The CRISPR/dCas9 system is a gene editing tool that has proven to be highly efficient and precise. By utilizing transcriptional activators, such as VP64, p65, and Rta, the system can effectively and stably activate target genes. Sox17, a transcription factor belonging to the SOX family, plays a crucial role in the differentiation of the germ layers and the determination of cell fates during the early stages of embryonic development. Sheep embryonic stem cells (sESCs) are characterized by their capacity for self-renewal and multidirectional differentiation, serving as a significant in vitro model for studying the mechanisms of cell differentiation during early embryonic development. However, the importing of exogenous genes into sESCs is challenging due to their unique growth characteristics. The objective of this study was to investigate the conditions necessary for successfully activating Sox17 in sESCs. To this end, we employed the CRISPR/dCas9 system along with liposome transfection, lentivirus invasion, and electroporation to activate Sox17 in sESCs. The expression of Sox17 was then determined by fluorescence quantitative PCR, on the basis of which the performance of different transfection methods was compared. The results indicated that the electroporation group had the best transfection effect and the highest Sox17 expression among the three transfection methods. The efficient and stable gene activation protocol will provide a reference for embryonic stem cell research in other species, especially livestock animals, and lay the foundation for the subsequent study of gene function and realization of precise cell fate regulation by regulating gene expression in sheep embryonic stem cells.
Animals ; CRISPR-Cas Systems/genetics* ; Sheep ; SOXF Transcription Factors/genetics* ; Embryonic Stem Cells/cytology* ; Genetic Vectors/genetics* ; Cell Differentiation/genetics* ; Transfection ; Gene Editing/methods*

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Establishing a new paradigm for high-quality development in public hospitals,embracing a comprehensive,systematic,and innovative strategic management philosophy,and instituting a modern hospital management system hold significant practical importance.It investigates the pathways through which the Guangzhou Women and Children's Medical Center has fulfilled its strategic commitments over its 17-year history.It delves into the meaning and value of constructing hospital strategic commitments,explores pathways for realization,analyzes effects.It aims to understand the normative role of strategic commitments in shaping the development paths of specialized women's and children's hospitals and to provide a reference for the high-quality development of similar hospitals across China.

Objective To explore the potential mechanism of action of paeoniflorin in diabetes mellitus,the related targets and pathways were preliminarily discussed,based on the network pharmacology and molecular docking technology.Methods Analyze the potential targets of paeoniflorin using the Swiss Target Prediction database.Genecards and OMIM databases yielded the genes of diabetes-related illnesses.After taking the intersection of the two,protein-protein interaction network(PPI)was established using STRING and Cytoscape programs to search for key genes with strong correlation and complete gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis.Use AutoDockTools and Pymol programs to complete protein molecule docking validation.Results The pharmacologically-related study revealed 63 targets associated with paeoniflorin,4 758 genes related to diabetes,and 50 intersection targets.15 key genes including vascular endothelial growth factor A(VEGFA),epidermal growth factor receptor(EGFR),V-Ha-ras harvey(HRAS),V-src sarcoma(SRC)and heat shock protein hs 90-alpha(HSP90AA1)were screened.RAs-associated protein 1,Ras,calcium and other signaling pathways were obtained by KEGG pathway analysis.Molecular docking results showed that paeoniflorin had good binding ability with key genes.Conclusion Paeoniflorin can treat diabetes through multiple targets and pathways,and this mechanism can provide a basis for the application of paeoniflorin in anti diabetes and drug research and development.