To address the constrains imposed by insufficient hydrogen peroxide (H2O2) and high glutathione (GSH) expression in tumor cells on the efficacy of chemodynamic therapy (CDT), zeolitic imidazolate framework-8 (ZIF-8) loaded with disulfiram (DSF) and 3-amino-1,2,4-triazole (3-AT) was synthesized via a one-pot approach. Subsequently, hyaluronic acid (HA)-modified cupric peroxide (CuO2) was in-situ grown on its surface through biomineralization to construct a multifunctional copper-based nanozyme ADZCH (3-AT/DSF@ZIF-8@CuO2-HA). This nanoplatform disrupts the intratumoral H2O2 homeostasis, depletes GSH, and synchronously delivers DSF and Cu2+ via cascade catalysis, thereby enhancing CDT and sensitizing tumors to DSF-based chemotherapy. The results of physicochemical characterization indicated that ADZCH presented a uniform core-shell structure with favorable dispersibility. Its particle size and Zeta potential were 196.5 nm and −19.5 mV, respectively. It possessed a microporous structure with a specific surface area of 81.8600 m2/g, and demonstrated efficient loading capacity for DSF and 3-AT, achieving drug loading efficiencies of 5.91% and 45.07%, respectively. Moreover, ADZCH can continuously and slowly release drugs in an acidic environment and maintain good stability under diverse physiological conditions. In vitro functional assays verified that ADZCH catalytically generated H2O2 and hydroxyl radicals while concurrently depleting GSH in a concentration- and incubation time-dependent manner. Cellular uptake experiments showed that HA modification significantly improved the uptake of nanoparticles by 4T1 cells. Cytotoxicity tests showed that 80 μg/mL ADZCH had a significant cytotoxic effect on 4T1 cells but no significant toxicity on L929 cells. DCFH-DA probe detection indicated that ADZCH could significantly induce intracellular reactive oxygen species (ROS) generation, thereby enhancing CDT efficacy. Live/dead staining experiments showed that ADZCH efficiently induced apoptosis, with the proportion of dead cells reaching 94.74%, demonstrating its promising potential for anti-tumor applications.This study provides new research ideas and experimental basis for overcoming the tumor microenvironment barrier and enhancing the anti-tumor effect of CDT combined with chemotherapy.

Objective To explore the effects and mechanism of Yijing Decoction containing serum on iron overload-induced oxidative stress of human ovarian granulosa cells.Methods SD rats were used to prepare medicated serum and blank serum.Iron dextran was used to induce oxidative damage of SVOG cells.The cells were divided into control group,model group,containing serum group,blank serum group,antioxidant group and iron chelating agent group.After corresponding intervention,cell viability was detected by CCK-8 method,estrogen(E2)and progesterone(P)content in supernatant were detected by ELISA,the contents of intracellular ferrous ion(Fe2+),malondialdehyde(MDA),adenosine triphosphate(ATP)and the activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),catalase(CAT)were detected by the kit,ROS content was detected by DCFH-DA fluorescence probe,the mitochondrial membrane potential was detected by JC-1 fluorescence probe,Western blot and RT-qPCR were used to detect the protein and mRNA expression of nuclear factor erythroid 2-related factor 2(Nrf2),transferrin(Tf),transferrin receptor 1(TfR1),ferritin heavy chain 1(FTH1)and ferritin light chain(FTL).Results Compared with the control group,the viability of SVOG cells decreased in the model group,the contents of E2 and P in cell supernatant decreased,the contents of Fe2+,MDA and ROS increased,the content of ATP and activities of SOD,GSH-Px and CAT decreased,the mitochondrial membrane potential decreased,the protein and mRNA expressions of Nrf2 and FTH1 decreased,and the protein and mRNA expressions of Tf,TfR1 and FTL increased,with statistical significance(P<0.05,P<0.01).Compared with the model group,the viability of SVOG cells in containing serum group,antioxidant group and iron chelating agent group increased,the contents of E2 and P in cell supernatant increased,the contents of Fe2+,MDA and ROS decreased,the content of ATP and activities of SOD,GSH-Px and CAT increased,the mitochondrial membrane potential increased,the protein and mRNA expressions of Nrf2 and FTH1 increased,and the protein and mRNA expressions of Tf,TfR1 and FTL decreased(P<0.05,P<0.01).Conclusion Yijing Decoction containing serum may relieve oxidative stress damage induced by iron overload,improve mitochondrial function,and restore granulosa cell function,thereby enhancing ovarian function,potentially through up-regulating Nrf2,FTH1 expression and down-regulating Tf,TfR1 and FTL expressions.

Objective: This study employs metabolomics to analyze the characteristic biomarkers and metabolic pathways in the serum of pregnant women with positive thyroid peroxidase antibodies(TPOAb) during early pregnancy. The objective is to explore the relationship between thyroid dysfunction and maternal-fetal health. Methods : Early-pregnancy women undergoing antenatal check-ups for thyroid function and antibody testing at our hospital were selected. According to the TPOAb results, participants were categorized into a TPOAb-positive group and a TPOAb-negative group. The serum metabolic profiles were analyzed using liquid chromatography-tandem mass spectrometry(LC-MS/MS) technology to identify differences between the two groups. The Kyoto Encyclopedia of Genes and Genomes(KEGG) database was utilized for metabolic pathway enrichment analysis of differential metabolites. Results : A total of 79 significantly different metabolites were identified in the serum of TPOAb-positive pregnant women compared to the control group, including 20 upregulated and 59 downregulated metabolites. KEGG enrichment analysis indicated that these differential metabolites were mainly involved in 21 key metabolic pathways. Among the metabolites associated with TPOAb, 31 were identified, with 6 showing positive correlation and 25 showing negative correlation. Metabolic pathway enrichment analysis revealed that these differential metabolites were closely related to Glycerophospholipid metabolism, Valine, leucine and isoleucine biosynthesis, Glycosylphosphatidylinositol(GPI)-anchor biosynthesis, and Glycerolipid metabolism pathways. Conclusion Significant differences in metabolites and their associated metabolic pathways are identified in the serum of TPOAb-positive pregnant women during early gestation, indicating that these metabolite alterations are closely linked to thyroid dysfunction and maternal-fetal health.

Objective To explore the feasibility of constructing a three-classification bone status screening radiomics model on chest CT images.Methods A total of 371 patients who underwent both chest and abdominal plain CT examinations were retrospec-tively selected and randomly divided into training set(296 cases)and test set(75 cases)in a ratio of 8︰2.Additionally,110 patients were included as external validation set using the same criteria.The 120 kVp abdominal images were transmitted to a quantitative compu-ted tomography(QCT)post-processing workstation to measure the bone mineral density(BMD)of the L1-L2 vertebral bodies.Patients were classified into osteoporosis(OP)group(BMD<80 mg/cm3),osteopenia group(80 mg/cm3≤BMD≤120 mg/cm3)and normal bone mass group(BMD>120 mg/cm3)based on QCT BMD results.The automatic segmentation model was used to segment T10-T12 vertebral trabecular bone on chest CT images and the radiomics models based on random forest(RF)and logistic regres-sion(LR)was established to evaluate BMD,enabling it to simultaneously distinguish OP,osteopenia,and normal bone mass.The diag-nostic performance of the two models were evaluated using metrics such as the area under the curve(AUC),sensitivity and specificity.The DeLong test was used to compare the differences between the two models.Results In the test set,the AUC for differentiating normal bone mass were 0.948 and 0.877 for the RF and LR models,respectively;the AUC for differentiating OP were 0.942 and 0.836,respectively;and the AUC for differentiating osteopenia were 0.871 and 0.688,respectively.The performance comparison results of the models showed that there was no statistically significant difference in AUC(0.966 vs 0.907,P>0.05)between RF model and LR model in the external validation set for distinguishing OP,while there was a statistically significant difference in AUC for distinguishing osteopenia(0.895 vs 0.749,P=0.009)and normal bone mass(0.975 vs 0.906,P=0.023).The RF model performance was superior to the LR model.Conclusion The radiomics model developed based on chest plain CT can be used for opportunistic OP screening with good diagnostic efficacy,and the the model based on the RF classifier outperforms the LR model.

Objective To explore the feasibility of spectral CT material decomposition technique Ca(Iodine)for opportunistic osteoporo-sis screening in enhanced scanning.Methods A total of 314 patients who underwent abdominal enhancement were selected.They were divided into group A and group B according to gender,and the groups were divided into three subgroups according to age(18-45 years,46-60 years,>60 years).The bone mineral density(BMD)values of L1-L3 vertebral were measured in the unenhanced images using quan-titative computed tomography(QCT)software,and Ca(Iodine)values were measured on three enhancement phases Ca(Iodine)based substance images.Pearson correlation analysis was performed between iodine intake and Ca(Iodine)values in three enhancement phases,Ca(Iodine)values and quantitative computed tomography bone mineral density(BMDQCT),and the diagnostic efficacy was analyzed.Results There was no significant difference in the Ca(Iodine)values of L1-L3 vertebral in three enhancement phases(F=0.001-0.018,P>0.05);there was no correlation between the Ca(Iodine)values of L1-L3 vertebral in three enhancement phases and iodine intake(r=0.073-0.105,P>0.05).Six groups of measured Ca(Iodine)values and BMDQCT both had a strong positive correlation(r=0.901-0.954,P<0.05).The Ca(Iodine)cutoff values for the diagnosis of osteoporosis and osteopenia were 830.41(2 mg/cm3)and 849.32(2 mg/cm3)respectively,with corresponding area under the curve(AUC)of 0.969[95%confidence interval(CI)0.943-0.985]and 0.973(95%CI 0.944-0.989),respectively.There were statistically significant differences in Ca(Iodine)values and BMDQCT in the>60 years age group between different genders(t=3.081-3.091,P<0.05).Conclusion Spectral CT material decomposition technique can be used for oppor-tunistic osteoporosis screening in enhanced scanning,with good diagnostic performance,which provides a new perspective for the clin-ical diagnosis of osteoporosis.

Prostaglandin E2(PGE2)regulates a variety of cells in tumor microenvironment(TME)and plays an important role in tumor growth,metastasis and immunity.PGE2 activates different downstream signaling pathways through its key receptor prostaglandin E receptor(EP4),regulating inflammation,immune cell function and angiogenesis to promote or inhibit tumor development.EP4 shows potential application values in adjuvant chemotherapy and immunotherapy as an emerging anti-tumor target.However,future studies on the roles of EP4 in tumor microenvironment,develo-pment of safer and more effective EP4 drugs,and its application in personalized immunotherapy are needed.

Objective·To investigate the changes in transcriptome and chromatin accessibility during the differentiation of human embryonic stem cells(hESCs)into neural progenitor cells(NPCs)using in vitro differentiation models and high-throughput multi-omics sequencing technologies.Methods·hESCs were first induced to differentiate into NPCs in vitro using the embryoid body formation method,and cells at both stages were collected.The cell phenotypes were identified by reverse transcription-quantitative real-time PCR(RT-qPCR)and immunofluorescence(IF)staining.Transcriptome sequencing(RNA-seq)was conducted to detect and analyze the differentially expressed genes(DEGs)between hESCs and NPCs.The assay for transposase-accessible chromatin with high-throughput sequencing(ATAC-seq)was employed to assess chromatin accessibility changes between hESCs and NPCs.Motif enrichment analysis was performed on differentially accessible chromatin regions to discover potential regulatory transcription factors.Finally,an integrated analysis of RNA-seq and ATAC-seq data and the protein-protein interaction(PPI)network were performed to identify key genes and regulatory pathways involved in the early stages of neural differentiation in vitro.Results·Both RT-qPCR and IF results indicated that the expression levels of pluripotency markers(NANOG and POU5F1)were high at the hESC stage but significantly decreased at the NPC stage,while early neural differentiation markers(PAX6,SOX1,and NES)were minimally expressed at the hESC stage but markedly upregulated at the NPC stage.RNA-seq analysis revealed that compared to the hESC stage,there were 5 597 genes upregulated and 3 654 genes downregulated at the NPC stage.Gene function enrichment analysis showed that the upregulated genes at the NPC stage were enriched in the functions related to neural development.ATAC-seq analysis demonstrated a total of 27 491 genomic regions had significant changes in chromatin accessibility during the differentiation from hESC to NPC,with 12 381 regions showing increased accessibility and 15 110 regions showing decreased accessibility.Motif enrichment analysis revealed that transcription factor genes such as DLX1 and LHX2 might play an important role in the differentiation process from hESCs into NPCs.Integrated analysis of RNA-seq and ATAC-seq data revealed that overlapping genes with high expression at the NPC stage were mainly enriched in axon guidance,forebrain development,and neuron migration.After neural differentiation,the expression levels of CTNND2 and LHX2 genes increased,and the chromatin accessibility of related genomic regions also increased.PPI network analysis indentified candidate downstream genes including PRKACA,CDH2,and ERBB4.Conclusion·The in vitro differentiation model of hESCs combined with high-throughput multi-omics sequencing technologies can be used to depict the changes in transcriptome and chromatin accessibility during the differentiation of hESCs into NPCs.In this process,the expression levels of genes related to axon guidance,forebrain development,and neuronal migration pathways increase and related chromatin accessibility is enhanced.

The p21-activated kinase 4 (PAK4), a key regulator of malignancy, is negatively correlated with immune infiltration and has become an emergent drug target of cancer therapy. Given the lack of high efficacy PAK4 inhibitors, we herein reported the identification of a novel inhibitor 13 bearing a tetrahydrobenzofuro[2,3-c]pyridine tricyclic core and possessing high potency against MIA PaCa-2 and Pan02 cell lines with IC₅₀ values of 0.38 and 0.50 μmol/L, respectively. This compound directly binds to PAK4 in a non-ATP competitive manner. In the mouse Pan02 model, compound 13 exhibited significant tumor growth inhibition at a dose of 100 mg/kg, accompanied by reduced levels of PAK4 and its phosphorylation together with immune infiltration in mice tumor tissue. Overall, compound 13 is a novel allosteric PAK4 inhibitor with a unique tricyclic structural feature and high potency both in vitro and in vivo, thus making it worthy of further exploration.

Venous system diseases mainly include varicose veins and venous malformations of lower limbs and the genital system. Most of them are chronic diseases that cause serious clinical symptoms to patients and affect their health and quality of life. Sclerotherapy has become the first-line therapy for venous system diseases. However, there are problems such as incomplete fibrosis and vascular recanalization after sclerotherapy, and improper operation will cause serious adverse consequences. Therefore, exploring a safe and effective sclerotherapy strategy is essential for developing clinically successful sclerotherapy. To solve the above problems, we proposed a new sclerotherapy strategy with a dual mechanism of "vascular damage and plasmin (PLA) system inhibition." We intended to construct a novel cationic surfactant (AEO_x-TA) by reacting tranexamic acid (TA), a parent structure, with fatty alcohol polyoxyethylene ether (AEO_x) by ester bonds. AEO_x-TA could damage vascular endothelium and initiate a coagulation cascade effect to induce thrombus. Furthermore, AEO_x-TA could be degraded by esterase and release the parent drug, TA, which could inhibit the PLA system to inhibit the degradation of thrombus and extracellular matrix and promote the process of vascular fibrosis. In addition, such surfactant-based sclerosants have foam-forming properties, and they can be blended with polyvinyl alcohol (PVA) to prepare a highly stable foam formulation (AEO_x-TA/P), which can achieve a precise drug delivery and prolonged drug retention time, thereby improving drug efficacy and reducing the risk of ectopic embolism. Overall, the novel cationic surfactant AEO_x-TA provides a new avenue to resolve the bottleneck: surfactant sclerosants' efficiency is relatively low in the current sclerotherapy.

Periodontitis is a common oral disease characterized by progressive alveolar bone resorption and inflammation of the periodontal tissues. Dimethyl fumarate (DMF) has been used in the treatment of various immune-inflammatory diseases due to its excellent anti-inflammatory and antioxidant functions. Here, we investigated for the first time the therapeutic effect of DMF on periodontitis. In vivo studies showed that DMF significantly inhibited periodontal destruction, enhanced mitophagy, and decreased the M1/M2 macrophage ratio. In vitro studies showed that DMF inhibited macrophage polarization toward M1 macrophages and promoted polarization toward M2 macrophages, with improved mitochondrial function, inhibited oxidative stress, and increased mitophagy in RAW 264.7 cells. Furthermore, DMF increased intracellular mitochondrial Tu translation elongation factor (TUFM) levels to maintain mitochondrial homeostasis, promoted mitophagy, and modulated macrophage polarization, whereas TUFM knockdown decreased the protective effect of DMF. Finally, mechanistic studies showed that DMF increased intracellular TUFM levels by protecting TUFM from degradation via the ubiquitin-proteasomal degradation pathway. Our results demonstrate for the first time that DMF protects mitochondrial function and inhibits oxidative stress through TUFM-mediated mitophagy in macrophages, resulting in a shift in the balance of macrophage polarization, thereby attenuating periodontitis. Importantly, this study provides new insights into the prevention of periodontitis.
Dimethyl Fumarate/pharmacology* ; Mitophagy/drug effects* ; Animals ; Mice ; Macrophages/metabolism* ; Periodontitis/prevention & control* ; RAW 264.7 Cells ; Oxidative Stress/drug effects* ; Peptide Elongation Factor Tu/metabolism* ; Mice, Inbred C57BL ; Male ; Mitochondria/drug effects*