Effect of MiR-155 Knockout Mediated by Dual sgRNAs on Drug Sensitivity of FLT3-ITD+AML / 中国实验血液学杂志

OBJECTIVE@#Two sgRNAs transfected FLT3-ITD+AML cell line MV411 with different binding sites were introduced into CRISPR/cas9 to obtain MV411 cells with miR-155 gene knockout. To compare the efficiency of miR-155 gene knockout by single and double sgRNA transfection and their effects on cell phenotypes.@*METHODS@#The lentiviral vectors were generated containing either single sgRNA or dual sgRNAs and packaged into lentivirus particles. PCR was conducted to measure gene editing efficiency, and miR-155 expression was evaluated by qPCR. CCK-8 assay was used to evaluate the cell proliferation, and calculate drug sensitivity of cells to adriamycin and quizartinib. Annexin V-APC/7-AAD staining was used to label cell apoptosis induced by adriamycin and quizartinib.@*RESULTS@#In the dual sgRNAs transfected cells, a cleavage band could be observed, meaning the success of gene editing. Compared with the single sgRNA transfected MV411 cells, the expression level of mature miR-155-5p was lower in the dual sgRNA transfected cells. And, dual sgRNA transfected MV411 were more sensitive to adriamycin and quizartinib with lower IC50 and higher apoptosis rate.@*CONCLUSION@#The inhibition rate of miR-155 gene expression transfected by dual sgRNA is higher than that by single sgRNA. Dual sgRNA transfection can inhibit cell proliferation, reverse drug resistance, and induce apoptosis more significantly. Compared with single sgRNA transfection, dual sgRNA transfection is a highly efficient gene editing scheme.

Radiomics in Antineoplastic Agents Development: Application and Challenge in Response Evaluation / 中国医学科学杂志(英文版)

The recent spring up of the antineoplastic agents and the prolonged survival bring both challenge and chance to radiological practice. Radiological methods including CT, MRI and PET play an increasingly important role in evaluating the efficacy of these antineoplastic drugs. However, different antineoplastic agents potentially induce different radiological signs, making it a challenge for radiological response evaluation, which depends mainly on one-sided morphological response evaluation criteria in solid tumors (RECIST) in the status quo of clinical practice. This brings opportunities for the development of radiomics, which is promising to serve as a surrogate for response evaluations of anti-tumor treatments. In this article, we introduce the basic concepts of radiomics, review the state-of-art radiomics researches with highlights of radiomics application in predictions of molecular biomarkers, treatment response, and prognosis. We also provide in-depth analyses on major obstacles and future direction of this new technique in clinical investigations on new antineoplastic agents.

Identify Myeloid Differentiation-Related MiRNAs Response to ATRA Induction by RNA Sequencing and CRISPR/Cas9 Gene Editing / 中国实验血液学杂志

OBJECTIVE@#To identify differentiation related miRNA and evaluate roles of miRNA during ATRA induced myeloid differentiation.@*METHODS@#The small RNA sequencing was used to analyze differential expressed miRNAs in ATRA induced NB4 cells. Then the several up or down-regulated miRNA were selected as the research candidates. SgRNAs targeting the genome of each miRNA were designed and NB4 cells with inducible expression of Cas9 protein were generated. After transduced sgRNA into NB4/Cas9 cells, the mutation level by PCR and surveyor assay were evaluated. The cell differentiation level was investigated by surface CD11b expression via flow cytometry.@*RESULTS@#A total of 410 mature miRNAs which expressed in NB4 cells were detected out after treated by ATRA, 74 miRNAs were up-regulated and 55 were down-regulated miRNAs with DNA cleavage generated by CRISPR/Cas9 was assayed directly by PCR or surveyor assay, quantitative PCR showed that the expression of miRNA was downregulated, which evaluated that gene edition successfully inhibitied the expression of mature miRNA. MiR-223 knockout showed the myeloid differentation of NB4 significantly inhibitied, while miRNA-155 knockout showed the myeloid differentation of NB4 cells significantly increased.@*CONCLUSION@#CRISPR/Cas9 is a powerful tool for gene editing and can lead to miRNA knockout. Knockouts of miR-223 and miR-155 have shown a differentiation-related phenotype, and the potential mechanism is the integrative regulation of target genes.

Imaging diagnosis and value of early gastric cancer / 中国实用外科杂志

Radiology is one of the common modalities in the diagnosis and evaluation of early gastric cancer(EGC), which include X-ray double contrast radiography, CT, MRI and PET.X-ray double contrast radiography is a classical method on the diagnosis of EGC. High concentration of barium, with the combination of double contrast, mucosal and thin-layer flow-coating methods, can dynamically observe the three-dimensional morphology of mucosal surface damage. Abdominal enhanced CT is one of the conventional methods for the staging of EGC.Hypotonic air-filling is recommended, and the standardized application of late arterial phase(40 seconds) enhancement and multiplanar reconstruction(MPR) techniques is emphasized. The demonstration of the continuous and complete lowintensity bands between the inner high enhanced cancer layer and outer intermediate enhanced muscularis propria layer of gastric wall, or the thickness of the inner high enhanced cancer layer less than 50% of the total thickness of gastric wall,which were the CT signs of EGC. The latter sign is of greater importance in the staging diagnosis of EGC originated from peptic ulcers. Dual-energy CT has the potential to improve the detection and staging accuracy of EGC. DW-MRI and DCEMRI imaging can highlight the display of EGC and improve the detection. PET is limited in the detecting ability of EGC, but it can assist in predicting the curative treatment of endoscopic mucosal resection(ESD) of EGC and monitoring the recurrence after ESD. Periodic follow-up should be conducted after the operation of EGC, especially ESD procedures. The followup plan can be formulated according to the relevant risk factors. Radiomics has the potential in the diagnosis of lymph node metastasis in EGC.