Objective To optimize the processing technology of Curculigo orchioides Gaertn.Methods On the basis of single factor experiment,the ratio of material to liquid,frying temperature and frying time were taken as the influencing factors,and the comprehensive scores of appearance traits,curculigoside,orcinol glucoside and orcinol gentiobioside were taken as the evaluation indexes.The optimal processing technology was optimized by Box-Behnken response surface method combined with analytic hierarchy process,and the process verification was carried out.Results The optimal process was as follows:the raw C.orchioides pieces were mixed with the same amount of Evodia rutaecarpa juice,placed in a closed container,moistened until the juice was completely absorbed,stir-fried at 130 ℃ for 6 min,taken out and dried at 60 ℃.For each 100 kg of Curculigo,10 kg of Evodia rutaecarpa was used.Evodia rutaecarpa juice preparation method:10 kg of Evodia rutaecarpa was soaked in 12 times the amount of water for 60 min,decocted for 50 min,extracted for 3 times,combined with the filtrate,and concentrated to 100 kg to obtain Evodia rutaecarpa juice.Conclusion The optimal processing technology of Curculigo orchioides Gaertn.is stable and feasible.

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.

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 explore the predictive value of serum high-mobility group box protein B1(HMGB1)and soluble receptor for advanced glycation end-products(sRAGE)in the occurrence and short-term prognosis of sepsis-associated encephalopathy(SAE).Methods Clinical data of 228 patients with sepsis were retrospectively analyzed.According to the presence of SAE,patients were divided into the SAE group(96 cases)and the non-SAE group(132 cases).General clinical data,laboratory test results,Acute Physiology and Chronic Health Evaluation Ⅱ(APACHEⅡ)scores,Sequential Organ Failure Assessment(SOFA)scores and serum HMGB1 and sRAGE levels were compared between the two groups.Multivariate Logistic regression analysis was performed to determine factors influencing SAE occurrence.Receiver operating characteristic(ROC)curves were plotted to evaluate the predictive ability of HMGB1,sRAGE and the HMGB1/sRAGE ratio to predict the occurrence and short-term prognosis of SAE.Kaplan-Meier survival curves were used to compare the 28-day mortality rates of SAE patients with different HMGB1 and sRAGE expression levels.Results Compared to the non-SAE group,patients in the SAE group exhibited elevated serum HMGB1 levels,decreased sRAGE levels and an increased HMGB1/sRAGE ratio(P＜0.05).The areas under the curve(AUC)for predicting SAE using HMGB1,sRAGE and the HMGB1/sRAGE ratio were 0.826(95%CI:0.770-0.872),0.682(95%CI:0.617-0.742)and 0.895(95%CI:0.848-0.932),respectively,indicating predictive value.Among the 96 SAE patients,52(54.2%)died within 28 days.There were no statistically significant differences in HMGB1,sRAGE and the HMGB1/sRAGE ratio between surviving and deceased patients(P＞0.05).Similarly,there were no significant differences in 28-day mortality rates between SAE patients with different HMGB1 or sRAGE expression levels.Conclusion Elevated serum HMGB1 and reduced sRAGE are of significant value in the auxiliary diagnosis of SAE,but have limited clinical predictive value for short-term prognosis.