In this study, RAW264.7 cells were employed to investigate the effects of honey-processed Astragalus on their energy metabolism and polarization, and explore the scientific connotation of the enhanced efficacy of honey-processed Astragalus on invigorating spleen-stomach and replenishing Qi. The medicated sera were prepared by intragastric administration of rats with water extracts of crude and honey-processed Astragalus, and the composition changes of medicated sera of crude and honey-processed Astragalus were analyzed by using LC-MS technology. The cell survival rates were detected and the concentrations of medicated sera were screened through CCK-8 assay. The differences of cell phagocytic rates, ATP energy metabolism, and NO secretion between crude and honey-processed Astragalus were evaluated by using neutral red phagocytosis assay, ATP detection kit, and NO detection kit. The effects of crude and honey-processed Astragalus on TNF-α secretion of RAW264.7 cells were detected by employing ELISA kit. The effects of crude and honey-processed Astragalus on polarization of RAW264.7 cells were evaluated by utilizing flow cytometry. The differential metabolites related to glycolysis in cell lysates and culture media were screened by using LC-MS technology. The experiment was approved by the experimental animal ethics committee from Shanxi University of Chinese Medicine (No. AWE202407352). The results showed that the contents of betaine, amino acids, and ononin in the prepared medicated sera of rats treated by intragastric administration with water extracts of honey-processed Astragalus increased compared to those in crude one. The results of CCK-8 experiment showed that there were no cytotoxic effects on RAW264.7 cells in the medicated sera of crude and honey-processd Astragalus at different concentration. The phagocytic index and ATP yield both increased to varying degrees after administration of the medicated sera to cells. The secretion of NO in normal and inflammatory cells increased and decreased respectively, and the effect of honey-processed Astragalus was better than that of crude one. The results of ELISA kit showed that the medicated sera of both crude and honey-processed Astragalus could promote the secretion of TNF-α in a concentration-dependent manner, and the promoting effect of honey-processed Astragalus was stronger than that of crude one. The results of flow cytometry showed that the medicated sera of both crude and honey-processed Astragalus could promote M1-type polarization and inhibit M2-type polarization of RAW264.7 cells, and the effect of honey-processed Astragalus was better than that of crude one. Compared to crude Astragalus, the metabolites related to glycolysis in the cell lysates and culture media of the medicated sera of honey-processed Astragalus were generally on the rise, indicating that the effect on promoting glycolysis of honey-processed Astragalus was better than that of crude one. In summary, honey-processed Astragalus can promote the polarization and energy metabolism of RAW264.7 cells, and participate in positive immune regulation, which is correlated with its enhanced effect of invigorating spleen-stomach and replenishing Qi.

OBJECTIVE To prepare tetrandrine （TET）-loaded chitosan（CS）-stearic acid （SA） nano micelles modified with folic acid （FA）（ FA-CS-SA/TET nano micelles）， characterize them and study the anti-inflammatory effect in vitro. METHODS FA- CS-SA/TET nano micelles were prepared by ultrasonic method； the preparation technology was optimized by orthogonal test and validation test was also performed with the mass ratio of FA-CS-SA to TET， ultrasound power and ultrasound times as the factors， using the comprehensive score of entrapment efficiency （EE）， drug loading （DL） and particle size as evaluation index. FA-CS-SA/ TET nano micelles prepared by the optimal technology were characterized， and their release performance in vitro was investigated. RAW264.7 cells were used as subjects to investigate their anti-inflammatory activity in vitro. RESULTS The optimal preparation technology included that the mass ratio of FA-CS-SA to TET was 2∶1， ultrasonic power was 200 W， and the ultrasonic frequency was 200 times. The parameters of FA-CS-SA/TET nano micelles prepared by optimized technology included that EE was （98.86± 0.30）%， DL was （28.57±0.34）%， the average particle size was （227.0±9.4） nm， polydispersity index was 0.42±0.04， and the Zeta potential was（12.6±2.3）mV， respectively. The nano micelles were uniform in appearance and round in shape. The nano micelles were released quickly in 0.5% sodium dodecyl sulfate solution， with a cumulative release rate of （79.49±3.43）% within 72 hours， and its anti-inflammatory effect was stronger than that of TET raw materials. CONCLUSIONS FA-CS-SA/TET nano micelles are prepared successfully in the study， with good drug loading performance， uniform particle size， and good in vitro anti-inflammatory activity.

Radiotherapy is an important means to treat lung cancer,but it is easy to cause lung injury and reduce the quality of life of patients.Flash radiotherapy(FLASH-RT)has attracted attention due to its extremely short radiation duration and high dose rate,which can reduce toxicity of normal tissue while ensures treatment intensity of tumor.Whether Flash-RT can reduce radiation-induced lung injury has become an important research topic in recent years.Based on the literature analysis method,this review systematically assessed the effects and mechanisms of Flash-RT and radiotherapy with conventional dose rate on lung injury through searching relevant literatures at home and abroad,so as to provide scientific basis for the treatment of patients with lung cancer by reviewing the comparisons about the effects and mechanisms between Flash-RT and radiotherapy with conventional dose rate on lung injury.Compared with radiotherapy with conventional radiation rate,Flash-RT can significantly reduce lung injury and improve quality of life of patients.It is still demanded to explore the Flash-RT mechanism in future,so as to develop the Flash-RT instrument that is suitable for different tumors and to conduct larger-scale clinical researches.

OBJECTIVE To explore in vitro dissolution and in vivo pharmacokinetics of Luteolin solid dispersion in Beagle dogs. METHODS The dissolution of Luteolin solid dispersion was investigated according to the second method （paddle method） of the “dissolution determination method” in the 2020 edition of Chinese Pharmacopoeia （Part Ⅳ）. UPLC-MS/MS method was established to determine the concentration of luteolin in the plasma of Beagle dogs. Twelve Beagle dogs were randomly divided into luteolin group and Luteolin solid dispersion group， with 6 dogs in each group. They were given relevant medicine orally at the dose of 10 mg/kg luteolin. Blood was collected before medication （0 h）， at 5， 10， 15， 30， 45 min and 1， 2， 4， 6， 8， 10， 12， 24， 48 h after administration. After protein precipitation with acetonitrile， the blood concentration of luteolin in Beagle dogs was determined by UPLC-MS/MS and the major pharmacokinetic parameters were calculated with non-compartmental models by using DAS 3.2.8 pharmacokinetic software. RESULTS The dissolutions of Luteolin solid dispersion in purified water and 0.1% sodium dodecyl sulfate solution was significantly higher than those of luteolin； the dissolution rate reached 95% in 0.1% sodium dodecyl sulfate solution for 120 minutes. The peak concentration （cmax） of luteolin in the Luteolin solid dispersion group of Beagle dogs was 5.62 times higher than the luteolin group， and the relative bioavailability was 348%. Compared with luteolin group， cmax and the area under the drug time curve of luteolin in the Luteolin solid dispersion group of Beagle dogs were significantly increased， while the apparent distribution volume was significantly reduced （P＜0.05）. CONCLUSIONS Luteolin solid dispersion can improve in vitro dissolution and bioavailability of luteolin in Beagle dogs.

Objectiveβ‑Site APP cleaving enzyme 1 (BACE1) is a rate-limiting enzyme involved in the formation of amyloid plaques in Alzheimer’s disease (AD), and its expression and activity play a crucial role in the development of AD. The interacting protein of BACE1 can directly or indirectly regulate BACE1 in the transcription, translation, modification, intracellular transport and other links of BACE1 by directly binding, indirectly binding, and participating in various cell signal transduction pathways, so as to participate in the occurrence of AD and the process of disease. This study aimed to screen and validate the interacting proteins of BACE1, providing new insights into the mechanisms of amyloid plaque formation. MethodsCo-immunoprecipitation (Co-IP) and mass spectrometry (MS) were used to enrich and identify BACE1 interacting proteins in the hippocampus of wild type (WT) mice and AD model mice. For candidate BACE1 interacting proteins, GO enrichment analysis and KEGG pathway enrichment analysis were used to explore the subcellular localization, molecular function, participating biological processes and participating signaling pathways of BACE1 interacting proteins. The protein-protein interaction (PPI) network of BACE1 was further constructed to explore the potential proteins interacting with BACE1. By searching the mouse genomeinformation (MGI) website and NCBI database, the more reliable proteins among the potential BACE1 interacting proteins were screened. Co-IP assay and immunofluorescence confocal technology were used to preliminarily verify the interaction between the proteins, and the changes in protein expression levels of the interacting proteins in AD cell models were explored. ResultsA total of 614 differentially expressed proteins interacting with BACE1 were identified in AD group. GO enrichment analysis showed that the BACE1 interacting proteins in the AD group were mainly located in membrane organelles such as Golgi apparatus, endoplasmic reticulum, endosome, lysosome and vesicles, which had molecular functions such as ion channel regulation, protein kinase activity, transcription factor binding and passive transmembrane transporter activity. It is mainly involved in the biological processes of immune response regulation cell surface receptor signaling pathway, targeting Golgi vesicles transport, circadian rhythm regulation, Purkinje cell layer development, etc. KEGG analysis showed that BACE1 interacting proteins in AD were mainly involved in the PI3K-Akt signaling pathway, mTOR signaling pathway and other neurodegenerative disease-related pathways. The PPI network of BACE1 showed that a total of 12 proteins were identified as high confidence binding proteins, including PRNP, APOE, SYP, NSF, NUMB, SNAP91, HSP90aa1, UCHL1, BIN1, SNX27, Rheb, Ap2m1, of which, NSF, NUMB, SNAP91, HSP90aa1 were newly identified candidate proteins. After further verification, we found that NSF not only interacts with BACE1, but also interacts with amyloid precursor protein (APP), the substrate of BACE1, and the expression level of NSF is up-regulated in the AD cell model constructed by Aβ₄₂ induction. ConclusionBACE1 binding proteins participate in multiple AD-associated biological processes and signal pathways. NSF is a newly identified BACE1 binding protein that interacts with BACE1, and the protein expression level of NSF is up-regulated in the AD cell model. It is predicted that the interaction between NSF and BACE1 is involved in regulating the course of AD, providing a new target and direction for the study of the mechanism of AD.

BACKGROUND:In recent years,with the development of biological scaffold materials and bioprinting technology,tissue-engineered bone has become a research hotspot in bone defect repair. OBJECTIVE:To summarize the current treatment methods for bone defects,summarize the biomaterials and bioprinting technology for preparing tissue-engineered bone scaffolds,and explore the application of biomaterials and printing technology in tissue engineering and the current challenges. METHODS:Search terms were"bone defect,tissue engineering,biomaterials,3D printing technology,4D printing technology,bioprinting,biological scaffold,bone repair"in Chinese and English.Relevant documents published from January 1,2009 to December 1,2022 were retrieved on CNKI,PubMed and Web of Science databases.After being screened by the first author,high-quality references were added.A total of 93 articles were included for review. RESULTS AND CONCLUSION:The main treatment methods for bone defects include bone transplantation,membrane-guided regeneration,gene therapy,bone tissue engineering,etc.The best treatment method is still uncertain.Bone tissue engineering technology is a new technology for the treatment of bone defects.It has become the focus of current research by constructing three-dimensional structures that can promote the proliferation and differentiation of osteoblasts and enhance the ability of bone formation.Biological scaffold materials are diverse,with their characteristics,advantages and disadvantages.A single biological material cannot meet the demand for tissue-engineered bone for the scaffold.Usually,multiple materials are combined to complement each other,which is to meet the demand for mechanical properties while taking into account the biological properties of the scaffold.Bioprinting technology can adjust the pore of the scaffold,build a complex spatial structure,and is more conducive to cell adhesion,proliferation and differentiation.The emerging 4D printing technology introduces"time"as the fourth dimension to make the prepared scaffold dynamic.With the synchronous development of smart materials,4D printing technology provides the possibility of efficient repair of bone defects in the future.

Objective:To investigate the early clinical characteristics of elderly patients with severe burns and the risk factors on prognosis.Methods:This study was a retrospective case series study. Clinical data of 124 elderly patients with severe burns who met the inclusion criteria and were admitted to the 12 hospitals from January 2015 to December 2020 were collected, including 4 patients from the Fourth People's Hospital of Dalian, 5 patients from Fujian Medical University Union Hospital, 22 patients from Guangzhou Red Cross Hospital of Jinan University, 5 patients from Heilongjiang Provincial Hospital, 27 patients from the First Affiliated Hospital of Naval Medical University, 9 patients from the First Affiliated Hospital of Nanchang University, 10 patients from Affiliated Hospital of Nantong University, 9 patients from Tongren Hospital of Wuhan University & Wuhan Third Hospital, 12 patients from the 924 th Hospital of PLA, 6 patients from Zhangjiagang First People's Hospital, 4 patients from Taizhou Hospital of Zhejiang Province, and 11 patients from Zhengzhou First People's Hospital. The patients' overall clinical characteristics, such as gender, age, body mass index, total burn area, full-thickness burn area, inhalation injury, causative factors, whether combined with underlying medical diseases, and admission time after injury were recorded. According to the survival outcome within 28 days after injury, the patients were divided into survival group (89 cases) and death group (35 cases). The following data of patients were compared between the two groups, including the basic data and injuries (the same as the overall clinical characteristics ahead); the coagulation indexes within the first 24 hours of injury such as prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time, D-dimer, fibrinogen degradation product (FDP), international normalized ratio (INR), and fibrinogen; the blood routine indexes within the first 24 hours of injury such as white blood cell count, platelet count, neutrophil-to-lymphocyte ratio, monocyte count, red blood cell count, hemoglobin, and hematocrit; the organ function indexes within the first 24 hours of injury such as direct bilirubin, total bilirubin, urea, serum creatinine, aspartate aminotransferase, alanine aminotransferase, total protein, albumin, globulin, blood glucose, triglyceride, total cholesterol, alkaline phosphatase, creatine kinase, electrolyte indexes (potassium, sodium, chlorine, calcium, magnesium, and phosphorus in blood), uric acid, myoglobin, and brain natriuretic peptide; the infection and blood gas indexes within the first 24 hours of injury such as procalcitonin, C-reactive protein, pH value, oxygenation index, base excess, and lactate; treatment such as whether conducted with mechanical ventilation, whether conducted with continuous renal replacement therapy, whether conducted with anticoagulation therapy, whether applied with vasoactive drugs, and fluid resuscitation. The analysis was conducted to screen the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns. Results:Among 124 patients, there were 82 males and 42 females, aged 60-97 years, with body mass index of 23.44 (21.09, 25.95) kg/m 2, total burn area of 54.00% (42.00%, 75.00%) total body surface area (TBSA), and full-thickness burn area of 25.00% (10.00%, 40.00%) TBSA. The patients were mainly combined with moderate to severe inhalation injury and caused by flame burns. There were 43 cases with underlying medical diseases. The majority of patients were admitted to the hospital within 8 hours after injury. There were statistically significant differences between patients in the 2 groups in terms of age, total burn area, full-thickness burn area, and inhalation injury, and PT, APTT, D-dimer, FDP, INR, white blood cell count, platelet count, urea, serum creatinine, blood glucose, blood sodium, uric acid, myoglobin, and urine volume within the first 24 hours of injury (with Z values of 2.37, 5.49, 5.26, 5.97, 2.18, 1.95, 2.68, 2.68, 2.51, 2.82, 2.14, 3.40, 5.31, 3.41, 2.35, 3.81, 2.16, and -3.82, respectively, P<0.05); there were statistically significant differences between two groups of patients in whether conducted with mechanical ventilation and whether applied with vasoactive drugs (with χ2 values of 9.44 and 28.50, respectively, P<0.05). Age, total burn area, full-thickness burn area, serum creatinine within the first 24 hours of injury, and APTT within the first 24 hours of injury were the independent risk factors for the mortality within 28 days after injury in elderly patients with severe burns (with odds ratios of 1.17, 1.10, 1.10, 1.09, and 1.27, 95% confidence intervals of 1.03-1.40, 1.04-1.21, 1.05-1.19, 1.05-1.17, and 1.07-1.69, respectively, P<0.05). Conclusions:The elderly patients with severe burns had the injuries mainly from flame burns, often accompanied by moderate to severe inhalation injury and enhanced inflammatory response, elevated blood glucose levels, activated fibrinolysis, and impaired organ function in the early stage, which are associated with their prognosis. Age, total burn area, full-thickness burn area, and serum creatinine and APTT within the first 24 hours of injury are the independent risk factors for death within 28 days after injury in this population.

Objective To explore clinical effect of vancomycin calcium sulfate combined with internal fixation on cal-caneal beak-like fracture secondary to calcaneal osteomyelitis caused by diabetic foot.Methods From April 2018 to October 2021,a retrospective analysis was performed on 5 patients with calcaneal bone osteomyelitis secondary to diabetic foot,includ-ing 2 males and 3 females,aged from 48 to 60 years old;diabetes course ranged from 5 to 13 years;the courses of diabetic foot disease ranged from 18 to 52 days;5 patients were grade Ⅲ according to Wagner classification.All patients were treated with debridement,vancomycin bone cement implantation,negative pressure aspiration at stage Ⅰ,vancomycin calcium sulfate and internal fixation at stage Ⅱ for calcaneal beak-like fracture.Surgical incision and fracture healing time were recorded,and the recurrence of osteomyelitis was observed.American Orthopedic Foot Andankle Society(AOFAS)score and exudation at 12 months after operation were evaluated.Results Five patients were successfully completed operation without lower extremity vascular occlusion,and were followed up for 16 to 36 months.The wound healing time after internal fixation ranged from 16 to 26 days,and healing time of fractures ranged from 16 to 27 weeks.AOFAS score ranged from 65 to 91 at 12 months after oper-ation,and 2 patients got excellent result,2 good and 1 fair.Among them,1 patient with skin ulcer on the back of foot caused by scalding at 5 months after operation(non-complication),was recovered after treatment;the wound leakage complication oc-curred in 2 patients,and were recovered after dressing change.No osteomyelitis or fracture occurred in all patients.Conclusion Vancomycin calcium sulfate with internal fixation in treating calcaneal osteomyelitis secondary to calcaneal osteomyelitis caused by diabetic foot could not only control infection,but also promote fracture healing,and obtain good clinical results.

Aim To explore the effect of ginsenoside Rg3 on the biological behavior of human gastric cancer SGC-7901 cells by regulating E2F1.Methods MTT assay was used to determine the effect of ginsenoside Rg3(0,80,160,320 μmol·L-1)on cell prolifera-tion.The effects of different concentrations of ginsen-oside Rg3 on apoptosis were measured by flow cytome-try.The effects of different concentrations of ginsen-oside Rg3 on cell migration and invasion were deter-mined by scratch healing experiment and Transwell ex-periment.The effects of different concentrations of gin-senoside Rg3 on the expression of E2F1,MMP-2,MMP-9,BCL-2 and Bax were determined by Western blot.Results Compared with the blank control group,the cell survival rate of 80,160 and 320 μmol ·L-1 ginsenoside Rg3 group was significantly lower,and it was concentration-dependent(P＜0.05).Com-pared with the blank control group,the apoptosis rate of 80,160 and 320 μmol·L-1 ginsenoside Rg3 group significantly increased in a concentration-dependent manner(P＜0.05).Compared with the blank control group,the number of cell migration in 80,160 and 320 μmol·L-1 ginsenoside Rg3 groups was significantly lower in a concentration-dependent manner(P＜0.05).Compared with the blank control group,the number of cell invasion in 80,160 and 320 μmol· L-1 ginsenoside Rg3 groups was significantly lower in a concentration-dependent manner(P＜0.05).The E2F1 mRNA and E2F1 protein expression in the 80,160,and 320 μmol·L-1 ginsenoside Rg3 groups were significantly reduced in a concentration-dependent manner compared with that in the blank control group(P＜0.05).The protein expression of MMP-2,MMP-9,and BCL-2 in the cells of 80,160,and 320 μmol ·L-1 ginsenoside Rg3 group significantly decreased compared with those of the blank control group,and BCL-2 significantly increased compared with that of the blank control group in a concentration-dependent man-ner(P＜0.05).Conclusions Ginsenoside Rg3 can reduce the proliferation,inhibit the migration and inva-sion of gastric cancer SGC-7901 cells,and promote the apoptosis of SGC-7901 cells in a concentration-depend-ent manner,and its mechanism may be related to the down-regulation of MMP-2,MMP-9,and BCL-2 ex-pression and up-regulation of Bax expression through E2F1.

Objective To develop a rapid and accurate Monte Carlo program(simplified code for dosimetry of carbon ions,SPEEDO)for carbon ion therapy.Methods For electromagnetic process,type Ⅱ condensed history simulation scheme and continuous slowing down approximation were used to simulate energy straggling,range straggling,multiple scattering,and ionization processes.For nuclear interaction,5 types of target nuclei were considered,including hydrogen,carbon,nitrogen,oxygen,and calcium.The produced secondary charged particles followed the same condensed history framework.The study simulated the transport of carbon ions in 4 materials(water,soft tissues,lung,and bone),and the calculated doses were validated against TOPAS(a Monte Carlo simulation software for radiotherapy physics),followed by a comparison with dose measurements in a water phantom from the HIMM-WW(a medical heavy-ion accelerator facility in Wuwei).Results SPEEDO's simulation results showed good consistency with TOPAS.For each material,in the voxel region where the physical dose was greater than 10％of the maximum dose point,the relative maximum dose error of both was less than 2％.At treatment energy of 400 MeV/u,SPEEDO's computation time was significantly less than that of TOPAS(13.8 min vs 105.0 min).SPEEDO's calculation results also showed good agreement with HIMM-WW measurements in terms of lateral dose distribution and integrated dose depth curve.Conclusion SPEEDO program can accurately and rapidly perform Monte Carlo dose calculations for carbon-ion therapy.