Objective To investigate the role of poly(rC)-binding protein 1(PCBP1)in cadmium(Cd)-induced ferroptosis in mouse neuroblastoma Neuro-2a(N2A)cells.Methods N2A cells were exposed to a concentration gradient of CdCl?(0,1,2,4 μmol/L)for 72 h.Cell viability was assessed by trypan blue staining.Western blotting was employed to detect the expression of ferroptosis-related proteins(GPX4,HMOX1,ACSL4)and PCBP1.Intracellular Fe2? level and lipid peroxidation were detected using FerroOrange and BODIPY581/591 C11 probes,respectively.Ferrostatin-1(Fer-1),a ferroptosis inhibitor,was applied to confirm the critical role of ferroptosis in Cd-induced cytotoxicity.Molecular docking was performed to elucidate the interaction between PCBP1 and ferritin,as well as the binding sites of Cd2?.PCBP1 overexpression plasmid was further constructed for functional validation.Results Cd exposure suppressed cell viability in N2A cells in a dose-dependent manner(P<0.01),significantly down-regulated GPX4 expression(P<0.05),up-regulated HMOX1 expression(P<0.01),and induced Fe2? overload and lipid peroxidation(P<0.01).Molecular docking revealed that Cd2? directly bound to the KH2 domain of PCBP1 and then co-localized on the outer surface of ferritin heavy chain.Overexpression of PCBP1 markedly reversed Cd-induced Fe2? accumulation,GPX4 down-regulation,lipid peroxidation,and cell death.Conclusion Cd exposure disrupts PCBP1-mediated iron homeostasis via transcriptional suppression and competitive displacement of metal ions,and then synergistically drives Fe2? overload-triggered ferroptosis cascades,ultimately leading to neurotoxicity.Targeting PCBP1-mediated iron homeostasis can effectively mitigate Cd-induced neurotoxicity,and may serve as a novel therapeutic strategy.

Objective To investigate the effect of long non-coding RNA(lncRNA)nuclear enriched abundant transcript 1(NEAT1)on the chondrocyte pyroptosis signaling axis in osteoarthritis(OA)and its potential mechanism.Methods Knee joint cartilage tissues were collected from 6 OA patients and 6 healthy individuals.The targeting relationship between miR-26a and lncRNA NEAT1 was predicted by TargetScan 7.2 and StarBase,and the targeting regulatory effect of lncRNA NEAT1 and miR-26a was verified by dual luciferase reporter assays.The cultured chondrocyte lines were divided into the control group(without drug intervention),IL-1β group(with 10 μg/L of IL-1β for inducing OA in vitro),IL-1β+NEAT1-EV group(with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-EV),IL-1β+NEAT1-OE group(with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-OE),IL-1β+NEAT1-OE+mimic-NC(with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of mimic-NC),and IL-1β+NEAT1-OE+mimic(with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of miR-26a mimic).The expression levels of lncRNA NEAT1 and miR-26a in OA cartilage tissues and cells were detected using qRT-PCR.The cell viability was determined using CCK-8 assay.The expression of Caspase1 and cleaved-Caspase1,as well as the pyroptosis markers[NOD-like receptor family pyrin domain containing 3(NLRP3),interleukin-18(IL-18)and cleaved-Gasdermin D]were detected using Western blot.Results In the OA cartilage tissue,the expression level of lncRNA NEAT1 was higher than that in the normal cartilage tissue,while the expression level of miR-26a was lower than that in the normal cartilage group(P<0.05).The dual luciferase reporter assays confirmed the targeted regulatory effect of lncRNA NEAT1 and miR-26a.Compared with the control group,the IL-1β group showed upregulation of lncRNA NEAT1,cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels(P<0.05),downregulation of miR-26a expression level(P<0.05),and a decrease of cell viability(P<0.05).Compared with the IL-1β+NEAT1-EV group,the IL-1β+NEAT1-OE group exhibited upregulation of lncRNA NEAT1,cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels(P<0.05),downregulation of miR-26a expression level(P<0.05),and a decrease of cell viability(P<0.05).Compared with the IL-1β+NEAT1-OE+mimic-NC group,the IL-1β+NEAT1-OE+mimic group showed downregulation of cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels,upregulation of miR-26a expression level(P<0.05),and an increase of cell viability(P<0.05).Conclusion lncRNA NEAT1 activates the chondrocyte pyroptosis signaling axis mediated by Caspase1 in OA by the targeted inhibition of miR-26a.

Biomolecular condensates are formed through phase separation of biomacromolecules such as proteins and RNAs. These condensates exhibit liquid-like properties that can futher transition into more stable material states. They form complex internal structures via multivalent weak interactions, enabling precise spatiotemporal regulations. However, the use of inconsistent and non-standardized terminology has become increasingly problematic, hindering academic exchange and the dissemination of scientific knowledge. Therefore, it is necessary to discuss the terminology related to biomolecular condensates in order to clarify concepts, promote interdisciplinary cooperation, enhance research efficiency, and support the healthy development of this field.

Objective: To investigate the differential expression of Yes-associated protein(YAP) in the healthy control group and in patients with rheumatoid arthritis(RA), and to explore its correlation with the progression of the disease. Methods: Three cases each of synovial membranes from the healthy control group and RA group were selected for RNA-sequence detection to identify differentially expressed genes. Data and laboratory indicators from 50 healthy control cases and 150 RA patients who met the criteria were collected. ELISA was used to detect the expression of YAP in the serum of each group, and then its correlation with other laboratory indicators in RA patients was analyzed. The receiver operating characteristic curve was applied to explore the diagnostic efficacy of the related risk factors for RA. Results: Expression of 14-3-3-gama, which regulated the disease process, was significantly down-regulated in the synovial membranes of RA patients compared to normal synovial membranes; YAP expression was significantly up-regulated in rheumatoid arthritis fibroblast-like synoviocytes compared to healthy controls and was correlated with haematological sedimentation, C-reactive protein, interleukin(IL)-1, IL-4, IL-6, IL-10, rheumatoid factor, anti-cyclic citrullinated polypeptide antibody, interferon alpha, tumour necrosis factor alpha, protein electrophoresis α1 globulin, protein electrophoresis alpha2 globulin, disease activity scores, disease activity index, and patient-reported outcome had correlations(P0.05); multifactor Logistic regression analysis showed that YAP expression was an independent influencing factor for the prognosis of RA patient. Conclusion YAP is differentially expressed between the healthy control group and RA group, which shows a significant positive correlation with the disease activity of RA, potentially becoming a new target for clinical treatment of RA.

Avulsion of permanent teeth is the most severe type of tooth injury.Replantation has been widely recognized as the optimal method for managing avulsed permanent teeth in teenagers.Due to significant damage to periodontal tissue and death of periodontal membrane cells,secondary progressive root resorption often occurs after tooth replantation.Managing progressive root resorption clini-cally poses a major challenge following tooth replantation.It is crucial to control root resorption and prolong tooth retention time.This article reviews recent research and treatment on progressive replacement absorption and inflammatory absorption after replantation.

Objective To investigate the effect of long non-coding RNA(lncRNA)nuclear enriched abundant transcript 1(NEAT1)on the chondrocyte pyroptosis signaling axis in osteoarthritis(OA)and its potential mechanism.Methods Knee joint cartilage tissues were collected from 6 OA patients and 6 healthy individuals.The targeting relationship between miR-26a and lncRNA NEAT1 was predicted by TargetScan 7.2 and StarBase,and the targeting regulatory effect of lncRNA NEAT1 and miR-26a was verified by dual luciferase reporter assays.The cultured chondrocyte lines were divided into the control group(without drug intervention),IL-1β group(with 10 μg/L of IL-1β for inducing OA in vitro),IL-1β+NEAT1-EV group(with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-EV),IL-1β+NEAT1-OE group(with 10 μg/L of IL-1β and transfected with 20 μg/L of NEAT1-OE),IL-1β+NEAT1-OE+mimic-NC(with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of mimic-NC),and IL-1β+NEAT1-OE+mimic(with 10 μg/L of IL-1β and co-transfected with 20 μg/L of NEAT1-OE and 50 μg/L of miR-26a mimic).The expression levels of lncRNA NEAT1 and miR-26a in OA cartilage tissues and cells were detected using qRT-PCR.The cell viability was determined using CCK-8 assay.The expression of Caspase1 and cleaved-Caspase1,as well as the pyroptosis markers[NOD-like receptor family pyrin domain containing 3(NLRP3),interleukin-18(IL-18)and cleaved-Gasdermin D]were detected using Western blot.Results In the OA cartilage tissue,the expression level of lncRNA NEAT1 was higher than that in the normal cartilage tissue,while the expression level of miR-26a was lower than that in the normal cartilage group(P<0.05).The dual luciferase reporter assays confirmed the targeted regulatory effect of lncRNA NEAT1 and miR-26a.Compared with the control group,the IL-1β group showed upregulation of lncRNA NEAT1,cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels(P<0.05),downregulation of miR-26a expression level(P<0.05),and a decrease of cell viability(P<0.05).Compared with the IL-1β+NEAT1-EV group,the IL-1β+NEAT1-OE group exhibited upregulation of lncRNA NEAT1,cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels(P<0.05),downregulation of miR-26a expression level(P<0.05),and a decrease of cell viability(P<0.05).Compared with the IL-1β+NEAT1-OE+mimic-NC group,the IL-1β+NEAT1-OE+mimic group showed downregulation of cleaved-Caspase1,NLRP3,IL-18,and cleaved-Gasdermin D expression levels,upregulation of miR-26a expression level(P<0.05),and an increase of cell viability(P<0.05).Conclusion lncRNA NEAT1 activates the chondrocyte pyroptosis signaling axis mediated by Caspase1 in OA by the targeted inhibition of miR-26a.

Avulsion of permanent teeth is the most severe type of tooth injury.Replantation has been widely recognized as the optimal method for managing avulsed permanent teeth in teenagers.Due to significant damage to periodontal tissue and death of periodontal membrane cells,secondary progressive root resorption often occurs after tooth replantation.Managing progressive root resorption clini-cally poses a major challenge following tooth replantation.It is crucial to control root resorption and prolong tooth retention time.This article reviews recent research and treatment on progressive replacement absorption and inflammatory absorption after replantation.

Objective To investigate the effect of osteoblast-derived exosome(Exo)mediating microRNA(miR)-494 on bone metabolism and bone remodeling balance in osteoporosis(OP)rats and its mechanism.Methods Exosomes was isolated and identified from MC3T3-E1 osteoblast cell line and transferred to Exo by electrical transfer.Forty SD rats were randomly divided into control,model,miR-494 mimic(miR-494),and miR-494 inhibitor group,10 rats in each group.The ovaries were removed to construct the OP model except the control group.After modeling,the miR-494 group and miR-494 inhibitor group received tail vein injections of exosomes containing the corresponding miRNA,at a dose of 3×109 particles.Four weeks later,bone parameters were detected in each group of rats by Micro-CT,serum bone markers were measured by ELISA,pathological changes in bone tissue were observed by HE staining,osteoclast numbers were detected by tartrate-resistant acid phosphatase(TRACP)staining,and the expression levels of bone remodeling-related proteins and toll-like receptor 4(TLR4)pathway-related proteins were determined by Western blotting.Results Typical cup-shaped or round exosomes were successfully isolated with a diameter of about 100 nm from MC3T3-E1 cells,which contained CD63,CD9,tumor susceptibility gene 101(TSG101),heat shock protein 70(HSP70)proteins and can be taken up by MC3T3-E1 cells.Compared with the model group,the bone parameters of the rats in the miR-494 mimic group decreased,serum bone markers bone Gla protein(BGP),TRACP,C-terminal telopeptide of type Ⅰ collagen(CTX-Ⅰ)increased,osteoprotegerin(OPG),procollagen type Ⅰ N-terminal propeptide(PⅠNP)decreased,bone trabeculae structure was disordered,osteoclasts increased,bone morphogenetic protein 2(BMP-2),Runt related transcription factor 2(RUNX2)in bone tissue downregulated,receptor activator of nuclear factor kappa-B ligand(RANKL)upregulated,TLR4,nuclear factor kappa-B p65(NF-κB p65)and myeloid differentiation primary response 88(MyD88)upregulated(all P＜0.05).In contrast,the situation of the miR-494 inhibitor group was opposite,bone parameters and OPG,PⅠNP increased,BGP,TRACP,CTX-Ⅰdecreased,bone structure returned to normal,osteoclasts decreased,BMP-2,RUNX2 in bone tissue upregulated,RANKL downregulated,TLR4,NF-κB p65 and MyD88 downregulated(all P＜0.05).Conclusion The transfer of miRNA-494 by Exo aggravates abnormal bone metabolism in OP rats and inhibits bone remodeling balance,suggesting that the mechanism of action may be related to the regulation of TLR4 pathway.

The core components of the Hippo signaling pathway encompass upstream regulatory molecules,core kinase cascade complexes,and downstream transcriptional regulation complexes.This pathway modulates cellular behaviors by influencing the effector molecules of its core components and plays a pivotal role in immune regulation.Effector molecules,such as Yes-associated protein(YAP),transcriptional coactivator with PDZ-binding motif(TAZ),transcriptional enhanced associate domain transcriptional factor(TEAD),monopolar spindle-one binder(MOB1),large tumor suppressor(LATS),can stimulate fibroblast-like synovial cell migration and invasion in rheumatoid arthritis,regulate osteoarthritis disease progression,promote pathological new bone formation in ankylosing spondylitis,sustain submandibular gland development while delaying Sjogren's syndrome progression,mediate alpha-smooth muscle actin in systemic sclerosis,and refine the regulation of target genes associated with pulmonary fibrosis.This article provides an overview of the regulatory mechanisms involving Hippo signaling pathway-related effector molecules in the pathogenesis and progression of rheumatic immune system diseases,to serve as a reference for exploring novel therapeutic targets of rheumatic immune system diseases.

OBJECTIVE: To observe the clinical significance of translocator proteins (TSPO) gene in the treatment of FLT3-ITD/DNMT3A R882 double-mutated acute myeloid leukemia (AML). METHODS: Seventy-six patients with AML hospitalized in the Department of Hematology of the Affiliated People's Hospital of Ningbo University from June 2018 to June 2020 were selected, including 34 patients with FLT3-ITD mutation, 27 patients with DNMT3A R882 mutation, 15 patients with FLT3-ITD/DNMT3A R882 double mutation, as well as 19 patients with immune thrombocytopenia (ITP) hospitalized during the same period as control group. RNA was routinely extracted from 3 ml bone marrow retained during bone puncture, and TSPO gene expression was detected by transcriptome sequencing (using 2-deltadeltaCt calculation). RESULTS: The expression of TSPO gene in FLT3-ITD group and DNMT3A R882 group at first diagnosis was 2.02±1.04 and 1.85±0.76, respectively, which were both higher than 1.00±0.06 in control group, but the differences were not statistically significant (P=0.671, P=0.821). The expression of TSPO gene in the FLT3-ITD/DNMT3A R882 group was 3.98±1.07, wich was significantly higher than that in the FLT3-ITD group and DNMT3A R882 group, the differences were statistically significant (P=0.032, P=0.021). The expression of TSPO gene in patients who achieved complete response after chemotherapy in the FLT3-ITD/DNMT3A R882 group was 1.19±0.87, which was significantly lower than that at first diagnosis, and the difference was statistically significant (P=0.011). CONCLUSION TSPO gene may be used as an indicator of efficacy in FLT3-ITD /DNMT3A R882 double-mutated AML.
Humans ; DNA (Cytosine-5-)-Methyltransferases/genetics* ; DNA Methyltransferase 3A ; Mutation ; Leukemia, Myeloid, Acute/drug therapy* ; Nucleophosmin ; Prognosis ; fms-Like Tyrosine Kinase 3/genetics* ; Receptors, GABA/therapeutic use*