1.Visualized and rapid detection of extracellular vesicles of glial cells in a simulated space environment
Tianyi ER ; Yu LAN ; Beiqin LIU ; Shuyue WANG ; Yali ZHAO ; Chengjia YANG ; Hong MA
Space Medicine & Medical Engineering 2024;35(1):26-30
Objective To rapidly visualize and detect the extracellular vesicles of glial cells in a simulated space environment.Methods By using 2,5 Gy irradiation and 12,24 h microgravity treatment,a damage model of glial cells was established in a simulated space environment.Exosomes extracted from conditioned media with reagent kits were transferred to neurons to elucidate the impact of glial cells on neurons.By performing live-cell fluorescence labeling of exosomes,a visualization monitoring scheme based on fluorescence intensity analysis was developed to characterize the release patterns of extracellular vesicles.The release patterns of exosomes were represented by the fluorescence intensity of the conditioned media.The effects of different storage conditions and duration on the quantity and size of exosomes were investigated.Results The exosomes released from damaged glial cells in the simulated space environment could to some extent protect neurons,with exosomes playing a decisive role in this process,and the neurosystem exosome visualization detection scheme was consistent with the traditional exosome validation scheme.An empirical curve for exosome quantity and size was established for semi-quantitative analysis,providing a new approach for rapid detection of exosomes in cell culture media.Furthermore,the optimal storage conditions for culture medium samples were clarified,laying the foundation for ground/space-based online analysis of culture medium samples after spaceflight.Conclusion Exosome rapid detection and analysis can be achieved through fluorescence labeling and utilized to investigate glial cell injury in a simulated space environment.
2.Clinical Characteristics and Influencing Factors of Rheumatoid Arthritis in Patients with Cold Dampness Obstruction Syndrome
Yanyu CHEN ; Yanqi LI ; Longxiao LIU ; Liubo ZHANG ; Tianyi LAN ; Nan ZHANG ; Cheng XIAO ; Yuan XU ; Qingwen TAO
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):140-146
ObjectiveTo study the clinical characteristics and influencing factors of rheumatoid arthritis (RA) in the patients with cold dampness obstruction syndrome. MethodsThe RA patients treated in the Department of Traditional Chinese Medicine and Rheumatology of the China-Japan Friendship Hospital from August 2022 to June 2024 were selected. The demographic information, clinical data, laboratory test results, and traditional Chinese medicine (TCM) symptom information were collected for syndrome differentiation, on the basis of which the characteristics and influencing factors of cold dampness obstruction syndrome were analyzed. ResultsA total of 258 RA patients were selected in this study, including 88 (34.1%) patients with cold dampness obstruction syndrome, 53 (20.5%) patients with dampness and heat obstruction syndrome, 31 (12.0%) patients with wind dampness obstruction syndrome, 29 (11.2%) patients with liver-kidney deficiency syndrome, 19 (7.4%) patients with Qi-blood deficiency syndrome, 14 (5.4%) patients with phlegm-stasis obstruction syndrome, 15 (5.8%) patients with stasis obstructing collateral syndrome and 9 (3.5%) patients with Qi-Yin deficiency syndrome. The patients were assigned into two groups of cold dampness obstruction syndrome and other syndromes. The group of cold dampness obstruction syndrome had lower joint fever, 28-tender joint count (TJC28), and 28-joint disease activity score (DAS28)-C-reactive protein (CRP) and higher central sensitization, cold feeling of joints, fear of wind and cold, cold limbs, and abdominal distention than the group of other syndromes (P<0.05). The binary logistic regression analysis showed that central sensitization (OR 5.749, 95%CI 2.116-15.616, P<0.001) and DAS28-CRP (OR 0.600, 95% CI 0.418-0.862, P=0.006) were the independent factors influencing cold dampness obstruction syndrome in RA. ConclusionCold dampness obstruction syndrome is a common syndrome in RA patients. It is associated with central sensitization, cold feeling of joints, abdominal distension and may be a clinical syndrome associated with central sensitization.
3.Astragali Radix Polysaccharide Inhibits Proliferation and Migration of Gastric Cancer Cells by Targeting ID1 and Akt
Peizheng SHI ; Shanshan XIAO ; Xinjiang ZHANG ; Yixiang NIE ; Xianchao WANG ; Jing HUANG ; Jie MEI ; Huaquan LAN ; Tuanyun JI ; Tianyi ZHANG ; Xiaoyong WEI ; Qiaohong YANG
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(15):96-105
ObjectiveTo explore the regulatory effects and mechanisms of Astragali Radix polysaccharide (APS) on inhibitor of differentiation1 (ID1) and protein kinase B (Akt) in gastric cancer. MethodsImmunohistochemical staining was used to detect the expression of ID1 and Akt in 61 gastric cancer tissue samples and 20 adjacent normal gastric tissue samples. Immunofluorescence was used to detect the localization of ID1 and Akt. The effects of APS at the concentrations of 0.625, 1.25, 2.5, 5, 10, 20 mg·L-1 on the proliferation of gastric cancer MGC-803 cells were examined by the cell counting kit-8(CCK-8) method and the colony formation assay. The target information of APS was retrieved from the Traditional Chinese Medicine Systems Pharmacology and Analysis Platform and Swiss Target Prediction. Keywords such as gastric cancer, gastric tumor, and stomach cancer were searched against GeneCards, UniProt, DisGeNET, and Online Mendelian Inheritance in Man (OMIM) for the screening of gastric cancer-related targets. The online tool jvenn was used to create the Venn diagram to identify the common targets, and STRING and Cytoscape were used to construct the protein-protein interaction network. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted via R 4.2.2 to predict the potential roles of APS in the development of gastric cancer. The cell scratch assay was employed to assess the effect of APS on the migration of MGC-803 cells. The protein and mRNA levels of ID1 and Akt in the cells treated with APS were determined by Western blot and Real-time PCR, respectively. ResultsCompared with the adjacent normal gastric tissue, the gastric adenocarcinoma tissue showed increased positive expression of ID1 (χ2 =81.00, P<0.01). Immunofluorescence detection showed that ID1 and Akt were mainly located in the cytoplasm of gastric adenocarcinoma cells. Bioinformatics analysis identified 14 common genes shared between APS and gastric cancer. The average degree of protein-protein interaction network nodes was 14.29. GO and KEGG pathway enrichment results showed that ID1 and Akt were significantly enriched in the Rap1 and phosphatidylinositol-3-kinase (PI3K) /Akt signaling pathways. Cell experiments demonstrated that 5-fluorouracil (0.1 mg·L-1) and APS (10, 20 mg·L-1) groups showed decreased cell proliferation, migration, and colony formation. Compared with the control group, 10, 20 mg·L-1 APS inhibited the proliferation of MGC-803 cells (P<0.01), with 10 mg·L-1 APS demonstrating stronger inhibitory effect. In addition, APS at 10, 20 mg·L-1 inhibited the migration (P<0.01) and colony formation (P<0.05, P<0.01) of MGC-803 cells. Compared with the control group, APS at 10, 20 mg·L-1 down-regulated the protein levels of ID1 (P<0.01) and Akt (P<0.05) and the mRNA levels of ID1 (P<0.05, P<0.01) and Akt (P<0.05, P<0.01) in MGC-803 cells. ConclusionID1 and Akt are highly expressed in the gastric adenocarcinoma tissue, which may be related to the development of gastric cancer. APS can down-regulate the protein and mRNA levels of ID1 and Akt to exert anti-tumor effects, which is expected to provide new therapeutic targets for gastric cancer treatment.