Search Results (635)

2-Amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is a major heat-induced contaminant in protein-rich foods, yet its effects on intestinal barrier homeostasis and luminal microecology remain insufficiently defined. In this study, adult zebrafish were exposed to dietary PhIP for 90 days at estimated intake doses of 0.006, 0.4, and 7.2 mg/kg bw/day to evaluate intestinal injury, microbial dysbiosis, and metabolic remodeling. PhIP exposure impaired growth-related indices and induced progressive intestinal lesions, accompanied by mucus barrier depletion, reduced goblet cell abundance, and downregulation of muc2. Tight junction integrity was disrupted, as indicated by decreased zo-1, occludin, and claudin1 expression, weakened ZO-1 and Claudin-1 immunofluorescence signals, and reduced tight junction-related protein levels. Serum LPS and intestinal pro-inflammatory cytokines were markedly elevated, whereas il-10 expression was suppressed, indicating increased endotoxin burden and inflammatory activation. 16S rRNA gene sequencing revealed Proteobacteria-enriched dysbiosis and exposure-associated shifts in candidate genera, including Chitinilyticum, Shewanella, Aeromonas, Acinetobacter, Microbacterium, and Reyranella. Untargeted metabolomics further identified luminal metabolic remodeling involving lipid-related compounds, organic acids, amino acid metabolism, arachidonic acid metabolism, the citrate cycle, and pathways related to choline and glycerophospholipid metabolism. Association analysis linked genus-level microbial variation and core pathway-related metabolites with LPS, inflammatory cytokines, and tight junction markers. These findings indicate that dietary PhIP exposure disrupts intestinal barrier homeostasis in parallel with Proteobacteria-related dysbiosis and luminal metabolic remodeling, providing an integrated microbiota-metabolite-barrier association framework for evaluating intestinal risks of heat-induced food contaminants. Full article

Background/Objectives: Diabetic cardiomyopathy (DCM) is largely driven by severe oxidative stress and calcium dyshomeostasis. We examined the targeted antioxidant and therapeutic effects of zinc sulfate (ZnSO₄) on contractile dynamics, oxidative damage, calcium turnover, and apoptosis/fibrosis in aged female rats with type 2 diabetes. Methods: Thirty-two aged female Wistar rats were divided into Control, Control + ZnSO₄, Diabetes (DM), and DM + ZnSO₄ groups. DM was induced via high-fat diet and 30 mg/kg streptozotocin. After a 4-week complication period, treatment groups received 10 mg/kg/day ZnSO₄ (i.p.) for 6 weeks. Left ventricular papillary muscle contraction, oxidative/antioxidant markers (MDA/GSH), and gene expressions (SIRT1, GLUT4, SERCA2a, RyR2, Cav1.2, PLN) were evaluated. Myocardial architecture, fibrosis, and apoptosis were analyzed immunohistochemically. In DM rats, contractile force (CF) and velocities (±dF/dtmax) significantly declined. Results: Concurrently, SIRT1, GLUT4, SERCA2a, RyR2, Cav1.2, and antioxidant GSH decreased, while oxidative lipid damage (MDA), PLN, Caspase-3 activity, Collagen I, and fibrosis increased (p < 0.001). ZnSO₄ treatment in diabetic rats acted as a potent antioxidant modulator; it restored redox balance, activated the SIRT1/GLUT4 pathway, protected calcium-handling proteins from oxidative degradation, and significantly improved contractile dynamics. It also preserved myocardial architecture by reducing apoptosis and fibrosis. In healthy rats, ZnSO₄ caused mild stress and early fibrosis. Conclusions: In conclusion, while inducing mild stress in healthy myocardium, zinc supplementation provides robust antioxidant protection in diabetic hearts. It activates SIRT1, suppresses oxidative damage, maintains calcium homeostasis, and restores contractile dynamics, demonstrating strong antioxidant therapeutic potential against DCM. Full article

Liancheng white ducks have a distinctive “white feathers, black beak, and green feet” phenotype, making them a useful model for studying pigmentation traits in waterfowl. The previous study found that the F1 generation of Liancheng white ducks crossed with white-feathered ducks and hemp-feathered ducks were all gray-black in color. This indicates the specificity and complexity of melanin deposition in Liancheng white ducks, which makes the selection and breeding of pigment traits through phenotyping difficult. The aim of this study was to investigate the candidate transcriptomic regulatory signals of melanogenesis in Liancheng white ducks. Skin, mouth skin, foot skin, liver, and muscle samples were collected from 130-day-old Liancheng white ducks. Morphological differences were observed via histological analysis, and extraction-based pigment levels were determined. The results showed that melanin granules were clearly observed in tissues other than the liver and were distributed mainly in the basal layer of the epidermis and around feather follicles; the pigment values in the tissues decreased in the order mouth skin > liver > foot skin > muscle and skin. However, the relatively high liver value should be interpreted cautiously because obvious melanin granule deposition was not observed histologically. Whole-transcriptome sequencing was performed on mouth skin and skin samples. In total, 3074 differentially expressed genes (DEGs) were screened; upregulated genes associated with melanogenesis included melanocyte inducing transcription factor (MITF) and tyrosinase (TYR); downregulated genes included agouti signaling protein (ASIP) and adenylate cyclase 2 (ADCY2). Eighteen differentially expressed microRNAs (DEmiRNAs) were identified. Based on target prediction and pathway enrichment analysis, novel_290 and apl-miR-11588-3p were identified as candidate miRNAs potentially associated with melanogenesis-related pathways, and their predicted target genes included phosphatidylinositol 3-kinase (PI3K) and Janus kinase 1 (JAK1). Additionally, 364 differentially expressed long noncoding RNAs (DElncRNAs) were identified; TCONS_00063335 and TCONS_00019814 were identified as candidate lncRNAs potentially associated with melanogenesis-related genes, including TYR and TYRP1. A putative ceRNA network was constructed based on the predicted miRNA–mRNA and miRNA–lncRNA relationships, and ENSAPLT00000025522–apl-miR-11588-3p–MAPK8IP3 was identified as a candidate network relationship associated with MAPK-related pigmentation pathways. However, because this relationship was inferred mainly from bioinformatic prediction and expression association analysis, further functional validation is required to confirm whether it contributes to melanogenesis regulation. These findings provide candidate transcriptomic and noncoding RNA information for the further investigation of tissue-specific pigmentation in Liancheng white ducks. Full article

In many university and healthcare projects, models are built for very different data types such as tables, institutional time series, and medical images, but they are deployed as separate applications. In this work, that separation made testing and maintenance difficult because each module had its own pipeline and runtime requirements. This paper presents an integrated AI lakehouse-style implementation that runs three model pipelines inside one containerized backend. For medical imaging, we used MRI datasets from IEEE DataPort: a four-class classification set with 7012 images (5708 train/1304 test) and a segmentation set with 3063 image–mask pairs. The classification model (ResNet50 transfer learning) is evaluated using a proper train–validation–test protocol across multiple splits (80/10/10, 70/10/20, 60/10/30, and 10/30/60), achieving a test accuracy of 99.00% under the standard 80/10/10 split. Additionally, a patient-level evaluation is conducted using an external glioma dataset to provide a more realistic assessment without data leakage. The segmentation model (DeepLabV3-ResNet50) achieved 83.09% validation mIoU and 88.79% Dice score. For university KPI forecasting, we used annual IPEDS and NSF HERD data from 2010 to 2023 for three universities (BSU, EOU, and UAB). To examine the effect of preprocessing on forecasting performance, two case studies are conducted. In the first case, linear interpolation is applied to generate semester-level data. In the second case, the original annual data is used directly without interpolation. Random Forest regression and ARIMA models are evaluated using MAE, RMSE, MAPE, and $R^2$. The results showed that interpolation improved apparent forecasting performance due to smoothing, while evaluation on the original annual data provided a more realistic assessment of model behavior. To further validate the framework on a larger dataset, an additional case study is conducted using a student dropout dataset. For water potability, we trained and compared multiple tabular classifiers on a large dataset (1,048,575 samples). A Random Forest model (100 trees, max depth 10) achieved 85.86% test accuracy and high recall for unsafe samples (0.8447). All modules are served via FastAPI and deployed together using Docker, with workflow automation routing requests to the correct endpoint. System-level benchmarking indicates that the backend maintains stable throughput and latency under concurrent requests. Full article

This narrative review synthesizes current knowledge on MADS-Box 27 (OsMADS27), a member of the AGL17 clade in rice that has emerged as a regulatory node linking nitrate signaling, root development, and abiotic stress tolerance. Because most functional and mechanistic studies on OsMADS27 to date have been conducted in rice, this review is centered on Oryza sativa, with cross-species comparisons used for evolutionary and comparative context. Specifically, we summarize the gene and protein structure, phylogenetic position, expression profile, upstream and downstream regulation, and emerging functional significance of OsMADS27. OsMADS27 is a typical MIKC-type MADS-box protein with root-preferential expression, and its activity is strongly influenced by nitrate availability and miR444-mediated regulation. Evidence from functional genomics, transcriptomics, ChIP-based studies, and transgenic analyses suggests that OsMADS27 contributes to the regulation of root architecture, nitrate uptake, hormonal crosstalk, and stress-responsive pathways. Notably, OsMADS27 enhances salt tolerance through nitrate-dependent activation of downstream targets such as OsHKT1;1 and OsSPL7, contributing to ion homeostasis and salinity tolerance. Recent findings also suggest roles in grain size regulation and yield improvement, expanding its significance beyond root biology. This review compares OsMADS27 with AGL17-clade genes and highlights its value for crop improvement aimed at salinity tolerance and nitrogen use efficiency. However, important research gaps remain, particularly the limited field-level validation, the absence of integrated multi-omics analyses, and the lack of functional studies of OsMADS27 orthologs in non-rice crops. Overall, OsMADS27 represents promising rice-centered target for future biotechnology applications, while its translational relevance to other cereals remains to be established through orthology analysis and field-level evaluation. Full article

Fatty liver disease represents a major metabolic disorder affecting domestic animals worldwide, with significant implications for animal health, welfare, and agricultural productivity. Disrupted communication between mitochondria and other organelles—particularly the endoplasmic reticulum, lipid droplets, and lysosomes—plays a critical role in disease pathogenesis. This review synthesizes knowledge on inter-organellar communication across domestic animals, with emphasis on species-specific adaptations. We address the “Dairy Cow Paradox”—periparturient dairy cows develop severe hepatic steatosis (>30% liver fat), yet under sterile conditions, they have a higher threshold for progressing to sterile steatohepatitis compared to rodents and humans. However, it is critical to note that severe fatty liver in dairy cows is indeed associated with impaired autophagy, inflammation, and liver damage, particularly when accompanied by ketosis or concurrent infections, and 39% of transition cows exhibit moderate to severe lymphocytic hepatitis. We propose that the tolerance to severe steatosis in dairy cows arises from three adaptations: (1) attenuated innate immune sensing via the cGAS-STING pathway; (2) enhanced lipid buffering from perilipin 5 (PLIN5) with a hypothesized ruminant-specific Val152 substitution that may stabilize lipid droplet–mitochondria contacts; and (3) dampened calcium signaling due to ER–mitochondria membrane lipid raft rigidity, elevated inositol 1,4,5-trisphosphate receptor 2 (IP₃R2) expression, and reduced mitochondrial calcium uniporter (MCU) conductance. We contrast this with the inflammatory steatohepatitis common in rodent models driven by calcium overload and mitochondrial DNA (mtDNA) release, and glucocorticoid-mediated mitofusin 1 (MFN1) suppression, causing mitochondrial fragmentation in poultry. We identify critical knowledge gaps, including the need to define bovine and avian mitochondria-associated endoplasmic reticulum membrane (MAM) proteomes and spatially resolve hepatic zonal communication patterns. Targeting organellar communication hubs with nutraceuticals or pharmacological agents offers promising therapeutic strategies. Full article

Efficient water resource management is critically important for arid regions such as southern Kazakhstan. This paper presents a hybrid Internet of Things (IoT) and Vision-based Internet of Things (VIoT) architecture for real-time monitoring of water levels in irrigation channels. The proposed system integrates an ultrasonic water-level sensor, an IP camera with edge-based computer vision processing on a Raspberry Pi, wireless communication, an autonomous solar power supply, and discharge estimation using Manning’s equation. The VIoT subsystem applies image processing techniques, including gauge calibration, Canny edge detection, and pixel-to-metric conversion, to automatically estimate water level from captured video frames. Water-level measurements obtained from IoT sensors and video-based analysis are combined through synchronised data fusion to improve monitoring accuracy and reliability. The hybrid approach leverages the complementary strengths of IoT and VIoT by combining continuous quantitative sensing with visual verification capabilities. Field experiments conducted on the Merke River in the Zhambyl region of Kazakhstan over a 14-day observation period demonstrated stable real-time operation with RMSE = 0.311 cm, MAE = 0.279 cm, and Pearson r = 0.99 between the ultrasonic sensor and the vision-based estimates. Sensitivity analysis indicated that water level is the most influential parameter in Manning-based discharge estimation, confirming the importance of accurate level detection. The proposed system improves reliability by cross-checking independent data sources, making it applicable to monitoring water levels in agricultural regions. Full article

Background: Hydroxysafflor Yellow A (HSYA), the major bioactive component from Carthamus tinctorius L., exerts significant protective effects against myocardial ischemia-reperfusion injury (MIRI). Mitophagy is pivotal in the pathological process of MIRI, yet the specific molecular mechanism underlying HSYA-mediated mitophagy regulation remains unclear. Objective: This study aimed to investigate the association between HSYA treatment and mitochondrial autophagy in murine MIRI and to explore the potential mechanistic role of the SIRT1-FOXO3-BNIP3 signaling pathway using functional loss-of-function and rescue experiments. These findings may provide preliminary evidence supporting the clinical translational potential in MIRI therapy. Methods: Mouse myocardial ischemia-reperfusion injury (MIRI) model and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced AC16 cardiomyocyte injury models were established. Metabolomics, molecular docking, and surface plasmon resonance (SPR) techniques were combined to screen the potential targets of HSYA. The SIRT1 inhibitor EX527 and SIRT1 siRNA were used to verify the underlying mechanism. Cardiac function, myocardial infarct size, mitochondrial function, the expression of autophagy-related proteins, and protein–protein interaction were detected and analyzed. Results: Compared with the MIRI group, HSYA significantly improved cardiac function in mice, as evidenced by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p < 0.01), attenuated ST-segment elevation, and improved myocardial perfusion. HSYA also markedly reduced myocardial infarct size (p < 0.01) and serum levels of CK-MB, LDH, and cTnI (all p < 0.01) and ameliorated myocardial histopathological damage and mitochondrial ultrastructural integrity. Mechanistic studies revealed that HSYA significantly upregulated the expression of SIRT1, FOXO3, BNIP3, Beclin-1, and the LC3II/I ratio while downregulating p62 expression (p < 0.01), consistent with enhanced mitophagy-related activity. Furthermore, these protective effects were markedly attenuated upon SIRT1 inhibition or siRNA-mediated silencing, whereas HSYA intervention partially reversed these alterations. Additionally, co-immunoprecipitation (Co-IP) and pull-down assays demonstrated that HSYA promoted protein–protein interactions between SIRT1-FOXO3, FOXO3-BNIP3, and BNIP3-LC3B. Conclusions: These findings highlight that HSYA is associated with improved cardiac function, enhanced mitophagy-related activity, and upregulated SIRT1-FOXO3-BNIP3 signaling, providing robust experimental evidence for its clinical translational application in MIRI treatment. Full article

Sulfane sulfur species are increasingly recognized as integral cellular components involved in signaling pathways and cytoprotection against oxidative stress in mammals. While their production in bacteria has been extensively studied, their functional role in bacterial oxidative stress defense remains poorly understood. Here, we demonstrate that sulfane sulfur generated by sulfide: quinone oxidoreductase decreases H₂O₂ sensitivity in Pseudomonas aeruginosa PAO1. Notably, this protective mechanism does not depend on sulfane sulfur acting as a direct H₂O₂ scavenger via nucleophilic reactions. Through persulfidation proteomic profiling, we reveal that persulfidation is a prominent post-translational modification in P. aeruginosa, reflecting the prevalence of deprotonated sulfane sulfur species. These species modify cysteine residues in proteins, including the well-known oxidative stress regulator OxyR. Specifically, sulfane sulfur modifies OxyR at Cys199 to form persulfidated OxyR C199-SSH, contributing to a single-Cys activated state that modulates promoter activity and DNA-binding affinity. Furthermore, sulfane sulfur-mediated persulfidation protects the critical cysteine residue of LpdG, a ROS-vulnerable dihydrolipoamide dehydrogenase, from irreversible overoxidation. Although LpdG is not part of the canonical H₂O₂-scavenging system, its preservation is essential for cell viability under oxidative stress. These findings establish endogenous sulfane sulfur species as key mediators of antioxidant defense in P. aeruginosa. Full article

Bone marrow-derived mesenchymal stem cells (BMSCs) maintain skeletal homeostasis by balancing adipogenic and osteogenic differentiation, yet clinically used drugs that bias this fate choice and their mechanisms remain incompletely defined. Here, we investigated whether dextromethorphan (DXM), a widely used antitussive, modulated lineage commitment in rat BMSCs and interrogated candidate upstream signaling modules. Rat BMSCs were induced with adipogenic medium or osteogenic medium in the presence of DXM (30 μM). Adipogenesis and osteogenesis were quantified using Oil Red O and Alizarin Red S staining with elution-based quantification, and lineage markers were measured by RT-qPCR. Intracellular Ca²⁺ and ROS were analyzed using flow cytometry, and the levels of p-AKT and p-ERK were assessed through Western blotting analysis. Under adipogenic induction, DXM increased lipid droplet accumulation and the mRNA levels of Pparγ and Fabp4. Although DXM elevated Ca²⁺ and ROS, the chelation of intracellular Ca²⁺ and pharmacological inhibition of Sig-1R/PLC–IP3R signaling, redox/ROS, NMDA receptors, AKT/ERK, Kv channels, bitter taste receptor-related signaling, and mTOR did not attenuate the DXM-enhanced adipogenesis. DXM reduced p-ERK without increasing p-AKT; U0126 lowered basal adipogenesis but did not block the DXM effect. Under osteogenic induction, DXM reduced matrix mineralization and downregulated Runx2 and Bglap mRNA levels, while Wwtr1 mRNA levels were not significantly changed. DXM also partially reversed the osteogenic induction-associated reduction in Mtor mRNA. Separately, under adipogenic induction, rapamycin attenuated baseline adipogenesis but did not prevent the additional lipid accumulation induced by DXM. Collectively, DXM shifted the osteogenic–adipogenic balance toward adipogenesis through a non-canonical mechanism. Full article

Pseudogenes are barely transcribed normally, but some of them are transcribed as long non-coding RNAs during tumorigenesis. The pathological features of pseudogenes in hepatocellular carcinoma (HCC) have not been illustrated clearly. Here, we engaged the pseudogene DUXAP8 into a feedback loop affecting the HCC indicator Piezo1 and concerning the HCC tumor microenvironment (TME). As we discovered, the DUXAP8 transcript was detectable in HCC. The transcriptional activity of DUXAP8 in HCC is associated with dismal HCC clinicopathological features. By depleting DUXAP8, the HCC cells presented the inhibition of cell proliferation along with significant cell apoptosis in vitro and potently suppressed the HCC tumorigenesis ability in vivo. Combined with the ChIP assay, the direct interaction of either the DUXAP8 transcript/miR-214-3p or miR-214-3p/KLF13 mRNA was verified, and the promoting effect of KLF13 on DUXAP8 transcription was also validated, which further illustrates a positive feedback loop of DUXAP8/miR-214-3p/KLF13. Moreover, KLF13 was found to facilitate Piezo1 transcription. As concluded, our findings suggest that the pseudogene DUXAP8 promotes HCC tumorigenesis through the feedback loop of DUXAP8/miR-214-3p/KLF13 and participates in HCC TME modulation by impacting Piezo1. Full article

Histone lactylation acts as a master regulator in tumor development, but its role in a noncoding RNA (ncRNA) network remains unclear. This study aims to reveal the interaction between H3K18la and the lncRNA-miRNA-mRNA regulatory network in hepatocellular carcinoma (HCC). Transcriptome sequencing and ChIP sequencing were performed in HCC and adjacent normal tissues. Cut&Run and qPCR were used to validate the H3K18la enrichment on LINC02732 and CD44 promoter. Dual luciferase reporter assay, qPCR and Western blotting were used to verify the LINC02732-miR-1291-ASF1B axis. Co-Immunoprecipitation was performed to validate ASF1B recruiting p300. CCK8 and mouse subcutaneous tumor formation were performed to demonstrate this axis promoting HCC. H3K18la enrichment on LINC02732 promoter elevates its expression in both HCC samples and cell lines, therefore enhancing ASF1B expression via sponging miR-1291. Moreover, ASF1B, a histone chaperone, promotes H3K18la by recruiting lactyltransferase p300, forming an ASF1B-H3K18la positive feedback loop. The axis upregulates CD44 expression and promotes HCC in vitro and in vivo. These findings demonstrated the influence of H3K18la on the LINC02732-miR-1291-ASF1B axis and the novel role of ASF1B in histone lactylation by recruiting p300, which together promoted HCC proliferation. Full article

Accurate prediction of Rate of Penetration (ROP) in carbonate formations remains constrained by the arbitrary selection of geomechanical input parameters in empirical drilling models. This study presents the first systematic field-based evaluation of sixteen geomechanical properties—grouped into three categories: strength parameters (uniaxial compressive strength (UCS), confined compressive strength (CCS), shear strength, thick-walled cylinder strength (TWC), friction angle, and cohesion), elastic moduli (Young’s modulus, shear modulus, bulk modulus, bulk compressibility, dynamic combined modulus (DCM), Poisson’s ratio, brittleness index), and in situ stress parameters (overburden pressure, minimum, and maximum horizontal stresses)—to identify optimal predictors for ROP modeling across PDC bit sizes of 12.25″ and 8.5″. Continuous wireline log data from two vertical carbonate wells in the Middle East (Well A: 1000–3370 m; Well B: 1945 to 3128 m; total intervals of 2370 m and 1183 m, respectively) penetrating formations comprising limestone, dolomite, sandstone, shale, anhydrite, and marly limestone were used. All sixteen geomechanical properties were computed using Interactive Petrophysics (IP) software with lithology-specific empirical correlations and validated against laboratory core measurements (R² = 0.79–0.95). Pearson and Spearman correlation analyses quantified parameter–ROP relationships, and the Al-Abduljabbar empirical model, recalibrated via multiple nonlinear regression, served as the evaluation framework. DCM consistently exhibited the strongest negative correlation with ROP across both bit sizes and achieved the highest model accuracy (R² = 0.54, AAPE = 25.33%), significantly outperforming the Bourgoyne and Young model (R² = 0.26, AAPE = 36.55%). A statistically validated scale-dependent effect was identified: Fisher’s Z-transformation tests confirmed that the correlation reversal between CCS and UCS across bit sizes is statistically significant (CCS: Z = −16.84, p < 0.001; UCS: Z = −6.75, p < 0.001), establishing CCS as the superior predictor at 12.25″ and UCS as the superior predictor at 8.5″—a finding not previously reported in the ROP literature. This reversal is attributed to the larger contact area of the 12.25″ bit, which promotes confinement-dominated rock failure better described by CCS, whereas the smaller bit produces localized stress concentration better represented by UCS. These results establish that (1) optimal geomechanical input selection is bit-size dependent, (2) nonlinear modeling outperforms linear frameworks for strength–ROP relationships, and (3) parameter relevance outweighs coefficient tuning in model robustness. DCM is recommended as the most operationally practical universal input, requiring only conventional compressional sonic and density logs. This study provides a systematic framework for geomechanical parameter selection with direct implications for drilling optimization in heterogeneous carbonate reservoirs. Full article

Pterostilbene (Pts), a small molecule stilbenoid and a dimethyl analogue of the star molecule resveratrol, exerts significant blood–brain barrier protection on cerebral ischemia-reperfusion injury and has received extensive attention. This study performed structural optimizations on Pts to obtain a series of derivatives and investigated their anti-ischemic activities both in vitro and in vivo, aiming to identify candidates with high safety and improved efficacy compared with Pts. The ADMET method was used to predict the drug-likeness of a series of Pts derivatives, and in vitro MTT cell viability analysis was conducted on neuroblastoma cells (SH-SY5Y) and brain microvascular endothelial cells (BMECs) after oxygen-glucose deprivation/reperfusion (OGD/R) injury. On the basis of the cytotoxicity results, four derivatives (NO. 1, NO. 3, NO. 5, and NO. 7) were selected for subsequent in vitro and in vivo biological activities evaluation. These compounds exhibited significantly higher TI values (18.29–30.61) in OGD/R-injured hBMECs compared with Pts (7.63) and effectively suppressed apoptosis, promoted cell migration, and enhanced tube formation capacity. In vivo, NO. 3 (5 mg/kg, ip., 7 d) demonstrated superior efficacy compared to Pts in improving cerebral blood flow, reducing infarction volume, enhancing neurological function, and modulating serum biomarker levels in middle cerebral artery occlusion/reperfusion (MCAO/R) rats, whereas NO. 1 and NO. 7 showed comparable efficacy to Pts. The acute intraperitoneal toxicity of NO. 3 was conducted and showed that the LD₅₀ of NO. 3 was estimated to be more than 300 mg/kg. In this study, the rational design and comprehensive evaluation of Pts derivatives were reported. Compound NO. 3 demonstrated superior pharmacological efficacy to Pts both in vitro and in vivo, and it may be a promising therapeutic candidate for ischemic stroke intervention. Full article

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive malignancy characterized by therapeutic resistance and poor prognosis, underscoring the need for new therapeutic strategies. Bufalin, a major bioactive constituent of Venenum bufonis, has shown antitumor activity in several cancer types; however, its mechanism of action in PDAC remains incompletely defined. In this study, we investigated the antitumor effects of bufalin in PDAC using in vitro assays, mouse tumor models, and integrative transcriptomic, proteomic, metabolomic, and bioinformatic analyses. Bufalin inhibited PDAC cell viability, clonogenic growth, migration, and tumor progression in vivo. Pharmacological rescue experiments indicated that ferroptosis contributes importantly to bufalin-induced cytotoxicity, although apoptosis- and pyroptosis-related pathways may also be involved. Multi-omics analyses revealed coordinated alterations in calcium homeostasis, endoplasmic reticulum (ER) stress/unfolded protein response (UPR) signaling, and ferroptosis-related metabolic pathways. Further experiments showed that bufalin was associated with disrupted intracellular Ca²⁺ homeostasis, IP₃R-linked ER Ca²⁺ release, activation of PERK/eIF2α/ATF4 signaling, increased ATF3 expression, reduced SLC7A11 and GPX4 expression, glutathione depletion, and enhanced lipid peroxidation. Molecular docking and surface plasmon resonance assays supported an in vitro physical interaction between bufalin and IP₃R1/IP₃R3, while inhibition of ER stress attenuated several bufalin-induced ferroptosis-related phenotypes. Bioinformatic analyses further showed that higher ER stress and ferroptosis signature scores were associated with improved overall survival in PDAC, and concurrent activation of both signatures was linked to the most favorable prognosis. Collectively, these findings support that bufalin suppresses PDAC progression through coordinated ER stress- and ferroptosis-related responses, highlighting ER stress-ferroptosis crosstalk as a potential therapeutic vulnerability in PDAC. Full article