Expression and clinical significance of lncRNA NORAD in patients with gestational hypertension

Objectives: Gestational hypertension (GH), the most common type of hypertensive disorders in pregnancy, often occurs in women during pregnancy. The purpose of this study was to investigate the expression and clinical significance of lncRNA NORAD in gestational hypertension, and to discuss the possibility of lncRNA NORAD as a diagnostic marker of gestational hypertension. Material and methods: A total of 219 participants were involved in the study. Basic clinical information of all participants was collected, and the expression of NORAD in serum was detected by RT-qPCR. ROC curves were drawn to evaluate the diagnostic value of NORAD expression for gestational hypertension. Multiple linear regression analysis was done to explore the relationship between NORAD and clinical variables. Logistic regression analysis was conducted to analyze the independent influence of different variables on the development of gestational hypertension into preeclampsia. Results: The expression level of NORAD in gestational hypertension was higher than that of healthy individuals, and the expression level of NORAD in preeclampsia was higher than that of gestational hypertension and healthy individuals. The ROC curve suggested that the expression of NORAD has a higher diagnostic value for gestational hypertension. Multiple linear regression analysis showed that systolic blood pressure (SBP) and diastolic blood pressure (DBP) were correlated with the expression of NORAD. SBP, DBP and NORAD were all factors that affect the development of gestational hypertension to preeclampsia, which were known by Logistic regression analysis. Conclusions: LncRNA NORAD may be used as a biomarker for gestational hypertension diagnosis and can influence its progression into preeclampsia

Optimization and Application of CRISPR/Cas9 Genome Editing in a Cosmopolitan Pest, Diamondback Moth

The CRISPR/Cas9 system is an efficient tool for reverse genetics validation, and the application of this system in the cell lines provides a new perspective on target gene analysis for the development of biotechnology tools. However, in the cell lines of diamondback moth, Plutella xylostella, the integrity of the CRISPR/Cas9 system and the utilization of this cell lines still need to be improved to ensure the application of the system. Here, we stabilize the transfection efficiency of the P. xylostella cell lines at different passages at about 60% by trying different transfection reagents and adjusting the transfection method. For Cas9 expression in the CRIPSPR/Cas9 system, we identified a strong endogenous promoter: the 217–2 promoter. The dual-luciferase and EGFP reporter assay demonstrated that it has a driving efficiency close to that of the IE1 promoter. We constructed pB-Cas9-Neo plasmid and pU6-sgRNA plasmid for CRISPR/Cas9 system and subsequent cell screening. The feasibility of the CRISPR/Cas9 system in P. xylostella cell lines was verified by knocking out endogenous and exogenous genes. Finally, we generated a transgenic Cas9 cell line of P. xylostella that would benefit future exploitation, such as knock-in and multi-threaded editing. Our works provides the validity of the CRISPR/Cas9 system in the P. xylostella cell lines and lays the foundation for further genetic and molecular studies on insects, particularly favoring gene function analysis