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Objective: The objective of this study was to screen lymphoma radiotherapy-resistant genes using CRISPR activation (CRISPRa). Methods: The Human CRISPRa library virus was packaged and then transfected into lymphoma cells to construct an activation library cell line, which was irradiated at the minimum lethal radiation dose to screen radiotherapy-resistant cells. Radiotherapy-resistant cell single-guide RNA (sgRNA) was first amplified by quantitative polymerase chain reaction (qPCR) in the coding region and then subject to next-generation sequencing (NGS) and bioinformatics analysis to screen radiotherapy-resistant genes. Certain radiotherapy-resistant genes were then selected to construct activated cell lines transfected with a single gene so as to further verify the relationship between gene expression and radiotherapy resistance. Results: A total of 16 radiotherapy-resistant genes, namely, C20orf203, MTFR1, TAF1L, MYADM, NIPSNAP1, ZUP1, RASL11A, PSMB2, PSMA6, OR8H3, TMSB4Y, CD300LF, EEF1A1, ATP6AP1L, TRAF3IP2, and SNRNP35, were screened based on the NGS results and bioinformatics analysis of the radiotherapy-resistant cells. Activated cell lines transfected with a single gene were constructed using 10 radiotherapy-resistant genes. The qPCR findings showed that, when compared with the control group, the experimental group had significantly up-regulated mRNA expression of MTFR1, NIPSNAP1, ZUP1, PSMB2, PSMA6, EEF1A1, TMSB4Y and TAF1L (p < 0.05). No significant difference in the mRNA expression of AKT3 or TRAF3IP2 (p > 0.05) was found between the two groups (p > 0.05). Conclusion: The 16 genes screened are potential lymphoma radiotherapy-resistant genes. It was initially determined that the high expression of 8 genes was associated with lymphoma radiotherapy resistance, and these genes could serve as the potential biomarkers for predicting lymphoma radiotherapy resistance or as new targets for therapy.
RESUMEN
PURPOSE: The two main strategies for managing osteoporosis using the Fracture Risk Assessment (FRAX®) are the fixed-probability threshold of the National Osteoporosis Foundation (NOF) and the age-dependent-probability threshold of the National Osteoporosis Guideline Group (NOGG), but there are no FRAX® Chinese-specific thresholds. This study examined the NOF and NOGG strategies for intervention thresholds using the Chinese FRAX® model for their appropriateness for Chinese postmenopausal women, and explored Chinese-specific thresholds. METHODS: Postmenopausal women (N = 264) >50 years old from community-medical centers in China were randomly selected. They completed a self-report questionnaire and underwent bone mineral density measurements and spinal X-rays. The 10-year risks for a major osteoporosis fracture and hip fracture were calculated using the Chinese FRAX® model. Using an osteoporosis diagnosis as the gold standard, we compared the abilities of the NOF and NOGG thresholds to detect osteoporosis by analyzing their sensitivity, specificity, accuracy, and positive and negative likelihood ratios. RESULTS: The 10-year risks for hip fracture and a major osteoporotic fracture increased with age. The NOF's accuracy in detecting osteoporosis was 83.33% and the NOGG's was 74.24%. The NOF thresholds showed higher accuracy and specificity than the NOGG thresholds. CONCLUSION: NOF thresholds are more appropriate for Chinese menopausal women.
