1.Isolation and characterization of normal mandibular periosteal stem cells from human and macaca mulatta and cross-species single-cell analysis
Zishuo WANG ; Yangyang LI ; Haitao WANG ; Duohong ZOU ; Zhiyuan ZHANG
Chinese Journal of Stomatology 2024;59(7):696-705
		                        		
		                        			
		                        			Objective:To investigate the presence of a distinct stem cell populations different from mesenchymal stem cells in the mandibular periosteum of both human and non-human primates (macaca mulatta), to explore its properties during intramembranous osteogenesis and to establish standard protocols for the isolation, culturing and expanding of mandibular periosteal stem cells (PSC) distinguished from other PSCs in other anatomical regions.Methods:Periosteum was harvested from the bone surface during flap bone removal in patients aged 18-24 years undergoing third molar extraction and from the buccal side of the mandibular premolar region of 6-year-old macaca mulatta respectively, and then subjected to single-cell sequencing using the Illumina platform Novaseq 6000 sequencer. Cross-species single-cell transcriptome sequencing results were compared using homologous gene matching. PSC were isolated from primary tissues using two digestion methods with body temperature and low temperature, and their surface markers (CD200, CD31, CD45 and CD90) were identified by cell flow cytometry. The ability of cell proliferation and three-lineage differentiation of PSC expanded to the third generation in vitro in different species were evaluated. Finally, the similarities and differences in osteogenic properties of PSC and bone marrow mesenchymal stem cells (BMSC) were compared. Results:The single-cell sequencing results indicated that 18 clusters of cell populations were identified after homologous gene matching for dimensionality reduction, and manual cellular annotation was conducted for each cluster based on cell marker databases. The comparison of different digestion protocols proved that the low-temperature overnight digestion protocol can stably isolate PSC from the human and m. mulatta mandibular periosteum and the cells exhibited a fibroblast-like morphology. This research confirmed that PSC of human and m. mulatta had similar proliferation capabilities through the cell counting kit-8 assay. Flow cytometry analysis was then used to identify the cells isolated from the periosteum expressed CD200(+), CD31(-), CD45(-), CD90(-). Then, human and m. mulatta PSC were induced into osteogenesis, adipogenesis, and chondrogenesis to demonstrate their corresponding multi-lineage differentiation capabilities. Finally, comparison with BMSC further clarified the oesteogenesis characteristics of PSC. The above experiments proved that the cells isolated from the periosteum were peiosteal cells with characteristics of stem cells evidenced by their cell morphology, proliferation ability, surface markers, and differentiation ability, and that this group of PSC possessed characteristics different from traditional mesenchymal stem cells.Conclusions:In this study, normal mandibular PSC from humans and m. mulatta were stably isolated and identified for the first time, providing a cellular foundation for investigating the mechanism of mandibular intramembranous osteogenesis, exploring ideal non-human primate models and establishing innovative strategies for clinically mandibular injury repair.
		                        		
		                        		
		                        		
		                        	
2.Characterization of paclitaxel-PLGA nanoparticles and their antitumor effects in vitro
Xiaojing WANG ; Zishuo GUO ; Haitong ZHANG ; Wanling CHEN ; Jialing LI ; Shouying DU ; Pengyue LI
China Pharmacy 2024;35(22):2721-2725
		                        		
		                        			
		                        			OBJECTIVE To characterize paclitaxel nanoparticles (PTX-PLGA-NPs) and evaluate their in vitro inhibitory effect on Lewis lung cancer cells. METHODS The PTX-PLGA-NPs prepared by the emulsion-solvent evaporation method were characterized in terms of particle size, polydispersity index (PDI), Zeta potential, microscopic morphology, encapsulation efficiency, drug loading, ultraviolet-visible absorption characteristics and stability. Using mouse Lewis lung cancer cells as the subjects and paclitaxel reference substance as the control, the cytotoxicity and in vitro killing activity of PTX-PLGA-NPs were detected using CCK-8 method and Calcein-AM/PI double staining method, respectively. The effects of PTX-PLGA-NPs on cell apoptosis and cell cycle were assessed by Annexin Ⅴ-FITC/PI staining method and PI staining method, respectively. RESULTS PTX-PLGA-NPs were spherical with an average particle size of (172.03±0.95) nm, PDI of 0.098±0.012, and Zeta potential of (-1.76±0.02) mV. The encapsulation efficiency and drug loading were (52.32±0.66)% and (7.07±0.18)%, respectively, and the ultraviolet-visible absorption characteristics were not affected by the carrier polylactic-co-glycolic acid. When stored in the dark at 4 °C for 7 days, no significant change was noted in particle size, and the average PDI (after 1, 2, 4 and 7 days of storage) was under 0.3. Compared with the paclitaxel reference substance group, the PTX-PLGA-NPs group had more cells in a state of death, the survival rate (at the PTX concentration of 11.2 μg/mL) was significantly decreased, and both the apoptosis rate and the proportion of G2 phase cells were significantly increased (P<0.05). CONCLUSIONS The prepared PTX-PLGA-NPs indicate homogeneity in particle size, uniform dispersion, stable properties, and stronger in vitro killing effect on lung cancer cells than PTX.
		                        		
		                        		
		                        		
		                        	
3. Memory and emotional disorders after thalamic stroke
Jiajia YANG ; Zishuo LIANG ; Qian WANG ; Dong MING
International Journal of Cerebrovascular Diseases 2019;27(10):760-764
		                        		
		                        			
		                        			 The thalamus plays an important role in the process of production of memory and emotion. Patients with thalamic stroke have memory and emotional disorders. Memory disorders are mainly manifested in the generation of hallucinatory memory and amnesia, while emotional disorders are mainly manifested in the effect of negative emotions. 
		                        		
		                        		
		                        		
		                        	
4.Clinical Features With In-hospital and Long-term Prognosis of Acute Myocardial Infarction in Patients Younger Than 40 Years by Different Genders
Wen HAO ; Huangtai MIAO ; Xiao WANG ; Jie CHANG ; Zishuo CHEN ; Shaoping NIE
Chinese Circulation Journal 2016;31(10):976-980
		                        		
		                        			
		                        			Objective: To evaluate clinical features with in-hospital and long-term prognosis of acute myocardial infarction (AMI) in patients ≤40 years of age by different genders and to analyze the predictors for major adverse cardiovascular event (MACE) occurrence. Methods: A total of 685 AMI patients ≤40 years treated in our hospital from 2012-01-01 to 2015-08-31 were consecutively enrolled. The patients were divided into 2 groups by gender: Male group,n=650 and Female group,n=35. The baseline data, clinical features, in-hospital MACE incidence were collected by telephone communication and compared between 2 groups; the long-term risk factors for MACE occurrence were analyzed. Results: The AMI ratio in male patients was 94.89%, in female was 5.11% and the onset age in Male group was higher than Female group (35.53±4.21) years vs (34.05±4.98) years,P=0.046. Compared with Female group, Male group showed the lower rates of coronary left main diseases (3.2% vs 11.4%,P=0.012) and in-hospital heart failure (8.3% vs 25.7%,P=0.001). The median follow-up time was of 727.0 (411.5, 1102.0) days and during that period, MACE occurrence rates in Male group was 46 (7.1%) cases and in Female group was 2 (5.7%) cases,P=0.758. Increased level of hs-TnI, (OR=1.003, 95% CI 1.001-1.006,P=0.020) and multi coronary artery disease (OR=1.964, 95% CI 1.018-3.790,P=0.044) were the independent predictors for long- term adverse event occurrence; while PCI (OR=0.475, 95% CI 0.241-0.936,P=0.031) was the protector for long-term prognosis in young male AMI patients. Conclusion: AMI patients≤40 years were mainly in male gender, the mean onset age in male was elder than female. Increased hs-TnI level and multi coronary artery disease were the predictors for MACE occurrence, while PCI was the protective factor for long-term prognosis in young male AMI patients.
		                        		
		                        		
		                        		
		                        	
            
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