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published before September 2024 in CNKI,PubMed,and Web of Science databases.Articles with poor relevance to the topic,outdated,or duplicated content were excluded,and finally,109 articles were included for review.RESULTS AND CONCLUSION:(1)This paper reviews recent methods for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

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）］

Segmental dystonia caused by levodopa and deep brain stimulation (DBS) in Parkinson′s disease (PD) has been described rarely and may be underrecognized in clinical practice. A case of peak-dose dyskinesia with segmental dystonia, including blepharospasm, jaw dystonia, torticollis, and dystonia in the left upper limb, after bilateral subthalamic nucleus DBS, is reported. The literatures on PD-related dystonia, including dystonia manifestations before and after treatment with medication and DBS, are reviewed to enhance clinicians′ recognition of movement disorders occurring during the treatment of PD and to improve patient outcomes.

Objective:To evaluate the feasibility of using the domestic ArcherQA system for fast and simplified independent verification of CyberKnife (CK) stereotactic radiotherapy (SRT) plans.Methods:SRT plans of 57 patients treated with CK at Tianjin Medical University Cancer Institute and Hospital from August 2021 to August 2022 were retrospectively analyzed, including 15 intracranial, 30 pulmonary, and 12 abdominal tumors cases. Point-dose and planar-dose verifications were performed using an ionization chamber and radiochromic films embedded in a homogeneous phantom, and the results were compared with those calculated by the treatment planning system (TPS). The localization CT images and corresponding SRT plans were imported into the ArcherQA system for independent dose verification and analysis. The correlation between ArcherQA results and phantom measurements was analyzed, with comparisons of target mean dose differences and γ pass rates.Results:Phantom measurement results showed, the measured point-dose differences for intracranial, lung, and abdominal plans were -0.94% ± 3.22%, 1.92% ± 2.05%, and 2.12% ± 0.77%, respectively. The mean dose differences in target dose calculation between ArcherQA and TPS: intracranial in the gross tumor volume (GTV) regions were 0.34% ± 2.21%, lung tumor GTV were -2.47% ± 2.46%, and abdominal tumor GTV were 0.80% ± 2.61%, respectively. Among them, the abdominal GTV region showed the highest correlation between ArcherQA and measured results ( r=0.78). The average two-dimensional γ pass rates (2 mm/2%, threshold=10%) measured using phantom films were 95.92% ± 2.35% for intracranial, 95.70% ± 2.74% for lung, and 96.74% ± 3.41% for abdominal tumors plans, respectively. The three-dimensional ArcherQA results showed comparable γ pass rates (1 mm/2%, threshold=10%) for lung and abdominal GTV and PTV regions, with similar medians and data dispersion to film measurements. Conclusions:The ArcherQA system enables rapid and efficient independent dose verification of CK SRT plans without the need for additional hardware. The verification results show good correlation with phantom measurements, supporting its potential as an auxiliary quality assurance tool in clinical CK SRT implementation.

OBJECTIVE The emergence of evolving variants of Coronavirus disease 2019(COVID-19)has fostered the need for change of newer and adaptive treatments for these infections.During the COVID-19 pandemic and persists,traditional Chinese medicine(TCM)herbs exhibit significant bioactivity and therapeutic effect.This study is aimed to evaluate the efficacy of four TCM preparations on 28-day mortality risk of patients and changes of the laboratory indicators.METHODS The retrospective cohort study included patients with COVID-19 who were admitted to the Jiangsu Province Hospital of Chinese Medicine from December 15,2022 to January 15,2023,and those died within 48 hours of admission or cannot be tracked for outcomes were excluded.The pri-mary outcome was survival status in 28 days(death or survival)starting from the day of admission.The second outcomes were labora-tory indicators,including absolute lymphocyte count,lactate dehydrogenase,creatinine,and blood urea nitrogen.Binary logistic re-gressions were used to estimate the effect of TCM preparations on the primary and secondary outcomes in main analysis.Meanwhile,heterogeneity and robustness of results from main analysis were assessed by subgroup analyses and multiple sensitivity analyses.RESULTS 1 816 eligible patients were included in analysis dataset,including 573 patients received standard care(control group)and 1 243 patients received TCM preparations(hospital preparation group).The 28-day mortality rate of hospital preparation group was lower than that of control group(4.75%vs.14.83%),and the difference was statistically significant(χ2=54.666,P<0.001).The risk of 28-day mortality was 0.535 times lower in the hospital preparation group as compared with the control group(OR=0.46,95%CI:0.305-0.708,P<0.001)showed by multivariable binary logistic regressions.Subgroup analyses showed that taking TCM preparations reduced the 28-day mortality risk.Sensitivity analyses demonstrated that the results of the main analysis for primary outcomes were robust.For secondary outcomes,the risk of abnormal absolute lymphocyte counts at discharge in the hospital prepara-tion group decreased by 0.284 times(OR=0.703,95%CI:0.515-0.961,P=0.027).CONCLUSION Compared with standard of care,taking four hospital preparations including Kanggan Heji,Feining Heji,Qishen Gubiao Keli,and Qianghuo Qushi Qingwen Heji decreased risk of 28-day mortality among hospitalized COVID-19 patients.TCM therapy achieves adequate therapeutic effects in COVID-19.

OBJECTIVE The emergence of evolving variants of Coronavirus disease 2019(COVID-19)has fostered the need for change of newer and adaptive treatments for these infections.During the COVID-19 pandemic and persists,traditional Chinese medicine(TCM)herbs exhibit significant bioactivity and therapeutic effect.This study is aimed to evaluate the efficacy of four TCM preparations on 28-day mortality risk of patients and changes of the laboratory indicators.METHODS The retrospective cohort study included patients with COVID-19 who were admitted to the Jiangsu Province Hospital of Chinese Medicine from December 15,2022 to January 15,2023,and those died within 48 hours of admission or cannot be tracked for outcomes were excluded.The pri-mary outcome was survival status in 28 days(death or survival)starting from the day of admission.The second outcomes were labora-tory indicators,including absolute lymphocyte count,lactate dehydrogenase,creatinine,and blood urea nitrogen.Binary logistic re-gressions were used to estimate the effect of TCM preparations on the primary and secondary outcomes in main analysis.Meanwhile,heterogeneity and robustness of results from main analysis were assessed by subgroup analyses and multiple sensitivity analyses.RESULTS 1 816 eligible patients were included in analysis dataset,including 573 patients received standard care(control group)and 1 243 patients received TCM preparations(hospital preparation group).The 28-day mortality rate of hospital preparation group was lower than that of control group(4.75%vs.14.83%),and the difference was statistically significant(χ2=54.666,P<0.001).The risk of 28-day mortality was 0.535 times lower in the hospital preparation group as compared with the control group(OR=0.46,95%CI:0.305-0.708,P<0.001)showed by multivariable binary logistic regressions.Subgroup analyses showed that taking TCM preparations reduced the 28-day mortality risk.Sensitivity analyses demonstrated that the results of the main analysis for primary outcomes were robust.For secondary outcomes,the risk of abnormal absolute lymphocyte counts at discharge in the hospital prepara-tion group decreased by 0.284 times(OR=0.703,95%CI:0.515-0.961,P=0.027).CONCLUSION Compared with standard of care,taking four hospital preparations including Kanggan Heji,Feining Heji,Qishen Gubiao Keli,and Qianghuo Qushi Qingwen Heji decreased risk of 28-day mortality among hospitalized COVID-19 patients.TCM therapy achieves adequate therapeutic effects in COVID-19.

Objective:To discuss the expression of Rh family C glycoprotein(RHCG)in the esophageal squamous cell carcinoma(ESCC)tissue and its effect on the malignant biological behavior of ESCC cells,and to clarify the value of RHCG as a diagnostic and prognostic marker for the ESCC patients.Methods:A total of 143 ESCC tissue samples and 105 adjacent normal tissue samples were collected.Using immunohistochemical staining method,141 ESCC samples were divided into two groups:RHCG low expression group(immunohistochemistry score≤6)and RHCG high expression group(immunohistochemistry score>6).Immunohistochemical method was used to detect the RHCG protein expression in 143 ESCC tissues and 105 normal tissues,and the relationship between the clinicopathological characteristics of the ESCC patients was analyzed.Receiver operating characteristic(ROC)curve and Kaplan-Meier survival analysis were used to evaluate the value of RHCG in diagnosis and prognosis of the ESCC patients;univariate and multivariate COX regression analysis were used to determine the independent risk factors affecting the prognosis of the ESCC patients.Gene Expression Profiling Interactive Analysis(GEPIA2)database was used to analyze the expression of RHCG mRNA in various tumor tissues.The ESCC TE-1 cells were cultured and transfected in to 6-well cell culture plates with different Lipofectamine2000∶RHCG ratios;the cells in RHCG transfection group were transfected with weights of 2.0,2.5,and 3.0 μg for 24 and 48 h,respectively,and the cells in NC group transfected with empty vector as control.Western blotting method was used to detect the RHCG protein expression level in the TE-1 cells in various groups after transfection at different concentrations and verify the optimal transfection conditions;cell counting kit-8(CCK-8)assay was used to detect the proliferation activities of the TE-1 cells;plate clone formation assay was used to detect the colony formation numbers of the TE-1 cells;Transwell chamber assay was used to detect the numbers of migrating TE-1 cells.Results:Compared with adjacent normal tissue,the RHCG gene expression level in various cancer tissues including ESCC,glioblastoma multiforme,and head and neck squamous cell carcinoma was significantly decreased(P<0.05).RHCG protein was mainly located on the cell membrane of normal esophageal squamous epithelial cells;the RHCG protein expression intensity in ESCC tissues was lower than that in adjacent normal esophageal tissue(χ2=109.373,P<0.001),and the patients in RHCG low expression group had poorer differentiation than those in RHCG high expression group(P=0.041).The area under the curve(AUC)value of RHCG for diagnosing ESCC was 0.86,with sensitivity and specificity of 95.1%and 75.0%,respectively;the Kaplan-Meier survival analysis results showed that compared with high RHCG expression group,the patients in low RHCG expression group had shorter survival time and poorer prognosis[harard ratio(HR)=0.269,95%confidence interval(CI):0.113-0.639,P=0.020];the COX regression analysis results showed that low RHCG expression could serve as an independent risk factor affecting the prognosis of ESCC[HR=4.569,95%CI=1.315-15.877,P=0.017)].The Western blotting results verified that the optimal transfection condition was 3.0 μg RHCG plasmid for 48 h,at which time RHCG overexpression was optimal and RHCG protein expression level was highest.The CCK-8 assay results showed that compared with control group,the proliferation activity in RHCG overexpression group was decreased on the 4th day after cell seeding(P<0.001).In the TE-1 cells,the colony formation number of the TE-1 cells in RHCG over-expression group was lower than that in control group(t=17.70,P<0.001).The Transwell chamber assay results showed that compared with control group,the number of migrating cells in RHCG over-expression group was decreased(t=23.74,P<0.001).Conclusion:RHCG expression is decreased in ESCC tissues and associated with poor prognosis in ESCC patients;overexpression of RHCG can inhibit the proliferation and migration of the TE-1 cells,providing a theoretical basis for RHCG as a novel diagnostic and prognostic marker and therapeutic target for ESCC.

Objective:To evaluate the feasibility of using the domestic ArcherQA system for fast and simplified independent verification of CyberKnife (CK) stereotactic radiotherapy (SRT) plans.Methods:SRT plans of 57 patients treated with CK at Tianjin Medical University Cancer Institute and Hospital from August 2021 to August 2022 were retrospectively analyzed, including 15 intracranial, 30 pulmonary, and 12 abdominal tumors cases. Point-dose and planar-dose verifications were performed using an ionization chamber and radiochromic films embedded in a homogeneous phantom, and the results were compared with those calculated by the treatment planning system (TPS). The localization CT images and corresponding SRT plans were imported into the ArcherQA system for independent dose verification and analysis. The correlation between ArcherQA results and phantom measurements was analyzed, with comparisons of target mean dose differences and γ pass rates.Results:Phantom measurement results showed, the measured point-dose differences for intracranial, lung, and abdominal plans were -0.94% ± 3.22%, 1.92% ± 2.05%, and 2.12% ± 0.77%, respectively. The mean dose differences in target dose calculation between ArcherQA and TPS: intracranial in the gross tumor volume (GTV) regions were 0.34% ± 2.21%, lung tumor GTV were -2.47% ± 2.46%, and abdominal tumor GTV were 0.80% ± 2.61%, respectively. Among them, the abdominal GTV region showed the highest correlation between ArcherQA and measured results ( r=0.78). The average two-dimensional γ pass rates (2 mm/2%, threshold=10%) measured using phantom films were 95.92% ± 2.35% for intracranial, 95.70% ± 2.74% for lung, and 96.74% ± 3.41% for abdominal tumors plans, respectively. The three-dimensional ArcherQA results showed comparable γ pass rates (1 mm/2%, threshold=10%) for lung and abdominal GTV and PTV regions, with similar medians and data dispersion to film measurements. Conclusions:The ArcherQA system enables rapid and efficient independent dose verification of CK SRT plans without the need for additional hardware. The verification results show good correlation with phantom measurements, supporting its potential as an auxiliary quality assurance tool in clinical CK SRT implementation.

Segmental dystonia caused by levodopa and deep brain stimulation (DBS) in Parkinson′s disease (PD) has been described rarely and may be underrecognized in clinical practice. A case of peak-dose dyskinesia with segmental dystonia, including blepharospasm, jaw dystonia, torticollis, and dystonia in the left upper limb, after bilateral subthalamic nucleus DBS, is reported. The literatures on PD-related dystonia, including dystonia manifestations before and after treatment with medication and DBS, are reviewed to enhance clinicians′ recognition of movement disorders occurring during the treatment of PD and to improve patient outcomes.