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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating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 analyze the differences in chemical components of Gardenia jasminoides before and after processing with ginger， and to evaluate the quality differences among different producing areas. METHODS Ultra-high performance liquid chromatography-tandem time-of-flight mass spectrometry was used to analyze the compositional differences of G. jasminoides before and after processing with ginger. The water content， total ash， and ethanol-soluble extract content of ginger- processed G. jasminoides were determined according to the 2020 edition of Chinese Pharmacopoeia. High performance liquid chromatography was adopted to determine the contents of genipin gentiobioside， geniposide， crocin Ⅰ and crocin Ⅱ in ginger- processed G. jasminoides. RESULTS A total of 49 chemical components were identified from raw G. jasminoides and ginger- processed G. jasminoides， including 14 flavonoids， 15 iridoids， 10 organic acids， 2 alkaloids and 8 other compounds. Among them， 42 components were detected in raw G. jasminoides， 28 in ginger-processed G. jasminoides， and 21 components were common to both. After processing with ginger， raw G. jasminoides lost 21 components （including iridoids， flavonoids， alkaloids， and others）， while 7 chemical components were added （including coumarins， organic acids， organic acid esters， and flavonoids）. For the 15 batches of ginger-processed G. jasminoides， the water content ranged from 5.64% to 7.11%， total ash from 2.92% to 4.87%， and ethanol-soluble extract from 40.61% to 58.02%. The average contents of genipin gentiobioside， geniposide， crocin Ⅰ and crocin Ⅱ were 0.108 7， 0.542 2， 0.565 0， and 0.012 5 mg/g， respectively. CONCLUSIONS After processing with ginger， G. jasminoides loses 21 components， while 7 new components are added. Differences are observed in the water content， total ash， ethanol-soluble extract， and the contents of genipin gentiobioside， geniposide， crocin Ⅰ， and crocin Ⅱ of ginger-processed G. jasminoides from different producing areas. Notably， samples from Fujian exhibit high contents of genipin gentiobioside and ethanol-soluble extract， while samples from Jiangxi have a high content of crocin Ⅰ.

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 investigate expression and clinical significance of T cell activation related membrane molecules in infertility patients with polycystic ovary syndrome(PCOS).Methods:A total of 80 PCOS infertility patients who visited Second Affiliated Hospital of Hainan Medical College from January 2020 to June 2021 were selected as observation group,and 80 cases of women who underwent physical examination at same time were selected as control group.Expressions of membrane molecules related to T cell acti-vation in peripheral blood of patients between two groups were compared.Patients with PCOS infertility were divided into pregnant group[66(82.5％)]and non pregnant group[14(17.5％)]according to different prognosis.General clinical data,peripheral blood inflammatory factors and T cell activation related membrane molecules expressions were compared between two groups,focusing on correlation between T cell activation related membrane molecules expressions and prognosis of PCOS infertility patients,ROC curve and decision-making curve of T cell activation related membrane molecule expressions were drawn to predict prognosis of PCOS infer-tility patients,whose prediction efficiency and net profitability were further analyzed.Pearson correlation analysis was used to explore correlation between T cell activation related membrane molecules expressions and clinical characteristics of PCOS infertility patients.Results:CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients in observation group were significantly higher than control group(P<0.05).Total testosterone(T),TNF-α,IFN-γ,CD4+CD45+T and CD4+CD63+T cells expressions of patients in non pregnancy group were significantly higher than pregnancy group(P<0.05).Multivariate Logistic regression analysis showed that CD4+CD45+T and CD4+CD63+T cells were independent predictors of prognosis of PCOS infertility patients(P<0.05),ROC analysis showed that AUC of CD4+CD45+T cells predicting prognosis of PCOS infertility patients was 0.756(0.721～0.826),and the best diag-nostic cut-off point was 48.1％,AUC of CD4+CD63+T cells was 0.746(0.711～0.834),and the best cut-off point for diagnosis was 31.2％,while AUC of CD4+CD45+T and CD4+CD63+T cells combination was 0.948(0.897～0.986).Decision curve analysis showed that combined prediction of CD4+CD45+T and CD4+CD63+T cells was higher than a single indicator.Correlation analysis showed that CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients with PCOS infertility was related to T,TNF-α,insulin resistance index(HOMA-IR),IFN-γ,IL-2 and IL-10(P<0.05).Conclusion:The higher the CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood,the worse the prognosis of patients with PCOS infertility.

Objective:To investigate expression and clinical significance of T cell activation related membrane molecules in infertility patients with polycystic ovary syndrome(PCOS).Methods:A total of 80 PCOS infertility patients who visited Second Affiliated Hospital of Hainan Medical College from January 2020 to June 2021 were selected as observation group,and 80 cases of women who underwent physical examination at same time were selected as control group.Expressions of membrane molecules related to T cell acti-vation in peripheral blood of patients between two groups were compared.Patients with PCOS infertility were divided into pregnant group[66(82.5％)]and non pregnant group[14(17.5％)]according to different prognosis.General clinical data,peripheral blood inflammatory factors and T cell activation related membrane molecules expressions were compared between two groups,focusing on correlation between T cell activation related membrane molecules expressions and prognosis of PCOS infertility patients,ROC curve and decision-making curve of T cell activation related membrane molecule expressions were drawn to predict prognosis of PCOS infer-tility patients,whose prediction efficiency and net profitability were further analyzed.Pearson correlation analysis was used to explore correlation between T cell activation related membrane molecules expressions and clinical characteristics of PCOS infertility patients.Results:CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients in observation group were significantly higher than control group(P<0.05).Total testosterone(T),TNF-α,IFN-γ,CD4+CD45+T and CD4+CD63+T cells expressions of patients in non pregnancy group were significantly higher than pregnancy group(P<0.05).Multivariate Logistic regression analysis showed that CD4+CD45+T and CD4+CD63+T cells were independent predictors of prognosis of PCOS infertility patients(P<0.05),ROC analysis showed that AUC of CD4+CD45+T cells predicting prognosis of PCOS infertility patients was 0.756(0.721～0.826),and the best diag-nostic cut-off point was 48.1％,AUC of CD4+CD63+T cells was 0.746(0.711～0.834),and the best cut-off point for diagnosis was 31.2％,while AUC of CD4+CD45+T and CD4+CD63+T cells combination was 0.948(0.897～0.986).Decision curve analysis showed that combined prediction of CD4+CD45+T and CD4+CD63+T cells was higher than a single indicator.Correlation analysis showed that CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood of patients with PCOS infertility was related to T,TNF-α,insulin resistance index(HOMA-IR),IFN-γ,IL-2 and IL-10(P<0.05).Conclusion:The higher the CD4+CD45+T and CD4+CD63+T cells expressions in peripheral blood,the worse the prognosis of patients with PCOS infertility.

Objective:To analyze the effect of precipitation on road traffic injuries (RTI) in Zhejiang Province.Methods:The RTI surveillance and meteorological data from 2009 to 2022 in Zhejiang Province were collected. Based on the time-stratified case-crossover design, the precipitation of case day and control day was compared, and the distributed lag nonlinear model was applied to analyze the correlation of precipitation and RTI. Stratified analyses were conducted to analyze the effect modification of gender, age, injury location, and temperature. An attributable fraction was used to assess the burden of RTI caused by precipitation.Results:A total of 239 970 RTIs were monitored in Zhejiang Province from 2009 to 2022, averaging 46 daily cases. The distributed lag nonlinear model showed that compared with no rain, the risk of RTI increased first and then decreased with the increase of precipitation. The risk of RTI was the highest when the precipitation was 30.99 mm ( OR=1.08, 95% CI: 1.05-1.11). The adverse effects on RTI mainly occurred on the day of precipitation, and it showed insignificant or protective effects with the extension of lag days. 1.34%(95% CI: 1.31%-1.36%) of RTI could be attributed to precipitation. Stratified analysis showed that gender, age, injury location, and temperature may modify the effect of precipitation on RTI. Precipitation caused a heavier burden on RTI in subgroups aged 18-64, females, and occurring on roads and in low temperatures. Conclusions:Precipitation can increase the risk of RTI. People aged 18-64 or females are the key groups for RTI prevention, and prevention and control efforts of precipitation-related RTI should be increased in road and low-temperature environments.

Background Prior epidemiological studies suggest that exposure to pyrethroid insecticides may adversely affect children’s respiratory health. However, only limited studies are currently available on this topic in China. Objective To explore the association between exposure to pyrethroid insecticides and pulmonary function in children in Shanghai. Methods From August 2019 to January 2020, a cross-sectional study was conducted, recruiting 163 healthy school-aged children (aged 5–12 years) from Shanghai Children’s Hospital, Shanghai Jiao Tong University School of Medicine. Basic information, including age, height, weight, and family income, was collected. Urine samples from the children were collected and were analyzed for the levels of three pyrethroid insecticide metabolites: 3-phenoxybenzoic acid (3-PBA), cis-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (CDCCA), and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane carboxylic acid (TDCCA). Gas chromatography-tandem mass spectrometry (GC-MS/MS) was used for the analysis. Spirometry was used to assess pulmonary function and recorded following parameters: peak expiratory flow (PEF), forced expiratory flow between the 25th and 75th percentiles of forced vital capacity (FEF_25-75), forced expiratory volume in one second (FEV₁), forced vital capacity (FVC), and FEV₁/FVC. Multiple linear regression and restricted cubic spline models were used to evaluate the associations between urinary pyrethroid insecticide metabolite levels and pulmonary function parameters. Results The study included 163 school-aged children, with an average age of (7.04 ± 2.08) years and an average body mass index (BMI) of (16.04 ± 2.72) kg·m⁻²; 75 (46.01%) of the participants were boys. The detection rates of 3-PBA, TDCCA, and CDCCA in urine were 85.28%, 17.79%, and 4.91%, respectively. The median creatinine-adjusted 3-PBA concentration was 0.150 μg·g⁻¹. After adjusting for confounders such as height, BMI, sex, age, delivery mode, annual family income, and maternal education level, the multiple linear regression model showed that urinary 3-PBA levels were negatively associated with both FVC [β=−0.030, 95% confidence interval (CI): −0.058, −0.003; P=0.031] and FEV₁ (β=−0.032, 95%CI: −0.064, 0.000; P=0.04998). The final restricted cubic spline model showed a nonlinear association between urinary 3-PBA levels and FVC, FEV₁, PEF, and FEF_25-75（P for nonlinear < 0.05；P for overall < 0.05）. Conclusion The level of pyrethroid insecticide exposure in school-aged children in Shanghai is relatively high. The urinary 3-PBA concentration is negatively associated with pulmonary function, indicating potential adverse effects of pyrethroid insecticide exposure on respiratory health of school-aged children.

Objective:Investigate the impact of calcification on the long-term outcomes of patients with coronary chronic total occlusion (CTO) after percutaneous coronary intervention (PCI).Methods:A retrospective cohort study was conducted. Patients who underwent PCI and had at least one CTO lesion at Fuwai Hospital between January 2010 and December 2013 were consecutively enrolled. Calcification was evaluated by coronary angiography, and patients were divided into two groups: moderate/severe calcification group and non/mild calcification group. Clinical follow-up was completed up to 5 years. Incidence of PCI-related complications and immediate procedural outcomes were compared between two groups, and the primary endpoint was the target lesion failure (TLF) at 5 years after PCI. Clinical follow-up endpoint events were analyzed using Kaplan-Meier survival analysis with log-rank test, and Cox multivariate regression model was used to evaluate the relationship between calcification and TLF.Results:The study included 2 659 CTO patients with an age of (57.2±10.5) years, of whom 442 (16.6%) were female, and among whom 13.5% (360/2 659) had moderate/severe calcification. Compared with the non/mild calcification group, the moderate/severe calcification group had a higher incidence of PCI-related complications (43.2% (156/361) vs. 32.5% (772/2 374), P<0.001) and procedural failure (34.3% (124/361) vs. 24.3% (577/2 374), P<0.001). Additionally, the moderate/severe calcification group showed a higher risk of the primary endpoint event (TLF) during the 5-year follow-up (19.8% vs. 15.3%, log-rank P=0.028). Higher incidence of cardiac death was observed in moderate/severe calcification group (5.7% vs. 2.7%, log-rank P=0.003). Cox multivariate regression analysis revealed that moderate/severe calcified plaques remained an independent risk factor for 5-year TLF after CTO-PCI ( HR=1.34, 95% CI: 1.01-1.79, P=0.043). Conclusion:Compared with CTO patients with non/mild calcification, those with moderate/severe calcification have higher procedural failure and complication rates, as well as poorer long-term prognosis, mainly due to an increase in cardiac death.