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*

Objective To test the reliability and validity of the Chinese version of the Caregiver Contri-bution to Self-Care of Chronic Illness Inventory in the caregivers of elderly patients with COPD.Methods The general information questionnaire and the Chinese version of the Caregiver Contribution to Self-Care of Chronic Illness Inventory were used to conduct questionnaire survey on 220 caregivers of elderly patients with COPD treated or hospitalized in the respiratory medicine department of a tertiary hospital in Hebei Province from October 2022 to February 2023,and the reliability and validity of the scale were tested.Results The i-tem-level content validity index was 0.833-1.000,the content validity index at the level of unanimity scale was 0.842 and the content validity index of the average scale level was 0.973.The confirmatory factor analysis showed that all the fitting indicators of the total scale and the caregiver's contribution to self-care mainte-nance,monitoring contribution and management contribution subscale were within the acceptable range.The Cronbach's α coefficients of the total scale and the three subscales were 0.918,0.865,0.830 and 0.897,re-spectively.The split-half reliabilities of the total scale and the three subscales were 0.843,0.855,0.842 and 0.906,respectively.The test-retest reliabilities of the total scale and the three subscales were 0.849,0.734,0.751 and 0.773,respectively.Conclusion The Chinese version of the Caregiver Contribution to Self-Care of Chro-nic Illness Inventory has good reliability and validity,and can be used as a reliable tool to measure the caregivers con-tribution to elderly COPD patients self-care.

Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

Objective To explore the pathogenic roles of miR-21, estrogen (E2), and estrogen receptor (ER) in adenomyosis. Methods We examined the expression levels of miR-21 in specimens of adenomyotic tissue and benign cervical lesions using qRT-PCR. In primary cultures of cells isolated from the adenomyosis lesions, the effect of ICI82780 (an ER inhibitor) on miR-21 expression levels prior to E2 activation or after E2 deprivation were examined with qRT-PCR. We further assessed the effects of a miR-21 mimic or an inhibitor on proliferation, apoptosis, migration and autophagy of the cells. Results The expression level of miR-21 was significantly higher in adenomyosis tissues than in normal myometrium (P<0.05). In the cells isolated from adenomyosis lesions, miR-21 expression level was significantly higher in E2 activation group than in ER inhibition+E2 activation group and the control group (P<0.05);miR-21 expression level was significantly lower in cells in E2 deprivation+ER inhibition group than in E2 deprivation group and the control group (P<0.05). The adenomyosis cells transfected with miR-21 inhibitor showed inhibited proliferation and migration, expansion of mitochondrial endoplasmic reticulum, increased lysosomes, presence of autophagosomes, and increased cell apoptosis, while transfection of the cells with the miR-21 mimic produced the opposite effects. Conclusion MiR-21 plays an important role in promoting proliferation, migration, and anti-apoptosis in adenomyosis cells by altering the cell ultrastructure, which may contribute to early pathogenesis of the disease. In addition to binding with E2, ER can also regulate miR-21 through other pathways to participate in the pathogenesis of adenomyosis, thus having a stronger regulatory effect on miR-21 than E2.

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.