One of the technical bottlenecks limiting the high yield of 1,4-butanediamine is the insufficient tolerance of strains to 1,4-butanediamine. Enhancing the tolerance of strains to 1,4-butanediamine is therefore a primary challenge that needs to be addressed for the construction of strains with high yields of 1,4-butanediamine. Staphylococcus pasteuri 326180 exhibits exceptional tolerance to high-concentration 1,4-butanediamine, serving as both an ideal model for studying the mechanism underlying the 1,4-butanediamine tolerance and a novel host for constructing strains capable of efficiently producing 1,4-butanediamine. However, for both the research on the tolerance mechanism and the modification of chassis strains, gene editing of S. pasteuri needs to be carried out at the molecular level. The research objective of this paper is to establish a genetic manipulation system for S. pasteuri, laying foundation for subsequent studies on tolerance mechanism and the modification of chassis strains. This study systematically optimized the electroporation conditions, including key parameters such as the growth phase of cells, electric field strength, electroporation buffer, and recovery medium, successfully establishing an electroporation method for S. pasteuri. Additionally, we constructed the gene editing plasmid pCpfOA by replacing the resistance expression cassette, optimized the selection markers for gene editing, and finally established a CRISPR/Cpf1-based gene editing technology for S. pasteuri, achieving an editing efficiency of 90%. The genetic manipulation system of S. pasteuri established in this study provides technical support for research into the tolerance mechanism of this bacterium and the genetic modification of chassis strains.
Staphylococcus/drug effects* ; Gene Editing/methods* ; Electroporation/methods* ; Plasmids/genetics* ; CRISPR-Cas Systems ; Genetic Engineering/methods*

AIM: To investigate the effects of agkis-trodon halys venom anti-tumor component (AHVAC-) on the biological behavior of gastric cancer MKN-28 cells. METHODS: Gastric cancer MKN-28 cells were treated with the experimental concentrations (5, 10, 15 μg/mL) of AHAVC- for 24 h. Cell proliferation and toxicity assay (cell counting kit-8, CCK-8) was used to detect the inhibition rates of the cells in different concentrations of AHVAC-. The migration ability of the cells was evaluated by wound-healing and Transwell assay. The apoptosis were observed by laser confocal microscopy with annexin V-mCherry/DAPI double staining, and the apoptosis rates were analyzed by flow cytometry with annexin V-FITC/PI double fluorescence staining. The protein level of Caspease-3 was determined by Western blot. RESULTS: Compared with normal control group, the results of AHVAC- concentration groups showed that with the increase of AHVAC- concentration, the proliferative activity of MN-28 cells decreased gradually (P<0.01), the cell migration ability decreased gradually (P<0.01), and the cell apoptosis rate increased (P<0.05). The expression of apoptosis-related protein Caspease-3 was up-regulated (P<0.01). CONCLUSION: AHVAC- inhibits proliferation and migration of gastric cancer MSN-28 cells and induces apoptosis.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.

The operating room was the core department of a hospital,and its operational efficiency had a significant impact on the high-quality development of a hospital.An analysis has revealed that low efficiency and irrational allocation in the operating room were mainly due to the lack of operational regulations and norms,the unreasonable arrangement of surgical specialties,and the unbalanced allocation of supporting resources.To address these issues,the First Affiliated Hospital of Zhengzhou University has taken into account the overall allocation of resources for the central operating room and the central operating room,and formulated strategies to improve operational efficiency,including adjusting the operational mechanism,optimizing the structure of surgical specialties,and providing corresponding supporting resources.Based on the adjustment of surgical structure,the implementation effect of the program was measured and evaluated,which provided practical strategies for optimizing operating room efficiency in hospitals.