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.

Objective:To investigate the efficacy and safety of endovascular treatment (EVT) in young patients with symptomatic intracranial atherosclerotic stenosis (sICAS).Methods:Young patients with sICAS admitted to the Department of Neurology, Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University, Medical School from January 2020 to July 2024 were included retrospectively. According to the therapeutic modalities, they were divided into a best medical treatment (BMT) group and an EVT group. The efficacy outcome was any stroke recurrence or death within 30 days and 1 year. The safety outcome was symptomatic intracranial hemorrhage (sICH) within 30 days and restenosis within 1 year.Results:A total of 113 patients were enrolled, including 85 males (75.2%), with a median age of 43 (interquartile range, 37-48) years; 44 patients (38.9%) received EVT, and 69 (61.1%) received BMT. Among the 44 patients who underwent EVT, 8 (18.2%) underwent balloon angioplasty and 36 (81.8%) underwent stenting. There was no significant difference in the incidence of stroke recurrence or death within 30 days (2.9% vs. 2.3%) and sICH incidence (0% vs. 2.3%) between the BMT group and the EVT group. However, the 1-year stroke recurrence or death rate in the EVT group was significantly lower than that in the BMT group (18.8% vs. 4.5%; P=0.029). Cox proportional hazards regression analysis showed that EVT was independently associated with a lower incidence of stroke recurrence or death within 1 year (hazard ratio 0.225, 95% confidence interval 0.051-0.996; P<0.05). The median age of the balloon angioplasty group was significantly lower than that of the stenting group (33.5 years vs. 46 years; P=0.007), while there were no significant differences in other demographic and baseline data. There was no significant difference in all efficacy and safety outcome between the balloon angioplasty group and the stenting group. Conclusions:For young patients with sICAS who have an unsatisfactory response to drug treatment, EVT can reduce the risk of stroke recurrence or death within 1 year without increasing the risk of sICH. The safety and efficacy of balloon angioplasty and stenting are similar.

Acute urinary retention(AUR)occurs frequently among astronauts on orbit.The current treatment is complex and easy to damage the urethra,which seriously affects the life and work of astronauts.In contrast,acupuncture,a traditional Chinese remedy,has shown promising results in managing urinary retention.However,the specific acupoints that could potentially prevent AUR remain uncertain due to the unique physiological conditions experienced by individuals in space compared to those on Earth.To address this gap,our research delved into the mechanisms of AUR and acupuncture within both traditional Chinese medicine and modern medical practices.We conducted a thorough literature review from Pubmed,Web of Science,CNKI,Wanfang database,VIP database and Chinese Medical Code database.A hierarchical evidence-scoring approach was utilized to analyze the included literatures,thus devised acupuncture protocols for the treatment of AUR.The outcomes of our study aim to establish a foundation for the application of acupuncture in managing AUR.

Under the influence of long-term space flights and the confined space environment,it is very easy to induce space depression syndrome,mainly manifested as decreased emotional stability,sleep disorders,mental fatigue,etc.,which seriously affect the living conditions and working abilities of astronauts.The current treatment methods mainly focus on psychological support and drug intervention.Acupuncture has a good effect in treating depression.Therefore,starting from the TCM pathogenesis and modern medical pathogenesis of aerospace depression syndrome,we conducted literature retrieval from databases such as the Chinese Medical Classic,China National Knowledge Infrastructure(CNKI),Wanfang,VIP,PubMed,and Web of science.For the included literature,we adopted the stratified evidence scoring method and combined the TCM mechanism and modern medical mechanism of the effect of acupuncture.A prescription for acupuncture points was constructed to provide a basis for selecting acupuncture points for the prevention of aerospace depression syndrome through acupuncture.

Objective To construct a conditional knockout mice model of cysteine-rich angiogenesis inducer 61(Cyr61,also known as Ccn1)gene in myocardium and skeletal muscle regulated by CRE recombinant enzyme.Methods C57BL/6 mice were used to create CKmm-Cre+/-Cyr61flox/flox mice by using CRISPR/Cas9 technology.The successful construction of conditional knockout mice was confirmed by identifying the Cyr61flox/flox,3'LoxP and Cre enzyme sequences in mice.The knockout efficiency of Cyr61 was confirmed by Western blot.The skeletal mus-cle function of Cyr61 knockout mice was evaluated and following related indicators in the myocardium and skeletal muscle were detected by Western blot:Aging(p53,p21),inflammatory response(TNF-α,IL-18,IL-1β),fibrosis(vimentin,TGF-β,α-SMA,COL1).Results The mice were successfully bred and identified.In comparison with the control group,Cyr61 protein level showed a significant decrease in both myocardium and skeletal muscle in the experimental group(P＜0.01).Compared with the control group,the experimental group mice showed increased skeletal muscle grip strength(P＜0.05),enhanced maximum single contraction force(P＜0.01),and increased average cross-sectional area of the anterior tibialis muscle(P＜0.05).The expression level of p21,TNF-α,IL-18,IL-1β,TGF-β,and COL1 proteins in the myocardium and skeletal muscle of the experimental group was all lower than those of the control group(P＜0.05).Conclusions Myocardial and skeletal muscle-specific Cyr61 gene condi-tional knockout mice were successfully constructed based on Cre-LoxP technology and heritable trait could be passed stably.

Objective To investigate the effects of body weight and length of SD rats on the carotid artery balloon injury-induced vascular stenosis model in order to provide a reference for replicating an ideal vascular stenosis mod-el.Methods Male rats were divided into three groups based on body weight and length.The CONQUEROR? SC PTCA balloon catheter was employed,with a fixed balloon inflation volume of 0.2 mL to induce injury in the left common carotid artery,while the right side served as a control.As soon as surgery operation,one rat from each group was selected for Evans Blue dye verification.Fourteen days later,the injured and contra lateral common ca-rotid arteries from remaining rats were harvested for HE staining to check the extent of stenosis.Based on these find-ings,six rats within the optimal range of body weight and length were selected for further validation.Results Rats with body weights ranging from 280 to 380 g(corresponding body lengths of 21.0-26.5 cm)underwent balloon catheter injury,resulting in endothelial detachment and varying degrees of stenosis in the common carotid artery.In rats weighing 280-300 g(body lengths of 21.0-22.5 cm)had severe stenosis or occlusion of the common carotid artery with thrombosis.In rats weighing 320-340 g(body lengths of 23.0-24.5 cm),the internal and external elastic plates of the common carotid artery were ruptured and the vascular morphology was abnormal.Conversely,rats weighing 360-380 g(body lengths of 25.0-26.5 cm)did not show any ruptured elastic laminae or thrombus formation in the common carotid artery,and the extent of vascular stenos in rats with a body weight of 360 g was moderate and uniform.The results of the repeated validation experiments were consistent.Conclusions Rats with a body weight range of 360 g(corresponding body length of 25.0-26.5 cm)are suitable for development of an ideal vascular stenosis model.

Objective To evaluate the acute toxicity and acute eye irritation of hydroquinone. Methods i) Acute toxicity test. Specific pathogen-free (SPF) Kunming mice were randomly divided into four dose groups, 10 mice per group with equal number of males and females. Hydroquinone was administered as a single exposure via oral gavage and intraperitoneal injection at doses of 0.00, 100.00, 250.00, and 500.00 mg/kg body weight. The mice were observed for 14 days. The toxic symptoms were recorded, and median lethal dose (LD₅₀) was calculated. ii) In vitro eye irritation test. Fertilized chicken embryos were randomly divided into four dose groups, with six embryos in each group. The chorioallantoic membrane (CAM) assay was used to evaluate the acute eye irritation potential of hydroquinone in vitro. iii) SPF Kunming mice were randomly divided into three doses groups, 10 mice per group with equal numbers of males and females. Hydroquinone was administered via tail vein injection at doses of 0.00, 25.00, and 50.00 mg/kg body weight. Blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), methemoglobin (MetHb), and cholinesterase levels were measured using colorimetric methods after one hour exposure. Organ coefficients of the heart, liver, spleen, lungs, and kidneys in mice were calculated. Results i) Following oral administration of 500.00 mg/kg body weight hydroquinone, the mice rapidly developed severe toxic symptoms, including agitation, cyanosis of the lips, eye closure, and limb convulsions. Trunk rigidity and curling occurred within 15-60 mins, ended up with death. After intraperitoneal injection at 500.00 mg/kg body weight hydroquinone, toxic reactions occurred more rapidly, with all mice died within five mins. The LD₅₀ values for acute oral and intraperitoneal exposure were 356.64 and 275.90 mg/kg body weight, respectively. Female mice had higher LD₅₀ values for acute intraperitioneal exposure than males (316.58 vs 276.38 mg/kg body weight). ii) The in vitro eye irritation test results showed an irritation score of 3.05 at a dose of 100.00 mg/kg body weight, indicating mild eye irritation, accompanied by vascular congestion and edema in the embryos. iii) Tail vein injection results showed that mouse serum ALT activity increased with increasing hydroquinone doses (all P<0.05), and ALT activity was higher in males than in females (P<0.01). Serum AST activity of mice in the 25.00 and 50.00 mg/kg body weight groups was higher than that in the 0.00 mg/kg body weight group (both P<0.05). With increasing hydroquinone-exposed doses, whole bood MetHb levels increased and cholinesterase activity decreased in both male and female mice (both P<0.05). Male mice had higher MetHb levels than females at corresponding doses among 25.00 and 50.00 mg/kg body weight groups (all P<0.05). Serum cholinesterase levels in male mice were higher than that in females at corresponding doses among 0.00 and 25.00 mg/kg body weight groups (both P<0.05). As the hydroquinone exposure dose increased, the liver organ coefficients decreased in both female and male mice (both P<0.05). The liver organ coefficient in male mice at the 50.00 mg/kg body weight group was higher than that in female mice (P<0.05). Compared to mice of the same sex in the 25.00 mg/kg body weight group, the kidney organ coefficients decreased in both female and male mice in the 0.00 mg/kg body weight group (all P<0.05), and decreased in the 50.00 mg/kg body weight group (all P<0.05). The male mice had lower kidney organ coefficient than female mice at 25.00 mg/kg body weight group (P<0.05). Conclusion Hydroquinone is classified as a moderately toxic substance. Increasing exposure doses result in pronounced eye irritation and affect hepatic and renal functions of mice.

It is of great significance to construct an information platform for performance appraisal of tertiary public hospitals to realize real-time monitoring and intervention of appraisal indicators, which is conducive to the transmission of responsibility for index optimization to departments and medical groups, and to promote the fine management of hospitals and promote high-quality development.This paper introduced the practice and effect of building a performance appraisal information platform for tertiary public hospitals in four aspects: accurate data filling, timely dynamic monitoring, visual display and safety management since 2022. At the same time, suggestions were put forward for platform optimization from four aspects: data quality control, co-construction and sharing, promotion and application, and system integration, in order to provide reference for other hospitals.

ObjectiveTo investigate the induction of ferroptosis by polyphyllin Ⅱ （PPⅡ） in hepatocellular carcinoma HepG2 cells and its underlying mechanism. MethodThe effect of PPⅡ （0， 1.5， 3.0， 4.5， 6.0， 9.0， 18.0 mg·L^-1） on the in vitro proliferation of HepG2 cells was assessed using the methyl thiazolyl tetrazolium （MTT） assay. Colony formation ability of HepG2 cells was evaluated through a colony formation assay. Cell migration ability was assessed via a scratch assay. Lactate dehydrogenase （LDH） content in HepG2 cells was measured using a kit. Reactive oxygen species （ROS） levels in HepG2 cells were observed using a fluorescence inverted microscope. Malondialdehyde （MDA）， glutathione （GSH）， and free Fe²⁺ content in HepG2 cells were detected using respective kits. The mitochondrial ultrastructure in HepG2 cells was observed by transmission electron microscopy. The expression of ferroptosis-related proteins p53， solute carrier family 7 member 11 （SLC7A11）， glutathione peroxidase 4 （GPX4）， long-chain acyl-CoA synthetase 4 （ACSL4）， and transferrin receptor 1 （TFR1） in HepG2 cells was detected using Western blot. ResultCompared with the control group， the PPⅡ treatment groups showed significantly decreased survival rate of HepG2 cells in a dose-dependent manner （P<0.01）， significantly reduced number of cell colonies （P<0.01）， significantly shortened scratch healing distance， inverse correlation of the migration distance with drug concentration （P<0.01）， significantly increased LDH leakage in cells （P<0.01）， significantly enhanced relative fluorescence intensity of intracellular ROS， and significantly increased accumulation of lipid peroxide MDA （P<0.01）， decreased intracellular GSH content with increasing drug concentration （P<0.01）， and significantly enhanced fluorescence intensity of FeRhoNox-1 in cells （P<0.01）. Moreover， cells exhibited vacuolation， and mitochondria showed significant shrinkage with reduced or even disappeared cristae. Compared with the results in the control group， the expression of p53， ACSL4， and TFR1 proteins significantly increased， while the expression of SLC7A11 and GPX4 proteins significantly decreased in the PPⅡ treatment groups （P<0.05）. ConclusionIn summary， PPⅡ induces ferroptosis in HepG2 cells by regulating the p53/SLC7A11/GPX4 signaling axis， promoting ACSL4 expression and Fe³⁺ uptake， leading to an imbalance in the antioxidant system.