1.Effect of Lycium barbarum polysaccharide on expression of vascular endothelial growth factor and hypoxia inducible factor-1α in ultraviolet B-radiated HaCaT cells
Hongchang ZHANG ; Lichun ZUO ; Haicai SUN ; Guangfu LYU ; Na LI ; Zhe LIN
Chinese Journal of Dermatology 2017;50(9):667-670
Objective To evaluate effects of Lycium barbarum polysaccharide (LBP) on expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1α (HIF-1α) in ultraviolet B (UVB)-radiated HaCaT cells.Methods Conventionally cultured HaCaT cells were divided into control group and LBP groups,which were firstly treated with DMEM,12.5,25.0,50.0 and 100 μg/ml LBP solution respectively for 4 hours,and then were irradiated by UVB at different intensity of 0,20,40,60 mJ/cm2 separately.After 24-hour continuing culture,CCK-8 assay was performed to determine the cell survival rate,and an enzymatic-biochemical method to estimate the activity of superoxide dismutase (SOD).RT-PCR and Western blot analysis were conducted to measure the mRNA and protein expression of HIF-1α and VEGF respectively.Results Compared with the control group at the same UVB radiation dose,the 12.5-,25.0-and 100.0-μ,g/ml LBP groups showed different extents of increase in survival rates of UVB-radiated cells (P < 0.05),and the 50.0-μg/ml LBP group showed the highest cell survival rate (P < 0.01).Among all the LBP groups,SOD activity was highest in the 50.0-μg/ml LBP group (P < 0.01).Along with the increase of UVB radiation dose,the mRNA and protein expression of HIF-1α and VEGF all gradually increased.Compared with the control group,the 50.0-μg/ml LBP group could effectively reduce the mRNA and protein expression of HIF-1α and VEGF in HaCaT cells (all P < 0.05).Conclusion LBP may play a role in protecting cells from UVB radiation-mediated damage,likely by influencing the mRNA and protein expression of HIF-1α and VEGF in HaCaT cells.
2.Genetic analysis of a weak D type61 sample from a blood donor, Jiangyin
Fang WANG ; Mengyao BIAN ; Qiurong YU ; Minglei WU ; Haiping ZHAO ; Ling SUN ; Buqiang WANG ; Hongjun GAO ; Haicai SHI ; Yi WU ; Ming GAO ; Yuping CHEN
Chinese Journal of Blood Transfusion 2022;35(7):701-704
【Objective】 To genetically analyze the Del sample from a blood donor in Jiangyin and make clear the molecular basis of the serological phenotype. 【Methods】 The EDTA anticoagulant blood were collected: buffy coat were used for nucleic acid extract and cDNA analysis; red blood cells for serological test. Tube method and microcolumn gel were used for serological test. Genotyping kit were used for exon analysis. Gene mutation was analyzed using the sequence analyzer. 【Results】 Serological analysis demonstrated the sample′s RhD phenotype was Del. The phenotype of RhCE was CCEe. Real-time fluorescence quota PCR result demonstrated the existence of all exones. Weak D15 and RHD* DEL1 [RHD(1227G>A)], which had a high frequency of occurrence in China, were excluded according to real-time fluorescence quota PCR result. Sequence analyzing result verified RHD(28C>T) SNP mutation in cDNA. The genotype of this sample was RHD*01 W. 61[RHD(28C>T)]. 【Conclusion】 A weak D61 was found among blood donors in our city, Jiangyin.