Objective: To detect the different proteomic characteristics of microvesicles (MVs) and exosomes (EXOs) released from apheresis platelets during storage, and to explore their role in mediating platelet storage damage lesion (PSL). Methods: Apheresis platelets were collected from the retention bag on the third day of storage. MVs and EXOs were isolated using differential centrifugation. Platelet, MVs and EXOs protein samples were extracted respectively, and the differentially expressed proteins were detected by quantitative proteomics technology. Further, the co-incubation model of MVs, EXOs and fresh platelets was adopted to evaluate the effect of extracellular vesicles on PSL. The aggregation response of platelets to collagen agonizers and the changes in ATP release rate were evaluated by optical turbidimetry. Flow cytometry was used to evaluate the changes of platelet early activation indicators (P-selectin and PAC-1) and mitochondrial membrane potentia. Western blot was used to detect the changes in the expression of key proteins for platelet activation and apoptosis (P-selectin, Integrin β3 and Bcl-xl). Results: Proteomic analysis revealed a significantly separation in protein expression profiles of platelet, MVs and EXOs samples within the latent variable space. Energy metabolization-related proteins such as mitochondrial respiratory chain complex and oxidative phosphorylation were enriched specifically, in MVs while EXOs were enriched with inflammation-related proteins. Co-incubation experiments confirmed that extracellular vesicles could significantly induce platelet responses to agonists (the maximum aggregation rate in the MVs group increased by 187.36%, P<0.001; 71.26%, in the EXOs group P=0.002). The maximum ATP release rate of platelets also increased (275.44% in the MVs group, P<0.001; 70.18% in the EXOs group, P=0.015). The expression of P-selectin increased (119.33% in the MVs group, P<0.001; 25.61% in the EXOs group, P=0.013), as detected by flow cytometry. The binding rate of PAC-1 increased (132.18% in MVs group, P<0.001; 21.41% in EXOs group, P=0.043), and the mitochondrial membrane potential decreased (20.49% in MVs group, P<0.001; 9.73% in EXOs group, P=0.044). In the MVs group, platelet P-selectin and Integrin β3 expression were significantly increased (100.83% and 395.64%, P<0.001), while Bcl-xl expression was lower than that in the control group (83.94%, P<0.001). Compared with the control group, P-selectin and Integrin β3 expression were also increased (27.89% and 181.91%, P=0.007和P=0.002), while Bcl-xl was decreased in the EXOs group (36.52%, P<0.001). Conclusion: MVs and EXOs derived from stored platelets show different proteomic characteristics. Compared with EXOs, MVs exhibits a stronger effect in inducing mitochondrial dysfunction. Mvs also promots PSL responses including platelet activation and apoptosis.

Objective Aiming at the problems of many items and long time to fill in the Constitution in Chinese Medicine Questionnaire(CCMQ)when evaluating individual constitution,the research uses artificial intelligence technology to select attributes,and to help construct a short version of the CCMQ.Methods Analyzing the constitution data provided by the Physical Examination Department of Jiangsu Province Hospital of Traditional Chinese Medicine,there are specific target variables as the classification of constitution types.Feature selection of genetic algorithm,cross-validation and KNN classification algorithm are used as filters to select problems,and the effect is evaluated by problem subset size,KNN classification accuracy and filling time.Results The method selected a short version of the CCMQ with 31 problems,and the average classification accuracy in the model was 86.16%,and the time was improved by 47.7%.Conclusion The algorithm can effectively find a better problem subset,achieve dimensionality reduction and have certain accuracy,thus helping to simplify the CCMQ.

Animals ; Bacterial Infections/metabolism* ; Glucose/metabolism* ; Homeostasis ; Insulin Secretion ; Mice ; Mice, Knockout ; Sirtuins/metabolism*

AIM: To study the effects of hypertonic NaCl-NaAc on microcirculation in hemorrhage-shocked rats.METHODS: SD rats were randomized into three groups of 7 5% NaCl(hypertonic saline, HS),5% NaCl-3 5% NaAc(hypertonic sodium acetate, HSA) and 0 9% NaCl(normal saline, NS) 4 mL/kg HS,HSA or NS was given intravenously following hemorrhagic shock The microcirculation of spinotrapezius muscle was observed RESULTS: HS increased mean aortic pressure more significant than HSA Variables including arteriolar and venular diameter,velocity and volumetric flow rate and open capillaries were increased and erythrocyte aggregation was decreased in 5 min after resuscitation with both HS and HSA solutions 5 min later,variables were deteriorated in HS group After local treatment, arteriolar and venular diameters were dilated significantly in HSA group CONCLUSION:HSA had superior effects to HS in improving microcirculation of hemorrhage-shocked rats