1.Effect of Serum Containing Zhenwutang on Apoptosis of Myocardial Mast Cells and Mitochondrial Autophagy
Wei TANG ; Meiqun ZHENG ; Xiaolin WANG ; Zhiyong CHEN ; Chi CHE ; Zongqiong LU ; Jiashuai GUO ; Xiaomei ZOU ; Lili XU ; Lin LI
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(3):11-21
ObjectiveTo explore the effect of serum containing Zhenwutang on myocardial mast cell apoptosis induced by angiotensin Ⅱ (AngⅡ) and the mechanism of the correlation between apoptosis and mitochondrial autophagy. MethodsIn this experiment, AngⅡ and serum containing Zhenwutang with different concentrations were used to interfere with H9C2 cardiomyocytes for 24 h, and the survival rate of H9C2 cardiomyocytes was detected by cell counting kit-8 (CCK-8) to screen the optimal concentration for the experiment. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of B-type natriuretic peptide (BNP) in cell culture supernatant, and immunofluorescence was used to detect the cell surface area to verify the construction of the myocardial mast cell model. Subsequently, the experiment was divided into a blank group (20% blank serum), a model group (20% blank serum + 5×10-5 mol·L-1 AngⅡ), low-, medium-, and high-dose (5%, 10% and 20%) serum containing Zhenwutang groups, an autophagy inhibitor group (1×10-4 mol·L-1 3-MA), and autophagy inducer group (1×10-7 mol·L-1 rapamycin). The apoptosis level of H9C2 cells and the changes of mitochondrial membrane potential were detected by flow cytometry. The lysosomal probe (Lyso Tracker) and mitochondrial probe (Mito Tracker) co-localization was employed to detect autophagy. Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), Bcl-2-related X protein (Bax), and cytochrome C (Cyt C) in apoptosis-related pathways and the relative mRNA expression of ubiquitin ligase (Parkin), phosphatase and tensin homolog (PTEN)-induced kinase 1 (PINK1), and p62 protein in mitochondrial autophagy-related pathways. Western blot was used to detect cleaved Caspase-3, cleaved Caspase-9, Bax, Bcl-2, and Cyt C in apoptosis-related pathways, phosphorylated ubiquitin ligase (p-Parkin), phosphorylated PTEN-induced kinase 1 (p-PINK1), p62, and Bcl-2 homology domain protein Beclin1 in mitochondrial autophagy-related pathways, and the change of microtubule-associated protein 1 light chain 3 (LC3) Ⅱ/Ⅰ ratio. ResultsCCK-8 showed that when the concentration of AngⅡ was 5×10-5 mol·L-1, the cell activity was the lowest, and there was no cytotoxicity. At this concentration, the surface area of cardiomyocytes was significantly increased (P<0.01), and the content of BNP in the supernatant of culture medium was significantly increased (P<0.05). Therefore, AngⅡ with a concentration of 5×10-5 mol·L-1 was selected for the subsequent modeling of myocardial mast cells. Compared with the blank group, the model group and the autophagy inhibitor 3-MA group had a significantly increased apoptosis rate (P<0.01) and significantly decreased mitochondrial membrane potential (P<0.01). The results of immunofluorescence co-localization showed that compared with the blank group, the model group had a significantly decreased number of red and green fluorescence spots. The results of Real-time PCR showed that compared with that in the blank group, the relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 in the model group was significantly up-regulated (P<0.01), while the relative mRNA expression of Bcl-2, Parkin, and PINK1 was significantly down-regulated (P<0.01). In addition, the relative protein expression of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 was significantly up-regulated (P<0.01). The LC3Ⅱ/Ⅰ was significantly decreased, and the relative protein expression of Bcl-2, p-Parkin, p-PINK1, and Beclin1 was significantly down-regulated (P<0.01). Compared with the model group, the serum containing Zhenwutang groups and the autophagy inducer group had significantly decreased apoptosis rate (P<0.01), and the decrease ratio of mitochondrial membrane potential is significantly lowered (P<0.01) in a dose-dependent manner. Additionally, both red and green fluorescence spots became more in these groups. In the 3-MA group, the number of red and green fluorescence spots decreased significantly. The relative mRNA expression of Bax, Caspase-3, Caspase-9, Cyt C, and p62 was significantly down-regulated (P<0.05, P<0.01), while that of Bcl-2, Parkin, and PINK1 was significantly up-regulated (P<0.01). In the serum containing Zhenwutang groups, the relative protein expression levels of Bax, cleaved Caspase-3, cleaved Caspase-9, Cyt C, and p62 were significantly down-regulated (P<0.05,P<0.01). The LC3Ⅱ/Ⅰ was significantly increased, and the relative protein expression levels of Bcl-2, p-Parkin, p-PINK1, and Beclin1 were significantly up-regulated (P<0.01). ConclusionThe serum containing Zhenwutang can reduce the apoptosis of myocardial mast cells and increase mitochondrial autophagy. This is related to the inhibition of intracellular Bax/Bcl-2/Caspase-3 apoptosis pathway and regulation of Parkin/PINK1 mitochondrial autophagy pathway.
2.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
3.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
4.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
5.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
6.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
7.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
8.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
9.Research on the Influence Mechanism of Primary Physicians'Salary Satisfaction on the Turnover Tendency in Wuhan
Mian XIA ; Yining GAO ; Haomiao LI ; Meizhou JIANG ; Mingou WANG ; Shuai JIANG
Chinese Health Economics 2024;43(5):13-17
Objective:To analyze the influence mechanism of primary physicians'salary satisfaction on the turnover tendency,explore the moderating role of professional identity,and to provide a basis for enhancing the stability of the primary care physician workforce.Methods:The questionnaire survey method was used to study the salary satisfaction,professional identity and turnover ten-dency among 298 primary care doctors in the main urban area of Wuhan,and 284 valid questionnaires were obtained.The data were statistically analyzed.Results:The propensity to leave scores of primary care physicians aged 40 to 50 years,with bachelor's de-gree and 11 to 15 years of work were(3.01±0.87),(3.02±0.90),and(3.03±0.86),respectively,with high were high turnover tendency;primary care physicians'salary satisfaction and professional identity had a negative effect on the propensity to leave(P<0.001);and professional identity had a negative effect on the salary satisfaction and turnover tendency(P<0.001).Conclusion:Primary physicians'propensity to leave is more pronounced in the high-quality human capital group,with moderate security but in-sufficient incentives for pay;higher levels of professional identity reduce the relationship between pay satisfaction and propensity to leave.
10.Research on Water Extraction Process of Flos Lonicerae-Fructus Forsythuae Based on Network Pharmacology and Design Space
Ting CUI ; Meizhou LI ; Lifan GAN ; Jiaming LIN ; Lijin LIANG ; Xingpeng HUANG ; Zhipeng ZHANG
Journal of Nanjing University of Traditional Chinese Medicine 2024;40(1):47-60
OBJECTIVE To optimize the water extraction process of Flos Lonicerae-Fructus Forsythuae and determine the range of water extraction process parameters.METHODS The active components were screened by network pharmacology,and the indica-tor ingredients were determined in combination with the quality markers under the relevant terms of Chinese Pharmacopoeia 2020 edition and the literature.Take extraction yield and the extraction rate of the indicative component as the critical quality attributes of the water extraction process to screen critical process parameters.The mathematical model was established by Box-Behnken experimental design to investigate the interaction between CQAs and CPPs and build the design space of the water extraction process of Flos Lonicerae-Fruc-tus Forsythuae.RESULTS The extraction percentages of phenolic acids,forsythoside A and forsythin were screened as the index components;specifications of medicinal slices,extraction time and water addition were the key process parameters.Based on the estab-lishment and optimization of the design space,the optimum water extraction process was obtained as follows:the medicinal slice of Lian-Qiao was broken into 0.8-1.2 cm,adding 12 times the amount of water in the first and extract for 30-50 min,10 times the a-mount of water in the second and extract for 25-30 min.CONCLUSION The verification results show that the measured value ob-tained by using the design space method to optimize the water extraction process is close to the predicted value,indicating that the method is stable and reliable,which can provide ideas for its further process development and quality control for the couple medicines of Flos Lonicerae-Fructus Forsythuae.

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