1.Quantitative measurement of human immunodeficiency virus type 1 RNA in seminal plasma with Nuclisens HIV-1 QT
Maofeng QIU ; Hao WU ; Yan JIANG
Chinese Journal of Blood Transfusion 1988;0(03):-
Objective To evaluate NucliSens HIV-1 QT(bioMerieux,Netherlands) in quantitating human immunodeficiency virus type 1(HIV-1) RNA in whole semen or seminal plasma from HIV-1-infected people.Methods Five levels of HIV-1 RNA were spiked to whole semen,seminal plasma and blood plasma samples from healthy people,and then measured by NucliSens HIV-1 QT.The same method was used to measure HIV-1 RNA in seminal plasma and blood plasma samples from 15 HIV-1 infected people.Results Nucleic acid amplification inhibitors were found in whole semen but not in seminal plasma when using NucliSens HIV-1 QT.No significant difference was found between normal seminal plasma and blood plasma samples spiked with HIV-1 RNA,and no false positive result was found in 10 normal seminal plasma samples.For 15 cases of HIV-1 infection,HIV-1 was detected in 80%(12/15) of the plasma samples and 40%(6/15) of the seminal plasma samples,with viral loads of
2.Effect of siRNA on hepatitis B virus X gene to HepG2.2.15 cells
Jinhong ZHU ; Chunmiao WANG ; Hong WU ; Meishuang LIN ; Xuelian ZHU ; Maofeng WU ; Xiaoman WU
The Journal of Practical Medicine 2016;32(13):2120-2123
Objective Using chemically synthesized small interfering RNA (siRNA) transfected HepG2.2.15 cells to construct a cell model in interfering hepatitis B virus (HBV) X gene, studying the inhibi-tion of HBV replication and antigen expression in vitro. Methods After transfection of HepG2.2.15 cell for 24 h, 48 h, 72 h, detecting the cell supernatant of HBsAg and HBeAg by chemiluminescence immunoassay, the cell supernatant HBxAg protein by ELISA , the HBx mRNA relative expression of transfected cell was detected by fluorescence quantitative polymerase chain reaction (PCR), the ability of cell proliferation was detected by CCK-8 assay. Results After HBx-siRNA transfected HepG2.2.15 cells, cell proliferation ability was inhibited. The cell of HBx mRNA and the cell supernatant of HBxAg expression decreased (P < 0.05); at the same time it in-hibited the expression of HBsAg and HBeAg. The suppressed peak and the inhibition rate were 66% and 58%respectively at 72 h. The fluorescence quantitative PCR confirmed that expression of HBV DNA in the super-natant was decreased. Conclusion The HepG2.2.15 cell interference model of HBV X gene has been success-fully constructed, which has an effect of inhibiting proliferation of HepG2.2.15 cells and replication and expres-sion of HBV gene in vitro.
3. Preliminary association of individual different plasma pazopanib concentration with CYP3A4 gene polymorphism
Maofeng WU ; Chang LIU ; Huihui DAI ; Zhangfeng MAI ; Danli HUANG ; Jingwei MIAO ; Lizhong LIU ; Yi FANG ; Yi FANG
Chinese Journal of Clinical Pharmacology and Therapeutics 2020;25(12):1376-1380
AIM: To study the pharmacokinetics (PK) of pazopanib tablets and explore the genetic mechanism of individual differences in drug metabolism primarily. METHODS: Fourteen healthy male subjects were respectively administrated with a single dose pazopanib tablet (200 mg) orally on the day of dosing, and their blood samples were collected from baseline to 96 hours. The serum concentration of pazopanib was measured by LC-MS/MS, the parameters of PK were calculated by winnonlin 6.3 software, and the gene polymorphism of cytochrome P450 3A4 (CYP3A4) was determined by snapshot method. RESULTS: The range of C