Analysis of Methods to Reduce Jogging Injuries from the Perspective of Risk Cognition.

Objectives: Injuries are common events that impair the function of joggers; it is important to take effective measures to reduce the incidence of jogging injuries to maximize its benefits. Methods: We collected questionnaires from 3468 Chinese exercisers using a stratified random sampling method. We analyzed these data using AMOS 22.0 (IBM) and SPSS 26.0 (IBM). Results: We constructed a relationship model of jogging risk cognition, jogging risk behaviors, and jogging injury incidence, and the indicators fit well. The path coefficient between jogging risk cognition and jogging risk behaviors of joggers was -0.64; the path coefficient between jogging risk behaviors and jogging injuries incidence was 0.44; and the path coefficient between jogging risk cognition and jogging injuries incidence was -0.23. Conclusions: The higher the level of jogging risk cognition of joggers, the lower their jogging risk behaviors, leading to a lower incidence of jogging injuries. Based on the generally low level of jogging risk cognition of joggers, the construction of a jogging risk cognitive education system that can be integrated into universities, middle schools and primary schools, as well as family, school and society, is conducive to comprehensively improve the level of jogging risk cognition of joggers.

Cas9 Protein Triggers Differential Expression of Inherent Genes Especially NGFR Expression in 293T Cells.

INTRODUCTION: CRISPR/CAS9 systems, which can be utilized in vitro biological experiments, have recently captured much attention for their important roles and benefits. However, full realization of the potential of CRISPR/CAS9 approaches requires addressing many challenges and side effects. The expression of genes and potential side effects of CRISPR/CAS9 in human cells remains to be elucidated. The aim of our study was to explore the effect of CRISPR/CAS9 on gene expression in 293T cells. METHODS: A Cas9-expressing PX458 plasmid and Cas9-deactivated PX458-T2A plasmid were used to study the role of CRISPR/CAS9 on regulating gene expression in 293T cells. Gene expression in 293T cells after transfection of the PX458 plasmid or PX458-T2A plasmid was detected by RNA sequencing and correlative statistical analysis. Differential gene expression in both PX458 transfected 293T cells and PX458-T2A transfected 293T cells compared with normal 293T cells was detected using quantitative reverse transcription polymerase chain reaction (RT qPCR). The mRNA and protein levels were measured using reverse transcription PCR and Western blot. Co-IP assay combined with shotgun LC-MS/MS were used to investigate the differences of NGFR-interaction proteins between PX458 transfected 293T cells and PX458-T2A transfected 293T cells. RESULTS: In this study, we observed that PX458 plasmid transfection and Cas9 expression can affect the expression of different genes, including FOSB (FBJ murine osteosarcoma viral oncogene homolog B), IL-11 (Interleukin-11), MMP1 (matrix metalloproteinase), CYP2D6 (CytochromeP4502D6), and NGFR (matrix metalloproteinase 1). Downregulation of NGFR after PX458 transfection was confirmed by RT qPCR and western blot analysis. NGFR expression was significantly lower in PX458 transfected 293T cells than in normal 293T cells and PX458-T2A transfected 293T cells. The co-IP dilutions analyzed by shotgun LC-MS/MS showed a total of 183 proteins interact with NGFR in PX458 transfected 293T cells while 221 proteins interact with NGFR were identified in PX458-T2A transfected 293T cells using the MASCOT engine. CONCLUSIONS: Cas9 expression by transfection of the PX458 plasmid was negatively correlated with the NGFR mRNA level and NGFR protein expression in 293T cells, while PX458-T2A, in which Cas9 is deactivated, did not affect NGFR expression. The decrease in NGFR expression also affects the amount of proteins that interact with NGFR. These results suggest that the effect of Cas9 on NGFR expression and the expression of other genes should be noticed when developing cell-based studies and therapies utilizing CRISPR/CAS9 systems.