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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating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.

Objective To provide scientific and normative guidance for capacity building and quality control standards for vision health management service institutions, thereby promoting the standardization and professional development of the industry, enhancing service quality, and improving the effectiveness of myopia prevention and control. Methods A multidisciplinary expert panel organized by Society of Vision Health Management, Chinese Health Association, based on standards such as the ^"Technical Specifications for Vision Health Management Services for Primary and Secondary School Students^" (T/CHAA 008-2019) and combined with field research data, revised and formed this consensus through multiple rounds of meetings and discussions. Results This consensus clarifies the positioning of service institutions, core service capability requirements (covering basic configuration, personnel and equipment, service processes, scope, and competency levels), and quality control points throughout the service process. It establishes an evaluation framework centered on ^"foundational support - service implementation - management effectiveness - outcome output.^" Conclusion This consensus provides systematic guidance for the standardized construction, capacity evaluation, and quality control of vision health management service institutions, which is of great significance for safeguarding the vision health of children and adolescents.

Based on spleen deficiency-phlegm dampness as the core pathogenesis of obesity， and integrating recent advances in modern medicine regarding the key role of immune inflammation in obesity， this paper proposes a multidimensional pathogenic network of "obesity-spleen deficiency-phlegm dampness-immune imbalance". Various traditional Chinese medicine （TCM） herbs that strengthen the spleen， regulate qi， and resolve phlegm and dampness can treat obesity by improving spleen-stomach transport and transformation， promoting water-damp metabolism， and regulating immune homeostasis. This highlights immune inflammation as an important entry point to elucidate the TCM concepts of "spleen deficiency-phlegm dampness" and the therapeutic principle of "strengthening the spleen and eliminating dampness to treat obesity". By systematically analyzing the intrinsic connection between "spleen deficiency generating dampness， internal accumulation of phlegm dampness" and immune dysregulation in obesity， this paper aims to provide theoretical support for TCM treatment of obesity based on dampness.

ObjectiveTo construct a stable monoclonal human embryonic kidney 293 （HEK293） cell line expressing recombinant human coagulation factor Ⅶ （rhFⅦ） and evaluate the expression level and procoagulant bioactivity of rhFⅦ. MethodsThe plasmid pCDNA3.1-EGFP-FⅦ was transfected into HEK293 cells to verify the effectiveness of the transfection system. The plasmid pCDNA3.1-FⅦ was transfected into HEK293 cells， and monoclonal stable transfected cell lines were selected using geneticin （G418）. The transcription of the FⅦ gene was identified by reverse transcription polymerase chain reaction （RT-PCR）. The expression level of rhFⅦ in the supernatant of the monoclonal stable transfected cell line was detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot. The concentration of rhFⅦ was determined by enzyme-linked immunosorbent assay （ELISA）， and the procoagulant activity of rhFⅦ was measured by human coagulation factor Ⅶ potency assay. ResultsHEK293 cells transfected with pcDNA3.1-EGFP-FⅦ showed green fluorescence， indicating that rhFⅦ was successfully expressed in the supernatant of HEK293 cells after transient transfection with pcDNA3.1-FⅦ. The monoclonal stable transfected cell line was obtained by G418 screening. RT-PCR identified that the FⅦ gene was integrated into the genome of the monoclonal stable transfected cell line. The cell viability was good as detected by Cell Counting Kit-8， and a single band of rhFⅦ was obtained by purification of the cell supernatant. The highest rhFⅦ expression was （1.27±0.09） mg/L， and the highest procoagulant activity was （380.29±13.80）%. ConclusionThe monoclonal HEK293 cell lines which can express rhFⅦ protein efficiently and stably with excellent procoagulant bioactivity is successfully screened.

AIM: To investigate the effect and potential mechanism of liraglutide(LIR)combined with neural stem cells(NSCs)in alleviating diabetic retinopathy(DR)in rats.METHODS:A DR rat model was established using a high-fat diet combined with a single intraperitoneal injection of streptozotocin(STZ). Rats in the control group were fed a normal diet and received an intraperitoneal injection of citric acid-sodium citrate buffer. After modeling, the rats were divided into the control group, DR group, DR+LIR group, DR+BMSCs group, DR+NSCs group, DR+BMSCs+LIR group, and DR+NSCs+LIR group. According to the grouping instructions, rats were given intraperitoneal injection of LIR at 200 μg/(kg·d)and intravitreal injection of bone marrow mesenchymal stem cells(BMSCs), or NSCs, or the combined therapy accordingly for 8 wk. Fasting blood glucose(FBG)and serum insulin levels, as well as retinal oxidative stress indexes and serum inflammatory indexes, were detected in each group. Retinopathy was evaluated by hematoxylin-eosin(HE)staining, retinal cell apoptosis was detected by TUNEL staining, SIRT1 expression in retinal tissues was detected by immunohistochemistry, and BDNF expression in retinal tissues was detected by immunofluorescence. The transcriptional levels of cZNF532, miR-29a-3p, SIRT1, TGF-β1, NRF2, Keap1, and BDNF in retinal tissues were detected by qRT-PCR, and the protein expression levels of SIRT1, TGF-β1, SMAD2/3, p-SMAD2/3, NRF2, Keap1, p65, and p-p65 in retinal tissues were detected by Western blot. RESULTS:Compared with the other groups of rats, the retinal morphology of rats in the DR+NSCs+LIR group was significantly improved, the levels of FBG, TNF-α, IL-1β, IL-6 and MDA were decreased(all P<0.05), the serum insulin level and the levels of SOD, CAT, and GSH-Px were increased(all P<0.05), the TUNEL-positive rate of retinal cells was reduced(all P<0.05). The expression levels of cZNF532, SIRT1, NRF2, and BDNF in retinal tissue were up-regulated(all P<0.05), whereas the expression levels of miR-29a-3p, TGF-β1, p-SMAD2/3, Keap1, and p-p65 were down-regulated(all P<0.05). CONCLUSION:LIR combined with NSCs alleviates DR by regulating the cZNF532-miR-29a-3p-SIRT1 network and its downstream TGF-β/SMAD, NF-κB, and Keap1/NRF2 signaling pathways.

Objective:To investigate the prevalence of asthenopia and dry eye,and to further explore the potential occupational hazard factors,so as to provide a theoretical basis for their prevention and con-trol.Methods:A cross-sectional survey was conducted on the selected respondents.For visual display terminal(VDT)workers in employing organizations such as banks,colleges,and government depart-ments,an online questionnaire independently developed by the research group was used for population surveys.Information including general information,work-related situations,work environment,visual health,and ergonomic factors was collected.The respondents were analyzed according to whether they suffered from asthenopia and dry eye.Relevant factors of asthenopia and dry eye were screened through t-test and Chi-square test.Subsequently,binary Logistic regression analysis was carried out to determine the risk factors of asthenopia and dry eye among the VDT workers.Results:The overall prevalence of as-thenopia was 52.5％(235/448)and dry eye was 36.8％(165/448).There were no significant diffe-rences in the prevalence of asthenopia and dry eye among different genders,age groups,and groups of length of service in VDT work.However,the highest prevalence of dry eye was observed in underweight individuals(42.9％),followed by normal weight(40.6％),overweight(28.0％),and obese indivi-duals(17.4％).There was a significant difference in the prevalence of dry eye among different body mass index(BMI)groups(x2=9.505,P=0.023).The lowest prevalence of asthenopia was observed among securities industry employees(22.6％),while higher rates were found in employees in companies(59.5％)and other employing organizations(68.8％).A significant difference in the prevalence of as-thenopia among different employing organizations(x2=14.832,P=0.022).The result of Logistic re-gression showed that a longer length of service in VDT work(OR=1.006,P＜0.001),a longer dura-tion of VDT after working hours(OR=1.002,P=0.032),a too-bright monitor(OR=2.875,P=0.022),glare during work(OR=1.500,P=0.038),a louder noise in work environment(OR=1.586,P=0.012),work-related musculoskeletal disorders(WMSDs)(OR=4.366,P＜0.001)and other factors were independent risk factors of asthenopia,while wearing frame glasses(OR=0.452,P=0.037)was an independent protective factor.Glare during work(OR=2.198,P＜0.001),WMSDs(OR=2.226,P=0.001)and other factors were independent risk factors of dry eye,while overweight(OR=0.448,P=0.006),obesity(OR=0.228,P=0.032)were independent protective factors of dry eye.Conclusion:The prevalence of asthenopia and dry eye among VDT workers is relatively high,and it is associated with multiple risk factors.During prevention and control,attention should be paid to taking reasonable breaks during work,controlling glare,and strengthening visual health training and promotion.

Objective:To explore the relationship of metacognitive regulation, self-efficacy, and motivation regulation with learning engagement among medical students in military academies.Methods:A total of 439 students from the Air Force Medical University were selected by convenience sampling in March to April 2023. The Metacognitive Self-Regulation Scale, Self-Efficacy Scale, Learning Engagement Scale, and Motivation Regulation Questionnaire were adopted for investigation. SPSS 25.0 was used for Pearson correlation analysis, and the Process procedure for analysis and testing of mediating effects.Results:A total of 436 usable questionnaires were collected. Metacognitive regulation, self-efficacy, motivation regulation, and learning engagement were significantly positively correlated ( r>0.477, P<0.01). The mediating effect of self-efficacy and motivation regulation and the chain mediating effect of self-efficacy-motivation regulation were significant between metacognitive regulation and learning engagement, and the effect sizes were 0.449, 0.244, and 0.130, accounting for 44.37%, 24.11%, and 12.85% of the total effect, respectively. The proportion of the total indirect effect was 81.32%. Conclusions:The metacognitive regulation of medical students in military academies directly affects learning engagement, which is also indirectly affected through the independent mediating effect of self-efficacy and motivation regulation or the chain mediating effect of self-efficacy-motivation regulation.

Objective:To study the impact of fluoride exposure through drinking water on the risk of hypertension among residents in Jishan County, Shanxi Province.Methods:From March to April 2023, a cluster sampling method was used to select permanent residents aged ≥18 years and residing for ≥10 years in 12 villages in drinking water-borne endemic fluorosis areas of Jishan County, Shanxi Province as the survey subjects. A questionnaire survey, physical examination, and morning urinary fluoride level testing were conducted. The least absolute shrinkage and selection operator (Lasso) regression were used to analyze the key influencing factors of hypertension. Restricted cubic spline was used to evaluate the linear relationship between urinary fluoride and hypertension. Logistic regression was used to analyze the impact of urinary fluoride on hypertension.Results:Finally, 2 453 survey subjects were included, aged (62 ± 10) years, including 1 565 patients (63.80%) with hypertension. There were significant differences in the distribution of age, gender, education level, annual household income, body mass index (BMI), and the level and distribution of urinary fluoride between hypertension group and normal blood pressure group ( P < 0.05). The Lasso regression results showed that age, education level, BMI, and urinary fluoride were the key influencing factors of hypertension, with coefficients of 1.04, - 0.12, 0.24 and 0.01, respectively. The results of the restricted cubic spline showed that there was a linear relationship between urinary fluoride and hypertension after adjusting for age, education level, and BMI ( Poverall = 0.018, Pnonlinear = 0.482). The logistic regression results showed that after adjusting for age, education level, and BMI, urinary fluoride > 4.68 mg/L was a risk factor for hypertension ( OR = 1.42, 95% CI: 1.10 - 1.84, P = 0.007). Conclusion:High urinary fluoride is a risk factor for hypertension in drinking water-borne endemic fluorosis areas of Jishan County, Shanxi Province.

Objective:To compare the clinical characteristics of severe Mycoplasma pneumoniae pneumonia（SMPP）with bronchiolitis and consolidation to raise awareness.Methods:Retrospective analysis was conducted on children hospitalized at the first affiliated hospital of Xiamen University diagnosed with（SMPP）from February 2023 to January 2024.The children were divided into bronchiolitis group and large consolidation group based on their high-resolution CT imaging findings.Data on basic information，clinical features，auxiliary examination results，and bronchoscopy findings were analyzed.Results:This study included 415 pediatric patients，including 203 with bronchiolitis and 212 with large consolidation.The proportion of males，atopy rate，and first-degree relative atopy rate in the bronchiolitis group were higher，and the age of the bronchiolitis group was younger（ P<0.05）.The fever peak，incidence of fever，and low breath sound in the bronchiolitis group were lower，and the incidence of hypoxemia，wheezing，shortness of breath，inspiratory tri-concave signs，moist rales were higher than those in the consolidation group（ P<0.05）.The white blood cell counts，platelet counts，and lactate dehydrogenase were higher and the percentage of neutrophils，C-reactive protein，ferritin，D-dimer，and fibrinogen degradation products were lower in the bronchiolitis group than those in the consolidation group（ P<0.05）.The rate of 2 or more bronchoscopy procedures，and the proportion of longitudinal plica，mucus plug，and plastic in the consolidation group were higher than those in the bronchiolitis group（ P<0.05）.After three months of follow-up，there were 8 cases of bronchiolitis obliterans，including 7 cases in the bronchiolitis group and 1 case in the large consolidation group. Conclusion:If atopy，wheezing，and dyspnea are present in young children and preschool children with confirmed SMPP，they should be closely monitored for signs of bronchiolitis SMPP.Children with significant consolidation on imaging should have the possibility of various intrapulmonary and extrapulmonary problems taken into account.Appropriate treatment options should be implemented during the acute phase based on the imaging findings.