1.Mechanism of Embryo Implantation Promotion via Exosomal miRNA-mediated Communication Network at Maternal-fetal Interface Based on Bushen Huoxue Therapy
Pei GUO ; Jiajun LIU ; Hang ZHOU ; Zeyi GUO ; Yili WANG ; Linwen DENG ; Qian ZENG ; Jinzhu HUANG
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(14):317-327
ObjectiveTo investigate whether Bushen Huoxue prescription improves embryo implantation through regulating exosomal miRNA to enhance maternal-fetal interface communication based on Bushen Huoxue therapy. MethodsIn the animal experiment, all the rats (except for the blank group) were administered hydroxyurea (450 mg·kg-1) via gavage for 10 d, as well as epinephrine (0.3 mg·kg-1) and mifepristone (5.5 mg·kg-1) via subcutaneous injection for 7 d to establish an implantation disorder model of kidney deficiency and blood stasis type. The Bushen Huoxue prescription (BSHX) groups were administered the prescription at different doses (7.30 g·kg-1 for the high-dose group, 3.65 g·kg-1 for the medium-dose group, and 1.83 g·kg-1 for the low-dose group) via gavage. The dydrogesterone group was administered the corresponding medicine (2.63 mg·kg-1) via gavage. After intervention for 10 days, uterine histopathological changes were observed via hematoxylin-eosin (HE) staining. Mucin (MUC1), forkhead box protein O1 (FoxO1), and homeobox A10 (HoxA10) expression levels were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) and Western blot. Cell experiment selected primary endometrial epithelial cells (EEC) and trophoblast cells (TC) as research subjects. Exosome-free medicated serum was prepared by ultracentrifugation and cultured in complete medium. Exosomes were isolated from cell supernatants by ultracentrifugation for cross-co-culture. After 48 h, migration and invasion abilities were assessed by scratch and Transwell assays. Sequencing was then performed on EEC-origin exosomal miRNA. ResultsThe model rats exhibited thin endometrium, along with reduced blood vessels, glandules, and pinopode numbers. BSHX improved endometrial morphology and increased pinopode numbers. MUC1, FoxO1, and HoxA10 expressions were downregulated in the model rats, while these parameters were upregulated after BSHX medium- and high-dose intervention. In the cell experiment, after exosome-free medicated serum intervention for 24 h, migration and invasion abilities were enhanced in the BSHX groups (P<0.01). In EEC-origin exosomal miRNA sequencing, gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed enrichment in biological processes (gastrulation, neuronal differentiation, alongside cell development and regeneration), involving the mitogen-activated protein kinase (MAPK), FoxO1, Wnt, mammalian target of rapamycin (mTOR), and tumor necrosis factor (TNF) signaling pathways. ConclusionBSHX promotes embryo implantation by improving endometrial receptivity via regulating exosomal miRNA. These findings provide potential targets for exosomal miRNA-based assisted reproductive strategies and a novel theoretical basis for infertility treatment by traditional Chinese medicine.
2.Application of artificial intelligence in quality control of mammographic images
Yunyun LYU ; Le FU ; Ruixin LI ; Zeyi ZHANG ; Xiaoli MU ; Hui WANG ; Huizhi CAO ; Jianli YU
Chinese Journal of Radiological Health 2026;35(2):173-179
Objective To evaluate the application value of artificial intelligence (AI) in the quality control of mammographic images and explore its feasibility for improving image quality. Methods A retrospective analysis was conducted on 500 mammographic images from 125 female patients. These images were acquired in December 2024 at the Department of Radiology, Obstetrics and Gynecology Hospital of Tongji University by two junior technologists with qualification certificate for junior radiologic technologists and ≤ 2 months of independent operation. The reference standard was the evaluation by a panel of senior experts comprising one associate chief radiologist and two intermediate radiologic technologists with over 10 years of experience in mammography. The evaluation was based on nine criteria within an AI quality control system. The correlation and consistency (Kappa test) of the assessment outcomes were compared among the junior technologist group, the AI quality control group, and the panel group. Additionally, the differences in the proportions of high-, medium-, and low-quality images rated by the three groups were analyzed. After one month of AI assistance, 200 images from 50 additional patients acquired by the same two junior technologists in January 2025 were collected. The area under the receiver operating characteristic curve and 95% confidence interval (95%CI) were calculated for the assessment performance of the junior technologist group before and after AI assistance. Differences were compared using the DeLong test. Changes in the disqualification rates for the nine criteria were analyzed. Results In consistency analysis, the AI quality control group showed high consistency with the panel group across multiple key criteria, with Kappa values ranging from 0.41 to 1.00. In contrast, the consistency between the junior technologist group and the panel group was generally low, with Kappa values ranging from 0.13 to 0.49. In comparison of image quality classification, no significant differences were observed in image quality classification between the AI group and the panel group (P>0.05). However, the proportion of images rated as high quality by the junior technologist group was significantly higher, while the proportions rated as medium and low quality were significantly lower, compared to those rated by the panel group (P<0.05). After AI assistance, the area under the receiver operating characteristic curve for the assessment performance of junior technologist group increased significantly from 0.56 (95%CI: 0.51-0.61) to 0.91 (95%CI: 0.87-0.94) (P<0.001). The disqualification rates for skin folds, incomplete inclusion of the pectoralis major muscle, and nipple not in profile decreased significantly. Conclusion AI demonstrates assessment capability comparable to the panel group in mammographic quality control. AI effectively enhances the quality of images acquired by junior technologists through real-time, objective feedback.
3.Application of artificial intelligence in quality control of mammographic images
Yunyun LYU ; Le FU ; Ruixin LI ; Zeyi ZHANG ; Xiaoli MU ; Hui WANG ; Huizhi CAO ; Jianli YU
Chinese Journal of Radiological Health 2026;35(2):173-179
Objective To evaluate the application value of artificial intelligence (AI) in the quality control of mammographic images and explore its feasibility for improving image quality. Methods A retrospective analysis was conducted on 500 mammographic images from 125 female patients. These images were acquired in December 2024 at the Department of Radiology, Obstetrics and Gynecology Hospital of Tongji University by two junior technologists with qualification certificate for junior radiologic technologists and ≤ 2 months of independent operation. The reference standard was the evaluation by a panel of senior experts comprising one associate chief radiologist and two intermediate radiologic technologists with over 10 years of experience in mammography. The evaluation was based on nine criteria within an AI quality control system. The correlation and consistency (Kappa test) of the assessment outcomes were compared among the junior technologist group, the AI quality control group, and the panel group. Additionally, the differences in the proportions of high-, medium-, and low-quality images rated by the three groups were analyzed. After one month of AI assistance, 200 images from 50 additional patients acquired by the same two junior technologists in January 2025 were collected. The area under the receiver operating characteristic curve and 95% confidence interval (95%CI) were calculated for the assessment performance of the junior technologist group before and after AI assistance. Differences were compared using the DeLong test. Changes in the disqualification rates for the nine criteria were analyzed. Results In consistency analysis, the AI quality control group showed high consistency with the panel group across multiple key criteria, with Kappa values ranging from 0.41 to 1.00. In contrast, the consistency between the junior technologist group and the panel group was generally low, with Kappa values ranging from 0.13 to 0.49. In comparison of image quality classification, no significant differences were observed in image quality classification between the AI group and the panel group (P>0.05). However, the proportion of images rated as high quality by the junior technologist group was significantly higher, while the proportions rated as medium and low quality were significantly lower, compared to those rated by the panel group (P<0.05). After AI assistance, the area under the receiver operating characteristic curve for the assessment performance of junior technologist group increased significantly from 0.56 (95%CI: 0.51-0.61) to 0.91 (95%CI: 0.87-0.94) (P<0.001). The disqualification rates for skin folds, incomplete inclusion of the pectoralis major muscle, and nipple not in profile decreased significantly. Conclusion AI demonstrates assessment capability comparable to the panel group in mammographic quality control. AI effectively enhances the quality of images acquired by junior technologists through real-time, objective feedback.
4.Application of artificial intelligence in quality control of mammographic images
Yunyun LYU ; Le FU ; Ruixin LI ; Zeyi ZHANG ; Xiaoli MU ; Hui WANG ; Huizhi CAO ; Jianli YU
Chinese Journal of Radiological Health 2026;35(2):173-179
Objective To evaluate the application value of artificial intelligence (AI) in the quality control of mammographic images and explore its feasibility for improving image quality. Methods A retrospective analysis was conducted on 500 mammographic images from 125 female patients. These images were acquired in December 2024 at the Department of Radiology, Obstetrics and Gynecology Hospital of Tongji University by two junior technologists with qualification certificate for junior radiologic technologists and ≤ 2 months of independent operation. The reference standard was the evaluation by a panel of senior experts comprising one associate chief radiologist and two intermediate radiologic technologists with over 10 years of experience in mammography. The evaluation was based on nine criteria within an AI quality control system. The correlation and consistency (Kappa test) of the assessment outcomes were compared among the junior technologist group, the AI quality control group, and the panel group. Additionally, the differences in the proportions of high-, medium-, and low-quality images rated by the three groups were analyzed. After one month of AI assistance, 200 images from 50 additional patients acquired by the same two junior technologists in January 2025 were collected. The area under the receiver operating characteristic curve and 95% confidence interval (95%CI) were calculated for the assessment performance of the junior technologist group before and after AI assistance. Differences were compared using the DeLong test. Changes in the disqualification rates for the nine criteria were analyzed. Results In consistency analysis, the AI quality control group showed high consistency with the panel group across multiple key criteria, with Kappa values ranging from 0.41 to 1.00. In contrast, the consistency between the junior technologist group and the panel group was generally low, with Kappa values ranging from 0.13 to 0.49. In comparison of image quality classification, no significant differences were observed in image quality classification between the AI group and the panel group (P>0.05). However, the proportion of images rated as high quality by the junior technologist group was significantly higher, while the proportions rated as medium and low quality were significantly lower, compared to those rated by the panel group (P<0.05). After AI assistance, the area under the receiver operating characteristic curve for the assessment performance of junior technologist group increased significantly from 0.56 (95%CI: 0.51-0.61) to 0.91 (95%CI: 0.87-0.94) (P<0.001). The disqualification rates for skin folds, incomplete inclusion of the pectoralis major muscle, and nipple not in profile decreased significantly. Conclusion AI demonstrates assessment capability comparable to the panel group in mammographic quality control. AI effectively enhances the quality of images acquired by junior technologists through real-time, objective feedback.
5.Outer membrane vesicles derived from Pasteurella multocida inhibit proliferation,invasion,and migration of bladder cancer cells and promote apoptosis
Yang WANG ; Zeyi WANG ; Xiangqian CAO ; Bing SHEN
Academic Journal of Naval Medical University 2025;46(8):1000-1008
Objective To investigate the biological effects of Pasteurella multocida(Pm)culture supernatant and Pm-derived outer membrane vesicle(OMV)on bladder cancer cells.Methods Pm was cultured and its supernatant was collected.The effects of the supernatant on proliferation,migration and invasion of bladder cancer cell lines(T24 and 5637)were assessed by cell counting kit 8(CCK-8),wound healing assay,and Transwell migration and invasion assays with phosphate-buffered saline(PBS)and brain heart infusion(BHI)broth as controls.Pm-OMV were isolated from the supernatant via ultracentrifugation,and the remaining components of the supernatant served as control.The effects of Pm-OMV on proliferation,migration and invasion of T24 and 5637 cells were assessed by CCK-8 and Transwell migration and invasion assays.Apoptosis was analyzed by flow cytometry.A nude mouse xenograft tumor model was established.After intratumoral multi-point injections of Pm-OMV or PBS,the tumor growth was evaluated and the effects of Pm-OMV on proliferation and apoptosis of bladder cancer cells in vivo were verified by Ki67(a proliferation marker)immunohistochemical staining and TUNEL assay.Results Pm culture supernatant significantly inhibited the proliferation,invasion,and migration of T24 and 5637 cells in vitro compared with PBS and BHI controls(all P<0.01).Pm-OMV not only inhibited the proliferation,invasion,and migration of T24 and 5637 cells,but also induced the apoptosis,and the differences were significant compared with the remaining components of the supernatant(all P<0.05).The nude mouse subcutaneous tumor transplantation experiment further confirmed that Pm-OMV inhibited the proliferation of bladder cancer cells and promoted apoptosis in vivo,and the differences were significant compared with the PBS control(all P<0.05).Conclusion Pm-OMV can inhibit the proliferation,invasion,and migration of bladder cancer cells and promote the apoptosis.It provides an experimental basis for studying the mechanism of microbial regulation of tumor progression and for developing new treatment strategies for bladder cancer.
6.Biomechanics characteristics during sitting up in knee osteoarthritis patients of different ages:a systematic review and meta-analysis
Ke WANG ; Zeyi ZHANG ; Liwen ZHANG ; Meizhen ZHANG
Chinese Journal of Tissue Engineering Research 2024;28(18):2939-2946
OBJECTIVE:There is no consensus on which sit-up strategy to adopt in knee osteoarthritis patients of different ages.Therefore,this study evaluated the biomechanical characteristics of sit-ups in knee osteoarthritis patients of different ages compared with healthy individuals by meta-analysis system and analyzed the sit-up movement patterns of patients of different ages to provide a reference for improving the sit-up function of patients. METHODS:By March 2023,observational studies of biomechanical characteristics of sitting up in patients with knee osteoarthritis and healthy population were retrieved on PubMed,Web of Science and CNKI.Subjects were required to be patients over 50 years of age with knee osteoarthritis who had Kellgren-Lawrence severity grading≥Ⅰ on knee imaging and who had regular knee pain.Subjects were analyzed by age(50-60 years vs.over 60 years)and severity(mild to moderate patients vs.severe patients)subgroups according to inclusion and exclusion criteria.Quality assessment was performed using the modified Down and black scale.Stata 16.0 software was used to perform subgroup analysis to determine the biomechanical characteristics of sitting up in patients with knee osteoarthritis of different ages and severities. RESULTS:A total of 14 randomized controlled trials(824 subjects)were included in the meta-analysis.The mean quality score of all included literature was 76.2,with a range of 66.7 to 86.7,all of which were of medium to high quality and representative.The included studies were of moderate to high quality and representative.Meta-analysis results found that(1)compared to healthy individuals,patients with knee osteoarthritis had longer total sitting up time(SMD=0.92,95%CI:0.76-1.09),P<0.001)and longer extension phase time(SMD=0.46,95%CI:0.18-0.74,P=0.001).Compared to mild to moderate patients,the total duration increased more significantly in severe patients(P<0.001)and the duration of the extension phase increased more significantly in patients over 60 years of age than in patients 50-60 years of age(P=0.001).(2)Compared to healthy individuals,patients with knee osteoarthritis had greater sitting-up trunk flexion motion range(SMD=0.64,95%CI:0.37-0.91,P<0.001);knee flexion motion range(SMD=-0.47,95%CI:-0.70 to-0.24,P<0.001)and ankle dorsiflexion motion range(SMD=-0.32,95%CI:-0.56 to-0.08,P=0.01)were smaller.And knee flexion motion range decreased more significantly in patients over 60 years of age than in patients 50-60 years of age(P<0.001).(3)The peak hip flexion moment(SMD=-0.57,95%CI:-0.83 to-0.31,P<0.001)and peak knee extension moment(SMD=-0.83,95%CI:-1.08 to-0.59,P<0.001)were smaller in patients with knee osteoarthritis. CONCLUSION:(1)Patients with knee osteoarthritis over the age of 60 years have a longer sit-up cushion and extension phase than patients aged 50 to 60 years.The increase in total sit-up duration was also more pronounced in patients with higher severity grades.The increased length of sitting up in patients with advanced age and knee osteoarthritis severity may increase the duration of cartilage loading,exacerbate knee pain symptoms,and increase the difficulty of sitting up in this population.(2)Patients with knee osteoarthritis exhibit limited knee and ankle flexion motion range.Knee mobility is more limited in patients over 60 years of age.(3)Patients with knee osteoarthritis have reduced peak hip flexion and knee extension moments,which may be a compensatory strategy for pain relief.
7.Effect Evaluation of Multidisciplinary Collaborative Diagnosis and Treatment Model for Children with Brain Injury
Xiongwu YU ; Yunli ZHOU ; Zhiyong DING ; Chaohong WANG ; Zeyi XIE ; Hongna LU ; Hua JIN
Journal of Kunming Medical University 2024;45(1):156-162
Objective To summarize the experience of multi-disciplinary team(MDT)in the pediatric department of Qujing Maternal and Child Health Hospital,and to evaluate the effectiveness of MDT on neonatal brain injury.Methods The clinical data of children with brain injury and treated in the pediatrics department of Qujing Maternal and Child Health Hospital from November 2019 to April 2023 were collected.The children with brain injury and treated from October 2019 to June 2020 were regarded as the non-MDT group,and the children with brain injury and treated from July 2020 to April 2023 were regarded as the MDT group for comparative analysis.Chi-square test/t-test was used to compare and analyze the clinical data of the two groups.Results Among the 890 cases of pediatric brain injury,there were 519 males and 371 females.The median and quartiles of the age distribution for the two groups were as follows:MDT group 2.00(0.82,5.00)years and non-MDT group 1.00(1.00,4.00)years.Craniocerebral injury was the main type of brain injury in both groups,in addition,among children with craniocerebral injury and intracranial hemorrhage,the cure rate of MDT group was higher than that of non-MDT group,and the difference was statistically significant(P<0.05).Among the 405 children in MDT group,154(38.0%)underwent the surgery,while among the 485 children in non-MDT group,121(24.9%)underwent the surgery.The difference was statistically significant(P<0.05).23.2% of children in MDT group were in critical condition during the hospitalization,which was significantly lower than that in non-MDT group(30.5%),and the difference was statistically significant(P<0.05).The unhealed rate of MDT group(2.0%)was also significantly lower than that of non-MDT group(5.6%),the cure rate of MDT group(40.5%)was significantly higher than that of non-MDT group(34.4%),and there was a statistically significant difference(P<0.05).The expense of treatment,medicine and sanitary materials in MDT group were lower than those in non-MDT group,and the differences were statistically significant(P<0.05).The multivariate Logistic regression model analysis of the cure rate of children with brain injury showed that the MDT model could effectively improve the cure rate of children with brain injury(RR = 1.513,95% CI = 1.134-2.020).The results of multiple linear regression model analysis showed that there was no statistical difference in the effect of MDT on the actual hospitalization days of children(P>0.05).Conclusion Using MDT model to diagnose and treat children with brain injury is helpful to improve the cure rate,reduce the risk of children's disease aggravation,and achieve the significant therapeutic effects in children with brain injury.MDT model is worth popularizing and applying in children with brain injury.
8.Postoperative autoantibody levels and clinical significance in non-small cell lung cancer
Zeyi WANG ; Yinyu MU ; Fuyi XIE
China Modern Doctor 2024;62(23):16-20
Objective To investigate the clinical significance of dynamic changes in the expression levels of tumor associated autoantibody(TAAB),including tumor protein 53(p53),protein gene product 9.5(PGP9.5),SRY-box containing gene 2(SOX2),G antigen 7 ATP-dependent RNA helicase(GAGE7),ATP-dependent RNA helicase(GBU4-5),melanoma antigen A1(MAGE A1),cancer/testis antigen(CAGE)in non-small cell lung cancer(NSCLC)patients before and after surgery.Methods Forty-one patients with NSCLC who underwent surgical treatment at the Department of Thoracic Surgery of Li Huili Hospital,Ningbo Medical Center,were selected from January 2019 to August 2022.Enzyme linked immunosorbent assay(ELISA)was used to detect the expression levels of seven TAAB markers in the serum of 41 NSCLC patients undergoing surgical treatment.Results The expression levels of PGP9.5 were found to be significantly higher in the squamous cancer group than in the adenocarcinoma group(P<0.05),while the expression levels of GAGE7 were found to be significantly higher in the adenocarcinoma group than in the squamous cancer group(P<0.05).The expression level of GBU4-5 was found to be significantly higher in the squamous carcinoma group than in the adenocarcinoma group at one week after surgery(Z=-2.095,P=0.036).Within the adenocarcinoma group,the expression levels of GBU4-5(P=0.027)and CAGE(P=0.042)were found to be significantly lower at one week after surgery compared with those before surgery.The expression levels of PGP9.5(P=0.004),GAGE7(P<0.001),and GBU4-5(P=0.014)were found to have decreased significantly at six months postoperatively in comparison to their preoperative levels.In the squamous cancer group,no statistically significant differences were observed in the expression levels at one week after surgery.The expression level of SOX2(P=0.011)exhibited a statistically significant decrease at six months postoperatively in comparison to the preoperative levels.Similarly,the expression level of GBU4-5 was found to be significantly decreased at one week postoperatively in comparison to the preoperative levels(P=0.009).The expression levels of PGP9.5(P=0.003),GAGE7(P=0.001),and GBU4-5(P=0.013)were found to have decreased significantly at six months postoperatively.Conclusion Surgical intervention can reduce the expression level of 7 TAABs in NSCLC patients.Monitoring 7 TAABs before and after surgical treatment is helpful to predict the surgical efficacy and postoperative disease progression of NSCLC patients.
9.Preliminary application of a novel distraction reductor in the surgical treatment of unstable distal radius fractures with metaphyseal volar comminution
Shengli XIA ; Meiqi QIAO ; Ziyuan MA ; Feng GAO ; Bin WANG ; Shaojun WANG ; Zeyi SUN ; Xiaoxiao ZHOU ; Cunguo YI ; Ribao SU ; Xiaoguang JIN ; Xiuhui WANG ; Xuhan CHEN
Chinese Journal of Orthopaedic Trauma 2023;25(8):663-669
Objective:To evaluate a novel distraction reductor in the surgical treatment of unstable distal radius fractures with metaphyseal volar comminution.Methods:From January 2019 to December 2020, 27 patients with unstable distal radius fracture complicated with metaphyseal volar comminution were treated at Department of Orthopaedics, Zhoupu Hospital Affiliated to Shanghai University of Medicine & Health Sciences. They were 6 males and 21 females, with an age of (69.4±9.4) years. All fractures were unilateral and closed, involving the right side in 17 cases and the left side in 10 cases. All patients were treated by internal fixation with an anatomical locking plate through the volar approach and the novel distraction reductor was used to reduce the fracture ends. Regular imaging examinations were performed to evaluate the reduction, maintenance and union of fractures after surgery. One year after operation, the curative efficacy was assessed by evaluation of the range of wrist motion, Disabilities of the Arm, Shoulder and Hand (DASH) score, Gartland-Werley score and Bartra radiology score.Results:All the operations went on successfully with a duration of (92.3±8.9) min. All the incisions healed primarily. The follow-up time was (15.9±2.9) months. The radial height, palmar tilt, ulnar inclination and articular surface step-off immediately after operation [(11.23±1.51) mm, 12.10°±3.44°, 20.54°±3.44°, and (0.95±0.42) mm] were not significantly lost compared with those one year after operation [(11.22±1.55) mm, 12.07°±3.44°, 20.51°±3.33°, and (0.93±0.40) mm] (all P>0.05). One year after operation, the range of wrist motion was good with dorsiflexion of 59.7°±5.5°, palm flexion of 63.0°±9.1°, pronation of 66.5°±5.5°, supination of 61.2°±5.6°, radial deviation of 22.7°±4.8°, and ulnar deviation of 30.3°±6.1°; DASH score was 13.5±5.5; Bartra radiology score was 88.6±6.5, giving an excellent and good rate of 88.9% (24/27);Gartland-Werley score was 2.7±2.1, giving an excellent and good rate of 92.6% (25/27). Follow-ups observed no poor fracture healing, internal fixation failure, tendon or nerve injury or traumatic arthritis. Conclusion:In the surgical treatment of unstable distal radius fractures with metaphyseal volar comminution, the novel distraction reductor can lead to ideal reduction of displaced fractures and effectively correct the shortening caused by volar cortex comminution to achieve satisfactory functional effects in clinic.
10.Detection Results Evaluation of Adipose Mesenchymal Stem Cells Derived Extracellular Vesicles by Nanoparticle Tracking Analysis and Nano-flow Cytometry
ZHANG Zeyi ; WANG Jing ; LI Ping
Chinese Journal of Modern Applied Pharmacy 2023;40(18):2543-2549
OBJECTIVE To explore the detection capability of nanoparticle tracking analysis(NTA) and nano-flow cytometry(nanoFCM) in detecting standard microspheres and adipose mesenchymal stem cells(AdMSCs) derived extracellular vesicles(EVs), provide experimental basis for the identification and quality control of EVs. METHODS Firstly, the detection capabilities of NTA and nanoFCM were verified by detecting standard microspheres. For the AdMSCs-EVs produced via PEG6000-based ultracentrifugation method, the general characterizations were performed with Western blotting and transmission electron microscope. The particle size distributions as well as the post-PKH67 labeling positive EV ratios were analyzed by NTA and nanoFCM respectively. RESULTS NTA and nanoFCM had similar capacity regarding to particle concentration detection, while nanoFCM had higher accuracy and better particle size differentiation ability. The EVs size distribution results showed that, samples analyzed by NTA presented a wider distribution while nanoFCM presented a narrower distribution. For detection of PKH67-positive EVs, the positive ratios detected by NTA were significantly lower than by nanoFCM. CONCLUSION NanoFCM has higher accuracy along with some application limitations. NTA is relatively less capable at particle size distinction, but has a wider particle size detection range. Therefore, for detection of EVs, NTA can satisfy the needs of size distribution characterization, while nanoFCM with high sensitivity for fluorescence detection is more suitable for analyzing the proportion of surface markers in EVs.


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