ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

ObjectiveTo explore the role of cucurbitacin B （CuB） in inducing ferroptosis in 4T1 cells and its mechanism. MethodsThe effects of CuB（0.2， 0.4， 0.8 μmol·L^-1）on the proliferation ability of 4T1 cells in vitro were detected using the methyl thiazolyl tetrazolium （MTT） assay. The clonogenic ability of 4T1 cells was detected by the plate cloning assay， and the levels of lactate dehydrogenase （LDH） in 4T1 cells were detected by the use of a kit. The mitochondrial membrane potential and reactive oxygen species （ROS） levels in 4T1 cells were detected by flow cytometry， and the mitochondrial ultrastructure of 4T1 cells was observed by transmission electron microscopy. The western blot was used to detect the expression of ferroptosis-related protein p53 in 4T1 cells， solute carrier family 7 member 11 （SCL7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， transferrin receptor protein 1 （TFR1）， and ferritin heavy chain 1 （FTH1）. ResultsCompared with that in the blank group， the survival rate of 4T1 cells in CuB groups was significantly decreased （P<0.05）， and the number of cell clones in CuB groups was significantly reduced （P<0.01）. In addition， compared with that in the blank group， the leakage of LDH in cells in CuB groups was significantly increased （P<0.01）， and the mitochondrial membrane potential of cells in CuB groups decreased significantly （P<0.01）. Cellular ROS levels were significantly elevated in CuB groups （P<0.01）. The mitochondria of cells in CuB groups were obviously wrinkled， and the mitochondrial cristae were reduced or even disappeared. Compared with that in the blank group， the protein expression of p53， ACSL4， and TFR1 were significantly up-regulated in CuB groups （P<0.05）， and that of SLC7A11， GPX4， and FTH1 were significantly down-regulated （P<0.05）. ConclusionCuB may inhibit SLC7A11 and GPX4 expression by up-regulating the expression of p53， which in turn regulates the p53/SLC7A11/GPX4 signaling pathway axis and accelerates the generation of lipid peroxidation substrate by up-regulating the expression of ACSL4. It up-regulates TFR1 expression to promote cellular uptake of Fe³⁺ and down-regulates the expression of FTH1 to reduce the ability of iron storage， resulting in an elevated free Fe²⁺ level. It catalyzes the Fenton reaction， generates excess ROS， imbalances the antioxidant system and iron metabolism， and then induces ferroptosis in 4T1 cells.

Ferroptosis， a new form of programmed cell death different from apoptosis， necrosis， and autophagy， is closely associated with a variety of physiological and pathological processes. Iron-mediated accumulation of reactive oxygen species is the main inducement of ferroptosis， the mechanism of which is related to intracellular lipid metabolism， iron metabolism， and antioxidant defense pathways. Multiple signaling axes and regulators jointly regulate the occurrence and disruption of ferroptosis. Studies have demonstrated that ferroptosis regulates the growth and proliferation of tumor cells. Inducing ferroptosis in tumor cells can control the growth， metastasis， and multi-drug resistance of tumors. Therefore， the effect and mechanism of ferroptosis on tumor cells have become a hot topic in anti-cancer research. As the research advances， a variety of ferroptosis inducers has been used in the clinical chemotherapy for cancers and demonstrate significant efficacy. Accordingly， the development of ferroptosis-inducing anticancer drugs has become a new research direction for tumor treatment. Some active ingredients such as lycorine， oleanolic acid， dihydroartemisinin， pseudolaric acid B， and ophiopogonin B of Chinese medicines can induce ferroptosis in tumor cells via lipid metabolism， iron metabolism， system X_c^-， and GPX4/GSH to regulate the development of tumors， demonstrating a promising prospect in clinical treatment. Based on the theory of the mechanism of ferroptosis， this paper reviews the research progress in ferroptosis induced by active ingredients of Chinese medicines in tumor cells and describes the metabolic regulatory network of ferroptosis from signaling pathways and regulatory factors， providing new strategies for applying active ingredients of Chinese medicines in the treatment of tumors.

Objective To explore the relationship between serum leucine 2-rich glycoprotein 1(LRG1),lactate dehydrogenase(LDH)with periodontal indicators and periodontal lesions degree in patients with type 2 diabetes mellitus(T2DM)and chronic periodontitis(CP).Methods 112 patients with T2DM combined with CP admitted to our hospital from July 2022 to July 2023(T2DM combined with CP group)were selected and catego-rized into mild,moderate,and severe groups according to the degree of periodontal pathology;112 patients with CP alone(CP group)were selected from our hospital during the same period,and then 112 patients with T2DM alone(T2DM group)were selected;and LRG1,LDH,and periodontal indexes were measured;Pearson method was applied to analyze the correlation between serum LRG1,LDH,and periodontal indicators.The influencing factors of severe T2DM combined with CP were analyzed using multivariate logistic regression.ROC curve was plotted to analyze the diagnostic value of serum LRG1 and LDH for severe T2DM combined with CP.Results The levels of LRG1 and LDH in the CP group,T2DM group,and T2DM combined with CP group increased sequentially(P＜0.05).The levels of serum LRG1,LDH,AL,PD,and BI increased sequentially in the mild,moderate,and severe groups(P＜0.05).According to Pearson correlation analysis,serum LRG1 was positively correlated with LDH(P＜0.05).Serum LRG1 and LDH were positively correlated with AL,PD,and BI(P＜0.05).According to logistic regression analysis,LRG1,LDH,AL,PD,and BI were all factors for severe T2DM combined with CP(P＜0.05).According to the ROC curve,the AUC for the combined diagnosis of serum LRG1 and LDH in severe T2DM with CP was 0.910,and the combination of the two was better than their individual diagnosis(Z combination vs LRG1=2.659,Z combination vs LDH=2.627,P＜0.05).Conclusion LRG1 and LDH are greatly elevated in the serum of patients with T2DM combined with CP,and they are related to periodontal indicators and the periodontal lesions degree.

Abstract OBJECTIVE To study the mechanism of NLRP3 gene’s regulation on inflammatory response in non-alcoholic fatty liver disease(NAFLD) mice induced by high-fat and high-fructose diet using NLRP3 gene knockout mice. METHODS Use male homozygous(NLRP3-/-) mice, and the high-fat and high-fructose diet was used to establish NAFLD model in NLRP3 knockout(KO) mice and wild-type(WT) mice, divided into KO high-fat and high-fructose diet(KO-HFD) group and WT high-fat and high-fructose diet(WT-HFD) group, while the WT and KO groups were also established. The body weight of mice in each group were observed. The changes of ALT, AST, TG, TC, MDA, SOD, lipidosis, apoptosis rate, IL-1β, IL-18, TNF-α, NF-κB, NLRP3, Caspase-1 and ASC in serum and tissue samples were tested to study the mechanism of the NLRP3 gene regulating the inflammatory response in NAFLD mice. RESULTS As time goes on, the mice weight of each group increased gradually, but the KO-HFD group increased less than the WT-HFD group. Each group’s level of ALT, AST, TG, TC in serum increased gradually, but the KO-HFD group increased less than the WT-HFD group. The level of MDA in liver tissues of each group was gradually increased and the level of SOD was gradually decreased, but the change range of KO-HFD group was smaller than that of WT-HFD group. The results of oil red O staining and Tunel section showed that the degree of lipid deposition and apoptosis in hepatocytes increased gradually in all groups, but the changes in KO-HFD group were less than that in WT-HFD group. The levels of serum and liver inflammatory factors IL-1β, IL-18, TNF-α and NF-κB increased in all groups, but the changes of KO-HFD group 20 weeks were less than those of WT-HFD group 20 weeks, and the difference was statistically significant. The expression levels of NLRP3 inflammasomes and related inflammatory cytokines NLRP3, Caspase-1, ASC, IL-1β and IL-18 in liver tissues of WT-HFD group were higher than those of KO-HFD group. The mRNA transcription levels of NLRP3, ASC, Caspase-1, IL-1β and IL-18 in WT-HFD group were gradually increased, while there was no change in KO-HFD group. CONCLUSION The NLRP3 gene may be activated in NAFLD mice model, resulting in increased expression of NLRP3 inflammasome-associated protein, promoting the synthesis and secretion of downstream inflammatory factors, resulting in significant inflammatory response and liver damage, and promoting the progression of NAFLD disease.

Talaromycosis marneffei has been increasing in recent years. Our understanding of this disease has gradually deepened through extensive basic and clinical research, but there are still many limitations. In this article, by incorporating the latest research advancements, we discuss important issues in managing Talaromycosis marneffei trends, aiming to guide effective prevention and control of the disease, improving public health, and reducing the healthcare burden.

Background: The public now imposes higher demands on the government than in the past, which has created the role overload faced by low-ranking government employees in China. This research investigates the relationship between role overload and health among low-ranking government employees and explores the mediating effects of burnout. Methods: It draws on a survey of 2064 low-ranking government employees by probability proportionate to size sampling in China's Shandong Province. Structural equation modeling (SEM) methods are used to analyze the data. Results: Both role overload and burnout were found to have negative effects on low-ranking government employees' health; however, the associations varied among the three age groups (less than 36, between 36 and 45, and over 45). Those over 45 reported the highest level of both physical and psychological health, while the youngest age group (less than 36) reported the lowest level of health. Role overload has a direct influence on health among government employees over 45 but not among those below 45. Burnout's mediating effects between role overload and health are significant among all age groups, but most significant among the youngest civil servants below 36. Conclusions The findings evidenced that both role overload and burnout affect low-ranking government employees' self-reported physical and psychological health. In addition, the effect of age differences in coping with role stressors and burnout should be considered.

Objective:To design a replaceable corneal scaler and explore its effect on improving the positive detection rate of corneal bacteria and the cure rate of patients with infectious keratitis.Methods:Totally 100 patients with infectious keratitis treated in the Eye&Ent Hospital of Fudan University in Shanghai from June 2020 to October 2021 were conveniently selected as the application object, 50 patients from June 2020 to January 2021 were selected as the control group, and 50 patients from February 2021 to October 2021 were selected as the experimental group. The control group used the traditional corneal scalpel for corneal scraping, and the experimental group used the replaceable corneal scalpel for corneal scraping. The positive detection rate of bacteria, the cure rate of patients as of the date of data collection, the comfort status of patients and the operation difficulty score of medical staff were compared between the two groups.Results:The 28 cases (56%) were positive in the experimental group, and 30 cases (60%) were cured as of the date of data collection; 16 cases (32%) were positive in the control group and 18 cases (36%) were cured as of the date of data collection. The difference between the two groups was statistically significant ( χ2=-2.47, -2.20, all P<0.05). The score of comfort in the experimental group was (83.92 ± 6.40) and that in the control group was (54.04 ± 10.69). The difference between the two groups was statistically significant ( t=17.14, P<0.01). The operation difficulty score of medical staff in the experimental group was (1.46 ± 0.54) and that of medical staff in the control group was (3.24 ± 0.72). The difference between the two groups was statistically significant ( t=-13.50, P<0.01). Conclusions:The use of replaceable corneal scalpel can not only improve the positive detection rate of patients, improve the treatment effect of patients, improve the comfort and satisfaction of patients, but also improve the work efficiency of medical staff and reduce the operational risk of medical staff, so as to improve the real professional value of medical staff.

Objective:To observe the therapeutic effect of quercetin (QUE) on triggering receptor expressed on myeloid cells (TREM-1) activated macrophage inflammation and lipopolysaccharide (LPS) induced acute lung injury (ALI) in mice, and explore its possible mechanism.Methods:In vitro cell experiment: The primary peritoneal macrophages of mice were collected by intraperitoneal injection of 3% calcium mercaptan acetate. The collected cells were divided into blank control group, dimethylsulfoxide (DMSO) vehicle group, TREM-1 agonist group (10 μg/ml), QUE group (10 μmol/L) and TREM-1 agonist + QUE group (cells were pretreated with 10 μmol/L QUE for 30 min before adding agonist). Enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of interleukin (IL)-1β, tumor necrosis factor (TNF)-α and IL-6 in the culture supernatant of primary macrophages; To observe the effect of QUE on LPS-induced TREM-1 protein levels, macrophages were divided into: normal control group, LPS group (100 ng/ml) and LPS+ QUE treatment group [macrophages were pretreated with 10 μmol/L QUE for 2 hours, and then incubated with LPS (100 ng/ml) for 16 hours]. Western blot was used to detect the expression of TREM-1 protein. In animal experiments: 80 male C57BL/6 mice were randomly divided into 4 groups (20 in each group): normal control group, ALI model group, QUE group and QUE treatment group (LPS+ QUE). In the ALI model group, the ALI model was established by intratracheal injection of 5 mg/kg LPS; The mouse ALI model was established by intratracheal injection of LPS 5 mg/kg in the QUE treatment group, and then intraperitoneal injection of 15 mg/kg QUE. The control group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of DMSO, and the QUE group was given the same amount of normal saline intratracheal followed by intraperitoneal injection of 15 mg/kg QUE. Hematoxylin-eosin (HE) staining was used to observe the pathological changes of lung tissue in each group; Inflammatory cells including IL-1β, TNF- α and IL-6 in bronchoalveolar lavage fluid (BLAF) of mice in each group were counted ; The expression of TREM-1 mRNA and protein in lung tissue of mice in each group was detected by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) and Western blot. Results:In vitro cell experiment: the secretion of IL-1β, TNF-α and IL-6 in the supernatant of primary macrophages in TREM-1 agonist group was higher than those in DMSO vehicle group, while the secretion of IL-1β, TNF-αand IL-6 in the supernatant of primary macrophages in TREM-1 agonist + QUE group were lower than that of TREM-1 agonist group (all P<0.001). The expression of TREM-1 protein in LPS group was higher than that in control group ( P<0.05), while the expression of TREM-1 protein in LPS + QUE group was lower than that in LPS group ( P<0.05). Animal experiments showed that compared with the control group, the ALI model group had higher lung pathological injury score, more total cells, macrophages and neutrophils in BALF and increased TNF-α, IL-6, IL-1β content (all P<0.001). The above indexes in QUE group were lower than those in ALI model group (all P<0.001). The results of qRT-PCR and Western blot showed that compared with the control group, the expression of TREM-1 mRNA and protein in the lung tissue of ALI model group was increased, while the expression of TREM-1 mRNA and protein in the lung tissue of QUE group was lower than that of ALI model group (all P<0.05). Conclusions:Quercetin can inhibit TREM-1 activation, reduce macrophage inflammatory response and LPS induced acute lung injury in mice.

Greenspace may bring benefits to human health. Evidence on greenness and health has accumulated in western countries, and several reviews have summarized such evidence. Researchers have also conducted some studies on greenspace and human health in Chinese population, but no prior review has pooled or summarized them. To provide more comprehensive evidence on this topic, we searched and summarized studies on greenspace and health that were conducted specifically in Chinese population. We found that a certain number of studies have been conducted in China, and the evidence indicates that greenspace exposure may reduce the odds of cardiovascular diseases, mental health disorders, adverse birth outcomes as well as mortality. However, most of the current reported studies were of cross-sectional design or randomized controlled trails targeting short-term effects, and prospective cohort studies were scarce. Moreover, greenness exposure was mainly assessed using greenness index like normalized difference vegetative index (NDVI), which were static and cannot distinguish construction and species of greenspaces. Future prospective studies with more precise greenness exposure assessment are warranted to evaluate the prior findings.