. Validation on rat chondrocytes and human cartilage showed that TNFAIP6 was highly expressed in OA and further increased in the presence of T2DM or high glucose. Our study identified several characteristic modules and hub genes in the pathogenesis of T2DM-induced OA, which may facilitate further investigation of its molecular mechanisms. Up-regulated TNFAIP6 may contribute to OA in patients with T2DM by the recruitment of pro-inflammatory CD14 + monocytes in the OA synovium, which provides a potential target for the diagnosis and treatment of T2DM-associated OA.

In order to study the effects of soybean isoflavones on the growth performance and lipid metabolism of juvenile Chinese mitten crabs, six experimental diets were formulated by gradient supplementation with 0%, 0.004% and 0.008% soybean isoflavones at different dietary lipid levels (10% and 15%). The groups were named as follows: NF-0 group (10% fat and 0% SIFs), NF-0.004 group (10% fat and 0.004% SIFs), NF-0.008 group (10% fat and 0.008% SIFs), HF-0 group (15% fat and 0% SIFs), HF-0.004 group (15% fat and 0.004% SIFs) and HF-0.008 group (15% fat and 0.008% SIFs). All crabs with an initial weight of 0.4 ± 0.03 g were fed for 8 weeks. The results showed that dietary supplementation with 0.004% or 0.008% SIFs significantly increased the weight gain and specific growth rate of crabs. Diets supplemented with 0.004% or 0.008% SIFs significantly reduced the content of non-esterified free fatty acids and triglycerides in the hepatopancreas of crabs at the 10% dietary lipid level. Dietary SIFs significantly decreased the relative mRNA expressions of elongase of very-long-chain fatty acids 6 (elovl6), triglyceride lipase (tgl), sterol regulatory element-binding protein 1 (srebp-1), carnitine palmitoyltransferase-1a (cpt-1a), fatty acid transporter protein 4 (fatp4), carnitine palmitoyltransferase-2 (cpt-2), Δ9 fatty acyl desaturase (Δ9 fad), carnitine palmitoyltransferase-1b (cpt-1b), fatty acid-binding protein 10 (fabp10) and microsomal triglyceride transfer protein (mttp) in the hepatopancreas of crabs. At the 15% dietary lipid level, 0.008% SIFs significantly increased the relative mRNA expressions of fatty acid-binding protein 3 (fabp3), carnitine acetyltransferase (caat), fatp4, fabp10, tgl, cpt-1a, cpt-1b and cpt-2 and significantly down-regulated the relative mRNA expressions of Δ9 fad and srebp-1. In conclusion, SIFs can improve the growth and utilization of a high-fat diet by inhibiting genes related to lipid synthesis and promoting lipid decomposition in juvenile Chinese mitten crabs.

Gliomas are one of the most challenging tumors to treat due to their malignant phenotype, brain parenchymal infiltration, intratumoral heterogeneity, and immunosuppressive microenvironment, resulting in a high recurrence rate and dismal five-year survival rate. The current standard therapies, including maximum tumor resection, chemotherapy with temozolomide, and radiotherapy, have exhibited limited efficacy, which is caused partially by the resistance of tumor cell death. Recent studies have revealed that ferroptosis, a newly defined programmed cell death (PCD), plays a crucial role in the occurrence and progression of gliomas and significantly affects the efficacy of various treatments, representing a promising therapeutic strategy. In this review, we provide a comprehensive overview of the latest progress in ferroptosis, its involvement and regulation in the pathophysiological process of gliomas, various treatment hotspots, the existing obstacles, and future directions worth investigating. Our review sheds light on providing novel insights into manipulating ferroptosis to provide potential targets and strategies of glioma treatment.

A-B, HeLa cells were infected with lentivirus expressing HA-c-ABL or shRNAs targeted to c-ABL. Cell lysates were analysed two days post-infection by immunoblotting. The mRNA levels of PLK1 were evaluated by qPCR ({plus minus} s.e.m., n=3). C, HeLa cells were treated with imatinib (10 μM) for indicated times, then PLK1 protein and mRNA levels were analyzed ({plus minus} s.e.m., n=3). D-F, HEK293 trasfected with indicated plasmids, the cells were incubated with 100 μg/mL CHX for indicated times.

Supplementary Materials and Methods, Figure Legend from TGFβ Induces “BRCAness” and Sensitivity to PARP Inhibition in Breast Cancer by Regulating DNA-Repair Genes

Background2021 World Health Organization (WHO) Central Nervous System (CNS) tumor classification increasingly emphasizes the important role of molecular markers in glioma diagnoses. Preoperatively non-invasive “integrated diagnosis” will bring great benefits to the treatment and prognosis of these patients with special tumor locations that cannot receive craniotomy or needle biopsy. Magnetic resonance imaging (MRI) radiomics and liquid biopsy (LB) have great potential for non-invasive diagnosis of molecular markers and grading since they are both easy to perform. This study aims to build a novel multi-task deep learning (DL) radiomic model to achieve preoperative non-invasive “integrated diagnosis” of glioma based on the 2021 WHO-CNS classification and explore whether the DL model with LB parameters can improve the performance of glioma diagnosis.MethodsThis is a double-center, ambispective, diagnostical observational study. One public database named the 2019 Brain Tumor Segmentation challenge dataset (BraTS) and two original datasets, including the Second Affiliated Hospital of Nanchang University, and Renmin Hospital of Wuhan University, will be used to develop the multi-task DL radiomic model. As one of the LB techniques, circulating tumor cell (CTC) parameters will be additionally applied in the DL radiomic model for assisting the “integrated diagnosis” of glioma. The segmentation model will be evaluated with the Dice index, and the performance of the DL model for WHO grading and all molecular subtype will be evaluated with the indicators of accuracy, precision, and recall.DiscussionSimply relying on radiomics features to find the correlation with the molecular subtypes of gliomas can no longer meet the need for “precisely integrated prediction.” CTC features are a promising biomarker that may provide new directions in the exploration of “precision integrated prediction” based on the radiomics, and this is the first original study that combination of radiomics and LB technology for glioma diagnosis. We firmly believe that this innovative work will surely lay a good foundation for the “precisely integrated prediction” of glioma and point out further directions for future research.Clinical trail registrationThis study was registered on ClinicalTrails.gov on 09/10/2022 with Identifier NCT05536024.

A, HeLa cells were transfected with c-ABL-specific siRNA or nonspecific control siRNA. After 12hrs transfection, HeLa cells were synchronized by a double thymidine treatment. Cell-cycle distributions were determined by flow cytometry analysis. B, HeLa cells were treated with the indicated shRNAs, then released into mitosis by using thymidine-nocodazol method. The p-H3S10-positive cells were counted at the indicated time points. Data are shown as mean {plus minus} s.e.m. (n=500). C, Representative immunofluorescence imagines of p-H3S10 staining cells were shown. Scale bar: 50 μm. D, HeLa cells were transfected with PLK1 shRNA together with Plk1 or Y425F mutant, and synchronized by a double-thymidine arrest. Cells were released into nocodazol, and collected at the indicated times after release. The levels of the indicated proteins were determined by western blot analysis.