BACKGROUND:Exosomes derived from mesenchymal stem cells play pivotal roles in cell communication and epigenetic regulation due to their low immunogenicity and targeted delivery effects,and have been clinically applied in the treatment of various diseases.OBJECTIVE:To review the isolation,purification,identification methods,and application progress of mesenchymal stem cell-derived exosomes,and to facilitate the development of large-scale preparation techniques and clinical translation of mesenchymal stem cell-derived exosomes.METHODS:The Chinese search terms"exosome,mesenchymal stem cells,isolation,purification,characterization,clinical application"and the English search terms"exosome,extracellular vesicles,mesenchymal stem cells,isolation,characterization,application"were used to search the literature published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating and purifying exosomes,comparing the characteristics of ultracentrifugation,ultrafiltration,size-exclusion chromatography,polymer precipitation,immunoaffinity,microfluidic methods,and other novel approaches based on their underlying principles.(2)Methods for identifying exosomes can be categorized into physical and biochemical analyses,characterizing exosomes based on their shape,size,and characteristic proteins.(3)Mesenchymal stem cell-derived exosomes have broad applications in multiple fields such as medical aesthetics,wound repair,and cancer treatment,due to their immune-regulatory properties and ability to cross biological barriers.(4)The clinical translation of exosomes faces challenges due to their complex structure,lack of universal isolation techniques,and poor stability,making it difficult to achieve in a short period of time.

BACKGROUND:Exosomes derived from mesenchymal stem cells play pivotal roles in cell communication and epigenetic regulation due to their low immunogenicity and targeted delivery effects,and have been clinically applied in the treatment of various diseases.OBJECTIVE:To review the isolation,purification,identification methods,and application progress of mesenchymal stem cell-derived exosomes,and to facilitate the development of large-scale preparation techniques and clinical translation of mesenchymal stem cell-derived exosomes.METHODS:The Chinese search terms"exosome,mesenchymal stem cells,isolation,purification,characterization,clinical application"and the English search terms"exosome,extracellular vesicles,mesenchymal stem cells,isolation,characterization,application"were used to search the literature published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating and purifying exosomes,comparing the characteristics of ultracentrifugation,ultrafiltration,size-exclusion chromatography,polymer precipitation,immunoaffinity,microfluidic methods,and other novel approaches based on their underlying principles.(2)Methods for identifying exosomes can be categorized into physical and biochemical analyses,characterizing exosomes based on their shape,size,and characteristic proteins.(3)Mesenchymal stem cell-derived exosomes have broad applications in multiple fields such as medical aesthetics,wound repair,and cancer treatment,due to their immune-regulatory properties and ability to cross biological barriers.(4)The clinical translation of exosomes faces challenges due to their complex structure,lack of universal isolation techniques,and poor stability,making it difficult to achieve in a short period of time.

ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes（TP-CX@TESs）， conduct its quality evaluation， and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators， Box-Behnken design-response surface methodology（BBD-RSM） was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide（LPS）-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention， the levels of inflammatory factors such as nitric oxide（NO）， interleukin-6（IL-6）， and tumor necrosis factor-α（TNF-α） in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2（JAK2）， signal transducer and activator of transcription 3（STAT3）， and α7 nicotinic acetylcholine receptor（α7nAChR）， and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was applied to measure the mRNA expression levels of JAK2， STAT3， the encoding gene of α7nAChR（CHRNA7）， and nuclear transcription factor-κB（NF-κB）. ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows：egg yolk lecithin content of 2.3%， ethanol volume fraction of 30%， and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of （105.60±3.85） nm， a polydispersity index of 0.19±0.03， and a Zeta potential of （-15.89±0.98） mV. The encapsulation efficiencies of triptolide， ferulic acid， and ligustilide were （76.88±4.40）%， （78.84±4.40）%， and （65.88±0.06）%， respectively. Both in vitro release and transdermal penetration of triptolide， ferulic acid， and ligustilide in TP-CX@TESs all followed the first-order kinetic model， showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO， TNF-α， and IL-6（P<0.01）， remarkably upregulated the protein expression levels of STAT3 and α7nAChR（P<0.01）， increased the mRNA expression level of CHRNA7， and significantly downregulated the mRNA expression level of NF-κB（P<0.05， P<0.01）. ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible， along with favorable in vitro release property， good transdermal permeability， and excellent in vitro anti-inflammatory activity， the mechanism is related to the inhibition of NF-κB.

ObjectiveTo prepare triptolide-Chuanxiong Rhizoma extract ethanol transfersomes（TP-CX@TESs）， conduct its quality evaluation， and investigate its in vitro anti-inflammatory efficacy and the underlying mechanisms. MethodsTP-CX@TESs was prepared via the ultrasonic injection method. With encapsulation efficiency and particle size as evaluation indicators， Box-Behnken design-response surface methodology（BBD-RSM） was employed to optimize the formulation process. The TP-CX@TESs prepared under the optimal process was characterized and evaluated for in vitro transdermal performance. A lipopolysaccharide（LPS）-induced RAW264.7 cell inflammation model was established. After 24 h of drug intervention， the levels of inflammatory factors such as nitric oxide（NO）， interleukin-6（IL-6）， and tumor necrosis factor-α（TNF-α） in the cell supernatant were detected. Western blot was used to determine the protein expression levels of Janus kinase 2（JAK2）， signal transducer and activator of transcription 3（STAT3）， and α7 nicotinic acetylcholine receptor（α7nAChR）， and real-time fluorescence quantitative polymerase chain reaction（Real-time PCR） was applied to measure the mRNA expression levels of JAK2， STAT3， the encoding gene of α7nAChR（CHRNA7）， and nuclear transcription factor-κB（NF-κB）. ResultsResults of BBD-RSM showed that the optimal formulation for preparing TP-CX@TESs was as follows：egg yolk lecithin content of 2.3%， ethanol volume fraction of 30%， and ratio of polysorbate-80 to egg yolk lecithin of 2∶5. Microscopic characterization revealed that TP-CX@TESs exhibited a spherical-like structure with a particle size of （105.60±3.85） nm， a polydispersity index of 0.19±0.03， and a Zeta potential of （-15.89±0.98） mV. The encapsulation efficiencies of triptolide， ferulic acid， and ligustilide were （76.88±4.40）%， （78.84±4.40）%， and （65.88±0.06）%， respectively. Both in vitro release and transdermal penetration of triptolide， ferulic acid， and ligustilide in TP-CX@TESs all followed the first-order kinetic model， showing a certain sustained-release property. Experimental results in RAW264.7 cells indicated that TP-CX@TESs significantly inhibited the release of NO， TNF-α， and IL-6（P<0.01）， remarkably upregulated the protein expression levels of STAT3 and α7nAChR（P<0.01）， increased the mRNA expression level of CHRNA7， and significantly downregulated the mRNA expression level of NF-κB（P<0.05， P<0.01）. ConclusionThe optimized formulation process of TP-CX@TESs is simple and feasible， along with favorable in vitro release property， good transdermal permeability， and excellent in vitro anti-inflammatory activity， the mechanism is related to the inhibition of NF-κB.

Objective To investigate the effects of dapagliflozin on inflammatory factors and prognosis in patients with type 2 diabetes mellitus (T2DM) and acute myocardial infarction (AMI). Methods In a randomized, double-blind trial, 146 patients with T2DM and AMI (within 7 days of onset) were divided into dapagliflozin (dapagliflozin 10 mg/d combining AMI standard therapy) and control (AMI standard therapy) groups, and were followed up for 12 months. Serum levels of interleukin-1β (IL-1β), IL-6, high-sensitivity C reactive protein (hs-CRP) at baseline, 1, 3, 6, and 12 months, and left ventricular ejection fraction (LVEF), brain natriuretic peptide (BNP), and major adverse cardiovascular events (MACE) rate at 12 months were compared between the two groups. Kaplan-Meier curves were used to analyze the cumulative incidences of MACE in the two groups. Results Three patients were withdrawn or dropped out. At 12 months, IL-1β, IL-6, and hs-CRP levels were significantly lower in dapagliflozin group (n＝71) than those in control group (n＝72, P＜0.01), approaching normal levels. Compared with the control group, LVEF was higher (P＜0.01), BNP was lower (P＜0.01), MACE incidence was lower (P＝0.047) in dapagliflozin group at 12 months. Generalized linear mixed models showed significant group-time interactions in IL-1β, IL-6, and hs-CRP (P＜0.001), and these factors declined faster in the dapagliflozin group. Kaplan-Meier curve showed the cumulative incidences of MACE and heart failure were lower in dapagliflozin group than those in non-dapagliflozin group (P＜0.05). Conclusions For patients with T2DM patients and AMI, dapagliflozin has good anti-inflammatory and cardioprotective effects.

Vascular cognitive impairment(VCI)is a type of cognitive dysfunction caused by cerebrovascular lesions,and its pathological mechanism includes mitochondrial damage.Therefore,maintaining mitochondrial homeostasis is crucial in the occurrence and progression of VCI.Mitochondrial homeostasis involves multiple levels,including biogenesis,autophagy,ion balance,protein quality control,membrane dynamics,and interactions with other organelles.This study outlines the regulatory mechanisms of mitochondrial homeostasis dysregulation at both the micro and macro levels,delves into the key role of mitochondrial homeostasis in VCI,analyzes potential therapeutic directions,and provides new scientific evidence and perspectives for the early diagnosis and treatment of VCI.

Objective:To analyze the comprehensive efficiency of resource allocation in urban public hospitals in 21cities of Guangdong Province from 2019 to 2023,aiming at providing empirical evidence for future medical policy makers and hospital managers in resource allocation and management.Methods:Using Data Envelopment Analysis and Malmquist index to evaluate the efficiency of health resource allocation in public hospitals in Guangdong.Results:From 2019 to 2023,the comprehensive efficiency of urban public hospitals in Guangdong Province was mildly ineffective,only 7 cities showing relative effectiveness,and there were significant regional differences in efficiency.Malmquist index analysis showed that the total factor productivity of health resource allocation in urban public hospitals in the province decreased from 2019 to 2020 and kept increasing from 2020 to 2023,mainly due to the increasing of the technological progress index and the scale efficiency index.Horizontally,the total factor productivity of 9 cities in the province has increased,while the total factor productivity of 12 cities has decreased,with significant regional differences.The input-output redundancy analysis shows that only 6 cities have no input-output redundancy.Conclusion:The overall efficiency of resource allocation in urban public hospitals in Guangdong Province from 2019 to 2023 is relatively low,with significant regional differences.In the future,it is necessary to coordinate and plan resource allocation,focus on refined management,strengthen talent training and technological improvement,improve the operational efficiency of existing resources,and promote high-quality development of hospitals.

Objective:To investigate the application value of optimized mesenteric defect closure technique in laparoscopic-assisted right hemicolectomy.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 26 patients who underwent laparo-scopic-assisted right hemicolectomy at Changzhi People′s Hospital Affiliated to Changzhi Medical College from May 2023 to June 2024 were collected. There were 11 males and 15 females, aged (65.7±1.8)years. All patients received optimized mesenteric defect closure using a combined extra-corporeal-laparoscopic suturing technique. Observation indicators: (1) surgical and intraoperative conditions; (2) postoperative conditions; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M( Q1, Q3) or M(range). Count data were described as absolute numbers. Results:(1) Surgical and intraoperative conditions. All 26 patients underwent laparoscopic-assisted right hemicolectomy with optimized mesenteric defect closure. The time of optimized mesenteric defect closure was 9.8(8.8,12.8)minutes, time of digestive tract reconstruction was 10.0(8.7,13.0)minutes, operation time was (164±4)minutes, volume of intraoperative blood loss was 50(50,100)mL. One of the 26 patients had intraoperative mesenteric hematoma, which required no specific intervention. The patient recovered uneventfully and was discharged from hospital. (2) Postoperative conditions. The visual analog scale pain score of 26 patients on postoperative day 1 and day 3 were 5(4,5) and 3(2,3), respectively. Time to postoperative first anal flatus and bowel movement were both 3(3, 4)days. Time to postoperative first intake of liquid diet was 2(2,3)days, duration of postoperative abdominal drainage was 4(3,5)days, and duration of postoperative hospital stay was 9(8,12)days. The number of lymph node dissected in 26 patients was 25(18,27) and the number of positive lymph node was 1(0,2). Cases in stage Ⅰ, stage Ⅱ and stage Ⅲ of tumor TNM staging were 5, 6, 15. (3) Follow-up. All 26 patients were followed up for 15(range, 6?20)months. Of the 26 patients, one case had incom-plete intestinal obstruction on postoperative day 25, which was diagnosed as adhesive intestinal obstruction based on imaging examination and classified as Clavien-Dindo grade Ⅱ. The patient recovered and was discharged after conservative treatment. None of the 26 patients had bleeding, infection of incision, anastomotic leakage, internal hernia, or delayed gastric emptying. There was no tumor recurrence, metastasis or death.Conclusion:The optimized mesenteric defect closure tech-nique in combined extracorporeal-laparoscopic suturing procedure can be used in laparoscopic-assisted right hemicolectomy.

Objective To explore the characteristics of acupoint selection and the law of acupoint compatibility in the treatment of Guillain-Barré syndrome(GBS)based on data mining.Methods Literature on acupuncture treatment of GBS was searched and screened from China National Knowledge Infrastructure,Wanfang Data Knowledge Service Platform,VIP,SioMed,PubMed,Embase,Web of Science,and Cochrane Library from the establishment of the database to December 1,2024 and to build database.The author information and acupuncture prescriptions were modified and imported according to the data entry requirements of the Traditional Chinese medicine inheritance computing platform V3.5,the relevant data were analyzed and integrated through the"Acupoint Analysis"module of the platform.Results A total of 140 related papers,145 valid prescriptions,involving 237 acupoints.The acupoints with higher frequency were Zusanli,Hegu,Quchi,Yanglingquan and etc.The analysis yielded 42 core acupoints,22 sets of strong association rules for core acupoints,and 5 new core acupuncture prescriptions.Conclusion Acupuncture treatment for GBS follows the principle of"treating impotence by taking Yangming alone",takes"benefiting Qi and draining heat,supporting the correctness and opening up the collaterals"as the main method,and attaches importance to"regulating the spirit and connecting the internal organs".

Objective To evaluate the prognostic value of monocyte human leukocyte antigen-DR(mHLA-DR),neutrophil-to-lymphocyte ratio(NLR),and CD4+T lymphocytes in sepsis.Methods A total of 29 patients with sepsis who were admitted to the department of critical care medicine of Qingdao Municipal Hospital from December 2023 to September 2024 were collected as the study subjects,and the patients were divided into survival group(20 cases)and death group(9 cases)according to the 28-day prognosis.Baseline data were collected from patients at the time of admission[including gender,age,acute physiology and chronic health evaluationⅡ(APACHEⅡ)score,sequential organ failure assessment(SOFA),white blood cell count(WBC),NLR,hemoglobin(Hb),platelet count(PLT),C-reactive protein(CRP),total protein(TP),alanine aminotransferase(ALT),aspartate aminotransferase(AST),creatinine(Cr),CD4+T lymphocyte count]and the mHLA-DR expression rate on the 1st,3rd,and 7th days of admission,and the difference between the mHLA-DR expression rate on the 3rd,7th and 1st days of admission and the 1st day of admission was calculated,which was recorded as ΔH3 and ΔH7.The receiver operator characteristic curve(ROC curve)was used to evaluate the predictive value of mHLA-DR expression,NLR,CD4+T lymphocyte count,SOFA score and APACHEⅡscore on the 28-day mortality risk of sepsis.Results Compared with the survival group,the APACHEⅡscore,SOFA score and NLR in the death group were significantly increased,and the ΔH7 and CD4+T lymphocyte counts were significantly decreased(all P<0.05).ROC curve analysis showed that ΔH7,NLR,CD4+T lymphocyte count,SOFA score and APACHEⅡscore were predictive of the 28-day prognosis of sepsis patients,and area under the curve(AUC)and 95%confidence interval(95%CI)were 0.817(0.635-0.999),0.789(0.611-0.966),0.786(0.588-0.985),and 0.853(0.685-1.000),0.844(0.659-1.000),all P<0.05.The combined detection of ΔH7 combined with NLR,ΔH7 combined with CD4+T lymphocytes,NLR combined with CD4+T lymphocytes,and ΔH7,NLR,and CD4+T lymphocytes also had predictive value for the 28-day prognosis of sepsis patients,with AUC and 95%CI of 0.867(0.735-0.998),0.878(0.752-1.000),0.883(0.760-1.000),and 0.928(0.837-1.000),respectively,all P<0.05.Conclusion The NLR and CD4+T lymphocyte count on the first day of admission to the hospital could predict the prognosis of sepsis patients,and the dynamic monitoring of mHLA-DR expression level in sepsis patients could also predict the prognosis of sepsis patients,but a single measurement of mHLA-DR expression level within 7 days was meaningless.In terms of single indicators,ΔH7 had the best predictor of the prognosis of sepsis patients among the 3 indicators of ΔH7,NLR and CD4+T lymphocyte count,and the combined detection of the 3 indicators was more advantageous in the prognosis of sepsis patients.