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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating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 establish the normal reference value of the left ventricular remodeling index（RI）in healthy adults，and to evaluate the phenotypes of RI in various left ventricular geometries and its relationship with left ventricular systolic and diastolic functions in asymptomatic hypertensive patients.Methods:A retrospective study design was employed，906 healthy Han Chinese volunteers were selected as the control group from the multicenter study "Echocardiographic Measurements in Normal Chinese Adults"（EMINCA），which was conducted in 2012. The normal reference range of left ventricular RI was established based on the 95% CI（ xˉ ± 1.96 s）. Statistical analyses were conducted to evaluate differences in RI across age groups and between genders. A total of 340 asymptomatic hypertensive patients in the Department of Health Management，Sichuan Provincial People's Hospital from July 2023 to March 2024 were prospectively included as the hypertension group. Conventional echocardiography and two-dimensional speckle tracking imaging were used to assess left ventricular structure and function. Parameters such as left ventricular mass index（LVMI），RI，relative wall thickness（RWT），the ratio of early diastolic mitral inflow velocity to the average early diastolic mitral annular velocity（E/e'），and the ratio of early diastolic to late diastolic mitral inflow velocities（E/A）were calculated. Differences of these parameters between the hypertension group and the healthy control group were compared. The hypertension group was stratified into 3 subgroups based on left ventricular posterior wall thickness at end-diastole（LVPWd）and RI：the non-LV hypertrophy group，the left ventricular hypertrophy with normal RI group，and the left ventricular hypertrophy with low RI group. Relevant parameters were compared among these subgroups，and the statistical significance of the differences was analyzed. Pearson correlation analysis and multiple linear regression analysis were performed to explore the relationships between left ventricular RI and GLS，LVEF，the average mitral annular peak systolic velocity（s'），e'，E/e'，E/A. Results:In the healthy control group，RI showed a declining trend with increasing age in both sexes. Furthermore，the RI was significantly higher in the female group compared to the male group（ P<0.001）. Compared with the healthy control group，LVMI and E/e' increased while RI，e' and s' decreased in the hypertension group（all P < 0.001）. Among the 3 subgroups of the hypertension group，compared with the other two groups，LVMI and Ｅ/e' increased while absolute GLS and s' reduced in the left ventricular hypertrophy with low RI subgroup（all P <0.001）. Pearson correlation analysis revealed that RI was negatively correlated with GLS and E/e'（ r=-0.457，-0.281；all P < 0.001），and positively correlated with LVEF，e'，s' and E/A（ r=0.229，0.394，0.150，0.172；all P < 0.05）. Multivariate linear regression analysis demonstrated that left ventricular RI was independently associated with GLS，LVEF，e'，s'，E/e' and E/A. Conclusions:The normal reference range of left ventricular RI tends to decrease with age and is typically higher in females than in males. In asymptomatic patients with primary hypertension，RI，systolic and diastolic functions of the left ventricular are lower，while LVMI is higher. Among these patients，functional impairment is more pronounced in patients with left ventricular hypertrophy and reduced RI. This indicates that left ventricular RI may offer an imaging basis for further classification and stratification of structural and functional abnormalities in the left ventricle of asymptomatic hypertensive patients.

Most adult cases of depression originate during adolescence.In recent years,the incidence and suicide rate of adolescent depression raised gradually,seriously endangering physical and mental health as well as the life safety of minors.Resting-state functional MRI(rs-fMRI)was a core tool for exploring neurocognitive and psychiatric-behavioral disorders.The progresses in rs-fMRI research of adolescent depression with suicidal thoughts or behaviors were reviewed in this article.

BACKGROUND: The available evidence regarding the benefits of percutaneous coronary intervention (PCI) on patients receiving dialysis with coronary artery disease (CAD) is limited and inconsistent. This study aimed to evaluate the association between PCI and clinical outcomes as compared with medical therapy alone in patients undergoing dialysis with CAD in China. METHODS: This multicenter, retrospective study was conducted in 30 tertiary medical centers across 12 provinces in China from January 2015 to June 2021 to include patients on dialysis with CAD. The primary outcome was major adverse cardiovascular events (MACE), defined as a composite of cardiovascular death, non-fatal myocardial infarction, and non-fatal stroke. Secondary outcomes included all-cause death, the individual components of MACE, and Bleeding Academic Research Consortium criteria types 2, 3, or 5 bleeding. Multivariable Cox proportional hazard models were used to assess the association between PCI and outcomes. Inverse probability of treatment weighting (IPTW) and propensity score matching (PSM) were performed to account for potential between-group differences. RESULTS: Of the 1146 patients on dialysis with significant CAD, 821 (71.6%) underwent PCI. After a median follow-up of 23.0 months, PCI was associated with a 43.0% significantly lower risk for MACE (33.9% [ n  = 278] vs . 43.7% [ n  = 142]; adjusted hazards ratio 0.57, 95% confidence interval 0.45-0.71), along with a slightly increased risk for bleeding outcomes that did not reach statistical significance (11.1% vs . 8.3%; adjusted hazards ratio 1.31, 95% confidence interval, 0.82-2.11). Furthermore, PCI was associated with a significant reduction in all-cause and cardiovascular mortalities. Subgroup analysis did not modify the association of PCI with patient outcomes. These primary findings were consistent across IPTW, PSM, and competing risk analyses. CONCLUSION This study indicated that PCI in patients on dialysis with CAD was significantly associated with lower MACE and mortality when comparing with those with medical therapy alone, albeit with a slightly increased risk for bleeding events that did not reach statistical significance.
Humans ; Percutaneous Coronary Intervention/methods* ; Male ; Female ; Coronary Artery Disease/drug therapy* ; Retrospective Studies ; Renal Dialysis/methods* ; Middle Aged ; Aged ; China ; Proportional Hazards Models ; Treatment Outcome