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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for isolating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 systematically evaluate the correlation between dynamic changes in inflammatory markers during treatment with epidermal growth factor receptor-tyrosine kinase inhibitors （EGFR-TKIs） in non-small cell lung cancer （NSCLC） patients and therapeutic efficacy， with the aim of providing evidence-based support for clinical prognosis assessment and treatment strategy adjustment. METHODS Databases including PubMed， Embase， Cochrane Library， CNKI， Wanfang Data， and CBM were searched from the inception to July 20， 2025. Following literature screening， data extraction and quality assessment， descriptive analysis was conducted on the outcomes of included studies. RESULTS A total of eight studies were included to analyze the correlation of 6 inflammatory markers before and after treatment with EGFR-TKIs with therapeutic efficacy. The risk of bias assessment identified six high-quality studies and two moderate-quality studies. Among these studies， seven studies demonstrated that lower levels of neutrophil-to-lymphocyte ratio （NLR）， derived neutrophil-to-lymphocyte ratio （dNLR）， platelet-to-lymphocyte ratio （PLR）， and monocyte-to-lymphocyte ratio （MLR）， higher lymphocyte-to-monocyte ratio （LMR） before treatment， as well as decreased NLR and MLR and increased LMR after treatment were associated with longer median progression-free survival. Five studies indicated that lower levels of NLR， dNLR， PLR， and interleukin-6 （IL-6）， higher LMR before treatment as well as decreased NLR and dNLR and increased LMR were associated with longer median overall survival. Three studies indicated that lower levels of IL-6 were associated with a higher objective response rate， while the association of these markers after treatment remained controversial； another study showed that an early decline in NLR， MLR， and PLR after treatment may be associated with objective response benefit. CONCLUSIONS Lower inflammatory levels during EGFR-TKIs therapy correlate with better therapeutic efficacy in NSCLC patients.

Objective To summarize the key precautions in the determination of nitrogen oxides (nitric oxide and nitrogen dioxide) in the workplace air and to explore potential optimization items in the national standard analytical method. Methods According to GBZ/T 160.29-2004 Methods for Determination of Inorganic Nitrogen Compounds in the Air Workplace, comparative experiments were conducted to evaluate and optimize critical technical parameters of the standardized method, including the oxidation efficiency of oxidation tubes and the preparation and storage of absorption solutions. The application details of the standard method were refined. Results The concentrations of nitrogen oxides (nitric oxide and nitrogen dioxide) were expressed as nitrogen dioxide equivalents. During calibration, the flow calibrator should be connected to the upstream of the air sampler, and the sampling system should undergo an air tightness check. Each batch of oxidation tubes should be validated before use. Before sampling, both end caps should be removed and the tube should be equilibrated for one hour in a clean environment with 30.00%-70.00% relative humidity. The prepared absorption stock solution in this method can be stored at 4 ℃ for up to 96 days. Commercial porous plate absorption tubes must be batch-validated before use. The sampling flow rate during sampling should be consistent with that specified in the standard method. After sampling, collected samples should be sealed in 10.00 mL amber glass bottles with screw caps and stored at 4 ℃ for up to 120 hours. Conclusion This study summarizes precautions for the sampling, detection, and calculation of nitrogen oxides (nitric oxide and nitrogen dioxide)in workplace air to strengthen quality control. Experimental optimizations of oxidation tube conditioning, absorption stock solution preparation and preservation, and sample storage conditions and durations may provide references for diversifying and simplifying the detection process, which facilitate the practical application in actual work.

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.

Objective The data related to adverse drug reaction of trastuzumab deruxtecan(T-DXd)were mined to provide references for its clinical management and adverse event handling.Methods Based on the Food and Drug Administration Adverse Event Reporting System(FAERS)database,the data reported from the post-marketing period of T-DXd until the fourth quarter of 2024 were extracted.The proportion imbalance method,specifically the reporting odds ratio(ROR)method,was used to mine and analyze the data related to adverse events of T-DXd.Results A total of 13 134 cases of adverse events related to T-DXd were obtained from the FAERS database.Gastrointestinal disorders were the most common,with 2348 adverse reaction signals,accounting for 17.88％.The most frequent adverse reactions were nausea(n=809),interstitial lung disease(n=732),and fatigue(n=579).The one with the highest ROR value was Listeriosis gastroenteritis(ROR=1228.74).The highest number of deaths were from interstitial lung disease(226),pneumonitis(90),and nausea(67).Conclusion By mining data on real-world adverse drug reaction related to T-DXd,suggests that pulmonary function,cardiac function,complete blood count and other parameters should be closely monitored during treatment for early diagnosis and treatment of corresponding complications.

Objective:To investigate the role of ubiquitin-specific protease 21 (USP21) in enterovirus group A type 71 (EV-A71) infection.Methods:Peripheral blood mononuclear cells (PBMC) were obtained from a cohort of 24 children infected with EV-A71 and 24 healthy children. Expression of USP21 was determined by real-time fluorescence quantitative PCR (qPCR). Additionally, the impact of USP21 overexpression or knockout on EV-A71 replication was evaluated using a combination of qPCR and western blot (WB) analysis. Furthermore, WB was employed to measure the levels of EV-A71 structural protein VP1, phosphorylated interferon regulatory factor 3 (IRF3) and other key molecules in the RIG-I-like receptor (RLR) signaling pathway. Co-immunoprecipitation (Co-IP) was utilized to investigate the effects of USP21 on the ubiquitin levels of EV-A71 nonstructural protein 2A protease (2A pro). Results:In comparison to healthy children, the expression of USP21 mRNA in PBMC of children infected with EV-A71 was notably elevated. The overexpression of USP21 significantly enhanced the cytopathic effects induced by EV-A71, upregulated levels of VP1 mRNA and protein, and facilitated EV-A71 replication, leading to a decrease in cell activity with increasing levels of USP21 transfection. Following the knockout of the USP21 gene, the VP1 mRNA levels were significantly declined in comparison to the control group. Furthermore, the overexpression of USP21 was found to have no impact on the transcriptional activity of EV-A71 2A pro. However, it was observed to enhance the expression of 2A pro protein, reduce the ubiquitination of 2A pro, suppress the protein levels of mitochondrial antiviral signaling protein (MAVS) and melanoma differentiation-associated gene 5 (MDA5), as well as decrease the phosphorylation of IRF3. Additionally, the induction of IFN-β mRNA by EV-A71 infection was downregulated. Conclusions:USP21 has been shown to enhance the replication of EV-A71 through the downregulation of 2A pro ubiquitination, suppression of MAVS and MDA5 protein expression, and inhibition of the interferon signaling pathway.

At present,the treatment level of cancer has made great progress.However,cancer therapy-related cardiovascular toxicity can adversely affect the prognosis of cancer patients.Studies have found that the high risk of cardiovascular toxicity has gradually become an important factor for non-tumor mortality.Therefore,there is a growing interest in oncologic cardiology.However,there is still a lack of consensus on the optimal strategies for the identification,diagnosis,and intervention of cancer therapy-related cardiovascular toxicity.This article proposes for the first time the concept of"four early"measures for cancer therapy-related cardiovascular toxicity,namely early assessment of risk factors,early identification of clinical manifestations,early diagnosis through modern medical methods,and personalized intervention in the early stage after diagnosis.Combined with relevant research at home and abroad,this article also discusses the shortcomings of research on cancer therapy-related cardiovascular toxicity and proposes future development directions,aiming to provide reference for the prevention and treatment of cardiovascular toxicity events in patients undergoing anti-tumor treatment.

Objective:To explore the efficacy and safety of mitotane in adrenal cortical carcinoma (ACC) at high risk of recurrence.Methods:A prospective observational study was designed from September 2022 to November 2023. ACC patients undergoing surgery with high recurrence risk (positive margin or Ki-67 index >10% or capsule rupture or large size or high-grade ACC) in Peking Union Medical College Hospital were enrolled in this study. All patients started mitotane treatment within 3 months after surgery, with a dose of 1.5 g/d, increased by 0.5 g per week. Once the dose reached 3 g/day, adjustments were made based on blood concentration levels. All patients received mitotane therapy for at least 1 year, and CT was performed every 12 weeks to evaluate the efficacy. The primary endpoint was 1-year progression-free survival (PFS) and safety. The efficacy was analyzed by Kaplan-Meier method for survival, and the occurrence of treatment-related adverse events was summarized.Results:A total of 12 ACC patients at high risk of recurrence were screened, comprising 6 males and 6 females. Tumors were located on the left side in 8 patients, on the right in 3, and bilaterally in 1. Five patients were classified as ENSAT stageⅡ, while 7 were classified as ENSAT stage Ⅲ. The maximum diameter of tumor was (9.07 ± 2.86) cm; the median age at diagnosis was 48 (35, 51) years, and the median Ki-67 index was (28.9 ± 16.1)%. The median time from surgery to initiation of mitotane therapy was 31 (23.0, 43.2) days, and 9 patients had blood drug concentrations of 14-20 mg/L. The median follow-up time was 16.7 (12.4, 25.2) months. At 1 year after mitotane therapy, 10 (83.8%) patients were still in disease-free survival state, with a median mitotane PFS of 27.6 months (95% CI 16.4-not reached). All ACC patients experienced 1-2 grade adverse events after taking mitotane. One patient (8.3%) experienced grade 3 adverse event, including the increasing of alanine aminotransferase and aspartate aminotransferase, as well as anorexia. No grade 4-5 adverse events occurred. The most common adverse events were gastrointestinal symptoms (10 cases), including nausea, vomiting, anorexia, and diarrhea, followed by liver function damage(9 cases) and neurotoxicity(4 cases). Conclusions:Mitotane has shown the prospect of improving the prognosis of ACC patients at high risk of recurrence after surgery. Because of its serious toxic and side effects, it is necessary to monitor its blood concentration to adjust the dosage, and take measures for adverse reactions to ensure the safety of patients.