RESUMEN
Objective To construct an shRNA lentiviral vector targeting the MEX3A gene and establish a bladder cancer cell line with stable MEX3A gene knockdown.Methods Real-time PCR was performed to detect the MEX3A gene expression.The recombinant lentiviral vector targeting the MEX3A gene was constructed using the GV115 plasmid.After identification and sequencing,the vectors were co-transfected with the packaging vector into 293T cells to produce lentiviral particles,which were then transduced into bladder cancer cells after viral titer determination.The cell line stably expressing the siRNA was established by antibiotic selection,and real-time PCR was carried out to detect the efficiency of the knockdown.Results Both bladder cancer cell lines,5637 and T24,expressed the MEX3A gene,and its expression was higher in 5637 than in T24.The identification and sequencing results showed that the MEX3A-shRNA lentiviral vector was successfully constructed,and the virus titer was observed to be higher after packaging.The results of the real-time PCR showed that MEX3A gene expression was stably inhibited in 5637 cells after lentiviral transduction.Conclusion Lentivirus-mediated RNAi technology could successfully establish a cell line with stable MEX3A gene knockdown.
RESUMEN
Objective To assess the feasibility and accuracy of ultrasound evaluation of facet joint in the lumbar spine by comparison of ultrasonography and CT measurements in normal and degenerative lumbar vertebra small joint.Methods Fifteen patients with chronic low back pain in Shengjing hospital of China Medical University were taken as the study group,and 20 healthy volunteers as the healthy controls from physical examination center of our hospital between January 2014 and December 2014.The ultrasonic and CT values were measured in L 1-S 1 small joints between normal and abnormal lumbar joints (height and width).The length and width of the left and right side small joints of healthy controls,ultrasonic measurement of healthy controls and patients with chronic low back pain,and value of ultrasonography and CT measurements were compared by using t test.Results The abnormal facet joint can be clearly showed by sonography,characterized by bone hyperplasia of edge joints,bone destruction,wear or thinning/thickening of articular cartilage.For the length and width of the lumber joints,the difference of ultrasonic measurement values between normal and abnormal lumbar joints were statistically significant [(1.26±0.03) cm vs (1.43±0.05) cm,t=2.626,P ＜ 0.05;(0.18±0.01) cm vs (0.15±0.02) cm,t=2.264,P ＜ 0.05].In patient group,the difference of length and width values in a total of 150 small joints between ultrasonography and CT were not statistically significant [(1.43 ± 0.17) cm vs (1.42 ± 0.16) cm],t=1.655,P ＞ 0.05;(0.15 ± 0.03) cm vs (0.14±0.03) cm,t=1.656,P ＞ 0.05].Conclusions The facet joint of the lumbar can be clearly shown by sonography and it is valuable in the diagnosis of small joint degeneration for lumbar vertebra.