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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for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

ObjectiveTo investigate the relationship between the vasodilation and hydrogen sulfide （H₂S） mechanism of total flavonoids of chrysanthemum （TFCC） and the large conductance Ca²⁺- activated K⁺ （BK_Ca） channels on vascular smooth muscle cells （VSMCs） of the middle cerebral artery in rats. In addition， this study will also investigate the effect of TFCC on the expression of BK_Ca channel alpha protein in rat middle cerebral artery VSMCs. MethodsThe primary method employed was acute digestion to isolate VSMCs from the middle cerebral artery of rats； whole-cell patch-clamp techniques were used to measure BK_Ca channel currents； primary tissue adherence culture was utilized to cultivate VSMCs from the middle cerebral artery of rats； Western blot were employed to determine protein expression levels. ResultsIn whole-cell patch-clamp experiments， both the H₂S donor NaHS （100 μmol/L） and endogenous H₂S enhanced BK_Ca channel currents. TFCC （30， 90， and 270 mg/L） also activatedBK_Ca channels and exhibited a certain concentration-dependent effect. Even after blocking endogenous H₂S production， TFCC （270 mg/L） still activated BK_Ca channels in VSMCs of the middle cerebral artery in rats. In Western blot experiments， the α-subunit of BK_Ca channel proteins was expressed in all groups of cells， but TFCC （30， 90， and 270 mg/L） and inhibitor IBTX group did not affect the expression of channel protein content.Conclusion TFCC can promote the opening of BK_Ca channels by promoting the generation of endogenous H₂S， or directly activate BK_Ca channels， thereby playing a role in relaxing cerebral blood vessels. However， TFCC had no significant effect on the expression of BK_Ca channel proteins.

Background: Malignant vasovagal syncope (VVS) is characterized by cardiac arrest lasting more than 3 seconds during a syncope episode or head-up tilt test (HUTT). We aim to conduct a risk assessment for potential malignant VVS before the HUTT by using economic, simple and convenient demographic data, in order to prevent adverse outcomes for pediatric VVS.Purpose: To explore the correlation between demographic factors and pediatric malignant VVS, and verify the value of these factors in early risk assessment for malignant VVS before HUTT, so as to optimize test safety and reduce adverse events. Methods: We conducted a retrospective analysis of the clinical data of 3,734 children who were initially diagnosed with VVS due to unexplained syncope and presyncope. Finally, 122 children who met the diagnostic criteria for malignant VVS were included in the malignant VVS group, and 661 children who did not meet the criteria during the same period were matched as the control group. By analyzing demographic data and other factors, we attempted to clarify the association between these factors and malignant VVS. Results: Linear relationship: age and body mass index (BMI) have independent protective effects on malignant VVS. For every 1-year increase in age and every 1 kg/m2 increase in BMI, the risk of malignant VVS decreases by 12% and 9%, respectively. Nonlinear relationship: When the age is <12.9 years old, for every additional year of age, the risk of malignant VVS decreases by 20%. For ages 12.9 years and above, the efficacy is not significant. There is no significant nonlinear relationship between BMI and malignant VVS. Conclusion Age and BMI are independent protective factors for pediatric malignant VVS. Before the age of 12.9 years, the incidence of malignant VVS gradually decreases with the increase in age, and thereafter there is no significant impact.

BackgroundThe mental health issues of medical staff have received increasing attention. Previous studies have mainly focused on the epidemiological characteristics and influencing factors， but there is a lack of sufficient understanding of the heterogeneity characteristics of the depression-anxiety symptoms network among medical staff. ObjectiveTo compare the differences in the depression-anxiety symptoms network between medical staff and community residents， so as to provide references for the promotion of the mental health of medical staff. MethodsFrom June 1， 2022 to December 31， 2024， a stratified random sampling method was used to recruit 1 959 medical staff from public hospitals and community health centers in 9 townships （industrial parks） of Dongguan City， Guangdong Province， as well as 2 412 community residents in this area， were selected as the research subjects. The general situation questionnaire， Patients' Health Questionnaire Depression Scale-9 item （PHQ-9）， and the Generalized Anxiety Disorder Scale-7 item （GAD-7） were used for investigation. The network visualization was conducted using the qgraph package of R 4.0.0. The centrality indicators of the network were calculated using the networktools package. The reliability of the network structure was evaluated using the bootnet package. The differences in the depression-anxiety symptoms network between medical staff and community residents were compared using the NetworkComparisionTest package. ResultsA total of 1 792 medical staff and 2 110 community residents completed the survey. The numbers of medical staff with mild， moderate and severe depressive symptoms were 381 （21.26%）， 217 （12.11%） and 169 （9.43%）， respectively， while the numbers of community residents were 322 （15.26%）， 98 （4.64%） and 66 （3.13%）， respectively. The numbers of medical staff with mild， moderate and severe anxiety symptoms were 411 （22.94%）， 126 （7.03%） and 93 （5.19%）， respectively， while the numbers of community residents were 320 （15.17%）， 73 （3.46%） and 18 （0.85%）， respectively. The comparison of the detection rates of depressive symptoms and anxiety symptoms between the two groups showed statistically significant differences （χ²=152.012， 205.953， P<0.01）. The network analysis results showed that the core symptom clusters of medical staff were "uncontrollable worry" "psychomotor retardation" "inability to relax"， and "depression"， while those of community residents were "uncontrollable worry" "inability to relax" "inferiority complex"， and "depression". There was a statistically significant difference in the network structure between the two groups （M=0.275， P<0.05）， and the overall connection strength of medical staff was lower than that of community residents （7.523 vs. 7.950， S=0.426， P<0.05）. ConclusionThe detection rates of depression and anxiety symptoms among medical staff were higher than those among community residents. The depression-anxiety symptoms network of medical staff was more concentrated， and the core symptoms were more prominent.［Funded by Social Development and Technology Project of Dongguan City （number， 20221800902552）］

ObjectiveTo observe the clinical efficacy and safety of Tianmeng Lingjiao Formula（天礞羚角方， TLF） as an adjunctive therapy for paroxysmal atrial fibrillation （PAF） with phlegm-heat harassing the interior syndrome. MethodsNinety patients with PAF and phlegm-heat harassing the interior syndrome were enrolled and randomly assigned in a 2∶1 ratio to test group （60 cases） and control group （30 cases）. The control group received conventional western medicine， while the test group received TLF in addition to the conventional treatment， with one dose administered daily. The treatment course for both groups was 4 weeks. The efficacy of traditional Chinese medicine （TCM） syndrome and the rate of atrial fibrillation （AF） conversion were compared between groups. Assessments included AF burden， frequency and duration of AF episodes， TCM syndrome scores， and European Heart Rhythm Association （EHRA） AF symptom classification before and after treatment. Safety indicators such as blood routine， urine routine， stool routine， liver function， kidney function， and electrolytes were also monitored. Adverse events were recorded. ResultsA total of 82 patients completed the study （54 in the test group and 28 in the control group）. After treatment， AF burden， the frequency and duration of AF episodes， and TCM syndrome scores were significantly reduced in both groups， and the test group showed lower indicators than the control group （P＜0.01 or P＜0.05）. The AF conversion rate was 25.93% （14/54） in the test group， significantly higher than 3.57% （1/28） in the control group （P＜0.05）. The total effective rate for TCM syndrome efficacy was 83.33% （45/54） in the test group， which was significantly higher than 67.86% （19/28） in the control group （P＜0.05）. After treatment， the EHRA atrial fibrillation symptom severity in the test group showed significant improvement， and the improvement in atrial fibrillation symptoms was superior to that in the control group （P＜0.01or P＜0.05）. No significant abnormalities were observed in safety indicators， and no adverse events occurred during the trial. ConclusionTLF as an adjunctive therapy can reduce AF burden， enhance the conversion rate of AF， decrease the frequency and duration of episodes， and improve clinical symptoms in patients with paroxysmal atrial fibrillation presenting with phlegm-heat harassing the interior syndrome， with a favorable safety profile.

Osteoporosis and cardiovascular calcification,two major age-related chronic diseases that China is confronting today,pose serious threats to public health.Previous studies have reported overlapping connections in the pathological processes and molecular mechanisms of these two diseases,particularly concerning inflammation,oxidative stress,and dysregulation of mineral metabolism,and that these two diseases tend to share common pathogenic factors.However,research exploring the comorbidity mechanisms of the two diseases remains limited in both depth and scope,largely due to the lack of widely accepted comorbidity animal models.Herein,we analyzed the latest research findings on the comorbidity mechanisms of vascular calcification and osteoporosis,focusing on summarizing the animal disease models currently in extensive use and the relevant evaluation criteria.We aim to provide new references for comorbidity research models and offer scientific evidence for future studies on pathological mechanisms and the development of new therapeutic strategies.

Objective To investigate the effects of secukinumab combined with E'xian Huaban decoction on traditional Chinese medicine symptom scores and nailfold microcirculation morphology in patients with plaque psoriasis.Methods A total of 120 patients with plaque psoriasis were selected and randomly divided into monotherapy group(n=60)and combination therapy group(n=60).The monotherapy group received secukinumab treatment,while the combination therapy group was treated with secukinumab plus E'xian Huaban Decoction.Clinical therapeutic efficacy,traditional Chinese medicine symptom scores(including skin lesions,pruritus,rough skin,scales and irritabili-ty),relevant scale[Psoriasisarea and Severity Index(PASI),Dermatology Life Quality Index(DLQI)and Visual Analogue Scale(VAS)]scores,inflammatory factors[interleukin-17(IL-17),interleukin-23(IL-23)and vascular endothelial growth factor(VEGF)]levels and nailfold microcircu-lation morphology(loop tube morphology,blood flow state and loop periphery state)were observed in both groups.Results The overall clinical efficacy rate in the combination therapy group was 95.00％,which was significantly higher than 83.33％ in the monotherapy group(P＜0.05).After treatment,traditional Chinese medicine symptom scores(skin lesions,pruritus,rough skin,scales and irritability),inflammatory factors(IL-17,IL-23 and VEGF)levels,scale(PASI,DLQI and VAS)scores as well as loop tube morphology scores,blood flow state scores and loop periphery state scores were signifi-cantly lower compared to pre-treatment values,and these parameters were significantly lower in the combination therapy group than the monotherapy group(P＜0.05).Conclusion Secukinumab combined with E'xian Huaban Decoction can significantly improve clinical symptoms and signs,and enhancing the clinical therapeutic effect.

Aralia taibaiensi, widely distributed in western China, particularly in the Qinba Mountains, has been utilized as a folk medicine for treating diabetes, gastropathy, rheumatism, and cardiovascular diseases. Saponins from A. taibaiensis (sAT) have demonstrated protective effects against oxidative stress and mitochondrial dysfunction induced by ischemia/reperfusion (I/R). However, the underlying mechanisms remain unclear. In vivo, middle cerebral artery occlusion/reperfusion (MCAO/R) induced inflammatory infiltration, neuronal injury, cell apoptosis, mitochondrial dysfunction, and oxidative stress in the ischaemic penumbra, which were effectively mitigated by sAT. sAT increased the mRNA and protein expression levels of apelin and its receptor apelin/apelin receptors (ARs) both in vivo and in vitro. (Ala13)-Apelin-13 (F13A) and small interfering RNA (siRNA) abolished the regulatory effects of sAT on neuroprotection mediated by adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK)/protein kinase B (Akt). Furthermore, sAT induced apelin/AR expression by simultaneously inhibiting P38 mitogen-activated protein kinase (P38 MAPK)/activating transcription factor 4 (ATF4) and upregulating hypoxia-inducible factor-1α (HIF-1α). Our findings indicate that sAT regulates apelin/AR/AMPK by inhibiting P38 MAPK/ATF4 and upregulating HIF-1a, thereby suppressing oxidative stress and mitochondrial dysfunction.
Animals ; Reperfusion Injury/prevention & control* ; Aralia/chemistry* ; Saponins/administration & dosage* ; AMP-Activated Protein Kinases/genetics* ; Male ; Apelin/genetics* ; Signal Transduction/drug effects* ; Neuroprotective Agents/administration & dosage* ; Brain Ischemia/genetics* ; Rats, Sprague-Dawley ; Rats ; Oxidative Stress/drug effects* ; Apelin Receptors/genetics* ; Humans ; Apoptosis/drug effects* ; Mice

Objective To evaluate the effects of Yiqi Huayu Jiedu Decoction combined with chemotherapy on disease-free survival rate and disease-free survival(DFS)of patients with stage Ⅱ and Ⅲ postoperative gastric cancer.Methods Totally 102 patients with stage Ⅱ and Ⅲ postoperative gastric cancer treated by Affiliated Hospital of Nanjing University of Chinese Medicine and Nanjing Drum Tower Hospital were selected.The patients were divided into control group(52 cases)and test group(50 cases)with block random method.The control group received conventional postoperative adjuvant chemotherapy for 4-6 cycles.The test group was treated with Chinese materia medica for 6 months on the basis of chemotherapy.Continue follow-up until the patients had recurrence,metastasis or death.DFS was analyzed by Kaplan-Meier method,and the average DFS of the two groups was estimated.Log Rank test was used to test the significance of differences in survival distribution between groups.Univariate and multivariate analyses of variables affecting prognosis were conducted using the Cox regression model.Adverse reactions of both groups were monitored.Results The recurrence and metastasis rate of the test group was 12.0%(6/50),while that of the control group was 34.6%(18/52),which was lower in the test group than in the control group(P<0.05).The 1-year and 2-year DFS rates and DFS of the control group were 86.5%,72.2%and 25.8 months respectively,while those of the test group were 95.6%,84.1%and 33.9 months respectively,with statistical significance(P=0.012).Cox-regression analysis suggested:The independent factors influencing the disease-free survival time of patients with gastric cancer were the test group(P=0.022),surgical methods of subtotal gastrectomy(P=0.038)and clinical stages Ⅱ(P=0.040).No adverse reactions related to Chinese materia medica were reported in the two groups.Conclusion Yiqi Huayu Jiedu Decoction combined with chemotherapy can improve the 1-year and 2-year DFS rate,prolong DFS,reduce the recurrence and metastasis rate,and improve the prognosis.