RESUMO
OBJECTIVE: To construct the RNA interference(RNAi) lentiviral vector of suppressor of variegation 3-9 homolog 1(Suv39 h1) and verify its interfering efficiency by transfecting it to the bone marrow-derived mesenchymal stem cells(BMSCs). METHODS: The oligonucleotides of RNA plasmid were designed and synthesized according to the gene sequence of Suv39 h1 and short hairpin RNA design principles. Three kinds of LV-Suv39 h1-RNAi recombinant plasmids with different lentivirus knockdown targets(KD1, KD2 and KD3) were constructed. After identification by restriction analysis and sequencing, the packaged lentivirus vectors with the three kinds of Suv39 h1 gene were transfected into rat BMSCs at logarithmic growth stage, and were named KD1, KD2 and KD3 transfection groups. The control group was transfected with the negative control virus. After 72 hours transfection, the transfection efficiency was evaluated, and the relative mRNA levels of Suv39 h1 were determined by quantitative real-time polymerase chain reaction(qPCR). RESULTS: Sequencing analysis demonstrated that three kinds of LV-Suv39 h1-RNAi recombinant plasmids were constructed correctly. The results of transfection efficiency evaluation showed that more than 80.00% green fluorescence was expressed in the BMSCs transfected with the three lentiviral vectors with a multiplicity of infection of 20. These results indicated that lentivirus was successfully constructed and transfection efficiency was high. The results of qPCR showed that the relative expression of Suv39 h1 mRNA in BMSCs of KD1, KD2 and KD3 transfection groups was lower than that in the control group(all P<0.05), and the relative expression of Suv39 h1 mRNA in KD1 and KD3 transfection groups was lower than that in KD2 transfection group(both P<0.05). However, there was no significant difference in the relative expression of Suv39 h1 mRNA between KD1 and KD3 transfection groups(P>0.05). CONCLUSION: The constructed lentiviral vector with low expression of Suv39 h1 was constructed successfully. This vector can be expressed in rat BMSCs, which lays a foundation to study the effect of Suv39 h1 gene in acute myeloid leukemia.
RESUMO
OBJECTIVE: To evaluate dual-energy spectral CT imaging in evaluating the degree of differentiation in colon cancer. METHODS: Forty-seven colon cancer patients underwent spectral CT during arterial phase (AP) and portal venous phase (PP), and were characterized pathologically differentiated to well-differentiated (A, n = 18) and poorly differentiated or undifferentiated carcinoma group (B, n = 29). Lesion iodine concentration (IC) was measured and normalized to that of aorta (NIC). CT numbers were measured and the slope (λ HU) of the spectral HU curve was calculated. These parameters were statistically compared between the two groups. ROC curves were used to evaluate their diagnostic efficacies. RESULTS: There were significant differences in IC (1.01 ± 0.20 vs. 1.59 ± 0.57 mg/ml), NIC (0.12 ± 0.03 vs. 0.19 ± 0.09), λ HU (1.41 ± 0.29 vs. 2.03 ± 0.85), and CT number at 70 keV (48.61 ± 9.03HU vs. 63.97 ± 15.86HU) between groups A and B in AP (p < 0.05), but no difference in PP. Using IC = 1.13 mg/ml in AP as the threshold, one obtained a sensitivity of 81.8% and a specificity of 71.4% for differentiating well-differentiated from poorly differentiated or undifferentiated carcinoma. These values were statistically higher than those (64.7% and 62.3%) using CT number at 70 keV. CONCLUSION: Spectral CT imaging parameters (IC, NIC, and λ HU) in AP provide improved accuracy for evaluating the degrees of differentiation in colon cancer than CT number at 70 keV.
