ObjectiveTo investigate the protective effect of genistein （Gen） on etoposide-induced chondrocyte senescence and its underlying mechanism. MethodsThe C28/I2 cell line was treated with different concentrations of Gen and etoposide， and the cell viability was detected by the CCK-8 assay. The senescence model of C28/I2 chondrocytes was induced by etoposide， with Gen intervention. Senescence-associated β-galactosidase （SA-β-gal） staining was performed to detect the senescence-positive rate and staining characteristics of chondrocytes. The expressions of peroxiredoxin 6 （Prdx6）， cyclin-dependent kinaseto clarify the functional necessity of Prdx6. ResultsCompared with the etoposide group， the C28/I2 chondrocyte viability significantly increased （P<0.01）， the expression ofsenescence-associated proteins p21 and p16 decreased （P<0.01， P<0.05）， the expression of senescence-associated genes p21 and p16 reduced （both P<0.01）， the fluorescence intensity of senescence-associated proteins p21 and p16 was diminished （P<0.05， P<0.01）， and the proportion of SA-β-gal-positive cells decreased （P<0.01） in the Gen+etoposide group. Compared with the Control group， the expression of Prdx6 was downregulated in the etoposide group （P<0.05）. Compared with the etoposide group， the expression of Prdx6 was upregulated in the Gen+etoposide group （P<0.01）. Compared with the Control group， the GPx activity significantly decreased in the si-Prdx6 group （P<0.01）. Furthermore， compared with the si-Prdx6 group， the GPx activity increased in the si-Prdx6+Gen group （P<0.05）. Molecular docking results revealed that Gen formed hydrogen bond interactions with the active site of Prdx6. After Prdx6 knockdown， the expression of senescence-associated genes p21 and p16 and the fluorescence intensity of senescence-associated proteins p21 and p16 both increased in the Gen+etoposide+si-Prdx6 group （both P<0.01）. ConclusionGen can inhibit etoposide-induced senescence of C28/I2 chondrocytes by upregulating the expression of Prdx6. This study provides potential drug targets and experimental basis for the prevention and treatment of chondrocyte senescence-related diseases.

ObjectiveTo investigate the effects and mechanisms of pulchinenoside A （PSA） on tumor necrosis factor-α （TNF-α） -induced abnormal proliferation， migration， and invasion of synovial fibroblasts. MethodsA synovitis model was established by stimulating SW982 synovial cells with TNF-α. The low-density lipoprotein receptor-related protein 1 （LRP1） was knocked down using LRP1-specific siRNA or inhibited by the receptor-associated protein （RAP）. Cells were treated with various concentrations of PSA， and cell viability was assessed by CCK-8 assay. Protein levels of LRP1， matrix metalloproteinase-3 （MMP3）， and N-cadherin were measured by Western blotting. Proliferation was evaluated using an EdU incorporation assay. Migration and invasion capacities were examined by Transwell and wound-healing assays. mRNA expression of LRP1， MMP3， N-cadherin， IL-1β， IL-6， and TNF-α was detected by reverse transcription quantitative real-time PCR （RT-qPCR）. Immunofluorescence staining was used to visualize the expression of MMP3 and N-cadherin. The involvement of LRP1 was further verified with the inhibitor RAP. ResultsCompared with the TNF-α-treated model group， PSA treatment significantly reduced SW982 cell viability （P<0.05）， downregulated MMP3 and N-cadherin expression （both P<0.01）， and decreased the proportion of EdU-positive cells （P<0.05）. Moreover， TNF-α stimulation upregulated LRP1 expression compared with normal controls （P<0.01）， whereas PSA treatment reversed this increase （P<0.05）. LRP1 knockdown or inhibition markedly decreased MMP3 and N-cadherin levels compared with the model group （both P<0.01）. Similarly， the combination of PSA and RAP led to significant reductions in MMP3 and N-cadherin expression compared with the model group （both P<0.01）. ConclusionPSA inhibits TNF-α-induced abnormal proliferation， migration， and invasion of synovial， at least in part， by downregulating LRP1 expression.

ObjectiveTo explore the clinical potential of magnetic resonance elastography （MRE） for assessing the stiffness of common intracranial tumors. MethodsA retrospective analysis was conducted on 47 patients with brain tumors who underwent surgical treatment at The First Affiliated Hospital of Sun Yat-sen University between April 2023 and September 2024. The cohort included 10 cases of glioma， 12 cases of vestibular schwannoma， and 25 cases of meningioma. All patients underwent multiparameter MRI and MRE preoperatively. After co-registering MRE images with contrast-enhanced 3D T1-weighted images， three regions of interest （ROIs） were placed in the areas of maximum and minimum tumor stiffness respectively to measure the shear stiffness values， and the mean was calculated as the average shear stiffness. Additionally， one ROI was placed in the white matter and one in the grey matter of the posterior horn of the lateral ventricle at the basal ganglia level to obtain normal reference values. A neurosurgeon， blind to the MRE results， graded the intraoperative tumor stiffness based on surgical videos using a 5-point scale （1： soft； 5： hard）. Intraclass correlation coefficient （ICC） was used to assess inter-observer consistency. Analysis of variance （ANOVA） and nonparametric tests were employed to compare the differences of shear stiffness values between groups. Spearman correlation analysis was used to examine the correlation between elasticity values and clinicopathological parameters. The discriminatory efficacy of shear stiffness values for tumor types was evaluated with receiver operating characteristic （ROC） curves. ResultsThe ICCs for elasticity values measured by two physicians in the hardest and softest tumor areas were 0.868 and 0.831， respectively （both P＜ 0.01）. The maximum， minimum， and average shear stiffness values of meningiomas［（1.91±0.28） m/s， （1.66±0.25） m/s， （1.78±0.26） m/s， respectively］ were significantly higher than those of gliomas［（1.59±0.23） m/s， （1.42±0.23） m/s， （1.50±0.22） m/s， respectively （P＜0.05）］. The shear stiffness values of vestibular schwannomas［（1.70±0.30） m/s， （1.51±0.23） m/s， （1.61±0.26） m/s， respectively） were lower than those of meningiomas and higher than those of gliomas， but these differences were not statistically significant （both P＞0.05）. The shear stiffness values of white matter were significantly higher than that of grey matter［（1.63±0.08） m/s vs. （1.39±0.12） m/s， P＜0.05）］. In the overall sample， the maximum， minimum， and average shear stiffness values were significantly positively correlated with the intraoperative hardness grade （r_s = 0.591， r_s = 0.541， r_s = 0.571， respectively； all P＜0.001）， negatively correlated with WHO grade （r_s = -0.458， r_s = -0.458， r_s = -0.480， respectively； all P＜0.01）， but showed no correlation with Ki-67 （all P＞0.05）. ConclusionMRE can be used to assess the stiffness of intracranial tumors and to some extent differentiate between tumor types， which aids in formulating more individualized preoperative surgical plans.

BACKGROUND:Ca2+expression in astrocytes has been found to be closely related to cognitive function,and the Ca2+signaling pathway regulated by inositol 1,4,5-trisphosphate receptors(IP3R2)and ryanodine receptor(RYR)2 receptors has become a hot spot in the study of cognitive disorder-related diseases. OBJECTIVE:To investigate the expression of Ca2+signals mediated by IP3R2 and RYR2 in hippocampal astrocytes in animal models of delayed encephalopathy after acute carbon monoxide poisoning,and to explore the possible pathogenesis of delayed encephalopathy after acute carbon monoxide poisoning. METHODS:C57BL mice with qualified cognitive function were selected by Morris water maze experiment and randomly divided into control group and experimental group.An animal model of delayed encephalopathy after acute carbon monoxide poisoning was established by static carbon monoxide inhalation in the experimental group,and the same amount of air was inhaled in the control group.Behavioral and neuronal changes,astrocyte specific marker glial fibrillary acidic protein,IP3R2,RYR2 receptor and Ca2+concentration in astrocytes of the two groups were detected using Morris water maze,hematoxylin-eosin staining,western blot,immunofluorescence double labeling and Ca2+fluorescence probe at 21 days after modeling. RESULTS AND CONCLUSION:In the Morris water maze,the escape latency of the experimental group was significantly longer than that of the control group(P<0.05).Hematoxylin-eosin staining results showed that in the experimental group,the number of hippocampal pyramidal cells decreased,the cell structure was disordered,and the nucleus was broken and dissolved.Immunofluorescence results showed that IP3R2 and RYR2 were co-expressed with glial fibrillary acidic protein in the hippocampus,and the expressions of IP3R2,RYR2 and glial fibrillary acidic protein were up-regulated in the hippocampus of the experimental group(P<0.05).Western blot analysis showed that the expressions of IP3R2,RYR2,and glial fibrillary acidic protein in the hippocampus of the experimental group were increased(P<0.05).Ca2+concentration in hippocampal astrocytes increased significantly in the experimental group(P<0.05).To conclude,astrocytes may affect Ca2+signals by mediating IP3R2 and RYR2 receptors,then impair the cognitive function of mice with carbon monoxide poisoning,and eventually lead to delayed encephalopathy after acute carbon monoxide poisoning.

Objective : To explore the effects and mechanisms of Kobophenol A ( KPA) on lipopolysaccharide ( LPS) -induced M1 macrophage polarization，and to provide a theoretical basis for the treatment of inflammatory immune diseases and the development of new drugs． Methods: The M1 macrophage polarization model of RAW264. 7 was established by LPS induction，and the peroxiredoxin 6 ( Prdx6) knockdown model was constructed using the Prdx6 inhibitor MJ33 and Prdx6-siRNA．RAW264. 7 cells，a mouse macrophage cell line，were treated with various concentrations of KPA． Cell viability was assessed using the CCK-8 assay．The expression levels of Prdx6 and M1 macrophage polarization-related proteins，including inducible nitric oxide synthase ( iNOS) and cyclooxygenase-2 ( COX-2) ，were detected by Western blot and immunofluorescence staining．The expression levels of Prdx6 and M1 macrophage polarization-related genes iNOS，interleukin-6 ( IL-6) ，and tumor necrosis factor α ( TNF-α) ，were measured by RT-qPCR. Flow cytometry was employed to detect the expression of cluster of differentiation 86 ( CD86) ，a marker of M1 macrophages． Results: Compared with the LPS-induced M1 macrophage polarization model ， KPA significantly reversed the morphological changes of M1 macrophage polarization in RAW264. 7 macrophages and decreased the expression of M1 macrophage polarization-related proteins iNOS，COX- 2，CD86 and related genes iNOS，IL-6，TNF-α ( all P＜0. 05) ．In addition，LPS significantly downregulated the expression of Prdx6 in RAW264. 7 macrophages，while KPA upregulated the expression of Prdx6．Moreover，treatment with the Prdx6 inhibitor MJ33 significantly upregulated the expression of iNOS，a marker of M1 macrophage polarization，in RAW264. 7 macrophages，whereas treatment with KPA significantly downregulated the expression of iNOS ( all P＜0. 05) ． Conclusion KPA inhibits LPS-induced M1 polarization of RAW264. 7 macrophages by upregulating the expression of Prdx6．

Stresses induced by the deficiency or excess of trace mineral elements, such as manganese (Mn), represent a common limiting factor for the production of crops like Brassica napus. To identify key genes involved in Mn allocation in B. napus and elucidate the underlying mechanisms, a member of the metal tolerance protein (MTP) family obtained in the previous screening of cDNA library of B. napus under Mn stress was selected as the research subject. Based on the sequence information and phylogenetic analysis, it was named as BnMTP10. It belongs to the Mn-cation diffusion facilitator (CDF) subfamily. Expression of BnMTP10 in yeast significantly improved the tolerance of transformants to excessive Mn and iron (Fe) and reduced the accumulation of Mn and Fe. However, the yeast transformants exhibited no significant changes in tolerance to excess cadmium, boron, aluminum, zinc, or copper. The qRT-PCR results demonstrated that the flowers of B. napus had the highest expression of BnMTP10, followed by roots and leaves. Subcellular localization studies revealed that BnMTP10 was localized in the endoplasmic reticulum (ER). Compared with wild-type plants, transgenic Arabidopsis overexpressing BnMTP10 exhibited enhanced tolerance to excessive Mn stress but showed no significant difference under Fe stress. Correspondingly, under excessive Mn stress, the Mn content in the roots of transgenic Arabidopsis increased significantly. However, under excessive Fe stress, the Fe content in transgenic Arabidopsis did not alter significantly. According to the results, we hypothesize that BnMTP10 may alleviate excessive Mn stress in plants by mediating Mn transport to the ER. This study facilitated our understanding of efficient mineral nutrients, and provided theoretical foundations and gene resources for breeding B. napus.
Brassica napus/genetics* ; Manganese/metabolism* ; Plants, Genetically Modified/genetics* ; Plant Proteins/physiology* ; Arabidopsis/metabolism* ; Gene Expression Regulation, Plant ; Phylogeny ; Cation Transport Proteins/metabolism* ; Stress, Physiological

Objective To summarize the medicinal properties of new foreign introducing Chinese materia medica Drimia maritima(L.)Stearn based on literature research;To further verify its five flavors by applying intelligent sense;To provide new ideas for the medicinal properties research of new foreign introducing Chinese materia medica.Methods Literature about Drimia maritima(L.)Stearn was retrieved from CNKI,VIP,Wanfang Data,PubMed,Web of Science,and combined with the TCM theories medicinal properties of Drimia maritima(L.)Stearn was summarized.PEN3 electronic nose and SA402B electronic tongue were used to obtain intelligent sensory information of Drimia maritima(L.)Stearn,and the principal component analysis(PCA)was used to identify its five flavors.Results Literature research summarize Drimia maritima(L.)Stearn with antioxidant,anti-tumor,strengthening cardiac functions and diuresis,reducing cough and asthma and other pharmacological effects.Combined with the theories,it explored that its property was mainly slightly warm,the tastes were mainly bitter,pungent,sweet,and slightly toxic,belonging to the lung,spleen,stomach and heart meridians.Based on intelligent sense,identification results of five tastes of Drimia maritima(L.)Stearn were bitter,slightly pungent,slightly sweet,salty,non-acid,basically consistent with the literature research.Conclusion Literature research combined with intelligent sense can better summarize and recognize the medicinal properties of Drimia maritima(L.)Stearn.This study can provide a certain reference for a more objective and adequate medicinal properties analysis of new foreign introducing Chinese materia medica.

Objective To explore the mechanism of Chaipo Decoction in intervening lipopolysaccharide(LPS)-induced acute lung injury(ALI)based on network pharmacology and experimental verification.Methods TCMSP database was used to screen the active components and targets of Chaipo Decoction.ALI disease targets were retrieved through GeneCards and OMIM databases,the intersection genes of drug and disease were obtained.They were imported into STRING platform and Cytoscape 3.9.1 software to construct protein interaction network and drug-active components-target network,core components and targets were screened,and GO and KEGG pathway enrichment analysis was performed on them.AutoDock 1.5.6 and PyMOL software were used to verify the molecular docking between the main active components and core targets.The ALI model of mice was induced by LPS,and was intervened by Chaipo Decoction.The pathological changes of lung tissue in mice were observed by HE staining.The contents of IL-6,IL-10 and TNF-α in serum of mice were detected by ELISA.Western blot was used to verify the expressions of target proteins in lung tissue.Results Totally 214 active components of Chaipo Decoction,such as norwogonin,baicalein,quercetin and ursolic acid were screened,with 1 101 targets and 271 potential targets for intervention in ALI,mainly SRC,STAT3,AKT1,EGFR,GRB2,PIK3CA,etc.It mainly affected PI3K/Akt signaling pathway,MAPK signaling pathway,apoptosis,Ras signaling pathway,etc.Molecular docking showed that the main active components of Chaipo Decoction had strong binding ability with core targets SRC,STAT3,AKT1,EGFR and GRB2.The results of animal experiments showed that Chaipo Decoction could alleviate the lung lesions in ALI mice,decrease the contents of IL-6 and TNF-α in serum,increase the content of IL-10,and significantly decreased the expressions of EGFR,PI3K,AKT and NF-κB p65 protein in lung tissue.Conclusion Chaipo Decoction can exert anti-ALI effect through multiple pathways and multiple targets,and its mechanism may be related to the inhibition of EGFR/PI3K/Akt signaling pathway.