OBJECTIVE: To explore the mechanism of crocin, a major active component of Crocus sativus (Zanghonghua), in regulating amyloid beta (Aβ) generation, endoplasmic reticulum (ER) stress, and autophagy in neuronal cells, with potential therapeutic applications in Alzheimer's disease (AD). METHODS: Mouse neuroblastoma Neuron2a (N2a) cells stably transfected with the human amyloid precursor protein (APP) Swedish mutant was used as a cellular model for AD (N2a/APP). Control cells were vector transfected (N2a/vector). The effects of 3 different doses of crocin on reactive oxygen species (ROS) generation, cytosolic calcium, and apoptosis were evaluated by flow cytometry. Aβ levels were determined by enzyme-linked immunosorbent assay. APP processing and ER stress proteins expressions were determined by Western blot. Autophagosome formation was evaluated by autophagy detection kit and confocal microscope. RESULTS: Crocin inhibited APP expression in N2a/APP cells and promoted α-cleavage of APP processing, while modestly reduced beta-secretase 1 (BACE1) and presenilin 1 (PS1, P<0.05 or P<0.01). ER stress markers, including the binding immunoglobulin protein/78-kD glucose-regulated protein (Bip/GRP78) and C/EBP homologous protein (CHOP), were elevated in N2a/APP cells compared to N2a/vector cells (P<0.05). Crocin could effectively reduce the levels of ER stress (P<0.05 or P<0.01). In addition, crocin enhanced autophagy by promoting formation of autophagosome (P<0.05 or P<0.01). CONCLUSION Crocin significantly inhibited Aβ generation by promoting α-cleavage of APP processing, inhibiting ER stress-associated unfolded protein response, and regulating autophagy.
Endoplasmic Reticulum Stress/drug effects* ; Autophagy/drug effects* ; Animals ; Endoplasmic Reticulum Chaperone BiP ; Mice ; Amyloid beta-Peptides/metabolism* ; Amyloid beta-Protein Precursor/metabolism* ; Carotenoids/pharmacology* ; Humans ; Cell Line, Tumor ; Reactive Oxygen Species/metabolism* ; Apoptosis/drug effects* ; Calcium/metabolism*

Tumor cell-intrinsic programmed death-ligand 1 (PD-L1) signals mediate tumor initiation, progression and metastasis, but their effects in ameloblastoma (AM) have not been reported. In this comprehensive study, we observed marked upregulation of PD-L1 in AM tissues and revealed the robust correlation between elevated PD-L1 expression and increased tumor growth and recurrence rates. Notably, we found that PD-L1 overexpression markedly increased self-renewal capacity and promoted tumorigenic processes and invasion in hTERT⁺-AM cells, whereas genetic ablation of PD-L1 exerted opposing inhibitory effects. By performing high-resolution single-cell profiling and thorough immunohistochemical analyses in AM patients, we delineated the intricate cellular landscape and elucidated the mechanisms underlying the aggressive phenotype and unfavorable prognosis of these tumors. Our findings revealed that hTERT⁺-AM cells with upregulated PD-L1 expression exhibit increased proliferative potential and stem-like attributes and undergo partial epithelial‒mesenchymal transition. This phenotypic shift is induced by the activation of the PI3K-AKT-mTOR signaling axis; thus, this study revealed a crucial regulatory mechanism that fuels tumor growth and recurrence. Importantly, targeted inhibition of the PD-L1-PI3K-AKT-mTOR signaling axis significantly suppressed the growth of AM patient-derived tumor organoids, highlighting the potential of PD-L1 blockade as a promising therapeutic approach for AM.
Ameloblastoma/metabolism* ; Humans ; B7-H1 Antigen/metabolism* ; Neoplasm Recurrence, Local/pathology* ; Signal Transduction ; Cell Proliferation ; Up-Regulation ; TOR Serine-Threonine Kinases/metabolism* ; Proto-Oncogene Proteins c-akt/metabolism* ; Telomerase/metabolism* ; Jaw Neoplasms/metabolism* ; Epithelial-Mesenchymal Transition ; Animals ; Cell Line, Tumor ; Female ; Male

Objective:To explore the relationship between international collaboration papers and academic impact in global health,using the member universities of the Chinese Consortium of Universities for Global Health(CCUGH)as a case study.Methods:The study focuses on journal articles in global health field published by 31 CCUGH member universities between 2014 and 2024.Descriptive statistical analysis of international and non-international collaboration publication volumes was conducted using Excel.Regression analysis and chi-square tests were performed using R to examine the relationship between international collaboration papers and academic impact,and the correlation between the breadth of collaboration and the academic impact of the papers.Results:From 2014 to 2023,the total number of publications,the number of non-international collaborationpublications,and the number of internationally collaborated publications all showed a consistent annual increase,with average annual growth rates of 56.7%,68.3%,and 41.4%,respectively.By the first half of 2024,the total number of publications had increased to 1.5 times that of the corresponding period in 2023.International collaboration positively influenced academic impact,with broader collaborative networks correlating with higher academic influence.Conclusion:The global health publication output of CCUGH member universities has steadily increased,but the volume of international collaboration papers and their proportion remain relatively low.Therefore,it is necessary for CCUGH member universities to strengthen international collaboration papers in global health.

Objective:To explore the impacts of carnosic acid(CA)on learning and cognitive function in epileptic rats and the mechanism of regulating PKA-CREB signaling pathway in this process.Methods:Epilepsy rat models were prepared by intraperi-toneal injection of 50 mg/kg PTZ.After successful modeling,the rats were randomly grouped into control group,model group,CA low-dose group,CA high-dose group,CA high+PKA inhibitor group(CA high+H-89 group),with 15 rats in each group.The epileptic be-havior of rats in each group was evaluated;Morris water maze experiment was applied to test the cognitive ability of rats;HE staining and Nissl staining were applied to detect the morphology of hippocampal tissue and neurons of rats in each group;ELISA was applied to detect the contents of IL-1β,TNF-α,MDA,and the SOD activity in the hippocampus of rats in each group;and Western blot was applied to detect the protein expressions of PKA,p-PKA,CREB,p-CREB and BDNF in the hippocampus of rats in each group.Results:Compared with control group,the hippocampal neurons of rats in model group were irregularly arranged,with pyknosis of the nuclei,and the number of neurons decreased,the epilepsy frequency,Racine grading,escape latency,time of first platform crossing,and levels of IL-1β,TNF-α and MDA all increased,the frequency of crossing the platform,SOD activity,p-PKA/PKA,p-CREB/CREB,and the expression of BDNF protein all reduced(P<0.05);compared with model group,the arrangement of neuronal cells in hippocampal tissue of rats in CA group gradually became orderly,nuclear pyknosis gradually decreased,and the number of neurons increased,the epilepsy frequency,Racine grading,escape latency,time of first platform crossing,and levels of IL-1β,TNF-α and MDA all reduced,the frequency of crossing the platform,SOD activity,p-PKA/PKA,p-CREB/CREB,and the expression of BDNF protein all increased(P<0.05);further injection of PKA inhibitors on the basis of high-dose CA treatment showed that the improvement effects of high-dose CA on cognitive dysfunction,neuronal damage,inflammation,and oxidative stress in epileptic rats were reversed(P<0.05).Conclusion:CA can improve the learning and cognitive function of epileptic rats by activating the PKA-CREB signaling pathway.

Objective:To analyze the differentially expressed messenger RNA(mRNA), microRNA(miRNA), and long non-coding RNA(lncRNA)during the browning of mouse subcutaneous adipose tissue, construct a competitive endogenous RNA(ceRNA)network, and provide a theoretical basis for investigating the regulatory mechanisms of white adipose tissue browning.Methods:A cold-stimulated mouse model was established for transcriptome sequencing.Bioinformatics tools were employed to screen for differentially expressed mRNAs, miRNAs, and lncRNAs.An integrated mRNA-miRNA-lncRNA analysis was performed to construct a ceRNA network.Gene Ontology(GO), Kyoto Encyclopedia of Genes and Genomes(KEGG), and Gene Set Enrichment Analysis(GSEA)were conducted on the differentially expressed mRNAs and ceRNA networks to explore transcriptional regulation during the cold-induced browning of subcutaneous adipose tissue.Results:Transcriptomic analysis of the cold-stimulated model identified 4, 256 differentially expressed RNAs, which include 3, 600 mRNAs, 588 lncRNAs, and 68 miRNAs.GO and KEGG analyses revealed that the browning of white adipose tissue involves immune-related processes, such as immune system processes, immune responses, adaptive and innate immune responses, and the positive regulation of T-cell activation.A ceRNA network associated with browning regulation was constructed, comprising 233 nodes(188 mRNAs, 34 miRNAs, and 11 lncRNAs)and 351 edges.Protein-protein interaction(PPI)analysis of the mRNAs within the ceRNA network highlighted pathways including apoptosis, intracellular signaling transduction, hypoxia-inducible factor-1(HIF-1), AMP-activated protein kinase(AMPK), Janus kinase-signal transducer and activators of transcription(JAK-STAT)signaling, carbon metabolism, glycolysis, and thyroid hormone pathways, all of which regulate lipid metabolism, hypoxia, and glycolysis.Cytohubba analysis identified the top 10 hub genes: Bcl2, Src, Cebpb, Creb1, Runx1, Foxo3, Ets1, Socs3, Slc2 a4, and Pkm. Conclusions:The ceRNA network that regulates the browning of white adipose tissue is involved in various pathways, including carbon metabolism, glycolysis, thyroid hormone signaling, growth hormone signaling, prolactin signaling, as well as the HIF-1, AMPK, and JAK-STAT pathways.Key regulatory miRNAs in this context include miR-30e-5p, miR-182-5p, miR-20b-5p, miR-144-3p, miR-363-3p, miR-141-3p, miR-203-3p, and miR-107-3p.These miRNAs may serve as critical targets for inducing browning in response to cold exposure.

AIM To explore the clinical effects of Supplemented Buyang Huanwu Decoction on postoperative patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern.METHODS One hundred and twenty patients were randomly assigned into control group(60 cases)for 6-week intervention of conventional treatment,and observation group(60 cases)for 6-week intervention of both Supplemented Buyang Huanwu Decoction and conventional treatment.The changes in clinical effects,TCM syndrome scores,spinal cord conduction signals(SEP amplitude,MEP amplitude),serum neurotrophic factors(NGF,IGF-1,BDNF),coagulation and inflammatory indices(PT,APTT,TNF-α,IL-1 β)and incidence of adverse reactions were detected.RESULTS The observation group demonstrated higher total effective rate than the control group(P＜0.05).After the treatment,the two groups displayed decreased TCM syndrome scores,TNF-α,IL-1β(P＜0.05),increased spinal cord conduction signals,coagulation and inflammatory indices(P＜0.05),and shortened PT,APTT(P＜0.05),especially for the observation group(P＜0.05).No significant difference in incidence of adverse reactions was found between the two groups(P＞0.05).CONCLUSION For the patients with lumbar vertebral fracture complicated with spinal cord injury due to Qi Deficiency and Blood Stasis Pattern,Supplemented Buyang Huanwu Decoction can safely and effectively promote neurological function recovery.

Glycoengineering was carried out in the mammalian cell line CHO for the production of pro-tein-based drugs.Firstly,the genome sequence of the Rosa26 locus of CHO cells was determined,the gRNA sequences were designed,and the landing pad was integrated into the Rosa26 locus of CHO cells by CRISPR/Cas9 technology.Three targeting vectors co-expressed by glycosyltransferases,which are β-1,4 galactosyltransferase(B4GALT1),α-2,6-sialyltransferase 1(ST6GAL1)and N-acetaminoglycosyl-transferase Ⅲ(GnT Ⅲ),were constructed by overlapping PCR and seamless ligation technology,and the three glycosyltransferase genes were integrated into the CHO Rosa26 locus by Cre enzyme-mediated cassette exchange technology.PCR confirmed that three glycosyltransferases had been successfully site-directed integrated into the Rosa26 site.The mRNA expression levels of the three glycosyltransferases were more than 50 000-fold by qRT-PCR,and the protein expression levels of the three glycosyltrans-ferases were more than 4-fold via western blotting(P＜0.001).A CHO-engineered cell line with three glycosyltransferases integrated into Rosa26 site was successfully constructed.

Background: and Purpose Symptomatic intracranial hemorrhage (sICH) following endovascular thrombectomy (EVT) is a severe complication associated with adverse functional outcomes and increased mortality rates. Currently, a reliable predictive model for sICH risk after EVT is lacking. Methods: This study used data from patients aged ≥20 years who underwent EVT for anterior circulation stroke from the nationwide Taiwan Registry of Endovascular Thrombectomy for Acute Ischemic Stroke (TREAT-AIS). A predictive model including factors associated with an increased risk of sICH after EVT was developed to differentiate between patients with and without sICH. This model was compared existing predictive models using nationwide registry data to evaluate its relative performance. Results: Of the 2,507 identified patients, 158 developed sICH after EVT. Factors such as diastolic blood pressure, Alberta Stroke Program Early CT Score, platelet count, glucose level, collateral score, and successful reperfusion were associated with the risk of sICH after EVT. The TREAT-AIS score demonstrated acceptable predictive accuracy (area under the curve [AUC]=0.694), with higher scores being associated with an increased risk of sICH (odds ratio=2.01 per score increase, 95% confidence interval=1.64–2.45, P<0.001). The discriminatory capacity of the score was similar in patients with symptom onset beyond 6 hours (AUC=0.705). Compared to existing models, the TREAT-AIS score consistently exhibited superior predictive accuracy, although this difference was marginal. Conclusions The TREAT-AIS score outperformed existing models, and demonstrated an acceptable discriminatory capacity for distinguishing patients according to sICH risk levels. However, the differences between models were only marginal. Further research incorporating periprocedural and postprocedural factors is required to improve the predictive accuracy.

This study aimed to explore the molecular mechanism of rubioncolin C(RuC) in inhibiting gastric cancer(GC). AGS and MGC803 cell lines were selected as cellular models. After treating the cells with RuC at different concentrations, the effects of RuC on the proliferation ability of GC cells were assessed using the CCK-8 method, real-time cellular analysis(RTCA), and colony formation assays. Transmission electron microscopy was used to observe subcellular structural changes. Immunofluorescence was applied to detect LC3 fluorescent foci. Acridine orange staining was used to evaluate the state of intracellular lysosomes. Western blot was employed to detect the expression of autophagy-related proteins LC3Ⅱ, P62, and lysosomal cathepsin D(CTSD). The SuperPred online tool was used to predict the target proteins that bound to RuC, and molecular docking analysis was conducted to identify the interaction sites between RuC and CTSD. The drug affinity responsive target stability(DARTS) assay was performed to detect the direct binding interaction between RuC and CTSD. The results showed that RuC significantly inhibited the proliferation and colony formation of GC cells at low concentrations, with 24-hour half-maximal inhibitory concentrations(IC_(50)) of 3.422 and 2.697 μmol·L~(-1) for AGS and MGC803 cells, respectively. After 24 hours of treatment with RuC at concentrations of 1, 2, and 3 μmol·L~(-1), the colony formation rates for AGS cells were 61.0%±1.5%, 28.0%±0.5%, and 18.2%±0.5%, respectively, while the rates for MGC803 cells were 56.0%±0.5%, 23.3%±1.0%, and 11.8%±1.0%, all of which were significantly reduced. Transmission electron microscopy revealed that RuC promoted an increase in autophagosome formation in GC cells. Immunofluorescence detection showed that LC3 fluorescent foci of GC cells increased with the increase in RuC dose. RuC up-regulated the expression of autophagy-related proteins LC3Ⅱ and P62 in GC cells. Acridine orange staining indicated that RuC altered the acidic environment of lysosomes. SuperPred online prediction identified CTSD as a potential target protein of RuC. Western blot analysis revealed that RuC induced the up-regulation of the inactive precursor of CTSD in GC cells. CTSD activity assays indicated that RuC reduced the activity of CTSD. Molecular docking simulations found that RuC bound to the substrate-binding region of CTSD, forming hydrogen bonds with the Tyr205 and Asp231 residues. Microscale thermophoresis and DARTS assays further confirmed that RuC directly bound to CTSD. In summary, RuC inhibits lysosomal activity by targeting and down-regulating the expression of CTSD, thereby inducing autophagosome accumulation in GC cells.
Humans ; Stomach Neoplasms/enzymology* ; Cathepsin D/chemistry* ; Cell Line, Tumor ; Molecular Docking Simulation ; Cell Proliferation/drug effects* ; Autophagosomes/metabolism* ; Autophagy/drug effects*

Ischemic stroke is a disease resulting from the cerebral ischemia and hypoxia caused by the blockage of brain vessels in the brain,and is characterized by the focal neurological signs.Pathologically,neuronal necrosis in the infarcted area and the neuronal degeneration or delayed death of neurons in the ischemic penumbra,contribute to the morphological basis of the disease.Ischemic stroke is regulated by multiple processes,including ferroptosis,apoptosis,and autophagy.Ferroptosis,a type of iron-dependent cell death,is closely associated with ischemic stroke.Nuclear factor erythroid 2-related factor 2(Nrf2),a key transcription factor,plays a critical role in maintaining cellular redox balance and regulating inflammatory responses.Nrf2 promotes the expression of solute carrier family 7 member 11(SLC7A11)and glutathione peroxidase 4(GPX4),thereby activating the Nrf2 signaling pathway to counteract ferroptosis and protect cells from damage.This article reviews and analyzes recent experimental studies on traditional Chinese medicine(TCM)therapy targeting the Nrf2/SLC7A11/GPX4 pathway to suppress ferroptosis.The studies have found that TCM therapy with herbal compounds,Chinese patent medicines,single herbal components and their active ingredients,and acupuncture and moxibustion can inhibit ferroptosis by activating the Nrf2/SLC7A11/GPX4 signaling pathway,which will provide novel strategies for the TCM intervention of ischemic stroke.