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1.
Article in Chinese | WPRIM | ID: wpr-880079

ABSTRACT

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.


Subject(s)
CRISPR-Cas Systems , Cell Differentiation , Gene Editing , MicroRNAs/genetics , Sequence Analysis, RNA , Tretinoin
2.
Article in Chinese | WPRIM | ID: wpr-816405

ABSTRACT

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.

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