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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for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

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.

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.

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.

Abstract To clarify the service targets, service contents, and safeguard mechanisms of the basic public service program for student vision health management, the study grounded in the current status of child and adolescent myopia prevention and control in China and integrated with the practical experience and innovative practices of Wuhan National Demonstration Area for Student Vision Health Management, takes education as the main thread and schools as the base, consolidating resources from multiple stakeholders including education administrative departments, schools, families, and professional technical service institutions. Centering on four core components—vision health education, monitoring and record keeping, risk prevention and control, and dynamic management, the study constructs a multi tiered basic public service system for vision health management of primary and secondary school students. It further proposes the use of an intelligent management platform and a "three tier color coded early warning" mechanism (red, yellow, blue) to prevent and control student myopia at the source, providing schools with replicable and operable implementation pathways to enhance the level of student vision health management.

Objective:To analyze the prediction of pathological classification of ground glass nodules based on artificial intelligence computed tomography (CT) quantitative parameters combined with histogram parameters.Methods:The clinical data of 268 suspected patients with ground glass nodules admitted to Foshan Fosun Chancheng Hospital from June 2021 to June 2023 were retrospectively selected as the research subjects. They were divided into pre invasive lesions group (100 cases) and invasive lesions group(168 cases) according to pathological classification. Basic data of patients with different pathological classifications and the CT characteristics were compared, the prediction of pathological classification of ground glass nodules based on CT quantitative parameters combined and histogram parameters were analyzed by receiver operating characteristic (ROC) curve.Results:The edge and boundary of the tumor, shape of the lesion, the peripheral signs of the lesion and the boundary between the two groups had statistical differences ( P<0.05). The CT quantitative parameters of maximum diameter, lesion volume, average CT value in the invasive lesions group and pre invasive lesions group had statistical differences: (15.29 ± 3.20) cm vs. (9.75 ± 2.14) cm, (1.54 ± 0.31) cm 3 vs. (0.51 ± 0.10) cm 3, (- 328.16 ± 46.35) HU vs. (-541.25 ± 100.30) HU, P<0.05. The CT histogram parameters of inproportion of solid components, entropy and maximum CT value in the invasive lesions group and pre invasive lesions group had statistical differences: (66.39 ± 13.25)% vs. (42.65 ± 11.20)%, 4.31 ± 0.52 vs. 3.32 ± 0.39, (-75.34 ± 21.27) HU vs. (-141.72 ± 32.43)HU, P<0.05. Compared with the single prediction of CT quantitative parameters and CT histogram parameters, the combined prediction of the two parameters had higher value in predicting different pathological subtypes of ground glass nodules (the area under the curve was 0.877, P = 0.001). Conclusions:The combined detection of CT quantitative parameters and histogram parameters based on artificial intelligence can effectively evaluate the invasion status of ground glass nodules, which is beneficial for improving the detection of different pathological types of ground glass nodules.

Objective:To evaluate the effect of transient receptor potential vanilloid type 1 (TRPV1) gene polymorphism on the postoperative analgesic efficacy of alfentanil in patients undergoing radical mastectomy.Methods:In this prospective study, adult female patients diagnosed with breast cancer who underwent surgical treatment at the First Affiliated Hospital of Zhengzhou University were selected as case group ( n=162), while age-matched healthy adult females were selected as control group ( n=80). All subjects had a body mass index of 18-30 kg/m 2. The genotype and allele frequencies of TRPV1 rs8065080 and rs222747 were compared between the two groups, and the consumption of alfentanil and requirement for rescue analgesia within 48 h after surgery were recorded in breast cancer patients with different TRPV1 rs222747 genotypes. Results:Three genotypes of CC, GC, and GG were detected at rs222747 locus of TRPV1 gene, and there was no significant difference in the genotype distribution between the two groups ( P>0.05). Three genotypes of CC, TC and TT were detected at rs8065080 locus of TRPV1 gene, and there was significant difference in the genotype distribution between the two groups ( P<0.05). There was no significant difference in the basic data of patients with different genotypes of TRPV1 gene rs222747 locus ( P>0.05). There were significant differences in the consumption of alfentanil during 0-24 h after surgery and during 24-48 h after surgery and rate of rescue analgesia within 48 h after surgery among patients with different genotypes of TRPV1 gene rs222747 locus ( P<0.05). Conclusions:The polymorphism at the TRPV1 gene rs222747 locus can influence the postoperative analgesic efficacy of alfentanil in breast cancer patients undergoing radical mastectomy, suggesting that genetic factors may be one of the reasons for the inter-individual variability in alfentanil efficacy.

Objective To compare the efficacy and adherence of rivaroxaban versus low-molecular-weight heparin for prophylactic anticoagulation in cancer patients with venous thromboembolism(VTE).Methods A total of 120 intermediate-to-high VTE risk patients with malignant tumors admitted to Depart-ment of Hematology and Oncology,West China Longquan Hospital of Sichuan University between September 2021 and December 2022,were randomly assigned to the rivaroxaban group(n=60)and the low-molecular-weight heparin group(n=60)using the random number table.The rivaroxaban group received oral Rivaroxa-ban,while the low-molecular-weight heparin group received subcutaneous injections of low-molecular-weight heparin sodium for prophylactic anticoagulation.All patients were followed up for 180 days.The primary end-point was medication adherence.The secondary endpoints included the incidence of VTE,bleeding events,and changes in coagulation parameters.Results The rate of good medication adherence was significantly higher in the rivaroxaban group than in the low-molecular-weight heparin group(95.00％vs.88.33％,P＜0.05).However,there were no statistically significant differences in the incidence of VTE or overall bleeding events between the two groups(P＜0.05).Following treatment,parameters including fibrinogen,prothrombin time(PT),and D-dimer levels showed significant improvement from baseline in both groups.Compared to the low-molecular-weight heparin group,the Rivaroxaban group demonstrated significantly higher fibrinogen levels,shorter PT,and lower D-dimer levels(P＜0.05).Stepwise logistic regression analysis identified the post-treatment platelet(PLT)count as a significant factor influencing bleeding events during prophylactic antico-agulation(P＜0.05).Khorana score≥3(high risk)was identified as a risk factor for bleeding events(P＜0.05).The incidence of clinically relevant non-major bleeding(CRNMB)was higher in the rivaroxaban group[11.67％(7/60)]compared to the low-molecular-weight heparin group[8.33％(5/60)],although the differ-ence was not statistically significant(P＜0.05).Kaplan-Meier curve analysis revealed no significant difference in the cumulative incidence of bleeding-free events between the two groups(P＜0.05).Conclusion Oral ri-varoxaban and subcutaneous low-molecular-weight heparin demonstrate comparable efficacy and safety for VTE prevention in cancer patients,but rivaroxaban significantly improves patient's adherence.