OBJECTIVE To identify the active components of Desmodium styracifolium （Osb.） Merr. extract （DME） against cholestatic liver injury （CLI）， and to verify its target and molecular mechanism， so as to provide scientific evidence for the clinical application of D.styracifolium （Osb.） Merr. and the development of new hepatoprotective agents.METHODS High-performance liquid chromatography was applied to establish fingerprints of 19 batches of DME， followed by similarity evaluation and identification of common peaks. An in vitro CLI model was constructed by lithocholic acid （LCA）-induced injury in HepG2 cells to evaluate the in vitro protective effect of DME against CLI. Spearman correlation analysis， grey relational analysis and partial least-squares regression analysis were adopted to investigate the spectrum-effect relationship of DME against CLI and preliminarily screen the core active components. Molecular docking and surface plasmon resonance technology were used to predict and verify the binding capacity between core active components and farnesoid X receptor （FXR）. qRT-PCR， Western blot and immunofluorescence staining tests were performed to validate the influences of core active components on the expression of molecules related to FXR/bile salt export pump （BSEP） pathway in CLI model cells.RESULTS A total of 12 common peaks were calibrated in the fingerprints of 19 batches of DME， and the similarity values were all above 0.990. Four common peaks were identified， namely peak 6 （schaftoside）， peak 7 （isoorientin）， peak 11 （isoschaftoside） and peak 12 （isovitexin）. DME at 80 μg/mL exerted the optimal protective effect on CLI-model cells， and the cell viability reached （75.66±4.60）% after 24 h of treatment. Spectrum-effect analysis revealed that schaftoside possessed the strongest correlation with the anti-CLI activity of DME and served as the core active component. Target prediction and validation results showed that the binding energy between schaftoside and FXR was －8.1 kcal/mol， and the equilibrium dissociation constant was 27.3 μmol/L. Mechanistic experiments demonstrated that 100 μmol/L schaftoside significantly elevated the expression levels of FXR and BSEP， and mRNA of their encoded genes Nr1h4 ， Abcb11 in CLI-model cells （ P ＜0.05）.CONCLUSIONS Schaftoside may be the core active component of DME against CLI， and its mechanism may be related to the activation of FXR/BSEP pathway.

BACKGROUND:Radiotherapy for head and neck tumors can easily cause xerostomia,seriously affecting the quality of life of patients.In recent years,engineered stem cells and their paracrine factors have shown therapeutic potential in the repair of salivary gland injury.However,there is currently no experimental study on the application of amniotic mesenchymal stem cell-derived exosome in radiation-induced salivary gland injury.OBJECTIVE:To preliminarily explore the repair effect of exosome derived from human amniotic mesenchymal stem cells on radiation-induced submandibular gland injury.METHODS:H uman amniotic mesenchymal stem cell exosomes were extracted and identified by ultrafiltration and ultracentrifugation.SD rats were randomly divided into a control group,a radiation injury group,and a radiation injury+exosome group.An in vitro model of radiation-induced submandibular gland injury was constructed using the submandibular gland tissue of SD rats irradiated with 18 Gy of radiation.One day after radiation modeling,exosome derived from human amniotic mesenchymal stem cells was injected into the submandibular gland in situ.Samples are taken at 1,3,7,and 14 days to detect the resting salivary flow rate.The structure of the submandibular gland tissue was observed by hematoxylin-eosin staining.The expression of glycogen particles in the submandibular gland tissue was observed by Periodic Acid-Schiff staining.Fibrosis in the submandibular gland tissue was observed by Masson staining.The secretion of salivary amylase was detected by enzyme-linked immunosorbent assay.The expression of aquaporin and tight junction proteins in submandibular gland tissue was observed by immunofluorescence staining.Real-time fluorescence quantitative PCR was used to detect the relative expression levels of aquaporins and salivary amylase mRNA in submandibular gland tissue.TUNEL assay was used to detect the apoptosis rate of submandibular gland tissues in each group.RESULTS AND CONCLUSION:After radiomodeling,compared with the radiation injury group,(1)hematoxylin-eosin staining observed that the submandibular gland tissue structure in the radiation injury+exosome group was restored,the nucleoli increased,the number of acinus increased,and the acinar atrophy improved.(2)Glycogen staining observed that the number and density of positive zymogen granules in the acinar cytoplasm of the radiation injury+exosome group gradually increased.(3)Masson staining results observed that the number and density of positive collagen fibers in the interstitium and around the ducts in the radiation injury+exosome group gradually decreased,the degree of fibrosis decreased,and the collagen deposition decreased.(4)The salivary flow rate in the radiation injury+exosome group increased(P＜0.05).The fluorescence intensity of aquaporin-5 was enhanced(P＜0.05)and the gene expression was significantly enhanced(P＜0.01).The fluorescence distribution of tight junction protein 4 was weakened and the fluorescence intensity decreased(P＜0.05,P＜0.01).The content of salivary amylase increased(P＜0.05)and gene expression were significantly increased(P＜0.01).The number of positive apoptotic cells decreased(P＜0.05,P＜0.01).It is indicated that local injection of exosome derived from human amniotic mesenchymal stem cells could improve the pathological morphology of submandibular gland tissue,promote saliva flow rate and amylase expression,and may play a functional repair role in radioactive submandibular gland injury by inhibiting acinar apoptosis.

Lung transplantation is a key therapeutic approach for patients with end-stage lung diseases. Although its clinical outcomes have significantly improved, multidimensional injuries sustained by donor lungs during procurement, preservation, and transplantation remain major challenges affecting graft survival and long-term prognosis. This article proposes the "Guangzhou Classification" for full-course management of donor lung injury, characterized by spatiotemporal dynamics. Based on the progression of disease stages, donor lung injuries are systematically divided into three types: primary injuries (including donor ICU-related lung injury, pathogen colonization, and cold ischemia injury), secondary injuries (such as ventilator-induced lung injury after transplantation, ischemia-reperfusion inflammatory storm, and early rejection), and accompanying injuries (organ toxicity caused by accumulation of postoperative sedatives, analgesics, and vasoactive drugs). Drawing on previous studies and the clinical experience of our center, this paper elaborates the temporal evolution, key risk factors, and prevention and treatment strategies of each injury category, and discusses future research directions. By targeting critical injury factors at each stage, this classification aims to optimize both short-term and long-term outcomes of lung transplantation.

The development of anticancer drugs to treat triple-negative breast cancer (TNBC) is an ongoing challenge. Immunogenic cell death (ICD) has garnered considerable interest worldwide as a promising synergistic modality for cancer chemoimmunotherapy. However, only few drugs or treatment modalities can trigger an ICD response and none of them exert a considerable clinical effect against TNBC. Therefore, new agents with potentially effective chemoimmunotherapeutic response are required. In this study, five new cyclometalated Ir(III) complexes containing isoquinoline alkaloid CˆN ligands were designed and synthesized. Among them, Ir-1 exhibited the highest in vitro cytotoxicity. Mechanistically, Ir-1 could trigger autophagy-dependent ferroptosis and a subsequent ferroptosis-dependent ICD response as well as indoleamine 2,3-dioxygenase (IDO) inhibition via reactive oxygen species (ROS)-mediated endoplasmic reticulum (ER) stress in MDA-MB-231 cells. When immunocompetent BALB/c mice were vaccinated with Ir-1-treated dying TNBC cells, antitumor CD8⁺ T-cell response and Foxp3⁺ T-cell depletion were induced, resulting in long-lasting antitumor immunity in TNBC cells. Moreover, combination therapy with Ir-1 and anti-PD1 could substantially augment in vivo therapeutic effects. Based on these results, Ir-1 is a promising candidate for chemoimmunotherapy against TNBC and its effects are mediated synergistically via ICD induction and IDO blockage.

OBJECTIVES: To investigate the prognostic significance of PDZ-binding kinase (PBK) in pan-cancer and its potential as a therapeutic target for pancreatic cancer. METHODS: PBK expression levels were investigated in 33 cancer types based on data from TCGA, GEO and CPTAC databases. RT-PCR and Western blotting were employed to examine PBK expression in clinical pancreatic cancer specimens and cell lines. The diagnostic and prognostic value of PBK in pancreatic cancer was evaluated using survival analysis, Cox regression analysis, ROC curve analysis, and clinical correlation studies. Gene enrichment and immune correlation analyses were conducted to explore the potential role of PBK in tumor microenvironment, and its correlation with drug sensitivity was investigated using GDSC and CTRP datasets. In pancreatic cancer BXPC-3 cells, the effects of lentivirus-mediated PBK knockdown on cell proliferation, migration, and invasion were examined using CCK-8, colony formation, and Transwell assays. The interaction between PBK and non-SMC condensin II complex subunit G2 (NCAPG2) was analyzed using co-immunoprecipitation and Western blotting. RESULTS: PBK was overexpressed in multiple cancer types, including pancreatic cancer. A high PBK expression was associated with a poor prognosis of the patients and correlated with immune infiltration and alterations in the tumor microenvironment. Elevated PBK expression was positively correlated with the sensitivity to MEK inhibitors (Trametinib) and EGFR inhibitors (Afatinib) but negatively with the sensitivity to Bcl-2 inhibitors (TW37) and niclosamide. In BXPC-3 cells, PBK knockdown significantly suppressed NCAPG2 expression and inhibited cell proliferation, migration, and invasion. Co-immunoprecipitation confirmed a direct binding between PBK and NCAPG2. CONCLUSIONS PBK is a key regulator of pancreatic cancer and interacts with NCAPG2 to promote tumor progression, suggesting its value as a potential biomarker and therapeutic target for pancreatic cancer.
Humans ; Pancreatic Neoplasms/genetics* ; Prognosis ; Biomarkers, Tumor/genetics* ; Cell Line, Tumor ; Cell Proliferation ; Adenocarcinoma/metabolism* ; Tumor Microenvironment ; Cell Movement ; Mitogen-Activated Protein Kinase Kinases

BACKGROUND:Recent studies have found that Nrf2/ARE signaling pathway activators have the characteristics of low toxicity and control,and have a protective effect against radiation tissue damage.OBJECTIVE:To investigate whether nitrooleic acid can protect submandibular gland epithelial cells from radiation injury by regulating the Nrf2/ARE signaling pathway.METHODS:Rat submandibular gland epithelial cells were cultured in vitro and CCK-8 assay was used to screen the optimal concentration and time of nitrooleic acid administration.Submandibular gland epithelial cells were divided into non-radiation group,radiation control group,nitrooleic acid group,nitrooleic acid+ML385(Nrf2/ARE signaling pathway specific inhibitor)group,and ML385 group.Submandibular gland cells were pretreated with nitrooleic acid and ML385 for 24 hous according to the experimental groups,and then irradiated with 5 Gy radiation to establish the models.At 48 hours after irradiation,CCK-8 assay was used to detect the cell proliferation rate.Real-time quantitative PCR was used to detect the expression of Nrf2,HO-1,and NQO1 mRNA in the cells.Real-time quantitative PCR and enzyme-linked immunosorbent assay were used to detect the cell secretion function and the expression of inflammatory factors.DCFH-DA fluorescent probe kit was used to detect the level of intracellular reactive oxygen species.RESULTS AND CONCLUSION:(1)Compared with the radiation control group,the proliferation rate of submandibular gland epithelial cells and the expression levels of secretion function related factors aquaporin 5 and α-amylase in the nitrooleic acid group of rats increased(P＜0.05),and the expression levels of Nrf2,HO-1,and NQO1 mRNA increased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-αdecreased(P＜0.05),and reactive oxygen species generation reduced(P＜0.01).(2)Compared with the nitrooleic acid group,the addition of nitrooleic acid and ML385 group resulted in a decrease in cell proliferation rate and expression levels of secretion function related factors aquaporin 5 and α-amylase(P＜0.05),and mRNA expressions of Nrf2,HO-1,and NQO1 were all decreased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-α increased(P＜0.05),and generation of reactive oxygen species increased(P＜0.05).(3)Results indicated that in the radiation environment,nitrooleic acid has a certain protective effect on the proliferation ability and secretion function of rat submandibular gland epithelial cells,reduces the expression of inflammatory factors,lowers intracellular reactive oxygen species levels,and alleviates the damage of rat submandibular gland epithelial cells caused by radiation.This function may be related to the activation of Nrf2/ARE signaling pathway.

Objective:To compare the effects of different appointment regimens on the daily waiting time, fixedness of treatment time and lateness rate of radiotherapy patients.Methods:Medical records of 5488 radiotherapy from 332 patients on the same linear accelerator in West China Hospital of Sichuan University from March to June 2022 were selected. Based on the radiotherapy information integration platform of MOSAIQ, all patients were randomly assigned to the morning class, afternoon class and evening class. Traditional manual appointment regimen was adopted for the morning class, 30 min appointment regimen for the afternoon class, and 15 min appointment regimen for the evening class, respectively. The differences in patient waiting time for treatment, fixedness of treatment time, and lateness rate under different appointment regimens were compared. The fixedness of treatment time and waiting time was determined by one-way ANOVA, and the 2×3 Chi-square test was adopted for the lateness rate. Results:The waiting time in the 15 min appointment, the 30 min appointment and manual appointment groups were (27.08 ± 17.21), (34.57± 19.12) and (41.50 ±20.94) min, respectively. There was statistical significance among three appointment regimens ( F=254.97, P<0.001). The waiting time was the shortest in the 15 min appointment group, followed by the 30 min appointment group, and the manual appointment group (all P<0.001 for two-group comparison). The fixedness of treatment time in the 15 min appointment, the 30 min appointment and the manual appointment groups were (15.60±7.87), (18.69±8.94) and (24.30±15.10) min, respectively. There was statistical significance among three groups ( F=25.23, P<0.001). Among them, the fixedness of treatment time in the 15 min appointment group was the highest, followed by the 30 min appointment group, and the manual appointment group (all P<0.001). The lateness rates in the 15 min appointment, the 30 min appointment and the manual appointment groups were 5.7%, 6.2% and 9.6%, respectively. The lateness rate in the manual appointment group was higher than those in the 15 min appointment and the 30 min appointment groups ( χ2=19.24、14.90, both P<0.001), but there was no statistical significance in the lateness rate between the 15 min appointment and 30min appointment groups ( χ2=0.39, P=0.535). Conclusion:In the clinical practice of conventional intensity-modulated radiotherapy technology carried out by conventional linear accelerator, the 15 min appointment regimen can shorten the waiting time for radiotherapy and improve the fixedness of daily radiotherapy time, which is worthy of clinical promotion.

BACKGROUND:Recent studies have found that Nrf2/ARE signaling pathway activators have the characteristics of low toxicity and control,and have a protective effect against radiation tissue damage.OBJECTIVE:To investigate whether nitrooleic acid can protect submandibular gland epithelial cells from radiation injury by regulating the Nrf2/ARE signaling pathway.METHODS:Rat submandibular gland epithelial cells were cultured in vitro and CCK-8 assay was used to screen the optimal concentration and time of nitrooleic acid administration.Submandibular gland epithelial cells were divided into non-radiation group,radiation control group,nitrooleic acid group,nitrooleic acid+ML385(Nrf2/ARE signaling pathway specific inhibitor)group,and ML385 group.Submandibular gland cells were pretreated with nitrooleic acid and ML385 for 24 hous according to the experimental groups,and then irradiated with 5 Gy radiation to establish the models.At 48 hours after irradiation,CCK-8 assay was used to detect the cell proliferation rate.Real-time quantitative PCR was used to detect the expression of Nrf2,HO-1,and NQO1 mRNA in the cells.Real-time quantitative PCR and enzyme-linked immunosorbent assay were used to detect the cell secretion function and the expression of inflammatory factors.DCFH-DA fluorescent probe kit was used to detect the level of intracellular reactive oxygen species.RESULTS AND CONCLUSION:(1)Compared with the radiation control group,the proliferation rate of submandibular gland epithelial cells and the expression levels of secretion function related factors aquaporin 5 and α-amylase in the nitrooleic acid group of rats increased(P＜0.05),and the expression levels of Nrf2,HO-1,and NQO1 mRNA increased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-αdecreased(P＜0.05),and reactive oxygen species generation reduced(P＜0.01).(2)Compared with the nitrooleic acid group,the addition of nitrooleic acid and ML385 group resulted in a decrease in cell proliferation rate and expression levels of secretion function related factors aquaporin 5 and α-amylase(P＜0.05),and mRNA expressions of Nrf2,HO-1,and NQO1 were all decreased(P＜0.05),while the expression levels of inflammatory factors interleukin-1β,interleukin-6,and tumor necrosis factor-α increased(P＜0.05),and generation of reactive oxygen species increased(P＜0.05).(3)Results indicated that in the radiation environment,nitrooleic acid has a certain protective effect on the proliferation ability and secretion function of rat submandibular gland epithelial cells,reduces the expression of inflammatory factors,lowers intracellular reactive oxygen species levels,and alleviates the damage of rat submandibular gland epithelial cells caused by radiation.This function may be related to the activation of Nrf2/ARE signaling pathway.

Objective: To establish and validate a whole blood (WB) supply model, thereby providing practical experience for the clinical application of WB in domestic trauma emergency care and informing the development of a wartime blood supply system for the military. Methods: A “10×24” WB supply model was established by formulating blood collection protocols, storage standards, and transfusion criteria. Multiple WB samples were tested under specific storage conditions to assess key indicators at different time points, including red blood cell (RBC), white blood cell (WBC), and platelet counts, hemoglobin concentration, coagulation parameters (PT, APTT, TT, FIB), coagulation factor activity, thromboelastography (TEG) parameters, and electrolyte levels. Additionally, clinical data from hemorrhagic patients who met the criteria for WB transfusion and were admitted between March and July 2024 were analyzed to evaluate WB transfusion volume. Results: RBC counts and hemoglobin levels remained stable in WB stored at 4℃ for up to 10 days. However, platelet counts and coagulation function (PT, APTT) significantly declined with prolonged storage, while potassium levels increased. From March to July 2024, the model was successfully applied to 23 patients with acute hemorrhage, with a median WB transfusion volume of 543 mL. A detailed case study of a severe traumatic hemorrhagic shock patient was reported, who was successfully treated with 5.5 units of refrigerated WB combined with component blood. Conclusion: The “10×24” WB supply model demonstrated acceptable changes in critical quality parameters under strict management and a 10-day rotation cycle. This model effectively supports the treatment of acute hemorrhage and holds promise for integration into the future wartime blood supply system of the military.

The NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammasome, a high-molecular-weight protein complex in the cytoplasm, is composed of three core components: the sensor protein NLRP3, the adaptor protein apoptosis-associated speck-like protein containing a caspase-recruitment domain (ASC) and the effector protein caspase-1. It plays a critical role in regulating host immune and inflammatory responses. Studies have shown that the NLRP3 inflammasome has increasingly become a focal point in tumor molecular biology field. A growing body of evidence indicates that the increased expression and activation of the NLRP3 inflammasome is closely associated with the pathogenesis of head and neck squamous cell carcinoma (HNSCC) and the tumor microenvironment (TME). It may promote tumor proliferation, invasion, migration, and other biological behaviors through various regulatory mechanisms while influencing tumor immune evasion and therapy resistance, which holds promise as a prognostic biomarker for patients. This review explores the current effect and mechanism of the NLRP3 inflammasome and its signaling pathways in head and neck cancer, providing insights into clinical targeted drug development and molecular immunotherapy.
Humans ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Inflammasomes/metabolism* ; Head and Neck Neoplasms/pathology* ; Squamous Cell Carcinoma of Head and Neck/metabolism* ; Tumor Microenvironment ; Signal Transduction ; Animals