KRAS mutations are a major driver of pancreatic ductal adenocarcinoma (PDAC). RASA2, a RAS GTPase-activating protein, modulates KRAS protein levels in wild-type contexts, suggesting it could play a potential role in PDAC. Here, we systematically investigated the biological function and molecular mechanisms of RASA2 in PDAC. Integrative analyses of multiple datasets and clinical samples demonstrated that RASA2 was consistently upregulated in KRAS-mutant PDAC and significantly associated with poor prognosis and metastatic progression. Gain- and loss-of-function studies revealed that RASA2 markedly enhanced PDAC cell migration and invasion in both KRAS-mutant and KRAS-wild-type models, suggesting that its pro-metastatic activity is largely independent of KRAS mutational status. Transcriptomic and mechanistic analyses revealed that RASA2 activated GLI1 through a TGFβ2-dependent, non-canonical Hedgehog pathway. Mechanistically, RASA2 interacted with RTF1 to promote H2BK120 ubiquitination at the TGFB2 promoter, thereby enhancing TGFβ2 transcription and activating downstream GLI1 signaling. Pharmacological inhibition of TGFβ signaling or genetic silencing of GLI1 effectively suppressed RASA2-driven migratory, invasive, and metastatic phenotypes in vitro and in vivo. Collectively, these findings reveal a mechanism by which RASA2-dependent epigenetic and transcriptional reprogramming promotes metastatic progression and nominate the RASA2- TGFβ2-GLI1 axis as a potential therapeutic target in PDAC.

BACKGROUND:Systemic inflammation, immune function, and nutritional status are closely linked to survival in gastric cancer (GC). The C-reactive protein-albumin-lymphocyte (CALLY) index integrates these dimensions; however, its dynamic perioperative changes remain unclear. This study evaluated the prognostic significance of the CALLY recovery ratio in patients undergoing curative gastrectomy. METHODS:We retrospectively analyzed 331 patients who underwent radical gastrectomy for gastric adenocarcinoma between January 2016 and December 2019. Preoperative CALLY (pre-CALLY) was calculated using laboratory values obtained within 7 days before surgery, and postoperative CALLY (POD30-CALLY) was assessed on postoperative day 30. The recovery ratio was defined as POD30-CALLY divided by pre-CALLY. Overall survival (OS) was evaluated using Kaplan-Meier analysis and Cox regression. Predictive performance was assessed by 36-month ROC curves. A nomogram incorporating independent prognostic factors was developed and validated. RESULTS:Preoperative inflammatory markers (NLR, PLR, mGPS, and pre-CALLY) were significantly associated with OS, but showed limited discrimination (36-month AUCs 0.573-0.605). POD30-CALLY alone was not significantly predictive (AUC 0.569, P = 0.087). In contrast, the recovery ratio demonstrated superior prognostic performance (AUC 0.704, P < 0.001). Multivariate analysis identified age (HR 1.022), TNM stage (HR 2.051), adjuvant chemotherapy (HR 0.401), and recovery ratio (HR 0.526) as independent predictors of OS. The nomogram showed good calibration and improved clinical net benefit. CONCLUSIONS:The CALLY recovery ratio is an independent and dynamic prognostic biomarker that improves risk stratification after curative gastrectomy.

Abstract Objective: This study investigates the effects of Celastrus orbiculatus Thunb extract (COE) on the metastasis of gastric cancer (GC) through MED12, aiming to elucidate the underlying mechanisms. Materials and Methods: Cell viability was evaluated via the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Adhesion and transwell assays were conducted to assess the adhesion, invasion, and migration capabilities of mediator complex subunit 12 (MED12)-deficient GC cells. Cell movement was tracked using the high-content imaging system. Western blotting was employed to examine the impact of COE on the expression of MED12 and the alterations in proteins related to invasion and migration in GC cells. Results: COE reduces MED12 expression in GC cells through a distinct mechanism. Experimental data demonstrate that COE and direct modulation of MED12 expression significantly impair the infiltration and movement of GC cells, with a notable reduction in movement distance. Furthermore, both treatments substantially downregulate matrix metalloproteinases expression, thereby inhibiting the invasive potential of tumor cells. In addition, COE treatment and MED12 deficiency reverse epithelial-mesenchymal transition-related protein alterations, providing new insights into the mechanisms driving tumor metastasis. Conclusions: MED12 deficiency serves as a critical barrier to GC invasion and metastasis, while COE modulation of MED12 expression offers a promising strategy for inhibiting GC progression. These results not only suggest new therapeutic strategies for GC but also propose possible routes for the creation of anti-tumor drugs.

Histopathological hematoxylin and eosin (H&E) slides contain valuable prognostic information for pancreatic ductal adenocarcinoma (PDAC), yet systematic feature extraction remains challenging. This multi-center study developed and validated an automated prognostic model using deep learning on digitized whole-slide images from 873 PDAC patients with surgical resection across three academic centers. The CrossFormer architecture achieved superior performance in external validation (area under the curve [AUC] = 0.774), significantly outperforming ResNet-18 (AUC = 0.716), ResNet-50 (AUC = 0.737), and DenseNet-121 (AUC = 0.729). Gradient-weighted Class Activation Mapping identified key prognostic features including desmoplastic stroma, high nuclear-to-cytoplasmic ratio, tumor necrosis, and immune cell infiltration. The pathomics signature effectively stratified patients into low-risk and high-risk groups with significant survival differences (p < 0.001). Critically, carbohydrate antigen 19-9 (CA19-9) retained prognostic value only in low-risk patients (hazard ratio [HR] = 2.70, p < 0.001) but not in high-risk patients (HR = 0.998, p = 0.990). High-risk patients derived substantial benefit from adjuvant chemotherapy (HR = 0.56, p = 0.038), whereas low-risk patients showed no significant benefit (HR = 0.83, p = 0.562). These findings provide actionable clinical insights: treatment intensification for high-risk patients and CA19-9-guided monitoring for low-risk patients. This validated, interpretable model transforms routine H&E slides into quantitative prognostic tools, enabling personalized treatment strategies without additional testing costs.

Cyclophosphamide (CP) is a widely prescribed chemotherapeutic and immunosuppressive agent known to cause significant reproductive toxicity. Ginseng effectively counteracts CP-induced reproductive toxicity, exhibiting pharmacological effects that resemble enhanced orexin signaling. In this study, molecular docking identified ginsenoside Rg1 as the candidate with the highest binding affinity for orexin receptor 1 (OX1R). In CP-treated male mice, Rg1 dose-dependently ameliorated reproductive toxicity, as evidenced by improved sexual behavior, sperm motility and DNA integrity, serum testosterone levels, and testicular histology. These protective effects were significantly attenuated by co-administration of the OX1R antagonist SB-334867. Notably, serum orexin-A levels did not differ significantly across all experimental groups, indicating that Rg1 acts directly on testicular OX1R rather than by elevating circulating orexin-A. Mechanistically, Rg1 restored testicular PI3K/AKT phosphorylation in an OX1R-dependent manner and preserved glucose transporter 3 and glucose transporter 8 immunoreactivity in germ cells. These findings demonstrate that Rg1 protects against CP-induced reproductive toxicity through OX1R-dependent activation of the PI3K/AKT pathway and maintenance of glucose transporter-mediated metabolism. Thus, OX1R represents a crucial mechanistic node and Rg1 is a promising adjunct therapy to mitigate chemotherapy-related male infertility.

The pharmacological effects of the natural product triptonoterpene have been scarcely documented. In our preliminary investigations, we identified that triptonoterpene possesses the capacity to impede the invasion and migration of gastric cancer (GC) cells. We are committed to further exploring the potential extensive effects of triptonoterpene on GC and to clarifying its exact mechanism of action. This study was designed to examine the influence and underlying mechanisms of triptonoterpene on apoptosis in GC cells. Treatment with triptonoterpene effectively inhibited the proliferation of GC cells. We noted that triptonoterpene was capable of inducing apoptosis in GC cells, a process mediated through the mitochondrial pathway. Additionally, triptonoterpene was found to inhibit the polymerization of the actin cytoskeleton. The presence of triptonoterpene also disrupted the ROCK/LIMK/cofilin signaling pathway in GC cells. We hypothesize that triptonoterpene may trigger the mitochondrial apoptosis pathway by modulating the actin cytoskeleton. By employing Latrunculin A (LAT-A, an actin filament disruptor) and BMS-5 (a LIMK inhibitor) to manipulate actin dynamics, we observed that both LAT-A and BMS-5 enhanced triptonoterpene-induced apoptosis and the accumulation of reactive oxygen species (ROS), and intensified the decline in mitochondrial membrane potential (MMP). Intriguingly, we discovered that GC cells treated with triptonoterpene exhibited mitochondrial fragmentation and colocalization of cofilin-1 with mitochondria. Western blot analysis further revealed that triptonoterpene facilitated the translocation of cofilin-1 from the cytoplasm to the mitochondria. Finally, the results of our in vivo experiments indicated that triptonoterpene significantly inhibited the proliferation of xenograft tumors and induced tumor cell apoptosis. This study provides experimental evidence for the development of triptonoterpene and explores new pathways for cancer treatment.

Background and objectiveRheumatoid arthritis (RA) is a chronic autoimmune disease characterized by the hyperproliferation and invasive behavior of rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS), which contributes to the degradation of articular cartilage and bone. Inhibition of RA-FLS proliferation, migration and invasion has become an important therapeutic strategy for RA. Triptolide (TPL), an epoxy diterpene lactone compound from the traditional Chinese medicine Tripterygium wilfordii Hook. f., has significant immunosuppressive and anti-inflammatory effects. However, the specific mechanisms of TPL-regulated effects on RA-FLS cytoskeleton and inhibition of invasive metastasis are not yet fully explored. The aim of this study was to investigate TPL-regulated effects on RA-FLS skeleton and reveal the specific mechanism of TPL-inhibition of RA-FLS migration and invasion.Materials and methodsIn vitro experiments were performed using RA-FLS cell line. Cell motility was evaluated by wound healing assay and Transwell assay as well as high content cell imaging system. Cytoskeletal remodeling was observed by cytoskeletal immunofluorescence staining and transmission electron microscopy (TEM). Network pharmacology predicted the targets of Triptolide. RhoA/Rho-associated kinase signaling pathway was detected by quantitative real-time PCR and Western blotting. Molecular docking and molecular dynamics simulations were used to validate the interaction of Triptolide with RhoA/Rho-associated kinase.ResultsTPL significantly inhibited RA-FLS cell motility, and reduced the displacement and cumulative distance of RA-FLS. Cytoskeleton staining assay and TEM observation showed cytoskeleton remodeling after TPL treatment. Network pharmacological prediction screened 45 targets associated with TPL intervention in RA via cytoskeleton, including TNF, KRAS, ESR1, RHOA, MAPK3 and CASP3. In the RhoA/Rho-associated kinase signaling pathway, TPL treatment inhibited protein expression and phosphorylation of RhoA, Rock, and Limk. TPL can enter RhoA, Rock1, and Rock2 target protein binding domains with stable binding activities, and may cause conformational changes of Rock1 related to molecular functions.ConclusionTPL inhibits RA-FLS in motility by regulating actin cytoskeleton remodeling through action on the RhoA/Rho-associated kinase signaling pathway.

BACKGROUND:Interleukin-33 (IL-33), a member of the IL-1 cytokine family, is constitutively expressed in barrier cells such as endothelial cells and fibroblasts. While the expression of IL-33 in regulatory T cells (Tregs) has been previously reported, its clinical significance in pancreatic ductal adenocarcinoma (PDAC) remains unclear. This study aims to investigate the clinical relevance and biological role of IL-33 + Tregs in PDAC. METHODS:Infiltration of IL-33 + Tregs was assessed by immunohistochemistry in 215 patients from our institute. The correlation between IL-33 + Tregs and clinical characteristics was analyzed. Additionally, the functional status of cytotoxic T cells in relation to IL-33 + Treg infiltration was examined. The impact of IL-33 + Tregs on the tumor microenvironment (TME) was further evaluated both in silico and in vitro . RESULTS:IL-33 + Tregs infiltration was confirmed in PDAC tissues, and its abundance was positively associated with microvascular invasion, perineural invasion, and elevated serum CA19-9 levels. Patients with higher tumor-infiltrating IL-33 + Tregs demonstrated poorer overall survival (OS) and recurrence-free survival (RFS) compared to those with lower infiltration levels. Multivariate analysis confirmed IL-33 + Tregs as an independent prognostic factor for both OS and RFS, with improved survival prediction when combined with tumor differentiation. Subgroup analyses indicated that serum CA19-9 was not a useful risk stratification tool in patients with high IL-33 + Treg infiltration, and these patients showed limited survival benefit from adjuvant chemotherapy. Furthermore, increased IL-33 + Treg infiltration was associated with more pronounced immunosuppressive TME, marked by a reduction in cytotoxic phenotypes and an upregulation of exhausted markers on CD8 + T cells. CONCLUSION:Our findings identify IL-33 + Tregs as a novel subtype of Tregs, with strong prognostic value for survival risk stratification and therapeutic response prediction in PDAC. IL-33 + Tregs exhibit more pronounced immunosuppressive capabilities, impairing CD8 + T cell function. With further investigation, IL-33 + Tregs may represent a promising immunotherapeutic target for PDAC.

Metabolomics and 16S rDNA sequencing have shown great potential in elucidating complex mechanisms associated with diseases. Currently, there is little research on the omics of gastric cancer and it lacks effective biomarkers. Objective Based on plasma metabolomics and 16S rDNA sequencing to evaluate the changes in metabolites and fecal microbiota of advanced gastric cancer. Method Firstly, plasma metabolomics was used to screen for differential metabolites and metabolic pathways in gastric cancer. Then, 16S rDNA sequencing was performed on fecal samples to study the differential intestinal microbiota in gastric cancer patients. Finally, conduct a correlation analysis between them. Result A total of 152 differential metabolites were identified, and we screened 10 of them. All metabolites were enriched into 42 differential metabolic pathways, of which 13 have P values less than 0.05. 16S rDNA sequencing showed significant differences in 4 microbial communities at the phylum level. There are significant differences in 23 communities at the genus level. We focus on Lactobacillales, Lactobacillus, Streptococcus, Veillonella, Bacilli and Megasphaera. Correlation analysis shows that the intestinal microbiota and plasma metabolites jointly affect the occurrence and development of gastric cancer. Conclusion For the first time, we comprehensively used plasma metabolomics and 16S rDNA sequencing to reveal the changes and correlations between metabolites and intestinal microbiota in advanced gastric cancer. We have discovered new potential biomarkers for gastric cancer. This deepens our understanding of the physiological and pathological mechanisms of advanced gastric cancer and helps to improve the diagnosis and treatment of advanced gastric cancer.

Lymphoma is a class of malignant tumors originating from the lymphatic system, with a complex tumor microenvironment playing a critical role in tumor initiation, progression, and therapeutic response. In recent years, the rapid development of organoid technology has provided a novel platform for lymphoma research, enabling effective in vitro simulation of the lymphoma microenvironment. Although methods for establishing lymphoma organoids have gradually matured and related studies are increasing, challenges such as the lack of standardized protocols and incomplete microenvironment reconstruction remain. This review aims to summarize the techniques, characteristics, and applications of lymphoma organoids in tumor microenvironment research, exploring their potential in immunotherapy and disease mechanism studies. It seeks to offer new insights and directions for future lymphoma research.

Phytochemical investigation of the ethyl acetate extract of the stems of Celastrus orbiculatus Thunb. led to the isolation of a new neolignan, celastorbiol A (1), and two new homoisoflavones, celastrones A and B (2 and 3), along with five known 3-benzylchroman derivatives, including 7,4'-dihydrohomoisoflavanone (4), 3-deoxysappanone B (5), 3'-deoxysappanone A (6), bonducelin (7), and sappanone A (8). Their structures were determined based on extensive spectroscopic and spectrometric data analyses, and computational studies. Compounds 1, 6, and 8 exhibited cytotoxic activities against human gastric cancer AGS and HGC-27 cell lines with IC50 values that ranged from 42.82 ± 0.98 to 98.04 ± 1.33 µM.

Tumor‐intrinsic programmed cell death 1 (PD‐1) has been shown to activate the mesenchymal epithelial transition factor (MET) pathway via its phosphorylation in pancreatic ductal adenocarcinoma (PDAC). However, the immunoregulatory consequences of MET activation remain poorly understood. Herein, a significant positive correlation between phosphorylated MET (p‐MET) and tumor‐intrinsic PD‐1 is verified, both of which are independently associated with adverse prognosis. Elevated p‐MET levels correlated with diminished CD8 + T cell cytotoxicity and increased regulatory T cell (Treg) infiltration. Single‐cell RNA sequencing revealed MET activation selectively drives the accumulation of intratumoral GITR⁺ Tregs—a distinct effector Treg subset with potent immunosuppressive function and high prognostic relevance. Compared to KLF2⁺ naïve Tregs, GITR⁺ Tregs exhibited an activated phenotype and enhanced expression of immunoregulatory markers. Subgroup analysis further demonstrated that elevated GITR⁺ Treg infiltration diminished the prognostic utility of serum CA19‐9, underscoring the immunosuppressive dominance of this Treg subset. Mechanistically, MET–IL‐23–STAT4 axis orchestrates GITR⁺ Treg‐mediated immune evasion in PDAC. In vivo, MET inhibition and GITR agonism synergize to enhance antitumor immunity in an orthotopic PDAC model. Collectively, these findings highlight MET signaling and GITR⁺ Tregs as actionable targets to counteract immune evasion and improve the efficacy of immunotherapeutic strategies in PDAC.

ETHNOPHARMACOLOGICAL RELEVANCE:With the rapid development of China's economic level, great changes have taken place in people's diet structure, gout has become a common disease that puzzles people's health, seriously affects the realization of China's "Healthy China" strategic goal. Gouty arthritis (GA) is a common joint disease caused by chronic purine metabolism disorder. Currently, drugs used to treat GA are allopurinol and colchicine. However, these drugs can only temporarily relieve the clinical symptoms of GA with significant side effects. More and more basic and clinical studies have confirmed that Traditional Chinese medicine has definite curative effect on GA.AIM OF THE STUDY:To elucidate the potential molecular mechanism of Tongfengkang (TFK) in the treatment of GA, and to provide experimental basis for the search and development of efficient and low-toxicity Chinese medicine for GA treatment.MATERIALS AND METHODS:Aqueous extract of TFK (AETFK) were determined by liquid phase high resolution mass spectrometry and the possible effective constituents were screened out. Acute GA model rats were established to detect the anti-inflammatory and detumification effects of AETFK on GA and explore the potential mechanism. The effect of AETFK on serum uric acid and urinary uric acid levels in acute GA rats was determined by automatic biochemical analyzer, and the effect of AETFK on the expression of acute GA-related immunoinflammatory factors were determined by protein thermal fluorescence chip. The effect of AETFK on the concentration of neutrophils in the joint fluid of acute GA rats were determined by Reichs-Giemsa staining. The effect of AETFK on macrophage activation was detected by ELISA. In order to further investigate the mechanism of AETFK in the treatment of GA, a rat model of hyperuricemia was established to detect the effect of AETFK on the level of uric acid in hyperuricemia model rats. Biochemical indexes of liver and kidney and hematoxylin-eosin staining (HE) were used to evaluate the effects of AETFK on the organs, and to preliminatively evaluate the safety of ventilation confufang.RESULTS:Compared with the model group, the joint swelling degree of GA rats in AETFK treatment group were significantly reduced, and the levels of blood uric acid and urine uric acid were also significantly decreased. Protein thermal fluorescence microarray results showed that the levels of gout - related inflammatory factors in GA rats in AETFK treatment group were significantly lower than those in control group. Reichsen-giemsa staining and ELISA showed that AETFK could reduce the activation of macrophages and the accumulation of neutrophils in the joint fluid. The results of liver and kidney biochemical indexes and HE staining showed that no obvious tissue damage was observed in the organs of rats treated with AETFK.CONCLUSIONS:AETFK not only has significant anti-inflammatory effects on GA, but also can significantly reduce the level of blood uric acid in GA rats, without obvious toxic and side effects. These effects may be related to AETFK's inhibition of neutrophil enrichment and macrophage activation during early inflammation.

Background and AimsPrevious studies have demonstrated that peptidylarginine deiminase 4 (PAD4) functions as a suppressor, promoter, or both in cancer pathogenesis and therapeutic outcomes. Although PAD4 expression has been proposed to be one of the molecular features of gastric cancer (GC), the biological basis of PAD4 in GC progression and chemotherapy has not been formally established.MethodsCell type-preferential expression of PAD4 was analyzed in both preclinical and clinical models. The colocalization of PAD4 expression and tumor-infiltrating neutrophils in GC patients was evaluated by immunofluorescence assay. The effects of forced expression of PAD4 on GC cell proliferation were evaluated both in vitro and in vivo. The effects of forced expression of PAD4 on the cytotoxicity of 5-fluorouracil and oxaliplatin were performed by EdU, Annexin V/PI, and comet assays. Co-immunoprecipitation assay was used to investigate the endogenous and exogenous interaction between PAD4 and p53. To investigate whether p53 participated in the chemopotentiating effects of PAD4, small interfering RNA (siRNA)-mediated knockdown of p53 was conducted in PAD4-overexpressing GC cells.ResultsContrary to the previous report, we initially observed that PAD4 was underexpressed in GC patients and presented as a favorable prognostic factor across the TCGA cohort. Interestingly, the normal gastric epithelial cell line GES-1 exhibited low-level expression of PAD4. In comparison, PAD4 was not detected in GC cell lines, including AGS, HGC-27, and MKN-45. Using an orthotopic mouse model of GC, we found that PAD4 was surprisingly abundant in HGC-27 cell line-derived xenografts. Immunofluorescence staining for PAD4 and neutrophil elastase indicated that PAD4 was mainly expressed in neutrophils but not in GC cells. PAD4 expression was upregulated in all-trans retinoic acid-induced neutrophil-like dHL-60 cells, which were used as a positive control for PAD4 expression. Surprisingly, the enforced expression of PAD4 suppressed the proliferation of GC cells in vitro and in vivo. In addition, cells harboring high PAD4 expression were more susceptible to G1/S boundary arrest, apoptotic death, and DNA damage by regulating p53 target proteins. Mechanistically, PAD4 might affect p53 function through physical interaction with p53, and the chemopotentiating effects of PAD4 could be compromised by p53 knockdown.ConclusionsPAD4 appeared to be constitutively expressed in tumor-infiltrating neutrophils but not in GC cells. Our findings highlight unique roles of PAD4, in which origin-dependent PAD4 might work complementarily on the progression and treatment vulnerability of GC.

Lung cancer is the most common oncological disease worldwide, with non-small cell lung cancer accounting for approximately 85% of lung cancer cases. α-Hederin is a monodesmosidic triterpenoid saponin isolated from the leaves of Hedera helix L. or Nigella sativa and has been extensively studied for its antitumor activity against a variety of tumor cells. It has been suggested that α-Hederin is a potential regulator of autophagy and has high promise for application. However, the specific mechanism and characteristics of α-Hederin in regulating autophagy are not well understood. In this study, we confirmed the potential of α-Hederin application in lung cancer treatment and comprehensively explored the mechanism and characteristics of α-Hederin in regulating autophagy in lung cancer cells. Our results suggest that α-Hederin is an incomplete autophagy inducer that targets mTOR to activate the classical autophagic pathway, inhibits lysosomal acidification without significantly affecting the processes of autophagosome transport, lysosome biogenesis, autophagosome and lysosome fusion, and finally leads to impaired autophagic flux and triggers autophagic damage in NSCLC.

ETHNOPHARMACOLOGICAL RELEVANCE:Celastrus orbiculatus Thunb. is an ancient traditional Chinese herb with a long history of medicinal use. The ethyl acetate extract of Celastrus orbiculatus Thunb. (COE) has been shown to have anti-tumor effects in various preclinical studies. However, the anti-invasive and metastatic efficacy of COE in non-small cell lung cancer (NSCLC) and the mechanism by which COE regulates cellular oxidation levels are yet to be elucidated.AIM:To study the anti-dissemination effect of COE on NSCLC and to elucidate the molecular mechanism of COE in regulating cellular oxidation levels and its effect on lung cancer invasion and metastasis.METHODS:CCK-8 assay was used to detect the toxic effects of COE on NSCLC. Transwell assay and high-content imaging was used to detect the Motility of NSCLC. Transmission electron microscopy and three-dimensional (3D) imaging of mitochondrial fluorescence were employed to detect the number and structure of mitochondria. JC-1 probe was used to detect the level of mitochondrial membrane potential. Firefly luciferase assay was used to detect the level of total intracellular ATP. MitoSox probe and DCFH-DA probe were applied to detect the level of reactive oxygen species (ROS) inside the mitochondria and the total intracellular ROS, respectively. Immunohistochemistry was used to detect protein expression in xenograft tumors.RESULTS:COE inhibited motility and induced DJ-1 downregulation in NSCLC at low toxic concentrations, and the antiseptic effect of COE was reduced significantly after the overexpression of DJ-1. COE induced structural disruption of mitochondria in NSCLC and accumulation of superoxide compounds, decreased the volume of membrane potential depolarization, and impaired energy production, ultimately leading to a large accumulation of ROS at the cellular level. The antioxidant acetylcysteine (NAC) significantly reversed the antiseptic capacity of COE. In a xenograft tumor model, protein expression of DJ-1, E-cadherin, N-cadherin, and MMP-2 in COE group was significantly changed compared to the model group.CONCLUSION:In the present study, COE inhibited NSCLC invasion and metastasis and was associated with the downregulation of DJ-1 and elevated ROS. COE-mediated downregulation of DJ-1 may be the primary cause of mitochondrial structural and functional dysfunction in NSCLC, eventually leading to ROS accumulation.

In order to create an electrochemical sensor for doxorubicin (DXB), an effective cancer treatment, multi-walled carbon nanotubes—a modified glassy carbon electrode made of mesoporous silica nanocomposite—were created in this work. Examining the mSiO2@MWCNTs nanocomposite's structure and composition using FTIR, XPS, and SEM revealed the combination of mSiO2 and MWCNTs nanostructure on the modified electrode surface. Studies on the electrochemical determination of DXB utilizing cyclic voltammetry (CV) and differential pulse voltammetry (DPV) methodologies showed that mSiO2@MWCNTs/GCE exhibited excellent sensitivity, selectivity, and precision. MWCNTs and mesoporous silica in the improved electrode work together to improve the sensor's overall performance by enhancing stability and sensitivity in the electrochemical oxidation of DXB. The sensor's linear range was found to be 30–750 μM, with a sensitivity of 0.05123 μA/μM and a detection limit of 14 nM, according to the results. The mSiO2@MWCNTs/GCE shows properties suitable for real sample analysis. The creation of this sensor represents a significant advancement in the field of electrochemical sensors and may open the door to the development of more precise and focused drug detection techniques.

The lymphoma incidence rate is on the rise, with invasive forms particularly prone to relapse following conventional treatment, posing a significant threat to human life and wellbeing. Numerous studies have shown that traditional Chinese botanical drug medicine offers promising therapeutic benefits for various malignancies, with previous experimental findings indicating that Celastrus orbiculatus extract effectively combats digestive tract tumors. However, its impact on lymphoma remains unexplored. This study aims to investigate the impact and underlying mechanisms of COE on the proliferation and apoptosis of Burkitt lymphoma cells. We diluted COE in RPMI-1640 medium to create various working concentrations and introduced it to human Burkitt lymphoma Raji and Ramos cells. To evaluate cell viability, we used the CCK-8 assay, and we observed morphological changes using HE staining. We also conducted Annexin V-PI and JC-1 staining experiments to assess apoptosis. By combining the cell cycle experiment with the EDU assay, we gained insights into the effects of COE on DNA replication in lymphoma cells. Using Western blotting, we detected alterations in apoptosis-related proteins. In vivo experiments revealed that following COE intervention, tumor volume decreased, survival time was prolonged, spleen size reduced, and the expression of tumor apoptosis-related proteins changed. Our findings indicate that COE effectively inhibits lymphoma cell proliferation and promotes apoptosis by regulating these apoptosis-related proteins.

目的:探讨茯苓中的三萜类化合物茯苓酸(PA),对肾癌侵袭转移的作用及机制.方法:采用细胞增殖与活性检测-8(CCK-8)试剂盒检测PA(0、20、40、80、160μmol·L-1)对细胞存活率的影响,并筛选PA浓度用于后续实验.采用克隆形成实验评估PA(0、20、40、80 μmol·L-1)对细胞增殖的作用.细胞黏附实验用来观察PA(0、20、40、80 μmol·L-1)对细胞黏附能力的效果.划痕实验和Transwell侵袭实验用来检测PA(0、20、40、80μmol·L-1)对细胞侵袭转移的影响.高内涵成像技术进一步动态观察和验证PA(0、20、40、80 μmol·L-1)对细胞运动的抑制作用.PA(0、20、40、80 μmol·L-1)对细胞中侵袭转移相关蛋白基质金属蛋白酶/基质金属蛋白酶组织抑制因子(MMP/TIMP)和相关通路关键蛋白Smads表达的影响通过蛋白免疫印迹法(Western blot)来检测.结果:CCK-8结果显示,与空白组比较,PA组细胞存活率显著降低(P＜0.01),24 h时PA对ACHN细胞的半数抑制浓度(IC50)为70.42 μmol·L1.克隆形成实验显示,与空白组比较,PA组细胞克隆群数目显著减少(P＜0.01).细胞黏附实验显示,与空白组比较,PA组细胞的黏附数量显著减少(P＜0.01).划痕实验表明,与空白组比较,PA组细胞的划痕愈合率降低(P＜0.05,P＜0.01).Transwell侵袭实验显示,与空白组比较,PA组细胞的穿膜数量减少(P＜0.01).高内涵成像技术表明,与空白组比较,PA组细胞累计迁移距离更短(P＜0.01).Westernblot结果表明,与空白组比较,PA组MMP-2、MMP-9蛋白的表达降低(P＜0.01),TIMP-1蛋白的表达升高(P＜0.01).此外,与空白组比较,PA组Smad2、Smad3蛋白的表达降低(P＜0.01).结论:PA可抑制肾癌细胞的增殖、侵袭转移,其机制可能是通过Smad2/3调控MMP/TIMP的稳态平衡.