1.Study on cell DNA damage induced by tetramine using comet assay
Yan LIU ; Chuanhong ZHU ; Libin DENG
Chinese Journal of Forensic Medicine 2002;0(05):-
Objective To detect DNA damage in rat lymphocytes and brain cells induced by tetramine and study the toxicological mechanism of tetramine.Methods Lymphocytes and brain cells were separated and collected from healthy rats.DNA damages of cells which were exposed to various doses of tetramine for 60 min was detected using the single cell gel electrophoresis (SCGE) or comet assay.Results These are different degree DNA damages of lymphocytes and brain cells exposed from doses 1/20 LD 50 doses of tetramine to 1/2 LD 50 doses of tetramine.The test groups are very significantly statistical different to the control group(P
2.Change of reference gene glyceraldehyde-3-phosphate dehydrogenase expression during 3T3-L1 adipocyte differentiation
Juan ZHANG ; Hongju TANG ; Xiao WANG ; Ning WANG ; Ruyuan DENG ; Fangfang JIAN ; Yun LIU ; Fengying LI ; Libin ZHOU
Chinese Journal of Endocrinology and Metabolism 2012;28(10):849-854
Objective To observe the change of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression during 3T3-L1 adipocyte differentiation as well as other reference gene expressions.Methods The mRNA expressions of several common reference genes were detected by real time-PCR on day 0,1,3,5,and 7 of 3T3-L1 adipocyte differentiation.Western blot was used to confirm the protein expressions of three common reference genes.Results (1) GAPDH and transferrin receptor(TFRC) mRNA expressions were significantly increased during adipocyte differentiation.GAPDH mRNA level was increased by 5.7,7.6,22.0,and 24.5 folds on day 1,3,5,and 7 after induction of adipocyte differentiation,but no apparent changes of β-actin,α-tubulin,peptidylprolyl isomerase A (PIPA),and 18S mRNA expressions were detected.The expression changes of key transcript factors for adipocyte differentiation such as PPARγ2,C/EBPα,and C/EBPβ were under-estimated by real time-PCR if GAPDH was chosen as the reference gene.Western blotting results showed that the GAPDH protein level increased gradually during adipocyte differentiation,especially on day 5 and 7 after adipocyte differentiation.There were no obvious changes of β-actin and α-tubulin protein expressions.(2) Berberine significantly inhibited mRNA and protein expressions of GAPDH in the process of adipocyte differentiation.GAPDH mRNA levels were reduced by 68.1% and 66.3% on day 5 and 7 after induction of adipocyte differentiation,but with no significant change in other reference genes.Conclusion It is not suitable for GAPDH to be used as an endogenous reference gene during 3T3-L1 adipocyte differentiation.
3. Etiological characteristics of an imported Chikungunya fever epidemic in Fujian province in 2018
Jinzhang WANG ; Naipeng KAN ; Yongjun ZHANG ; Libin YOU ; Yuping WANG ; Yanqin DENG ; Kuicheng ZHENG ; Yuwei WENG
Chinese Journal of Experimental and Clinical Virology 2019;33(3):253-256
Objective:
To study the etiological characteristics of an imported Chikungunya fever (CHIK) epidemic in Fujian province in 2018.
Methods:
Serum samples collected at different days after the onset of the two CHIK cases were detected by real-time RT-PCR and ELISA. Structural protein E1 gene was amplified by RT-PCR and sequenced for nucleotide characteristics analysis and phylogenetic tree analysis.
Results:
RNA of Chikungunya virus (CHIKV) was detected in the 4 serum samples collected on the first 5 days of the disease, and the earliest IgM antibodies were detected in specimens on the 5th day of the disease, however, IgG antibodies were only detected in specimen on 10th day. Compared with the S27-African prototype strain, 12 mutant points were found in the amino acids of E1 genes in this study. The E1 genes of the two CHIK cases were exactly the same, and they were closest to the evolutionary relationship with the strain isolated in the Philippines in 2014. Their genotype was Asian genotype.
Conclusions
This epidemic was confirmed to have been imported from the Philippines after the infection with the Asian genotype CHIKV, which suggests that Fujian province should strengthen the monitoring of persons entering from the CHIK epidemic area, so as to prevent imported cases from causing local outbreaks.