AIM: To investigate the relationship between vascular smooth muscle cell (VSMC) injury, organelle stress response and autophagic cell death (autophagy) and ferroptosis induced by the chemical hypoxia inducer cobalt chloride (CoCl2) through the bioinformatics analysis and in vitro cell experimentation. METHODS: The dataset GSE119226 of VSMC treated with cobalt chloride was acquired from the gene expression database (GEO). The R language was used to investigate the relationship between CoCl2 treatment and organelle stress response (Golgi stress, endoplasmic reticulum stress) and two forms of cell death (ferroptosis and autophagic cell death). With primary cultured rat VSMC (rVSMC) and CoCl2-induced anoxia model, the changes in cell viability were detected by CCK-8 method, and reactive oxygen species (ROS) levels were measured using DCFH-DA method. The expression levels of HIF-1α (a key molecule in hypoxia), Golgi stress markers GM130 and p115, endoplasmic reticulum stress markers GRP78 and CHOP, autophagy markers LC3-II / LC3-I and Beclin1, and ferroptosis markers GPx4 and xCT were detected by Western blot. The effect of inducing or inhibiting organelle stress and cell death on the CoCl2-induced cell damage was also observed. RESULTS: Differentially expressed genes analysis of GSE119226 dataset showed that CoCl2 treatment of VSMCs had significant effects on organelle function and stress response, autophagy and ferroptosis-related genes, in which endoplasmic reticulum stress, protein processing in endoplasmic reticulum, regulation of Golgi to plasma membrane protein transport, autophagy / autophagic cell death, and ferroptosis pathways were remarkably enriched. The results of in vitro experiment showed that compared with normal rVSMC, cell viability was significantly decreased after CoCl2 treatment, as well as HIF-1α protein expression and ROS levels in rVSMCs were increased. In rVSMC treated with Co-Cl2, the expression levels of Golgi structural proteins GM130 and p115 (reflecting the occurrence of Golgi stress) were decreased, while the markers GRP78 and CHOP (reflecting the occurrence of endoplasmic reticulum stress) were increased. At the same time, CoCl2 treatment also reduced the expression of autophagy markers LC3-II/LC3-I and Beclin1 (indicating the decrease levels of autophagy), while the expression of ferroptosis markers GPx4 and xCT were decreased (indicating the occurrence of ferroptosis). Compared with CoCl2 treatment group, induced Golgi stress, endoplasmic reticulum stress, or ferroptosis could further reduce cell viability, while inhibition of these processes could improve cell viability. On the other hand, increasing the level of autophagy can improve the cell viability. CONCLUSION: Hypoxia induced by cobalt chloride can lead to VSMC injury. Golgi stress, endoplasmic reticulum stress, ferroptosis, and the reduction of autophagy level play an important role in it. Inhibition of organelle stress response and ferroptosis, or increase of autophagy level can improve VSMC injury caused by cobalt chloride.

Objective To observe the improved effect of propofol on vascular hyporeactivity in septic rats and its underlying mechanism.Methods A total of 96 SD rats(12 weeks old,both genders,weighing 200～220 g)were randomly divided into sham group(n=16),sepsis group(n=16,cecal ligation and puncture),propofol group(n=16),propofol+ROCK inhibitor Y-27632 group(n=16),propofol+PKCαinhibitor GO6976 group(n=16),propofol+IP3 inhibitor 2-APB group(n=8)and propofol+gap junction inhibitor metoclopramide sodium(Movens)group(n=8).In vitro vascular ring reactivity and vascular calcium sensitivity were measured to observe the improved effects of propofol on vascular hyporeactivity in septic rats and its relationships with RhoA/ROCK,PKCα,IP3 and cell gap junction.Results Determination of in vitro vascular ring and calcium sensitivity showed that the contractile reactivity to norepinephrine(NE)and to calcium sensitivity were significantly decreased in the arterial rings isolated from the septic rats compared with those from the sham group,with the dose-response curve shifting to the right,and most significant decrease by 51.42％in the superior mesenteric artery(SMA,P＜0.05).Propofol treatment significantly improved the hyporeactivity and calcium sensitivity of the vessels isolated from the septic rats,especially those of the femoral artery with a recovery rate of 89.57％(P＜0.05).In comparison with the propofol group,the dose-response curves of the propofol+Y-27632 group and the propofol+GO6976 group were shifting to right,indicating that Y-27632 and GO6976 could significantly inhibit the amelioration of propofol on calcium sensitivity of SMA in severely septic rats with an inhibitory rate of 146.95％and 88.63％(P＜0.05),respectively.Isolated vascular reactivity measurement demonstrated that Y-27632 and Movens treatment significantly antagonized the ameliorated role of propofol on hyporeactivity of blood vessels from the septic rats with an inhibitory rate of 40.79％and 169.90％(P＜0.05),separately,while no such effect was observed in the propofol+GO6976 and propofol+2-APB groups.Conclusion Propofol treatment can significantly improve vascular hyporeactivity of septic rats,which may attribute to the increase of vascular calcium sensitivity through RhoA/ROCK pathway.

Objective To investigate the clearance capacity of cardiac resident macrophages in post-sepsis and its underlying mechanism.Methods A mouse model of sepsis was established using cecum perforation ligation.Thirty male C57BL/6 mice(8 weeks old,weighing 20～25 g)were randomly and equally divided into a sham operation group(sham group)and a model group(sepsis group).Immunofluorescence assay was employed to label the cardiomyocytes and macrophages to observe the apoptosis of cardiomyocytes and the phagocytosis by cardiac resident macrophages.Cardiac resident macrophages were extracted for transcriptomic sequencing to determine the functional changes of the cells after sepsis.Cardiac resident macrophage cell lines were established at the cellular level and served as the normal group(RAC group),and the RAC cells treated with LPS were subjected as the sepsis group(RAC+LPS group).Then the differences in the ability to clear apoptotic cardiomyocytes between the 2 groups were observed.Then DQ-BSA-RED lysosomal activity detection probe,Lyso-Sensor yellow/bule dye,ELISA,and Western blotting were applied to detect the lysosomal function of cardiac resident macrophages,activity and expression of important lysosomal hydrolases,changes in contents and related subunits of vacuolar-type adenosine triphosphatases(V-ATPase).Results Compared with the sham group,the sepsis group had larger number of apoptotic cardiomyocytes(P＜0.05)and increased phagocytosis of cardiomyocytes by cardiac macrophages(P＜0.05).The results of transcriptomic sequencing revealed a significant dysfunction of lysosome-associated functions of cardiac-resident macrophages after sepsis.In in vitro experiments,the RAC+LPS group had a reduced fragmentation capacity of apoptotic cardiomyocytes,reduction in the intensity of yellow fluorescence of lysosomes(P＜0.05),and decrease in lysosomal hydrolase activity(P＜0.05)when compared with the RAC group.In addition,LPS treatment significantly decreased the activity and expression of V-ATPase and its major subunit ATP6V1A in cardiac resident macrophages(P＜0.05).Conclusion Cardiac resident macrophages show reduced clearance of apoptotic cardiomyocytes after sepsis,which may be related to a decrease in the activity of ATP6V1A,an important subunit of its lysosomal V-ATPase,and reduced activity of lysosomal hydrolases.

Objective:To analyze the hotspots and trends of the researches on artificial intelligence (AI) in the diagnosis and treatment of traumatic brain injury (TBI).Methods:Based on the core database of Web of Science, the studies over AI in the diagnosis and treatment of TBI published from January 2000 to June 2024 were obtained by searching with the subject headings. VOSviewer software was used to analyze the publication year trend, country publication volume, country cooperation network, author publication volume, author citation frequency and author cooperation network. CiteSpace software was also used to identify key words with a significant rise in frequency over a short period of time to obtain the research trends.Results:A total of 2 662 relevant studies were retrieved, from which 677 related with AI in the diagnosis and treatment of TBI were finally enrolled. The number of published studies per year generally showed a rapid growth from 2018 to 2023. The United States had the highest number of publications as a country (362 studies). The author Camarillo had the most publications (9 studies). Rehabilitation was the keyword with the highest frequency (133 times) and the clustering topics containing the three largest number of keywords were virtual reality (VR), mild TBI, and deep learning. The keywords of mobile application, mobile health and intracranial pressure showed a significant increase in frequency from January 2022 to June 2024.Conclusions:VR technology, mild TBI and deep learning technology are the research hotspots of AI in TBI diagnosis and treatment. Mobile apps, mobile health, and intracranial pressure may be new research trends for AI in the diagnosis and treatment of TBI.

Objective:To discuss the effect of fluid resuscitation on the occurrence of ferroptosis in vascular tissue and the structure of vascular cells in the rats with blast injury complicated with hemorrhagic shock,and to clarify its mechanism.Methods:A total of 54 healthy adult SD rats were randomly divided into normal group,blast injury complicated with hemorrhagic shock(model)group,and the fluid resuscitation(treatment)group,and there were 18 rats in each group.Among them,10 rats were randomly selected to observe the surival status and another 8 rats were selected to detect the other indexes.The average survival time(ST),24 h and 72 h survival rates of the rats in various groups were observed;the blood pressure(BP),heart rate(HR),and respiratory rate(RR)of the rats in various groups were observed;the levels of serum creatinine(Scr),blood urea nitrogen(BUN),lactate(LAC),glucose(GLU),iron ions,glutathione(GSH),and malondialdehyde(MDA)and the activities of aspartate aminotransferase(AST),alanine aminotransferase(ALT)and lactate dehydrogenase(LDH)in serum of the rats in various groups were detected;Western blotting method was used to detect the expression levels of ferroptosis marker proteins glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11),and heme oxygenase 1(HO-1)proteins in superior mesenteric artery tissue of the rats in various groups;the pathomorphology of the superior mesenteric artery of the rats in various groups was observed.Results:All the rats in normal group survived for 72 h,while the longest ST of the rats in model group did not exceed 9 h.Compared with model group,the ST and 24 h survival rate(SR)of the rats in treatment group were significantly increased(P＜0.05).Compared with normal group,the BP,HR,and RR of the rats in model group were significantly decreased(P＜0.01).Compared with model group,the BP,HR,and RR of the rats in treatment group were significantly increased after fluid resuscitation(P＜0.05).Compared with normal group,the activities of AST and ALT,and the levels of Scr and BUN in serum of the rats in model group were significantly increased(P＜0.01).Compared with model group,the serum levels of LAC and GLU of the rats in treatment group were significantly decreased(P＜0.01).Compared with normal group,the concentration of iron ion,GSH level,MDA level,LDH activity in serum of the rats in model group were significantly increased(P＜0.05);compared with model group,the concentration of iron ion and LDH activity in serum of the rats in treatment group was significantly decreased(P＜0.01).Compared with normal group,the expression levels of GPX4 and SLC7A11 in superior mesenteric artery tissue of the rats in model group were significantly decreased(P＜0.05);compared with model group,the expression levels of GPX4 and SLC7A11 in superior mesenteric artery tissue of the rats in treatment group were significantly increased(P＜0.05).Compared with normal group,the expression level of HO-1 protein in superior mesenteric artery tissue of the rats in model group was increased(P＜0.01);compared with model group,the expression level of HO-1 protein in superior mesenteric artery tissue of the rats in treatment group was increased(P＜0.01).The microscopic pathology results showed that the cell arrangement in the layers of the superior mesenteric artery tissue of the rats in model group was disordered,the swelling was significant and the thickness was increased;the pathological changes in superior mesenteric artery tissue of the rats in treatment group was alleviated.The ultramicroscopic pathology results showed that the endothelial cell structure of blood vessels of the rats in normal group was intact,and there was no swelling in the subendothelial matrix;the vascular endothelial cell membrane of the rats in model group was damaged,there were cytoplasmic dissolution and fragmentation,and the swelling of the subendothelial matrix was significant;the swelling of the vascular endothelial cells in treatment group was alleviated.Conclusion:Ferroptosis occurs in vascular tissue of the rats with blast injury complicated with hemorrhagic shock,and fluid resuscitation can alleviate the structural damage of the vascular cells by inhibiting the vascular tissue ferroptosis.

The endocannabinoid system(ECS)consists of a variety of long-chain unsaturated fatty acid analogs,mainly anandamide(AEA)and 2-arachidoniyl glycerol(2-AG),along with their specific binding G protein-coupled receptors,cannabinoid receptor 1(CB1R)and cannabinoid receptor 2(CB2R).It affects the life process and biological activity of almost all cells in the body by influencing cell material and energy metabolism.In the liver,the physiological expression of ECS is at a low level.The expression and secretion of ECS in the liver can be strongly stimulated by liver injury factors.ECS acts as a trigger in multiple liver diseases.It is known to be related to the process of hepatocyte steatosis and promote the formation and development of non-alcoholic fatty liver disease(NAFLD)and alcoholic liver disease(ALD).It is involved in the inflammatory processes of liver diseases and greatly affects the immune-inflammatory response in liver tissue.It is also involved in the formation of liver fibers and promotes the occurrence and development of liver fibrosis and cirrhosis.Finally,the role and mechanisms of ECS in the occurrence and development of liver diseases are elaborated in detail by listing lipid metabolism-related liver diseases(NAFLD and ALD)and other liver diseases.

OBJECTIVE To investigate the protective effects and mechanism of ziyuglycoside Ⅰ on acute lung injury in sepsis rats based on network pharmacology， and conduct experimental verification. METHODS The network pharmacology was used to predict the potential target of ziyuglycoside Ⅰ in the treatment of acute lung injury following sepsis. The rat model of sepsis was reproduced by cecum ligation and puncture for experimental verification. Totally 192 SD rats were randomly divided into the sham operation group （Sham group）， sepsis group （Sep group）， conventional therapy group （CT group） and ziyuglycoside Ⅰ group （Zg Ⅰ group）， respectively. Sham group and Sep group were given sterile normal saline， and CT group and ZgⅠ group were given relevant volume of Ringer’s solution and ziyuglycoside Ⅰ. The arterial blood gas， serum inflammatory factors， lung wet/dry mass ratio， pathological changes of lung tissue， pulmonary vascular permeability， the expressions of pulmonary vein tight junction protein 1 （ZO-1） and vascular endothelial cadherin （VE-cadherin） protein and 72-hour survival were observed in each group. RESULTS Results of network pharmacology showed that there were 47 potential targets of ziyuglycoside Ⅰ in the treatment of sepsis. The results of gene ontology function enrichment analysis and Kyoto encyclopedia of genes and genomes pathway enrichment analysis showed that the mechanism could 598486924@qq.com be correlated with biological processes such as positive regulation of reactive oxygen species metabolism， wound healing， regulation of endothelial cell proliferation， cell activation， blood vessel development， response to oxidative stress， etc.， and with signaling pathway such as apoptosis， tight junction， HIF-1 signaling pathway， etc. The results of experimental verification showed that compared with Sham group， pH value and the level of partial arterial oxygen pressure were decreased significantly in Sep group （P＜0.05）， while the level of partial pressure of carbon dioxide， serum levels of tumor necrosis factor α， interleukin 6 were increased significantly （P＜0.05）； the ratio of lung wet/dry mass was increased significantly （P＜0.05）； the protein expressions of ZO-1 and VE-cadherin were decreased significantly （P＜0.05）； 72 h survival rate decreased，the survival time was significantly shortened （P＜0.05）； the results of pathological observation of lung tissue showed that the rats’ alveoli were extensively ruptured， the alveolar wall was thickened and accompanied with edema， and there was obvious inflammatory cell infiltration； the results of pulmonary vascular permeability observation showed that the lung surface of rats was dark， with a large amount of Evans blue exudation， and the left lower lung was obviously dark blue. Compared with Sep group， the levels of above indexes almost were reversed significantly in CT group and ZgⅠ group （P＜0.05）； the lung histopathology and pulmonary vascular permeability were significantly improved， and the recovery degree of ZgⅠ group was greater than that of CT group， which was close to the results of Sham group. CONCLUSIONS Ziyuglycoside Ⅰ can significantly reduce inflammatory reaction and acute lung injury in septic rats， which is related to vascular function and tight junction signal pathway.

Macrophages are a heterogeneous cell population involved in tissue homeostasis, inflammation, and various pathophysiological processes. Selective clearance of macrophages in vivo is a widely accepted method used to investigate whether macrophages are involved in any specific biological regulatory mechanism. Understanding the different methods and principles of macrophage depletion helps explore appropriate modeling solutions to investigate the relationship between macrophages and diseases. Hepatic macrophages play a crucial role in hepatotoxicity. This article reviews the methods of macrophage depletion and the regulatory role of macrophage depletion in liver injury and repair.

Helicobacter pylori (Hp) infection is closely related to the occurrence and development of chronic gastritis, peptic ulcer, and gastric cancer. At present, the treatment of Hp is mainly the bismuth-containing quadruple therapy as the first-line treatment to eradicate Hp infection. However, the eradication treatment still faces the challenges related to the rising antibiotic resistance, the decrease in eradication rate year by year, the adverse events, the poor patient's compliance and the dysregulation of gastrointestinal microbiome. Therefore, more and more researches are focusing on finding an effective treatment with the use of natural therapy. This article reviewed the research progress of pathogenic mechanism and treatment of Hp infection.

Objective:To explore and establish the scoring method of injury assessment in rats with trauma combined with seawater immersion, so as to provide a reference for injury assessment in the special environment of trauma combined with seawater immersion.Methods:Sixty-four SD rats were divided into two groups according to the random number table, including hemorrhagic shock group and compound injury group, with 32 rats per group. Each group was divided into trauma combined with seawater immersion group and simple trauma group, with 16 rats per group. In trauma combined with seawater immersion group, the hemorrhagic shock model was placed in 15℃ seawater for 1 hour to start bleeding, and the blood loss was 30% of the total blood volume. The composite injury model caused 10% Ⅱ degree burns and was incised along the vental midline with a length of about 2 cm, and then placed in 15℃ seawater for 1 hour. The death and survival time were recorded.The survival time significantly longer than 4 hours out of water was recorded as survival, and significantly shorter than 4 hours out of water was recorded as death. Data were observed within 9 hours after injury, including the changes of physiological indexes (respiration, blood pressure, anal temperature) and arterial blood gas (blood glucose, pH value, blood lactic acid, arterial oxygen partial pressure, arterial carbon dioxide partial pressure, bicarbate, sodium ion, chloride ion, calcium ion, potassium ion). Each index were compared between trauma combined with seawater immersion group and simple trauma group. According to the survival situation of all the trauma combined with seawater immersion group at 4 hours out of water, the rats were divided into survival group and death group. The indicators affecting survival were screened, and then the scatter plot of each index corresponding to the mortality rate was established. According to the trend of each index in different interval in the scatter chart, the score table of injury condition was established.Results:The total mortality was 28% (9/32) in trauma combined with seawater immersion group, and was 6% (2/32) in simple trauma group ( P<0.05). The survival time in trauma combined with seawater immersion group [(8.1±3.7)hours] was shorter than that in simple trauma group [(11.3±4.8)hours] ( P<0.05). In trauma combined with seawater immersion group, the respiratory rate[(58.8±2.9)times/min] was slower than that in simple trauma group [(100.4±7.2)times/min], blood pressure [(80.0±25.1)mmHg] was lower than that in simple trauma group [(89.8±18.1)mmHg], and anal temperature [22.4(20.1, 25.0)℃] was significantly lower than that in sample trauma group [31.7(30.5, 33.2)℃], pH value (7.1±0.1) was lower than that in simple trauma group (7.3±0.1), and arterial oxygen partial pressure [(196.3±34.1)mmHg], arterial carbon dioxide partial pressure [45.5(35.1, 51.1)mmHg], serum sodium [145(142, 148)mmol/L], serum chlorine [120(115, 125)mmol/L], serum calcium [(1.3±0.1)mmol/L]as well as serum potassium [(3.6±0.8)mmol/L] were higher than those in simple trauma group [(149.4±22.6)mmHg, 29.7(25.6, 34.5)mmHg, 142(139, 144)mmol/L, 118(114, 121)mmol/L, (1.2±0.1)mmol/L, (3.3±0.6)mmol/L] (all P<0.05). There were no significances in other indexes between the two groups ( P>0.05). In death group, the breathing[36(30, 36)times/min], blood pressure [(43.1±21.8)mmHg], anal temperature [(20.0±1.9)℃], pH value (7.1±0.1), and bicarbonate [(12.3±2.2)mmol/L] were significantly inhibited or suppressed compared with survival group [60(48, 78)times/min, (86.6±19.3)mmHg, (23.0±3.1)℃, 7.2±0.1, (14.6±2.3)mmol/L (all P<0.05). While the two groups showed no significant differences in other indices ( P>0.05). Therefore, the respiration, blood pressure, rectal temperature, pH value and bicarbonate that significantly affect the survival of rats were screened. According to the death rate corresponding to different intervals, a score value was assigned to the interval as the weight of its impact on survival, namely on the severity of the injury, and an injury score table for trauma combined with seawater immersion in rats was established. The injury scoring scale <6 points indicated no death, 6-9 points indicated the mortality of 50%, ≥9 points indicated the mortality of 71%. The 6 points and 9 points were cutoff value of the scale. It can be considered that the scale of <6 points was classified as minor injury, 6-9 points as moderate injury, and ≥9 points as severe injury. Conclusions:The seawater immersion can result in reduced survival time and increased early mortality, manifested as respiratory depression, more serious blood loss, severe hypothermia, severe metabolic acidosis, water and electrolyte disorders (high sodium, high chlorine, high calcium, and high potassium), etc. According to the respiration, blood pressure, anal temperature, pH value and bicarbate, which affect the survival of rats, the injury rating scale of rats with trauma combined with seawater immersion can be established by using the scatter chart. The predicted mortality rate by using the rating scale was roughly consistent with the actual mortality rate, so the injury rating scale basically had a good prediction and hint for the trauma rats combined with seawater immersion.