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.

AIM To optimize the processing technology of red Notoginseng Radix et Rhizoma and evaluate its blood tonifying activity.METHODS On the basis of a single factor experiment,with steaming temperature,steaming time,drying temperature,and drying time as influencing factors,the total contents of notoginsenoside R1,ginsenoside Rg1,Rb1,Rk3,Rh4,and 20(R)-ginsenoside Rg3 as evaluation indicators,Box-Behnken response surface method ology was used to optimize the processing technology.Upon the anemic mouse models jointly induced by 1-acetyl-2-phenylhydrazine(APH)and cyclophosphamide(CTX),the investigation of the blood tonifying activity of red Notoginseng Radix et Rhizoma was carried out in contrast to that of the steamed Notoginseng Radix et Rhizoma.RESULTS The optimal conditions,contributing saponin content of 8.326%and RSD of 0.087%,were determined as follows:steaming temperature of 130℃,steaming time of 4 hours,drying temperature of 60℃,and drying time of 48 h.The pharmacological activity revealed that the different processing techniques were responsible for the different blood enriching activity of notoginseng,with red Notoginseng Radix et Rhizoma displaying a better efficacy than that of steamed Notoginseng Radix et Rhizoma.CONCLUSION This stable and feasible method can be used to control the production of red Notoginseng Radix et Rhizoma.

This study constructed a LHCGR-CRE-luc-HEK293 transgenic cell line according to the activation of the cAMP signaling pathway after recombinant human chorionic gonadotropin binding to the receptor. The biological activity of recombinant human chorionic gonadotropin was assayed using a luciferase assay system. The relative potency of the samples was calculated using four-parameter model. And the method conditions were optimized to validate the specificity, relative accuracy, precision and linearity of the method. The results showed that there was a quantitative potency relationship of human chorinonic gonadotropin (hCG) in the method and it was in accordance with the four-parameter curve. After optimization, the conditions were determined as hCG dilution concentration of 2.5 μg·mL^-1, dilution ratio of 1∶4, cell number of 10 000-15 000 cells/well, and induction time of 6 h. The method had good specificity, relative accuracy with relative bias ranging from -8.9% to 3.4%, linear regression equation correlation coefficient of 0.996, intermediate precision geometric coefficient of variation ranging from 3.3% to 15.0%, and linearity range of 50% to 200%. This study successfully established and validated a reporter gene method to detect hCG biological activity, which can be used for hCG biological activity assay and quality control.

Objective To develop and validate a deep learning model for automatic identification of liver CT contrast-enhanced phases.Methods A total of 766 patients with liver CT contrast-enhanced images were retrospectively collected.A three-phase classification model and an arterial phase(AP)classification model were developed,so as to automatically identify liver CT contrast-enhanced phases as early arterial phase(EAP)or late arterial phase(LAP),portal venous phase(PVP),and equilibrium phase(EP).In addition,221 patients with liver CT contrast-enhanced images in 5 different hospitals were used for external validation.The annotation results of radiologists were used as a reference standard to evaluate the model performances.Results In the external validation datasets,the accuracy in identifying each enhanced phase reached to 90.50%-99.70%.Conclusion The automatic identification model of liver CT contrast-enhanced phases based on residual network may provide an efficient,objective,and unified image quality control tool.

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.