1.Effects of recombinant antisense c-myc adenovirus in induction of differentiation of HL-60 cells
Long CHEN ; Jieping CHEN ; Chen LIN ;
Journal of Third Military Medical University 2003;0(17):-
Objectives To evaluate the potential of antisense c myc gene therapy for acute promyelocytic leukemia by investigating the biological effects and molecular mechanisms in induction of differentiation of HL 60 cell line using recombinant antisense c myc adenovirus (Ad AS c myc). Methods Cultured HL 60 cells treated with Ad AS c myc or Ad LacZ with polybrene and protamine sulfate were analyzed by X gal staining, morphology, MTT, flow cytometric analysis, RT PCR, and immunocytochemical techniques in vitro . Results HL 60 cells could be transfected effectively by Ad LacZ+protamine sulfate (79.8%). The level of c myc transcription and the version could be strongly inhibited by Ad AS c myc in the transfected HL 60 cells. Ad AS c myc could strongly inhibit the cell growth in HL 60 cells (51%). Ad AS c myc could reduce the ratio of nuclear/cytoplasm, and increase the activity of peroxidase in HL 60 cells. Ad AS c myc could lead to the blocking of G 0/G 1 phase in HL 60 cells. Ad AS c myc could also increase the expression of c fos in HL 60 cells. Conclusion The expression of Ad AS c myc can inhibit the growth and induce differentiation of HL 60 cells in vitro . The biological effects of Ad AS c myc may be closely associated with the activity of peroxidase and c fos gene in HL 60 cells. Ad AS c myc is of clinical potential in gene thera py for acute promyelocytic leukemia.
2.Application value of CMIA in HCV infection validation in low risk population
Linfeng WU ; Jinfeng ZENG ; Yuanzhang SUN ; Jieping LONG ; Lilin WANG
International Journal of Laboratory Medicine 2017;38(10):1301-1303
Objective To detect the reactive samples of enzyme-linked immunosorbent assay (ELISA1) by chemiluminescence microparticle immunoassay (CMIA),and to analyze the application value of CMIA in HCV infection validation of blood donors.Methods Nucleic acid 3-item combined testing (NAT),another ELISA2,HCV antibody supplementary test(Western Blot test,WB) and CMIA test supplemented in blood samples of 102 ELISA1 anti-HCV reactive blood donors were retrospectively analysed.Results Among 102 blood donors of anti-HCV positive,32 cases (31.37%,32/102) were HCV RNA reactive samples,50 cases (49.02%,50/102) were ELISA2/WB reactive simultaneously.With CMIA NAT results as the reference standard,CMIA was poorly correlated with HCV RNA (Spearman correlation coefficient rs=0.395,P<0.01),and the consistency between them was weak by Kappa test (Kappa=0.270,P<0.01).With ELISA2/WB detection results as the reference standard,CMIA was highly correlated with the results(Spearman correlation coefficient rs=0.713,P<0.01),and which showed high consistency by Kappa test (Kappa=0.674,P<0.01).Conclusion CMIA as a detection method of protein label after HCV infection has great value in the HCV infection confirmation in low-risk population.
3.Analysis on the results of domestic HCC RNA genotype diagnostic kit
Jingfeng ZENG ; Liang LU ; Yuanzhang SUN ; Jieping LONG ; Xinghui GU ; Xiaoxuan XU ; Tong LI ; Dongmei NIE ; Lilin WANG
International Journal of Laboratory Medicine 2017;38(2):157-159,162
Objective To evaluated the HCV genotyping results which obtained by genotype diagnostic kit in Shenzhen area. Methods 158 samples which ELISA test of anti-HCV were positive were collected from voluntary blood donors from 2014 to 2015,and were tested by PCR fluorescence probe method for viral load.The samples which viral load were greater than 1.0 ×103 IU/mL were then tested by HCV RNA genotype diagnostic kit.To analysis the proportion of different genotypes and the correla-tion between genotypes with vrial load.Results 54 HCV RNA reactive sample were quantity by PCR fluorescence probe method from 158 anti-HCV positive samples.The genotyping data for 45 cases which vrial load greater than 1.0×103 IU/mL were obtained by HCV RNA genotype diagnostic kit.The frequencies HCV genotype 1b,2,3 and 6 were 57.78%(26/45),6.67%(3/45),8.89%(4/45)and 26.67%(12/45),respectively.One-way ANOVA analysis showed that significant difference in viral loads was found be-tween different HCV genotype 1b and 2(F =2.861,P <0.05),and there was a significant difference in viral loads and anti-HCV S/CO by sex(P <0.05).Fisher′s exact test showed the significance difference between age and genotypes(P <0.05 ).Conclusion HCV 1b and 6 were the most predominant genotypes due to the higher viral load than the other subtypes among volunteer blood do-nors in Shenzhen,while the proportion of HCV 2,3 declined.