Objective:To screen host cell proteins interacting with rotavirus nonstructural protein 2(NSP2) and provide a theoretical basis for the discovery of antiviral targets.Methods:E. coli BL21(DE3) was transformed with a recombinant plasmid NSP2-pGEX-6P-1 and induced by IPTG to express NSP2-GST. GST affinity chromatography was used to purify NSP2-GST and Western blot was performed for verification. NSP2-GST and GST proteins were used as targets to capture interacting proteins in MA104 cells via GST pull-down. After silver staining, differential proteins were screened by in-gel enzymatic hydrolysis and mass spectrometry. Protein pilot platform was used to filter peptides, and the names and the biological functions of the proteins were obtained by Paragon algorithm. The potential connection between the interacting proteins was demonstrated by protein interaction network diagrams and GO functional annotation analysis. Moreover, the molecular docking of the top three proteins with NPS2 was predicted using HDOCK server and verified by docking and confidence scores, so as to investigate the visual docking model between interacting proteins. Results:SDS-PAGE and Western blot showed the successful purification of the recombinant protein NSP2. Ten host proteins including PKM2, which might interact with NSP2, were identified by GST pull-down and protein profiling. GO analysis and interaction diagrams revealed that RPS4X, EZR, SUPT16H and EIF2S3 mediated molecular expression; PKM2, LDHA and ATP5A1 participated in energy metabolism; HSP90, ACTB and ANXA2 were involved in biological movement. Besides, there were functional connections and interaction networks among them. Molecular docking further verified PKM2, HSP90 and RPS4X did interact with NSP2, and the interaction force were strong enough to form a stable structure.Conclusions:This study successfully discovered several host proteins including PKM2, HSP90 and RPS4X that could interact with NSP2, providing reference for investigating the process of rotavirus infection and making related prevention and control strategies.

Pancreatic β-cell failure due to a reduction in function and mass has been defined as a primary contributor to the progression of type 2 diabetes (T2D). Reserving insulin-producing β-cells and hence restoring insulin production are gaining attention in translational diabetes research, and β-cell replenishment has been the main focus for diabetes treatment. Significant findings in β-cell proliferation, transdifferentiation, pluripotent stem cell differentiation, and associated small molecules have served as promising strategies to regenerate β-cells. In this review, we summarize current knowledge on the mechanisms implicated in β-cell dynamic processes under physiological and diabetic conditions, in which genetic factors, age-related alterations, metabolic stresses, and compromised identity are critical factors contributing to β-cell failure in T2D. The article also focuses on recent advances in therapeutic strategies for diabetes treatment by promoting β-cell proliferation, inducing non-β-cell transdifferentiation, and reprograming stem cell differentiation. Although a significant challenge remains for each of these strategies, the recognition of the mechanisms responsible for β-cell development and mature endocrine cell plasticity and remarkable advances in the generation of exogenous β-cells from stem cells and single-cell studies pave the way for developing potential approaches to cure diabetes.

Objective : To screen the RNA binding proteins interacting with NBV nonstructural protein σNS in host cells and to analyze their bioinformatics functions. Methods : In this study , the eukaryotic expression vector pEF⁃HA⁃MB⁃S3 of NBV σ NS was constructed and transfected into HEK293T cells after verification. After RNase A treatment , the obtained protein lysate was enriched by immunoprecipitation to enrich σNS binding proteins , identified and analyzed by LC⁃MS/MS mass spectrometry , and the properties and specific functions of proteins were discovered with the help of related Bioinformatics tools. Results : In this study , 32 candidate RNA binding proteins interacting with NBV σNS proteins were successfully screened , and the results of bioanalysis showed that these proteins were mainly located in cytoplasm and nucleus , and were mainly involved in biological processes such as cell metabolism , biological regulation , virus translation and transcription. Conclusion This study preliminarily analyzes the function of RNA binding proteins interacting with NBV σNS , which lays a foundation for further study on the mechanism of σNS protein in NBV life cycle.