Protein liquid-liquid phase separation (LLPS), a pivotal phenomenon intricately linked to cellular processes, is regulated by various other proteins. However, there is still a lack of high-throughput methods for screening protein regulators of LLPS in target proteins. Here, we developed a CRISPR/Cas9-based screening method to identify protein phase separation regulators by integrating bimolecular fluorescence complementation (BiFC) and fluorescence-activated cell sorting (FACS). Using this newly developed method, we screened the RNA-binding proteins that regulate PABPN1 phase separation and identified the tumor suppressor QKI as a promoter of PABPN1 phase separation. Furthermore, QKI exhibits decreased expression levels and diminished nuclear localization in colorectal cancer cells, resulting in reduced PABPN1 phase separation, which, in turn, promotes alternative polyadenylation (APA), cell proliferation, and migration in colorectal cancer.
Humans ; Colorectal Neoplasms/genetics* ; RNA-Binding Proteins/genetics* ; Poly(A)-Binding Protein I/genetics* ; CRISPR-Cas Systems ; Flow Cytometry ; Cell Proliferation ; Cell Line, Tumor ; Cell Movement

Objective To investigate the safety of training using a lower-limb rehabilitation robot and its effectiveness in terms of stroke survivors' cardiopulmonary function.Methods Thirty-two hemiplegic stroke survivors were randomly divided into a robot group and a control group,each of 16.Both groups received conventional rehabilitation medication and training,120 min/d,5 d/week for 8 weeks.The robot group was additionally trained with a Flexbot robotic gait training apparatus for 30 min/d,5 d/week for the same 8 weeks.The subjects' maximum oxygen consumption (VO2max),heart rate,blood pressure,ventilation (VE) and rate of perceived exertion (RPE) were quantified before and after the training.Results After the 8 weeks there was no significant difference in average heart rate,blood pressure,VE or RPE between the two groups.The average VO2max of the robot group was,however,significantly higher than that of the control group.Conclusion Robotic gait training is safe and can improve the cardiopulmonary function of stroke survivors.

objective To investigate the multiple early variables that influenced the 6th months prognosis of comprehensive function in patients with stroke.Methods Two hundreds and eleven patients of primary cerebral infarction and primary cerebral hemorrhage in Shanghai were divided into treated group and controlled group randomly.Patients in the treated group were given standardized tertiary rehabilitation,while those in the controlled group received normal internal medicine treatments but without standardized tertiary rehabilitation.Twenty seven early variables were collected with regard to the patients'medical history,physical examination,scores with CNFD,S-FMA and MBI at the time of enrollment.The patients'FCA scores at the ends of the 6th months after stroke were used as the comprehensive functional outcome.Stepwise multiple regression analysis applied to analyze the data.Results Stepwise multiple regression analysis revealed that therapeutic regimen,CNFD scores,age,diabetes mellitus,SFMA scores,smoking and labour intensity predicted comprehensive functional outcome at the ends of the 6th months after stroke.Conclusion Certain early variables predict the long-term comprehensive functional prognosis of stroke patients.Therapeutic regimen,CNFD scores and age were significant predictors.