1.Study on determination conditions for lymphocytic proliferation by CCK-8 method in mice
Yuanyuan WANG ; Xiaoyuan TAN ; Minghua HU ; Fangli MA ; Zhiying HUANG ; Jinqiang LIANG
Drug Evaluation Research 2017;40(2):206-209
Objective To study the optimum determination conditions for lymphocytic proliferation by CCK-8 method in mice.Methods To study the different influence factors of spleen cell proliferation experiment stimulated by mitogen concanavalin A (ConA) or lipopolysaccharide (LPS),including cell preparation method,lymphocytic density,FBS and stimulating agent concentration in culture medium,and stimulating immediately or 24 h after preparing cell,with cross design or two factor completely randomized design.Results Spleen lymphocytic proliferation rate of preparation method by light suppression was higher than that of the light grind.The appropriate concentration of spleen cells was 5 × 106/mL.The proliferation rate has no significant difference after being stimulated for 48 or 72 h by ConA (2,5,or 1 0 μg/mL) or LPS (10,20,or 50 μg/mL) under 10%,15%,or 20% FBS concentration in culture medium.The proliferation rate of stimulating immediately after preparing cell was higher than that of 24 h after preparing cell.Conclusion The optimum conditions of Balb/C mouse spleen cell proliferation assay stimulated by ConA and LPS are as follows:preparation of spleen cells with light pressure,spleen cell concentration of 5 × 106/mL,direct stimulation with 2-10 μg/mL ConA or 10-50 μg/mL LPS in the day of preparation.
2.Low-temperature condensation deposition method for 3D printing of bone tissue engineering poly-L-lactic acid/pearl powder composite scaffold
Fangli GANG ; Rui SHI ; Chunyang MA ; Yi XIAO
Chinese Journal of Tissue Engineering Research 2024;28(17):2702-2707
BACKGROUND:The repair of large-scale bone defects is still facing serious challenges.It is of great significance to develop personalized,low-cost,and osteogenic-inducing tissue engineering scaffolds for bone repair. OBJECTIVE:To explore the process of 3D printing bone tissue engineering scaffold containing pearl composite material by low-temperature condensation deposition method,and further test the physicochemical properties and in vitro biological functions of the composite scaffold. METHODS:Pearl powder was prepared by grinding and sieving.The pearl powder of different qualities was added into the poly-L-lactic acid ink,so that the mass ratio of pearl powder to poly-L-lactic acid was 0,0.1,0.2,0.3,and 0.5,respectively.The 3D-printed poly-L-lactic acid/pearl powder scaffolds were prepared using the low-temperature condensation deposition method.The microstructure,compressive properties,water contact angle,cytocompatibility,and in vitro bone differentiation ability of the printed poly-L-lactic acid/pearl powder composite scaffolds were detected. RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the five groups of scaffolds all had micropores with a diameter of 2 μm or even smaller,irregular shapes and interconnectivity.(2)All the five groups had good compressive properties.The compressive strength of the pearl powder 0.5 group was higher than that of the other four groups(P<0.05).The water contact angle of the pearl powder 0.2 group and the pearl powder 0.5 group was smaller than that of the pearl powder 0 group(P<0.01,P<0.001).(3)Bone marrow mesenchymal stem cells were co-cultured with five groups of scaffolds for 1,3,and 5 days,respectively.The cell proliferation in pearl powder 0.1,0.2,0.3,and 0.5 groups cultured for 3 and 5 days was faster than that in pearl powder 0 group(P<0.05).After 1 day of culture,live-dead staining exhibited that the number of cells on the scaffold was small,but all of them were living cells.(4)Bone marrow mesenchymal stem cells were inoculated on the scaffold surface of the pearl powder 0 group and pearl powder 0.1 group respectively for osteogenic differentiation.The alkaline phosphatase activity induced for 4 and 6 days in the pearl powder 0.1 group was higher than that in the pearl powder 0 group(P<0.05).(5)The results showed that the poly-L-lactic acid/pearl powder composite scaffold had good compressive strength,hydrophilicity,cytocompatibility,and osteogenic properties.
3.Comparison of the application value of magnetic-controlled capsule endoscopy and traditional capsule endoscopy in the diagnosis of intestinal diseases
Fangli WU ; Jing LI ; Guifang LU ; Jiahui YANG ; Wenhui MA ; Shuixiang HE ; Mudan REN
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(2):247-251
【Objective】 To compare the clinical value of magnetic-controlled capsule endoscopy (MCE) and traditional capsule endoscopy (CE) in the diagnosis of intestinal diseases in hospitalized patients. 【Methods】 A single-center retrospective study was conducted in 263 inpatients who underwent MCE and CE in The First Affiliated Hospital of Xi’an Jiaotong University from March 2016 to March 2020. The information included the patients’ general data, chief complaints, and results of capsule endoscopic examination. 【Results】 ① The overall detection rate in small intestinal diseases was 74.45% in MCE group and 73.81% in CE group, respectively (P=0.905). The three most common diseases in the two groups were erosive/ulcerative lesions, vascular lesions, and lymphangiectasia. ② The endoscopic auxiliary rate was significantly lower in MCE group than in CE group (0% vs. 9.49%, P<0.001). ③ There was no significant difference in the rate of intestinal incompletion between the two groups (7.94% vs. 13.87%, P=0.185). 【Conclusion】 MCE is similar to CE in the diagnostic value for intestinal diseases. Currently, it can be used as one of the methods of small intestinal examination, but this needs to be supported by more multicenter and sizable simple studies.
4.Genetic characteristics of influenza A H3N2 virus influenza season in Xiangyang City in 2022-2023
Jing SHI ; Fangli TONG ; Shengyang ZHU ; Yunxia GAN ; Lu MA ; Narenqimuge TONG ; Bin FANG ; Peng CHEN ; Gang YANG
Journal of Public Health and Preventive Medicine 2024;35(3):32-36
Objective To analyze the prevalence and genetic characteristics of influenza A(H3N2) viruses in the city of Xiangyang in 2022-2023, and to provide a scientific basis for predicting the epidemic and mutation of influenza virus. Methods Throat swab specimens of the influenza like cases were collected from national influenza monitoring sentinel hospitals in Xiangyang every week. RNA was extracted from the specimens for influenza diagnosing using real-time RT-PCR.Viruses were isolated from H3N2 positive specimens, and HA and NA genes were amplified and sequenced.3D modeling analyses were conducted. Results The gene phylogenetic tree showed that the H3N2 isolates in 2022-2023 belonged to 3C.2a1b.2a1 and 3C.2a1b.2a2 branches, respectively. The A(H3N2) influenza virus strains all had amino acid point mutation sites on important antigenic determinants of HA protein. The epitope mutations of the 2022 A(H3N2) strain mainly occurred in regions B, C, and D. The epitope mutations of the A(H3N2) strain in 2023 mainly occurred in regions C and D. Different glycosylation sites of HA gene were found in 2022-2023 strains. No variation was found in key amino acid sites associated with neuraminidase inhibitor resistance. The difference of overall structure was not obvious in the three-dimensional simulation structure diagram. Conclusion The A(H3N2) influenza strains isolated in this study have shown antigenic drift, especially the mutation of HA, which may affect the protective effect of the vaccine on the local population and lead to influenza epidemic. The variations of HA and NA suggest that close attention should be paid to the epidemic and genetic variation of H3N2 subtype influenza virus, to provide a scientific basis for the selection of influenza virus vaccine strains and the prevention and control of influenza.