BACKGROUND: Oral squamous cell carcinoma (OSCC) is a prevalent type of cancer with a high mortality rate in its late stages. One of the major challenges in OSCC treatment is the resistance to epidermal growth factor receptor (EGFR) inhibitors. Therefore, it is imperative to elucidate the mechanism underlying drug resistance and develop appropriate precision therapy strategies to enhance clinical efficacy. METHODS: To evaluate the efficacy of the combination of the Ca 2+ /calmodulin-dependent protein kinase II (CAMK2) inhibitor KN93 and EGFR inhibitors, we performed in vitro and in vivo experiments using two FAT atypical cadherin 1 ( FAT1 )-deficient (SCC9 and SCC25) and two FAT1 wild-type (SCC47 and HN12) OSCC cell lines. We assessed the effects of EGFR inhibitors (afatinib or cetuximab), KN93, or their combination on the malignant phenotype of OSCC in vivo and in vitro . The alterations in protein expression levels of members of the EGFR signaling pathway and SRY-box transcription factor 2 (SOX2) were analyzed. Changes in the yes-associated protein 1 (YAP1) protein were characterized. Moreover, we analyzed mitochondrial dysfunction. Besides, the effects of combination therapy on mitochondrial dynamics were also evaluated. RESULTS: OSCC with FAT1 mutations exhibited resistance to EGFR inhibitors treatment. The combination of KN93 and EGFR inhibitors significantly inhibited the proliferation, survival, and migration of FAT1 -mutated OSCC cells and suppressed tumor growth in vivo . Mechanistically, combination therapy enhanced the therapeutic sensitivity of FAT1 -mutated OSCC cells to EGFR inhibitors by modulating the EGFR pathway and downregulated tumor stemness-related proteins. Furthermore, combination therapy induced reactive oxygen species (ROS)-mediated mitochondrial dysfunction and disrupted mitochondrial dynamics, ultimately resulting in tumor suppression. CONCLUSION Combination therapy with EGFR inhibitors and KN93 could be a novel precision therapeutic strategy and a potential clinical solution for EGFR-resistant OSCC patients with FAT1 mutations.
Humans ; ErbB Receptors/metabolism* ; Mouth Neoplasms/metabolism* ; Cell Line, Tumor ; Animals ; Drug Resistance, Neoplasm/genetics* ; Cadherins/metabolism* ; Carcinoma, Squamous Cell/metabolism* ; Mice ; Mutation/genetics* ; Mice, Nude ; Protein Kinase Inhibitors/therapeutic use* ; Cetuximab/pharmacology* ; Afatinib/therapeutic use* ; Cell Proliferation/drug effects* ; Signal Transduction/drug effects*

Acute liver failure (ALF) is a life-threatening condition associated with macrophage-mediated inflammatory responses. Effective therapies and drugs are still lacking to date. Here, we reveal that a derivative of xanthohumol, CAM12203, alleviates lipopolysaccharide (LPS) + d-galactosamine (D-GalN)-induced ALF through limiting macrophage-mediated inflammation, with the most significant impact on interleukin-1β (IL-1β) transcription. Through biotin labeling-mediated pull-down and LC-MS/MS analysis, diacylglycerol kinase ζ (DGKζ), a lipid-metabolizing kinase, is identified as the direct target of CAM12203. Mechanistically, DGKζ is induced in macrophages upon inflammatory stimuli and is upregulated observed on clinical liver failure samples. Its product phosphatidic acid (PA) boosts phospholipase C (PLC)-inositol 1,4,5-trisphosphate (IP₃)-Ca²⁺ signaling and subsequent janus kinase 2 (JAK2)-signal transducer and activator of transcription 3 (STAT3) cascade, ultimately promoting IL-1β production and liver failure. DGKζ knockdown/ablation or inhibition significantly impairs the DGKζ-STAT3-IL-1β pathway along with ALF progression. Finally, CAM12203 is confirmed to be a new DGKζ inhibitor and acts against inflammation in a DGKζ-reliant manner. Taken together, CAM12203 inhibits IL-1β transcription in macrophages by binding to DGKζ and blocking the DGKζ-STAT3 axis, thereby exerting an ameliorative effect on ALF. These results not only highlight CAM12203 as a promising lead compound for ALF treatment, but also define DGKζ as a novel therapeutic target.

T-cell acute lymphoblastic leukemia (T-ALL) is a highly aggressive hematologic malignancy with a poor prognosis, despite advancements in treatment. Many patients struggle with relapse or refractory disease. Investigating the role of the super-enhancer (SE) regulated gene ubiquitin-specific protease 20 (USP20) in T-ALL could enhance targeted therapies and improve clinical outcomes. Analysis of histone H3 lysine 27 acetylation (H3K27ac) chromatin immunoprecipitation sequencing (ChIP-seq) data from six T-ALL cell lines and seven pediatric samples identified USP20 as an SE-regulated driver gene. Utilizing the Cancer Cell Line Encyclopedia (CCLE) and BloodSpot databases, it was found that USP20 is specifically highly expressed in T-ALL. Knocking down USP20 with short hairpin RNA (shRNA) increased apoptosis and inhibited proliferation in T-ALL cells. In vivo studies showed that USP20 knockdown reduced tumor growth and improved survival. The USP20 inhibitor GSK2643943A demonstrated similar anti-tumor effects. Mass spectrometry, RNA-Seq, and immunoprecipitation revealed that USP20 interacted with hypoxia-inducible factor 1 subunit alpha (HIF1A) and stabilized it by deubiquitination. Cleavage under targets and tagmentation (CUT&Tag) results indicated that USP20 co-localized with HIF1A, jointly modulating target genes in T-ALL. This study identifies USP20 as a therapeutic target in T-ALL and suggests GSK2643943A as a potential treatment strategy.

OBJECTIVES: To develop an E-selectin-targeting nanomedicine delivery system that competitively inhibits E-selectin-neutrophil ligand binding to block neutrophil adhesion to vessels and suppress their recruitment to the lesion sites. METHODS: Doxorubicin hydrochloride (DOX)-loaded liposomes (IEL-Lip/DOX) conjugated with E-selectin-affinity peptide IELLQARC were developed using a post-insertion method. Two formulations [2-1P: Mol(PC): Mol(DPI)=100:1; 2-3P: 100:3] were prepared and their modification density and in vitro release characteristics were determined. Their targeting efficacy was assessed in a cell model of LPS-induced inflammation, a mouse model of acute lung injury (ALI), a rat femoral artery model of physical injury-induced inflammation, and a zebrafish model of local inflammation. RESULTS: The prepared IEL-Lip/DOX 2-1P and 2-3P had peptide modification densities of 4.76 and 7.57 pmoL/cm², respectively. Compared with unmodified liposomes, IEL-Lip/DOX exhibited significantly reduced 48-h cumulative release rates at pH 5.5. In the inflammation cell model, IEL-Lip/DOX showed increased uptake by activated inflammatory endothelial cells, and 2-1P exhibited a higher trans-endothelial ability. In ALI mice, the fluorescence intensity of IEL-Lip/Cy5.5 increased significantly in lung tissues by 53.71% [Z-(2-1P)] and 93.41% [Z-(2-3P)], and 2-1P had an increased distribution by 24.19% in the inflammatory lung tissue compared to normal mouse lung tissue. In rat femoral artery models, 2-1P had greater injured/normal vessel fluorescence intensity contrast. In the zebrafish models, both 2-1P and 2-3P showed increased aggregation at the site of inflammation. CONCLUSIONS This E-selectin-targeting nanomedicine delivery system efficiently targets activated inflammatory endothelial cells to increase drug concentration at the inflammatory site, which sheds light on new strategies for treating neutrophil-mediated inflammatory diseases and practicing the concept of "one drug for multiple diseases".
Animals ; Liposomes ; Rats ; Nanomedicine ; E-Selectin ; Drug Delivery Systems ; Inflammation/drug therapy* ; Mice ; Doxorubicin/analogs & derivatives* ; Zebrafish ; Acute Lung Injury/drug therapy*

Comfortable supragingival scaling uses ultrasonic cleaning with sedation and minimally invasive techniques to minimize the negative emotions of patients and improve patient compliance. At present, there is still much room for optimization of the environment, equipment, and operation steps for the development of comfortable supragingival scaling. On this basis, the Department of Periodontology, College of Stomatology, Xi'an Jiaotong University proposed a 16-step new model of comfortable supragingival scaling. The new model incorporates adjustments to the previous model concerning the environment, equipment, operational procedures, process links and services, comprising 16 steps for optimization and innovation. Clinical practice has confirmed that the 16-step new model of comfortable supra-gingival scaling can significantly improve patient satisfaction and adherence to medical treatment, and it has good promotional value.
Humans ; Dental Scaling/instrumentation* ; Patient Satisfaction ; Patient Compliance ; Gingiva

Objective To investigate the effect of mulberry anthocyanin on the morphology and func-tion of vascular endothelial cells(VEC)and its possible mechanism at the level of autophagy.Methods VEC were cultured in vitro and divided into 5.5/0 mmol/L control group,11/0.125,22/0.250,33/0.500,44/1.000,55/2.000 mmol/L high glucose/high fat groups.The cell functional levels and autophage levels in each group were detected.The optimal high glucose/high lipid concentration was selected as the VEC function inju-ry model group.10,25,50 μmol/L mulberry anthocyanins were added into the model group for processing and divided into the low,middle and high concentrations of mulberry anthocyanin groups.The effects of mulberry anthocyanin on the morphology,function and autophagy of VEC in high sugar and high fat stress were ob-served.Then the autophagy regulator(rapamycin)was added to study its action mechanism.HE staining was used to observe the number and morphology of cells,the CCK-8 assay was used to detect the viability of VEC,the flow cytometry was used to detect the level of reactive oxygen species(ROS)in VEC,the Western blot was used to detect the expression level of Beclin-1 and p62 proteins in VEC,and the kit method was used to detect the level of nitric oxide(NO)and endothelin-1(ET-1)in VEC.Results Compared with the control group,the number and morphology of cells in the middle and high concentrations of high glucose/high fat groups were changed,the cell viability,p62,NO levels were significantly decreased(P<0.05),while the ROS,ET-1,and Beclin-1 expression levels were increased significantly(P<0.05).Compared with the model group,the number of cells in the mulberry anthocyanin groups was increased,morphology was improved,the cellular viability,p62 and No levels were significantly increased,and ROS,Beclin-1 and ET-1 levels were significantly decreased(P<0.05).Compared with the high concentration mulberry anthocyanin group,the cells number was decreased af-ter adding rapamycin,the morphology changed to be irregular,the NO level was decreased,and the ET level was increased(P<0.05).Compared with the control group,the p-phosphatidylinositol 3-kinase(PI3K)and p-protein kinase B(Akt)relative expression levels in the model group were significantly decreased(P<0.05);compared with the model group,the p-PI3K and p-Akt relative expression levels in the high concentration mulberry anthocyanin group were significantly increased(P<0.05).Conclusion Mulberry anthocyanin could in-hibit autophagy by activating the PI3K/Akt/mammalian target of rapamycin(mTOR)signaling pathway and improve the morphology and function of VEC under high glucose and lipid stress.

Objective:To explore the mechanism of Coicis Semen in treating non -small cell lung cancer combined with network pharmacology and in vitro cell experiment.Methods:The components and potential targets of Coicis Semen were obtained from TCMSP, and NSCLC related targets were collected from Genecards and Drugbank. They were intersected to get common targets. An intersection target PPI network was constructed using STRING database. GO function and KEGG pathway enrichment analysis were performed using DAVID database. A "disease-drug component-target" network was constructed using Cytoscape 3.8.0. The CCK-8 method was used to screen the concentration of Coicis Semen, and the cells were randomly divided into control group, Coicis Semen low-concentration group (0.125 mg/ml), Coicis Semen high-concentration group (0.25 mg/ml), Coicis Semen low concentration+FoxO1 inhibitor group, and FoxO1 inhibitor group (50 μmol/L) using a random number table. Cell proliferation ability was determined using CCK-8 method, and cell migration and invasion ability were detected using cell scratch assay and Transwell cell invasion assay. Western blot was used to detect the expression of FoxO1, cofilin-1, and p-cofilin-1 proteins.Results:A total of 9 active components and 246 targets were screened from Coicis Semen, 1 074 targets related to non-small cell lung cancer, and 11 intersecting targets were identified. The key targets of Coicis Semen in the treatment of non-small cell lung cancer were MAPK1, MAPK8, MAPK14, CDK2, INSR, AR, FGFR2, PGR, DHFR, and PLAU; GO functional enrichment analysis showed that Coicis Semen had an impact on the cellular, molecular, and biological functions of A549 cells; KEGG pathway enrichment analysis showed that the pathways of Coicis Semen in treating non-small cell lung cancer mainly focus on FoxO pathway, MAPK pathway, prostate cancer pathway, Ras pathway, etc. Compared with the control group, the proliferation rate, migration rate, and invasion number of A549 cells in the low and high concentration groups of Coicis Semen decreased ( P<0.05), p-cofilin-1 expression decreased ( P<0.05), and FoxO1 expression increased ( P<0.05). Conclusion:Coicis Semen can treat non-small cell lung cancer by regulating MAPK1, MAPK8, and MAPK14 targets, and acting on pathways such as the FoxO and MAPK pathways.

Osteoarthritis(OA),a common age-related chronic disease,is characterized by degenerative changes in the joints and surrounding tissues.Traditionally,research on OA has primarily focused on the pathological changes in articular cartilage and its repair.However,with the advancements in animal disease modeling in recent years,especially the widespread use of spatiotemporally specific transgenic mouse models,scholars have gradually come to realize that the subchondral bone also plays an important role in the occurrence and development of OA.That is,the pathological changes in articular cartilage and bone mutually affect and promote each other,jointly driving the progression of OA,involving such pathological processes as vascular invasion,ectopic calcification,nerve growth,and the occurrence of pain.Given the complexity of cartilage-bone pathological relationship,it is difficult to conduct in-depth research on subchondral bone pathology using clinical human samples,or to simulate the pathological processes of OA through in vitro cell experiments.Therefore,animal models play an irreplaceable role in investigating the pathological mechanisms of OA and developing clinical drugs.This review,in addition to providing an overview of OA animal models,synthesizes the latest progress in animal experimental research on OA,highlighting the active role of the cartilage-bone pathological relationship in OA progression.These new findings provide references for future in-depth investigations and also provide a theoretical basis for developing fundamental strategies for OA prevention and treatment.

Osteoporosis and cardiovascular calcification,two major age-related chronic diseases that China is confronting today,pose serious threats to public health.Previous studies have reported overlapping connections in the pathological processes and molecular mechanisms of these two diseases,particularly concerning inflammation,oxidative stress,and dysregulation of mineral metabolism,and that these two diseases tend to share common pathogenic factors.However,research exploring the comorbidity mechanisms of the two diseases remains limited in both depth and scope,largely due to the lack of widely accepted comorbidity animal models.Herein,we analyzed the latest research findings on the comorbidity mechanisms of vascular calcification and osteoporosis,focusing on summarizing the animal disease models currently in extensive use and the relevant evaluation criteria.We aim to provide new references for comorbidity research models and offer scientific evidence for future studies on pathological mechanisms and the development of new therapeutic strategies.

[Abstract] [Objective] To establish a new centrifugal-membrane hybrid plasmapheresis (CMHP) model, and observe its clinical efficacy, safety and advantages in the treatment of hyperlipidemia by comparing with centrifugation therapeutic plasma exchange (cTPE). [Methods] A retrospective analysis was performed on 47 patients with hyperlipidemia treated in our department from August 2021 to September 2023, with a total of 60 treatments. They were divided into two groups: CMHP group with 37 patients and 46 treatments; cTPE group included 10 patients with 14 treatments. In the CMHP group, the plasma was separated by a blood cell separator and the plasma components were separated by a two-stage membrane plasma component separator. In the cTPE group, the plasma was separated by a blood cell separator for therapeutic plasma exchange. The clinical efficacy, safety and advantages of the two groups were compared. [Results] The reduction ratios of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) before and after treatment in CMHP group were 68.56%[59.81%, 73.42%], 65.80%[55.55%, 75.98%] and 46.57%[36.02%, 54.83%] and 66.67%[43.48%, 76.24%] respectively, with statistically significant difference (P<0.001). The decrease ratios of TC, TG and HDL-C before and after treatment in cTPE group were 42.52%[29.67%, 49.85%], 52.32%[38.43%, 67.07%] and 22.36%[8.51%, 33.65%], respectively, and the difference was statistically significant (P<0.05). The plasma treatment multiple was 1.48 (0.29) in CMHP group, which was significantly higher than 0.87 (0.26) in cTPE group, with a statistically significant difference (P<0.001), resulting in higher TC and HDL-C reduction ratios in CMHP group than in cTPE group, with a statistically significant difference (P<0.001), while there was no significant difference in TG reduction rate between the two groups (P>0.05). At the same time, the cTPE group required 1 520.00 mL[1 462.50 mL, 2 000.00 mL] plasma input, while the CMHP group achieved zero blood input. The adverse reaction ratio was 6.52%(3/46) in CMHP group and 7.14%(1/14) in cTPE group, and the difference was not statistically significant (P>0.05). [Conclusion] Compared with cTPE, CMHP can better reduce blood lipid levels without any blood products, avoid the spread of blood infectious diseases, and have a low incidence of adverse reactions, so it has a good clinical application prospect.