ObjectiveTo evaluate the antitumor activity of Periplaneta americana extract（PAE） against human-derived lung adenocarcinoma organoids（LUAD-PDOs） and to elucidate its potential mechanism based on transcriptomics. MethodsFresh tumor and adjacent normal tissues from patients with LUAD were collected to construct LUAD-PDOs and normal lung organoid（Nor-PDOs） models using 3D organoid culture technology. The effective intervention concentration of PAE was determined using the cell counting kit-8（CCK-8） assay. Experimental groups included the model group（LUAD-PDOs）， normal group， model administration group（LUAD-PDOs+PAE）， and normal administration group（Nor-PDOs+PAE）. Hematoxylin-eosin（HE） staining was used to observe the pathological structures of PDOs， immunohistochemistry（IHC） was performed to detect the expressions of the proliferation marker Ki-67 and lung adenocarcinoma differentiation markers cytokeratin-7（CK-7） and Napsin A， TUNEL staining was applied to detect cell apoptosis. RNA sequencing（RNA-Seq） was conducted to identify differentially expressed genes（DEGs）， followed by Gene Ontology（GO）， Kyoto Encyclopedia of Genes and Genomes（KEGG）， and Gene Set Enrichment Analysis（GSEA）， alongside protein-protein interaction（PPI） network analysis to screen core mechanisms. Finally， key targets were validated by integrating external database analysis with immunofluorescence（IF）. ResultsNor-PDOs and LUAD-PDOs that highly recapitulated the pathological characteristics of the primary tissues were successfully established. The CCK-8 assay determined that the effective intervention concentration of PAE was 16 g·L^-1. Morphological observation showed that Nor-PDOs exhibited lumen-forming structures， whereas LUAD-PDOs displayed dense， solid structures. CCK-8 and TUNEL assays revealed that， compared with the model group， PAE intervention inhibited the proliferation of LUAD-PDOs and promoted apoptosis in LUAD cells， while showing no significant effect on the viability of Nor-PDOs. Transcriptomic analysis identified 719 DEGs that were significantly reversed after PAE intervention（347 up-regulated and 372 down-regulated）（P<0.05）. GO enrichment analysis indicated that DEGs in the model administration group were significantly enriched in biological processes related to cell cycle regulation compared to the model group. KEGG pathway analysis revealed that PAE affected pathways related to proliferation and metabolism， including pathways in cancer and the p53 signaling pathway. GSEA further confirmed that PAE significantly enhanced the activity of the p53 signaling pathway（P<0.05）. PPI network analysis indicated that breast cancer type 1 susceptibility protein（BRCA1） and checkpoint kinase 1（CHEK1） were the core down-regulated targets in the p53 pathway. IF verified the high expression of BRCA1 and CHEK1 in LUAD-PDOs and their significant downregulation after PAE intervention（P<0.05）. Furthermore， survival analysis based on The Cancer Genome Atlas（TCGA） database indicated that low expression of BRCA1 and CHEK1 was significantly associated with prolonged overall survival in patients with LUAD（P<0.05）. ConclusionPAE effectively inhibits proliferation of LUAD-PDOs and promotes their apoptosis， its anti-tumor mechanism is potentially associated with the activation of the p53 signaling pathway， with BRCA1 and CHEK1 genes likely serving as key downstream targets for the effects of PAE.

BACKGROUND: T-cell lymphoblastic lymphoma/acute lymphoblastic leukemia (T-LBL/ALL) is an aggressive form of hematological malignancy associated with poor prognosis in adult patients. Histone deacetylases (HDACs) are aberrantly expressed in T-LBL/ALL and are considered potential therapeutic targets. Here, we investigated the antitumor effect of a novel HDAC inhibitor, chidamide, on T-LBL/ALL. METHODS: HDAC1, HDAC2 and HDAC3 levels in T-LBL/ALL cell lines and patient samples were compared with those in normal controls. Flow cytometry, transmission electron microscopy, and lactate dehydrogenase release assays were conducted in Jurkat and MOLT-4 cells to assess apoptosis and pyroptosis. A specific forkhead box O1 (FOXO1) inhibitor was used to rescue pyroptosis and upregulated gasdermin E (GSDME) expression caused by chidamide treatment. The role of the FOXO1 transcription factor was evaluated by dual-luciferase reporter and chromatin immunoprecipitation assays. The efficacy of chidamide in vivo was evaluated in a xenograft mouse. RESULTS: The expression of HDAC1, HDAC2 and HDAC3 was significantly upregulated in T-LBL/ALL. Cell viability was obviously inhibited after chidamide treatment. Pyroptosis, characterized by cell swelling, pore formation on the plasma membrane and lactate dehydrogenase leakage, was identified as a new mechanism of chidamide treatment. Chidamide triggered pyroptosis through caspase 3 activation and GSDME transcriptional upregulation. Chromatin immunoprecipitation assays confirmed that chidamide led to the increased transcription of GSDME through a more relaxed chromatin structure at the promoter and the upregulation of FOXO1 expression. Moreover, we identified the therapeutic effect of chidamide in vivo . CONCLUSIONS This study suggested that chidamide exerts an antitumor effect on T-LBL/ALL and promotes a more inflammatory form of cell death via the FOXO1/GSDME axis, which provides a novel choice of targeted therapy for patients with T-LBL/ALL.
Humans ; Pyroptosis/drug effects* ; Forkhead Box Protein O1/genetics* ; Aminopyridines/pharmacology* ; Animals ; Mice ; Benzamides/pharmacology* ; Cell Line, Tumor ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Phosphate-Binding Proteins/metabolism* ; Histone Deacetylase Inhibitors/pharmacology* ; Jurkat Cells ; Histone Deacetylases/metabolism* ; Apoptosis/drug effects* ; Gasdermins

This study aims to systematically characterize the targeted effects of Quanduzhong Capsules on cartilage lesions in knee osteoarthritis by integrating spatial transcriptomics data mining and animal experiments validation, thereby elucidating the related molecular mechanisms. A knee osteoarthritis model was established using Sprague-Dawley(SD) rats, via a modified Hulth method. Hematoxylin and eosin(HE) staining was employed to detect knee osteoarthritis-associated pathological changes in knee cartilage. Candidate targets of Quanduzhong Capsules were collected from the HIT 2.0 database, followed by bioinformatics analysis of spatial transcriptomics datasets(GSE254844) from cartilage tissues in clinical knee osteoarthritis patients to identify spatially specific disease genes. Furthermore, a "formula candidate targets-spatially specific genes in cartilage lesions" interaction network was constructed to explore the effects and major mechanisms of Quanduzhong Capsules in distinct cartilage regions. Experimental validation was conducted through immunohistochemistry using animal-derived biospecimens. The results indicated that Quanduzhong Capsules effectively inhibited the degenerative changes in the cartilage of affected joints in rats, which was associated with the regulation of Quanduzhong Capsules on the thioredoxin-interacting protein(TXNIP)-NOD-like receptor family pyrin domain containing 3(NLRP3)-bone morphogenetic protein receptor type 2(BMPR2)-fibronectin 1(FN1)-matrix metallopeptidase 2(MMP2) signal axis in the articular cartilage surface and superficial zones, subsequently inhibiting cartilage matrix degradation leading to oxidative stress and inflammatory diffusion. In summary, this study clarifies the spatially specific targeted effects and protective mechanisms of Quanduzhong Capsules within pathological cartilage regions in knee osteoarthritis, providing theoretical and experimental support for the clinical application of this drug in the targeted therapy on the inflamed cartilage.
Animals ; Osteoarthritis, Knee/metabolism* ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Rats ; Male ; Humans ; Capsules ; Female ; Disease Models, Animal

Objective To explore the feasibility and safety of non-anticoagulation veno-venous extracorporeal membrane oxygenation(VV-ECMO)in patients with severe chest trauma.Methods A retrospective cohort study method was used.A total of 19 patients with severe chest trauma who received VV-ECMO with a delayed anticoagulation strategy at Zhengzhou Central Hospital Affiliated to Zhengzhou University from January 2018 to October 2021 were included in the delayed anticoagulation group,and 20 patients with severe chest trauma who received VV-ECMO with a non-anticoagulation strategy from November 2021 to October 2024 were included in the non-anticoagulation group.The overall clinical characteristics of the patients were statistically analyzed,including gender,age,injury severity score(ISS),acute physiology and chronic health evaluationⅡ(APACHEⅡ),reason for VV-ECMO,use of vasoactive drugs,oxygenation index(PaO2/FiO2),and interval from injury to VV-ECMO.The primary outcomes were hemorrhagic and thrombotic complications.The secondary outcomes were blood transfusion during VV-ECMO,VV-ECMO time,mechanical ventilation time,intensive care unit(ICU)length of stay,and 28-day mortality.Results There was no significant difference in gender,age,ISS score,APACHEⅡscore,reason for VV-ECMO,use of vasoactive drugs,PaO2/FiO2,and interval from injury to VV-ECMO between the non-anticoagulation group and the delayed anticoagulation group.There was no significant difference in overall incidence of hemorrhagic and thrombotic between the two groups[incidence of hemorrhagic complications:15.0%(3/20)vs.31.6%(6/19),incidence of thrombotic:15.0%(3/20)vs.5.3%(1/19),both P>0.05].The infusion rate of 4 or more paked red blood cell(PRBC)within 24 hours during VV-ECMO in the non-anticoagulation group was significantly lower than that in the delayed anticoagulation group[5.0%(1/20)vs.31.6%(6/19),P<0.05].The amount of PRBC and platelet transfusion and the time on VV-ECMO in the non-anticoagulation group during VV-ECMO were significantly lower than those in the delayed anticoagulation group[PRBC(U):5.8±3.8 vs.8.1±3.1,platelets(U):1(0,1)vs.2(1,3),time on VV-ECMO(hours):71.55±24.37 vs.114.21±34.08,all P<0.05].There were no statistically significant differences in the amount of plasma and cryoprecipitate transfusion during VV-ECMO,mechanical ventilation time,ICU hospitalization time,and 28-day mortality between the two groups.Conclusion For patients with severe chest trauma receiving VV-ECMO withholding routine systemic anticoagulation did not result in thrombotic complications or higher mortality and required less PRBC and platelet transfusions.Non-anticoagulant VV-ECMO is safe and feasible for patients with severe chest trauma with high risk of bleeding.

Objective To reveal the scientific output and trends in pulmonary nodules/early-stage lung cancer prediction models. Methods Publications on predictive models of pulmonary nodules/early lung cancer between January 1, 2002 and June 3, 2023 were retrieved and extracted from CNKI, Wanfang, VIP and Web of Science database. CiteSpace 6.1.R3 and VOSviewer 1.6.18 were used to analyze the hotspots and theme trends. Results A marked increase in the number of publications related to pulmonary nodules/early-stage lung cancer prediction models was observed. A total of 12581 authors from 2711 institutions in 64 countries/regions published 2139 documents in 566 academic journals in English. A total of 282 articles from 1256 authors were published in 176 journals in Chinese. The Chinese and English journals which published the most pulmonary nodules/early-stage lung cancer prediction model-related papers were Journal of Clinical Radiology and Frontiers in Oncology, respectively. Chest was the most frequently cited journal. China and the United States were the leading countries in the field of pulmonary nodules/early-stage lung cancer prediction models. The institutions represented by Fudan University had significant academic influence in the field. Analysis of keywords revealed that multi-omics, nomogram, machine learning and artificial intelligence were the current focus of research. Conclusion Over the last two decades, research on risk-prediction models for pulmonary nodules/early-stage lung cancer has attracted increasing attention. Prognosis, machine learning, artificial intelligence, nomogram, and multi-omics technologies are both current hotspots and future trends in this field. In the future, in-depth explorations using different omics should increase the sensitivity and accuracy of pulmonary nodules/early-stage lung cancer prediction models. More high-quality future studies should be conducted to validate the efficacy and safety of pulmonary nodules/early-stage lung cancer prediction models further and reduce the global burden of lung cancer.

Objective:Explore the protective effect and mechanism of methyl badosolone (CDDO-Me) on rats with traumatic brain injury (TBI).Methods:A total of 72 SPF-grade SD rats aged 8 weeks were randomly (random number) divided into 4 groups ( n=18) using the random number table method: Sham, TBI, TBI+Vehicle, and TBI+CDDO-Me. The rat TBI model was established using the hydraulic impact head injury method. The TBI+CDDO-Me group was administered CDDO-Me (dissolved in 1% DMSO, at a dose of 10 mg/kg) via intraperitoneal injection 30 minutes after modeling, twice a day for a total of 3 days. On the third day after modeling, brain tissue was collected for pathological and water content detection after mNSS scoring. Immunofluorescence double staining was used to detect the expression of nuclear factor erythroid2 related factor 2 (Nrf2); immunohistochemical staining was used to detect the expression of ionized calcium binding adapter molecule-1(Iba-1); ELISA was used to detect the levels of tumor necrosis factor-α(TNF-α), interleukin (IL)-1β, and IL-18 in serum; kits were used to detect the levels of malondialdehyde (MDA) and reactive oxygen species (ROS); Western blot was used to detect the expression of the Nrf2 pathway, B-cell lymphoma-2 (BCL-2), and BCL-2 associated X protein (BAX). Results:(1) Compared with the Sham group, the mNSS scores and water content in the injured cortex of the TBI group rats were significantly increased (both P<0.05), and both significantly decreased after CDDO-Me intervention (both P<0.05). (2) Compared with the Sham group, the proportion of Nissl-stained injured neurons and apoptotic positive cells in the TBI group rats were significantly increased (both P<0.05), and both significantly decreased after CDDO-Me intervention (both P<0.05), accompanied by a decrease in BAX protein expression and upregulation of BCL-2 protein expression (both P<0.05). (3) Immunofluorescence and Western blot results showed that compared with the Sham group, the expression of total Nrf2, nuclear Nrf2, HO-1, and NQO1 proteins in the TBI group were all increased (all P<0.05), and the increase was more significant after CDDO-Me intervention (all P<0.05). (4) Immunohistochemistry and ELISA results showed that compared with the Sham group, the levels of MDA, ROS, Iba-1 in brain tissue and the levels of TNF-α, IL-1β, and IL-18 in serum in the TBI group rats were all significantly increased (all P<0.05), and all significantly decreased after CDDO-Me intervention (all P<0.05). Conclusion:CDDO-Me helps to reduce oxidative stress and inflammatory responses in TBI rats, and the mechanism may be related to the activation of the Nrf2/HO-1 antioxidant stress pathway.

AIM:To investigate the mechanism of action of Tamarix chinensis Lour.on streptozotocin-induced type 2 diabetes mellitus(T2DM)through network pharmacology,molecular docking and ex-perimental validation.METHODS:Using the TCSMP database and Swiss Target Prediction tools screen the active components and predict potential tar-gets in Tamarix chinensis Lour..Retrieving potential disease targets associated with T2DM from data-bases such as GeneCards,OMIM,and DisGeNET.The intersection targets of Tamarix chinensis Lour.and T2DM disease was obtained through Venny platform.The STRING database was used to con-structed PPI network,and Cytoscape 3.8.0 soft-ware was use to visualized.GO function enrich-ment and KEGG pathway enrichment analysis were performed through the Metascape database.Dock-ing of important target proteins and compounds was carried out by AutoDock software.SPF grade male rats were randomly divided into normal group,model group,MET group(88.5 mg/kg),TE high-dose(800 mg/kg)group,TE medium-dose(400 mg/kg)group and TE low-dose(200 mg/kg)group(n=10).High-fat and high sugar feed com-bined with low dose STZ(45 mg/kg)was used to in-duce T2DM rat model,and the rats were adminis-tered orally for 5 weeks.Fasting blood glucose(FBG),insulin(FINS)level and HOMA-IR index,bio-chemical indicators[superoxide dismutase(SOD),malondialdehyde(MDA),glycosylated hemoglobin A1c(HbA1c)and inflammatory factor[interleukin-1β(IL-1β),tumor necrosis factor α(TNF-α),vascu-lar cell adhesion molecular(VCAM-1)levels of the rats were also observed;morphological changes of renal tissue was observe by HE staining.RESULTS:Based on the screening conditions of oral bioavail-ability(OB)≥ 30％and drug like properties(DL)≥0.18,a total of 19 main active ingredients with po-tential therapeutic effects on T2DM were screened from Tamarix chinensis Lour.,including ergosta-5,24(28)-dien-3,7,16-triol,quercetin-3,3'-dimethyl ether,kaempferol,quercetin,and others.By analyz-ing the potential targets of Tamarix chinensis Lour.for treating T2DM,a total of 185 potential target genes were screened,including SRC,EGFR,HSP90AA1,AKT1,ESR1,H1F1A,TNF,PIK3R1,etc,in-volving cancer signaling pathways,insulin resis-tance,MAPK signaling pathways,PI3K Akt signaling pathways,etc.Molecular docking results showed that the binding energies were all less than-5.0 kcal/mol,indicating that a strong binding abili-ty between the active ingredients screened by Tam-arix chinensis Lour.and the potential targets for the treatment of T2DM.The animal experiment re-sults showed that compared with the model group,the weight loss of rats in the MET and TE groups was slowed down,and the levels of FBG,FINS,MDA,HbA1c,IL-1β,TNF-α,VCAM-1,HOMA-IR in-dex were reduced,the SOD level was increased,and the differences were statistically significant(P＜0.05,P＜0.01),Renal tissue cellular morphology also showed notable improvement.Most importantly,all these results demonstrating dose-dependent ef-fects.CONCLUSION:Tamarix chinensis Lour.dis-plays a significant therapeutic effect on T2DM through multi-component,multi-target,and multi-pathway synergistic actions to improve blood glu-cose levels.The findings of this study provide a the-oretical basis for the clinical application of Tamarix chinensis Lour.in the treatment of T2DM.

Objective:To preliminarily explore whether sirtuin1 (SIRT1) activation can inhibit neuronal ferroptosis in rats after traumatic brain injury (TBI) by regulating hypoxia-inducible factor-1α (HIF-1α)-mediated glycolysis.Methods:(1) Six SD rats were randomly divided into sham-operated group and TBI group, with 3 rats in each group; TBI model in the TBI group was established by hydraulic impact method, and rats in the sham-operated group underwent same surgery without impact. Cortical tissues of the two groups were sent for tandem mass tag (TMT) labeled quantitative proteomics detection to analyze the differential expression proteome; Kyoto encyclopedia of genes and genomes (KEGG) and gene set enrichment analysis (GSEA) were used to detect pathway enrichment of the screened differential proteins. (2) Twelve SD rats were randomly divided into sham-operated group and 1-day, 3-day and 7-day post-TBI groups, with 3 rats in each group. Treatment methods were the same as above; Western blotting was used to detect SIRT1 protein expression. (3) Forty-eight rats were randomly divided into sham-operated group, TBI group, TBI+vehicle group and TBI+SIRT1 agonist group, with 12 rats in each group; rats in the sham-operated group and TBI group accepted treatment as above; rats in the TBI+SIRT1 agonist group were intraperitoneally injected with SRT1720 (dissolved in ≤ 5% dimethyl sulfoxide, at a dose of 20 mg/kg) within 30 minutes after modeling, twice a day (with an interval of 12 hours); and rats in the TBI+vehicle group were injected with same dose of dimethyl sulfoxide at the same time. One d after modeling, neurological deficit was assessed using modified Neurological severity score (mNSS), brain water content was measured by dry-wet weight method, histopathological changes in the cortical lesions were observed by HE staining, mitochondrial ultrastructure was examined by transmission electron microscopy, malondialdehyde (MDA) content and superoxide dismutase (SOD) activity in the brain tissues were detected by colorimetry, and protein expressions of SIRT1, HIF-1α (key protein in the glycolytic pathway), glutathione peroxidase 4 (GPX4, key protein in the ferroptosis pathway), and acyl-CoA synthetase long-chain family member 4 (ACSL4, key protein in the ferroptosis pathway) were evaluated by Western blotting.Results:(1) KEGG analysis revealed that the glycolysis pathway and HIF-1 signaling pathway were obviously enriched in the cortical tissues of rats in the TBI group compared with the sham-operated group; GSEA showed that the HIF-1 signaling pathway (mmu04066) and ferroptosis pathway (mmu04216) gene sets in the cortical tissues of rats in the TBI group exhibited enrichment trends compared with those in the sham-operated group. (2) Compared with the sham-operated group, the 1-day, 3-day, and 7-day post-TBI groups had significantly decreased SIRT1 protein expression ( P<0.05), with the most prominent decline in 1-day post-TBI group. (3) Compared with the TBI+vehicle group, rats in the TBI+SIRT1 agonist group showed significantly reduced mNSS score and brain tissue water content (9.83±1.17 vs. 7.66±1.21; [83.62±0.91]% vs. [80.09±0.68]%, P<0.05). HE staining indicated clearer structure of the cortical area at the injury sites, and improved neuron morphology in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group; and transmission electron microscopy showed reduced mitochondrial shrinkage and partial restoration of cristae structures in the TBI+SIRT1 agonist group compared with those in the TBI+vehicle group. Compared with the TBI+vehicle group, the TBI+SIRT1 agonist group exhibited significantly decreased MDA content ([62.72±9.20] nmol/g vs. [39.34±3.48] nmol/g), increased SOD activity ([1.95±0.23] U/mg vs. [2.48±0.14] U/mg), elevated GPX4 protein expression (0.37±0.04 vs. 0.46±0.03), and decreased HIF-1α and ACSL4 protein expressions (1.16±0.15 vs. 0.81±0.12; 1.14±0.06 vs. 1.29±0.04), with significant differences ( P<0.05). Conclusion:SIRT1 activation can exert neuroprotective effect by inhibiting HIF-1α-mediated glycolysis and reducing neuronal ferroptosis after TBI.

Objective:To establish a quality control method for Aiye standard decoction.Methods:The ultra performance liquid chromatogrphy (UPLC) column Waters ACQUITY HSS T3 C18 (2.1 mm×150 mm,1.8 μm) was used to gradient elution by acetonitrile and 0.1% formic acid in water. 16 batches of Aiye standard decoction fingerprints were established by UPLC and the common peaks were determined in the fingerprints. The contents of 12 components were determined. The 16 batches of Aiye standard decoction were analyzed by similarity calculation, hierarchical cluster analysis (HCA), principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) for analysis of differential components of Artemisiae Argyi Folium from different origins.Results:A total of 13 common peaks were marked in the fingerprints of 16 batches of Aiye standard decoction, 12 of which were identified by comparison with reference substance, including chlorogenic acid, sochlorogenic acid A, neochlorogenic acid, cryptochlorogenic acid, caffeic acid,1,3-O-Dicaffeoylquinic acid, schaftoside, isochlorogenic acid B,1,5-O-Dicaffeoylquinic acid, isochlorogenic acid C, jaceosidin and eupatilin. Similarity evaluation, PCA and HCA all classified the 16 batches of Aiye standard decoction into 2 categories. Orthogonal partial least squares discriminant analysis screened 5 differential biomarkers from 13 common peaks. The content determination results showed that the phenolic compounds and flavonoids in samples from Hubei were significantly higher than that in samples from other areas.Conclusion:This method can effectively analyze the differences in the quality of Aiye standard decoction from different origins, and provide reference for the formulation of quality standards for Aiye standard decoction and related preparations.

Objective:To establish a method for the determination of triterpenes and nucleosides in Poria based on HPLC; To accurately determine the various bioactive components in Poria.Methods:Similarity evaluation, clustering analysis and principal component analysis were used to analyze the similarities and differences of different medicinal parts of Poria, and the key chromatographic peaks that could reflect the characteristics were found.Results:The Poricoic acid A and dehydroeburiconic acid could be used as the identification basis for Poriae Cutis and White Poria; at the same time, Polyporenic acid C, dehydropachymic acid and dehydrotrametenolic acid could be used to evaluate Rubra Poria and Poriae Cutis; uridine, guanosine and adenosine may be essential ingredients for evaluating the quality of White Poria, Poriae Cutis and Rubra Poria. In different medicinal parts of Poria, the triterpenes were showed significant differences; by contrary, there were little differences among the same medicinal parts.Conclusion:This study reveals the quality differences between different medicinal parts of Poria, which can provide a scientific basis for the rational application and pharmacodynamic standardization of Poria.