Objective To investigate the potential mechanism of miR-21-5p in alleviating cerebral ischemia-reperfusion injury by targeting STAT3.Methods The HT22 cells were induced by OGD/R to construct a cell model of cerebral ischemia reperfusion injury.The expression of miR-21-5p was detected by RT-qPCR.The CCK-8 assay,TUNEL staining and flow cytometry were respectively used to detect the cell viability and apoptosis.ELISA assay was used to determine the contents of inflammatory factors IL-6,IL-10 and TNF-α in the cell supernatant.Western blot was used to detect the expression levels of p-STAT3/STAT3,Cleaved-Caspase-3,Bax and Bcl-2 proteins.The TargetScan database was used to predict the binding sites of miR-21-5p and STAT3.The dual-luciferase reporter gene assay was used to verify the targeting relationship between miR-21-5p and STAT3.Results The relative expression level of miR-21-5p was down-regulated in HT22 cells which induced by OGD/R(P<0.001).The cell viability(P<0.0001)was decreased and the apoptosis rate(P<0.001)was increased in OGD/R induced-HT22 cells.The contents of pro-inflammatory factors IL-6(P<0.001)and TNF-α(P<0.001)was increased,while the content of anti-inflammatory factor IL-10(P<0.001)decreased.After transfection with miR-21-5p mimic,cell viability was enhanced,apoptosis rate was reduced and neuroinflammation was inhibited.MiR-21-5p could target and bind to STAT3.After miR-21-5p inhibitor transfection,cell viability decreased,apoptosis was promoted,and neuroinflammation occurred;STAT3 inhibitor Stattic could reverse the effect of miR-21-5p inhibitor.Conclusion MiR-21-5p could specifically bind to STAT3 and reduce the neuroinflammation and apoptosis of OGD/R induced-HT22 cells.

AIM:To investigate the effects of puerarin(PU)on fecal metabolomics in atherosclerosis(AS)model mice.METHODS:Apolipoprotein E gene knockout(ApoE-/-)mice were fed a high-fat,high-cholesterol diet to es-tablish an AS model.The mice were randomly divided into control,model,and PU treatment groups,with five mice in each group.Body weight changes were recorded weekly.After 12 weeks,plasma inflammatory cytokines,lipid profiles,and aortic plaque area were measured,and fecal metabolomic analysis was performed using untargeted metabolomics.RE-SULTS:(1)The high-fat,high-cholesterol diet successfully induced the AS model,with significant increases in body weight(P<0.01),plasma levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β(P<0.01),as well as higher levels of low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),and triglycerides(TG),and reduced levels of high-density lipoprotein cholesterol(HDL-C)(P<0.01).The plaque area in both the whole aorta and the aortic sinus was significantly increased(P<0.01).(2)PU treatment significantly reduced body weight(P<0.05)and inflammatory cytokine levels(P<0.01),and improved lipid metabolism disorders.Specifically,TC,TG,and LDL-C lev-els significantly decreased(P<0.05 or P<0.01),while HDL-C levels significantly increased(P<0.01).In addition,the plaque area in both the whole aorta and the aortic sinus was significantly reduced in the PU-treated group(P<0.01).(3)Metabolomic analysis showed that PU regulated the levels of L-valine,L-phenylalanine,and L-histidine,restoring abnor-mal amino acid metabolism pathways.Additionally,PU downregulated abnormal levels of phosphatidylcholine and ce-ramide,promoting lipid metabolism balance.PU also modulated bile acid metabolism and oxidative stress mechanisms,improving purine and pyrimidine metabolism and related energy metabolism pathways.CONCLUSION:PU significantly ameliorates metabolic abnormalities in AS mouse models by regulating lipid metabolism,amino acid metabolism,bile acid metabolism,and gut microbiota-related metabolic pathways.

AIM:To investigate the effects of puerarin(PU)on fecal metabolomics in atherosclerosis(AS)model mice.METHODS:Apolipoprotein E gene knockout(ApoE-/-)mice were fed a high-fat,high-cholesterol diet to es-tablish an AS model.The mice were randomly divided into control,model,and PU treatment groups,with five mice in each group.Body weight changes were recorded weekly.After 12 weeks,plasma inflammatory cytokines,lipid profiles,and aortic plaque area were measured,and fecal metabolomic analysis was performed using untargeted metabolomics.RE-SULTS:(1)The high-fat,high-cholesterol diet successfully induced the AS model,with significant increases in body weight(P<0.01),plasma levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),and IL-1β(P<0.01),as well as higher levels of low-density lipoprotein cholesterol(LDL-C),total cholesterol(TC),and triglycerides(TG),and reduced levels of high-density lipoprotein cholesterol(HDL-C)(P<0.01).The plaque area in both the whole aorta and the aortic sinus was significantly increased(P<0.01).(2)PU treatment significantly reduced body weight(P<0.05)and inflammatory cytokine levels(P<0.01),and improved lipid metabolism disorders.Specifically,TC,TG,and LDL-C lev-els significantly decreased(P<0.05 or P<0.01),while HDL-C levels significantly increased(P<0.01).In addition,the plaque area in both the whole aorta and the aortic sinus was significantly reduced in the PU-treated group(P<0.01).(3)Metabolomic analysis showed that PU regulated the levels of L-valine,L-phenylalanine,and L-histidine,restoring abnor-mal amino acid metabolism pathways.Additionally,PU downregulated abnormal levels of phosphatidylcholine and ce-ramide,promoting lipid metabolism balance.PU also modulated bile acid metabolism and oxidative stress mechanisms,improving purine and pyrimidine metabolism and related energy metabolism pathways.CONCLUSION:PU significantly ameliorates metabolic abnormalities in AS mouse models by regulating lipid metabolism,amino acid metabolism,bile acid metabolism,and gut microbiota-related metabolic pathways.

Objective To explore the application effect of the Mini-CEX evaluation model based on the OBE concept in the clinical Practice teaching of neurology.Methods We Selected 100 students who will Practice in the Department of Neurology from 2022 to 2023 as the research objects,and divided them into the experimental group(n=50)and the control group(n=50).Under the guidance of the OBE concept,the experimental group was guided by learning outcomes,refined the teaching objectives,and applied the Mini-CEX evaluation mode for evaluation and feedback.In contrast,the control group adopted the traditional teaching mode.Combined with the observation data,we analyzed and compared the data of various indicators of the two groups of students at the beginning and end of the internship.Results At the end of the internship,the scores of clinical consultation,Physical examination,humanistic medicine,clinical diagnosis,health consultation,organizational effect,and overall evaluation of the experimental group were significantly improved and were higher than those of the control group.After the Practice,in terms of skill test scores,the experimental group scored higher than the control group,the difference was statistically significant(P<0.05),and the experimental group also scored higher in satisfaction evaluation than the control group.Conclusion The Mini-CEX evaluation teaching model based on the concept of OBE is applied to the clinical practice teaching of the neurology department,which can enhance the training effect of students'clinical practice skills.

OBJECTIVE To evaluate the correlation between the solid dispersion(SD) powder properties and the relative crystallinity(RC) during SD recrystallization. METHODS Andrographolide was used as model drug, polyvinylpyrrolidone K30, polyethylene glycol 8000, Poloxam 188, Soluplus® as carrier materials, 12 SD powders were prepared by three preparation processes, and then the SD powder properties were determined. Afterwards, the principle component analysis and partial least squares analysis(PLS) were used to evaluate the correlation between the SD powder properties and RC during SD recrystallization. RESULTS The correlation model was successfully established by partial least square method between the SD powder properties and the RC. The VIP value of particle size parameter D(0.5) was >1.2, which indicated that the particle size was the key factor affecting the recrystallization of SD powder. CONCLUSION In practical work, SD powder with different particle size parameters can be obtained by different preparation methods or by adjusting the process parameters of the selected preparation method, so as to improve the recrystallization stability of SD.

Growth hormone deficiency (GHD) has become a serious healthcare burden, and presents a huge impact on the physical and mental health of patients. Here, we developed an actively separated microneedle patch (PAA/NaHCO₃-Silk MN) based on silk protein for sustained release of recombinant human growth hormone (rhGH). Silk protein, as a friendly carrier material for proteins, could be constructed in mild full-water conditions and ensure the activity of rhGH. After manually pressing PAA/NaHCO₃-Silk MN patch to skin for 1 min, active separation is achieved by absorbing the interstitial fluid (ISF) to trigger HCO₃ ^‒ in the active backing layer to produce carbon dioxide gas (CO₂). In rats, the MN patch could maintain the sustained release of rhGH for more than 7 days, and produce similar effects as daily subcutaneous (S.C.) injections of rhGH in promoting height and weight with well tolerated. Moreover, the PAA/NaHCO₃-Silk MN patch with the potential of painless self-administration, does not require cold chain transportation and storage possess great economic benefits. Overall, the PAA/NaHCO₃-Silk MN patch can significantly improve patient compliance and increase the availability of drugs, meet current unmet clinical needs, improve clinical treatment effects of GHD patients.

Bacillus spp. are probiotics and can secrete a variety of natural antimicrobiol active substances, of which lipopeptides are an important class. Up to now, about 90 lipopeptides have been identified, and most of them are cyclic lipopeptides. surfactin, iturin, fengycin, bacillomycin and polymyxins are widely studied, and the first three have huge potential for application due to their properties of surfactants and anti-fungal, anti-bacterial, anti-viral, anti-tumor and anti-inflammatory functions. In this paper, the research progress in the structure, function, synthesis regulation, separation, purification and production of surfactin, iturin and fengycin was reviewed. Synthetic biology is a vital means to increase the yield of lipopeptides, and in the future, lipopeptides can be used in crop cultivation, animal farming, food, medicine and petroleum industries as well as environmental protection. Future research should be strengthened on the discovery of new lipopeptides, synthesis of high-activity lipopeptides, economical production of lipopeptides on a large scale and their safety evaluation.
Anti-Bacterial Agents ; Anti-Infective Agents/pharmacology* ; Bacillus ; Bacillus subtilis ; Lipopeptides/pharmacology* ; Peptides, Cyclic/pharmacology*

Objective:To analyze the mistakes in the application of tuina therapy in the rehabilitation of stroke,in order to elevate the clinical effects in the treatment of stroke by tuina.Methods:The issues are found out by searching and analyzing the relevant articles on tuina treatment of stroke.Results:In the tuina treatment of stroke,mistakes exist in the intervening time of the treatment,in the emphasis of tuina with negligence of rehabilitation,in the assessment of therapeutic effects,in the selection of the manual techniques and acupoints,and in the negligence of health education.Conclusion:In the tuina treatment of stroke,it is advisable to start treatment in the early stages,use the manual techniques in different phases,to cooperate with rehabilitation at the proper time and to emphasize health education,in order to avoid or reduce the occurrence of misuse syndrome,and eliminate the appearance of disuse syndrome,for displaying the unique advantage of tuina therapy fully and promoting the innovation and development of TCM rehabilitative techniques.

Clopidogrel was believed to be superior to aspirin by the well-known CAPRIE trial. However, no other large clinical trials demonstrated the same results, but all focused on the combination use of clopidogrel with aspirin, and combination therapy in CREDO was called the "Emperor's New Clothes". However, no one overturned the results of these clinical trials by quantitatively analyzing them. We reviewed ten large-scale clinical trials about clopidogrel. On the basis of results of CAPRIE, CREDO and CHARISMA trials, we re-estimated their minimal sample sizes and their powers by three well-established statistical methodologies. From the results of CAPRIE, we inferred that the minimal sample size should be 85 086 or 84 968 but its power was only 30.70%. A huge gap existed. The same was also true of CREDO and CHARISMA trials. Moreover, in CAPRIE trial, 0 was included in the 95% confidence interval and 1 was included in the 95% confidence interval for the relative risk. There were some paradoxical data in CAPRIE trial. We are led to conclude that the results in CAPRIE, CREDO, and from the subgroup analysis in CHARISMA trials were questionable. These results failed to demonstrate that clopidogrel was superior to aspirin or that clopidogrel used in combination with aspirin was better than aspirin alone. The cost-effectiveness analyses by some previous studies were not reliable.