1.Relationship between polymorphism site rs5744168 of TLR5 and AAV
Linlin HE ; Chao XUE ; Yunhua LIAO ; Suzhen WEI ; Aimei GONG ; Yangfei LAI
The Journal of Practical Medicine 2016;32(17):2821-2825
Objective To investigate the relationship between putative rs5744168 of Toll-like receptors 5 (TLR5)and ANCA associated small vasculitis (AAV) in Guangxi Han nationality. Methods Polymorphism was analyzed by polymerase chain restricted fragments length polymorphism in 120 cases with AAV and 212 controls. Results (1)There were two genotypes of CC and CT in AAV group and control group. The frequencies distribution of CC and CT in 120 AAV patients were 82.50% and 17.50% respectively and the frequencies of allele C and T 91.25% and 8.75%,respectively. In controls,the genotypefrequencies of CC and CT were 88.68% and 11.3%, and frequencies of allele C and T 94.34% and 5.66%, respectively. No significant difference was found in either genotype distribution or allele frequencies between the patients and the controls ( P > 0 . 05 ) . ( 2 ) Significant reductions in the incidence of BUN, uric acid, quantitative test of 24 h urinary protein and erythrocyte sedimentation rate(ESR) were found in CC genotype (P < 0.05). (3) Binary regression model with a logit link function found total cholesterol was related with AAV. Conclusion The susceptibility of AAV in Guangxi Han population has nothing to do with the polymorphism of rs5744168.In AAV patients, polymorphism of rs5744168 may be associated with ESR, BUN, uric acid and quantitative test of 24 h urinary protein levels.
2.Cloning, expression and characterization of gene encoding human stem cell growth factor-alpha and its synergetic effect with rhGM-CSF on proliferation of human umbilical cord mesenchymal stem cells.
Xinlei PENG ; Yanyan MA ; Jing RONG ; Zhenling ZHAO ; Bo HAN ; Wei CHEN ; Yangfei XIANG ; Qiuying LIU ; Yifei WANG ; Zhe REN ; Xiangrong ZHOU ; Haijia CHEN
Chinese Journal of Biotechnology 2011;27(11):1667-1676
To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation
;
drug effects
;
Cells, Cultured
;
Cloning, Molecular
;
Drug Synergism
;
Escherichia coli
;
genetics
;
metabolism
;
Granulocyte-Macrophage Colony-Stimulating Factor
;
pharmacology
;
Humans
;
Mesenchymal Stromal Cells
;
cytology
;
metabolism
;
Recombinant Fusion Proteins
;
biosynthesis
;
genetics
;
pharmacology
;
Stem Cell Factor
;
biosynthesis
;
genetics
;
Umbilical Cord
;
cytology
3.Inhibitory Effect of Hederasaponin B on Gastric Cancer HGC-27 Cells and Its Mechanism Based on Bioinformatics
Yangfei WEI ; Shuo WANG ; Tao CHEN ; Shuping YAN ; Peipei LI ; Hongmei LI ; Cheng SHEN ; Yumei MA ; Zhibo SONG ; Aijing LI ; Yulin LI
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(16):50-58
ObjectiveTo investigate the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell and the mechanism. MethodMethyl thiazolyl tetrazolium (MTT) assay, hematoxylin-eosin (HE) staining, 4',6-diamidino-2-phenylindote (DAPI) staining, colony formation assay, scratch assay, and flow cytometry were employed for the analysis of apoptosis and cell cycle. Thereby, the inhibitory effect of hederasaponin B on gastric cancer HGC-27 cell was investigated. Then the Pharm Mapper, UniProt, Swissdock, STRING, and Metascape were used for target screening, gene annotation, molecular docking, protein-protein interaction (PPI) network construction, Gene Ontology (GO) term and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis to explore the mechanism. ResultHederasaponin B (15, 30, 60, 120 μmol·L-1) can significantly reduce the survival rate of HGC-27 cell (P<0.01) in a time-dependent and dose-dependent manner compared with the blank group. It had no significant toxicity to normal GES-1 cell at concentration below 120 μmol·L-1. Compared with the blank group, hederasaponin B (30, 60, 120 μmol·L-1) induced cytoplasmic vacuolization, and nuclear deformation and karyopyknosis, inhibited the migration of HGC-27 cell (P<0.01), and brought about the apoptosis (P<0.05, P<0.01) and cell cycle arrest of HGC-27 cell (P<0.05, P<0.01). Hederasaponin B (10, 20, 30 μmol·L-1) also suppressed the independent survival ability and proliferation ability of HGC-27 cell (P<0.01). The possible action targets were kinesin-like protein KIF11, cGMP-specific 3,5 cyclic phosphodiesterase, caspase-3, serine/threonine protein kinase Chk1, proto-oncogene tyrosine protein kinase, epidermal growth factor receptor, and mitogen-activated protein kinase (MAPK) 8. The mechanism may be related to MAPK signaling pathway (pathways in cancer), adhesion connection, focal adhesion and proteoglycans in cancer (epithelial cell signaling pathways in Helicobacter pylori infection). ConclusionHederasaponin B exerts significant inhibitory effect on gastric cancer HGC-27 cell through multiple targets and multiple pathways.