1.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.
2.The median effective dose of ciprofol combined with sufentanil to in-hibit tracheal intubation reaction in patients undergoing intracranial aneurysm embolization
Fangzhou LU ; Meijuan LIU ; Qiong ZENG ; Wenbin ZHANG ; Jun LU
Chinese Journal of Clinical Pharmacology and Therapeutics 2025;30(8):1092-1098
AIM:To investigate the median effec-tive dose of ciprofol combined with sufentanil to in-hibit tracheal intubation reaction in patients under-going intracranial aneurysm embolization.METH-ODS:Forty-five patients who underwent emboliza-tion for intracranial aneurysms were divided into two groups according to age:non-elderly group(aged 18-64 years)and elderly group(aged>65 years).Patients in the two groups were first given intravenous ciprofol,of which the initial dose of cip-rofol was 0.4 mg/kg and sufentanil 0.3 μg/kg and rocuronium 0.6 mg/kg were respectively injected according to BIS value and modified observer's as-sessment of alertness and sedation(MOAA/S)score.Then,endotracheal intubation was defined as 3 minutes after the induction of the study drug.Dixon sequential method was adopted,and the dose of ciprofol for the next patient was deter-mined according to whether the tracheal intuba-tion reaction was positive(Positive reaction of tra-cheal intubation was defined as the patient's kino-motor reaction such as coughing during tracheal in-tubation or the increase of MAP or HR within 2 minutes after intubation was greater than 20%of the basic value).When the tracheal intubation indi-cated a positive response,the next patient was raised by one gradient,otherwise,it was lowered by one gradient until the study ended at 7 crosses.The common dose ratio of adjacent patients was 1∶1.1.The Probit probability method was used to cal-culate the 50%effective dose(ED50),95%effective dose(ED95)and the corresponding 95%confidence interval(CI)of ciprofol.Perioperative adverse reac-tions were recorded in both groups.RESULTS:In the non-elderly group,the ED50 of ciprofol inhibit-ing positive tracheal intubation reaction was 0.472 mg/kg(95%CI 0.419-0.565 mg/kg),and the ED95 was 0.567 mg/kg(95%CI 0.513-1.293 mg/kg).In the elderly group,the ED50 of ciprofol inhibiting tra-cheal intubation reaction was 0.409 mg/kg(95%CI 0.383-0.434 mg/kg)and the ED95 was 0.452 mg/kg(95%CI 0.430-0.591 mg/kg).CONCLUSION:The ED50 of ciprofol combined with sufentanil inhibiting positive tracheal intubation reaction was 0.472 mg/kg in non-elderly patients and 0.409 mg/kg in elder-ly patients.
3.Feasibility analysis of wide-detector CT coronary angiography using three-low scanning technology
Zixiong ZHANG ; Wenjun QIAO ; Jingjing ZHANG ; Manman YANG ; Jiayi CHEN ; Meijuan ZHOU
Chinese Journal of Medical Physics 2025;42(3):374-380
Objective To analyze various scanning parameters and contrast agent injection schemes,and explore the feasibility of three-low scanning technology characterized by lower radiation dose,slower injection rate and reduced contrast agent dosage in coronary angiography with wide-detector CT.Methods A total of 210 patients who underwent coronary angiography were recruited and randomly divided into group A(routine group,n=105)and group B(three-low group,n=105),and all of them were examined using the Revolution CT from the American GE company.Group A and group B adopted 120 and 100 kV tube voltages,respectively.The contrast injection rate and contrast agent dosage were calculated using formulas based on body mass index and beat per minute.Image quality was objectively evaluated using CT number,signal-to-noise ratio and contrast-to-noise ratio,and subjectively scored using Likert 5-point method.Results No statistical differences were found in the basic data between two groups(P>0.05).The CT numbers of the aortic sinus orifice in coronary artery images obtained from group A and group B were(442.70±58.26)and(454.11±62.36)HU,respectively.The differences between two groups in CT number,image score,signal-to-noise ratio,and contrast-to-noise ratio were trivial(P>0.05),while statistically significant differences were noted in injection rate,contrast agent dosage,tube current,and effective radiation dose(P<0.05).Group B reduced the effective radiation dose by 33.89%as compared with group A.Conclusion Based on the patient's body mass index and beat per minute,a personalized contrast agent injection scheme can be developed.Both routine group and three-low group can obtain image quality that meets imaging diagnosis requirements,and the latter has better image quality,demonstrating that the three-low scheme benefits the patients more for it can reduce the patient's radiation dose and contrast agent dosage while achieving higher image quality.
4.The median effective dose of ciprofol combined with sufentanil to in-hibit tracheal intubation reaction in patients undergoing intracranial aneurysm embolization
Fangzhou LU ; Meijuan LIU ; Qiong ZENG ; Wenbin ZHANG ; Jun LU
Chinese Journal of Clinical Pharmacology and Therapeutics 2025;30(8):1092-1098
AIM:To investigate the median effec-tive dose of ciprofol combined with sufentanil to in-hibit tracheal intubation reaction in patients under-going intracranial aneurysm embolization.METH-ODS:Forty-five patients who underwent emboliza-tion for intracranial aneurysms were divided into two groups according to age:non-elderly group(aged 18-64 years)and elderly group(aged>65 years).Patients in the two groups were first given intravenous ciprofol,of which the initial dose of cip-rofol was 0.4 mg/kg and sufentanil 0.3 μg/kg and rocuronium 0.6 mg/kg were respectively injected according to BIS value and modified observer's as-sessment of alertness and sedation(MOAA/S)score.Then,endotracheal intubation was defined as 3 minutes after the induction of the study drug.Dixon sequential method was adopted,and the dose of ciprofol for the next patient was deter-mined according to whether the tracheal intuba-tion reaction was positive(Positive reaction of tra-cheal intubation was defined as the patient's kino-motor reaction such as coughing during tracheal in-tubation or the increase of MAP or HR within 2 minutes after intubation was greater than 20%of the basic value).When the tracheal intubation indi-cated a positive response,the next patient was raised by one gradient,otherwise,it was lowered by one gradient until the study ended at 7 crosses.The common dose ratio of adjacent patients was 1∶1.1.The Probit probability method was used to cal-culate the 50%effective dose(ED50),95%effective dose(ED95)and the corresponding 95%confidence interval(CI)of ciprofol.Perioperative adverse reac-tions were recorded in both groups.RESULTS:In the non-elderly group,the ED50 of ciprofol inhibit-ing positive tracheal intubation reaction was 0.472 mg/kg(95%CI 0.419-0.565 mg/kg),and the ED95 was 0.567 mg/kg(95%CI 0.513-1.293 mg/kg).In the elderly group,the ED50 of ciprofol inhibiting tra-cheal intubation reaction was 0.409 mg/kg(95%CI 0.383-0.434 mg/kg)and the ED95 was 0.452 mg/kg(95%CI 0.430-0.591 mg/kg).CONCLUSION:The ED50 of ciprofol combined with sufentanil inhibiting positive tracheal intubation reaction was 0.472 mg/kg in non-elderly patients and 0.409 mg/kg in elder-ly patients.
5.Feasibility analysis of wide-detector CT coronary angiography using three-low scanning technology
Zixiong ZHANG ; Wenjun QIAO ; Jingjing ZHANG ; Manman YANG ; Jiayi CHEN ; Meijuan ZHOU
Chinese Journal of Medical Physics 2025;42(3):374-380
Objective To analyze various scanning parameters and contrast agent injection schemes,and explore the feasibility of three-low scanning technology characterized by lower radiation dose,slower injection rate and reduced contrast agent dosage in coronary angiography with wide-detector CT.Methods A total of 210 patients who underwent coronary angiography were recruited and randomly divided into group A(routine group,n=105)and group B(three-low group,n=105),and all of them were examined using the Revolution CT from the American GE company.Group A and group B adopted 120 and 100 kV tube voltages,respectively.The contrast injection rate and contrast agent dosage were calculated using formulas based on body mass index and beat per minute.Image quality was objectively evaluated using CT number,signal-to-noise ratio and contrast-to-noise ratio,and subjectively scored using Likert 5-point method.Results No statistical differences were found in the basic data between two groups(P>0.05).The CT numbers of the aortic sinus orifice in coronary artery images obtained from group A and group B were(442.70±58.26)and(454.11±62.36)HU,respectively.The differences between two groups in CT number,image score,signal-to-noise ratio,and contrast-to-noise ratio were trivial(P>0.05),while statistically significant differences were noted in injection rate,contrast agent dosage,tube current,and effective radiation dose(P<0.05).Group B reduced the effective radiation dose by 33.89%as compared with group A.Conclusion Based on the patient's body mass index and beat per minute,a personalized contrast agent injection scheme can be developed.Both routine group and three-low group can obtain image quality that meets imaging diagnosis requirements,and the latter has better image quality,demonstrating that the three-low scheme benefits the patients more for it can reduce the patient's radiation dose and contrast agent dosage while achieving higher image quality.
6.Catheter-associated and non-catheter-associated urinary tract infection in hospitalized patients in Suzhou City:a multicenter study on epidemiologi-cal characteristics
Jingxue LIU ; Xiuzhen WANG ; Meizhen QIAO ; Junji ZHANG ; Wei DING ; Shu-kai ZHU ; Meijuan JIN ; Xiaochao SONG
Chinese Journal of Infection Control 2025;24(8):1056-1065
Objective To explore the epidemiological characteristics and differences in antimicrobial resistance be-tween catheter-associated urinary tract infection(CAUTI)and non-CAUTI of healthcare-associated infection(HAI),and provide scientific basis for precise clinical prevention and control.Methods Based on the regional HAI surveillance platform in Suzhou City,urinary tract infection(UTI)surveillance data reported by 61 member units from January 2020 to December 2024 were analyzed retrospectively.Pathogen distribution,detection rate of multi-drug-resistant organisms(MDROs),and antimicrobial resistance spectrum characteristics of patients in the CAUTI group and non-CAUTI group were compared.Results The incidence of CAUTI in patients in CAUTI group was 0.99‰,the incidence of healthcare-associated UTI in patients in non-CAUTI group was 0.14%.There was statis-tically significant difference in the distribution of UTI pathogens between the two groups(P<0.05).The patho-gens of the CAUTI group were mainly Gram-negative bacteria(56.1%),with high proportions of Escherichia coli(19.6%)and Klebsiella pneumoniae(15.0%).In the non-CAUTI group,the proportion of Gram-negative bacteria was higher(64.7%).Antimicrobial susceptibility testing results showed that the resistance rates of Escherichia co-li to tobramycin,cephalosporins,and carbapenems in the CAUTI group were all higher than those in the non-CAU-TI group(all P<0.05).Except for tigecycline,the resistance rates of Klebsiella pneumoniae to other antimicrobial agents in the CAUTI group were all significantly different from the non-CAUTI group(all P<0.05).The resis-tance rates of Acinetobacterbaumannii to ticarcillin/clavulanic acid,quinolones,most cephalosporins,carbapenems,and aminoglycosides in the CAUTI group were higher than those of the non-CAUTI group(all P<0.05).The de-tection rates of MDROs were higher in the CAUTI group,especially that of carbapenem-resistant Klebsiella pneu-moniae,accounting for 57.8%.Conclusion There are significant differences in pathogen distribution and antimi-crobial resistance of UTI between the CAUTI group and the non-CAUTI group.It is necessary to establish a re-gional antimicrobial resistance surveillance system for pathogens in UTI,and provide basis for the rational use of an-timicrobial agents in clinical practice.
7.To study the relationship between lymphocyte subsets and renal clinicopathological features and prognosis in patients with IgA nephropathy
Shenglei ZHANG ; Ruicong TIAN ; Jingjing JIN ; Fan LU ; Meijuan CHENG ; Yaling BAI ; Jinsheng XU
The Journal of Practical Medicine 2025;41(3):352-357
Objective To examine the association between lymphocyte subsets and renal clinicopathological characteristics as well as prognosis in patients with IgA nephropathy(IgAN).Methods The retrospective analysis included general clinical data and pathological examination results of IgAN patients diagnosed by renal biopsy at the Fourth Hospital of Hebei Medical University from January 2018 to January 2022.Correlation tests were conducted to examine the relationship between lymphocyte subsets and other significant clinicopathological parameters.The optimal cut-off value of CD4+T determined using the Youden index,and patients were grouped accordingly.Kaplan-Meier survival curves and Cox regression analyses were employed to compare the low and high CD4+T lymphocyte groups among IgAN patients,identifying factors influencing renal function progression.The endpoint event was defined as a decrease in estimated glomerular filtration rate(eGFR)of≥30%from baseline,progression to end-stage renal disease(ESRD)[eGFR<15 mL/(min·1.73 m2)or initiation of renal replacement therapy],or all-cause mortality.Results Low CD4+T lymphocytes were significantly positively correlated with blood IgA levels and the proportion of glomerular crescents in IgAN patients(all P<0.05).This study included a total of 53 IgAN patients,divided into two groups based on CD4+T lymphocyte counts:20 patients in the low CD4+T lymphocyte group and 33 patients in the high CD4+T lymphocyte group.In the low CD4+T lymphocyte group,there was a higher proportion of males and a lower proportion of glomerular crescents(P<0.05).Kaplan-Meier survival analysis revealed that patients with low CD4+lymphocytes had a significantly lower cumulative renal survival rate(Log-Rank test χ2=4.188,P=0.041).Cox regression analysis indicated that low CD4+lymphocytes were an independent risk factor for the progression of renal function decline in IgAN patients(HR=2.614,95%CI:1.006~6.788,P=0.048).Conclusions Patients with higher levels of CD4+T lymphocytes exhibit a lower risk of adverse renal outcomes.In contrast,patients with IgA nephropathy and low CD4+T lymphocyte counts tend to have poorer renal survival rates.
8.Identification of associated factors and construction of a predictive model for membranous nephropathy patients with IgM deposition
Lei HE ; Yunhui ZHANG ; Jingjing JIN ; Meijuan CHENG ; Shenglei ZHANG ; Yaling BAI ; Jinsheng XU
Chinese Journal of Nephrology 2025;41(7):489-497
Objective:To explore the associated factors for membranous nephropathy (MN) patients with IgM deposition, and to construct a prediction model.Methods:This study was a retrospective cohort study. Patients diagnosed with MN with IgM deposition by renal biopsy in the Fourth Hospital of Hebei Medical University from February 2017 to December 2023 were retrospectively included. Clinical and pathological data were collected. The study population was randomized into a training set and a validation set at a 7:3 ratio. The endpoint event was defined as the remission of MN, and the patients were divided into remission group and non-remission group to compare the clinical and pathological examination results. Least absolute shrinkage and selection operator regression analysis and Cox regression analysis were used to explore the associated factors of poor prognosis of MN patients with IgM deposition. Internal validation was conducted using the validation set data. The clinical efficacy of the predictive model was evaluated by calculating the area under the receiver operating characteristic (ROC) curve and generating calibration curves. The total nomogram score for each patient was calculated based on the training set data, and the predictive performance was assessed by plotting the ROC curve. Patients were then stratified into low-risk and high-risk groups according to the optimal cut-off value derived from the ROC analysis of the total nomogram score. Kaplan-Meier survival analysis was performed to compare the remission rate between the two groups. Model performance was evaluated using the validation set.Results:A total of 200 MN patients with IgM deposition were included, and 49.0% of them achieved clinical remission. In the training set, statistically significant differences were observed in 24-hour urine protein quantification ( Z=-2.638, P=0.008), renal arteriolar wall thickening ( χ2=6.891, P=0.009), the proportion of patients receiving immunosuppressive therapy ( χ2=21.381, P<0.001), and the proportion of patients treated with corticosteroids combined with cyclophosphamide ( χ2=10.107, P=0.001). Through least absolute shrinkage and selection operator regression and Cox regression, 2 factors associated with clinical remission in MN patients with IgM deposition were simultaneously identified from 16 potential associated factors, including the use of immunosuppressants ( HR=3.823, 95% CI 2.055-7.113, P<0.001), and renal arteriolar wall thickening ( HR=0.428, 95% CI 0.221-0.831, P=0.012). Incorporating the clinical measurement of phospholipase A2 receptor (PLA2R) antibodies, a predictive model was established. The performance of the model was evaluated using the training dataset, yielding an area under the ROC curve of 0.731 (95% CI 0.648-0.814), with a sensitivity of 88.7% and a specificity of 55.1%. The optimal cut-off value was a total nomogram score of 41.7 points. The Kaplan-Meier survival analysis showed that the remission rate was significantly higher in the low-risk group than that of the high-risk group (Log-rank test, χ2=33.525, P<0.001). Model validation was performed using the validation dataset, which showed an AUC of 0.715 (95% CI 0.591-0.839), sensitivity of 70.4%, and specificity of 63.6%. Similarly, the Kaplan-Meier survival analysis demonstrated a significantly higher remission rate in the low-risk group than in the high-risk group (Log-rank test, χ2=8.467, P=0.004). Conclusion:A nomogram predictive model for remission of MN patients with IgM deposition, based on serum PLA2R antibody levels, the use of immunosuppressive therapy, and renal arteriolar wall thickening is developed. The model demonstrates a moderate clinical applicability.
9.Identification of endothelial cell key genes associated with pathogenesis and invasion of human venous malformations using single-nucleus RNA sequencing-based co-expression network analysis
Wenbo LIU ; Junjie LIN ; Meijuan ZHANG ; Chunjie YUAN ; Xiaojuan FENG ; Wenting JIAO ; Junbo QIAO ; Wenqiu WANG ; Bin FANG ; Changkuan CHEN
Chinese Journal of Preventive Medicine 2025;59(4):458-467
Objective:This study aimed to identify key genes in endothelial cell (EC) associated with the pathogenesis and progression of human venous malformations (VMs) through bioinformatics analysis, providing potential biomarkers for early screening and targeted therapy of VMs.Methods:A case-control study was conducted using surgically resected tissue specimens from VMs patients at the Third Affiliated Hospital of Zhengzhou University (from September 2021 to September 2023), with malformed venous tissues as the experimental group and distal normal venous tissues as controls. Single-nucleus RNA sequencing (snRNA-seq) was performed on paired experimental and control samples from four VM patients. High-dimensional weighted gene co-expression network analysis (hdWGCNA), combined with gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein-protein interaction (PPI) network analysis, identified critical genes. Validation experiments included 15 additional VM cases and controls using reverse transcription quantitative polymerase chain reaction (RT-qPCR), immunohistochemistry (IHC), and Western blot.Results:A total of 55 430 nuclei were captured using snRNA-seq, with 30 391 nuclei from the experimental group and 25 039 nuclei from the control group. Cluster analysis identified 22 distinct cell populations, which were annotated into 8 cell types. hdWGCNA revealed four modules associated with invasion, which were enriched in angiogenesis, integrin signaling, and cell adhesion according to GO analysis. KEGG pathway analysis indicated that the PI3K-AKT signaling pathway and focal adhesion are key regulatory mechanisms. PPI network analysis combined with cytoscape identified EGFL7, TEK, and FLT1 as key genes. RT-qPCR results demonstrated that the relative mRNA expression levels of these three genes in the experimental group (6.66±2.31, 1.86±0.62, 3.49±0.58) were significantly higher than those in the control group (1.05±0.14, 1.00±0.14, 1.06±0.25), with statistically significant differences ( t=9.37, 4.27, 11.20, P<0.05). Immunohistochemical analysis showed that the relative protein expression levels of these three genes in the cytoplasm of the experimental group (0.84±0.15, 0.68±0.14, 0.85±0.12) were also significantly higher than those in the control group (0.19±0.05, 0.23±0.06, 0.30±0.05), with statistically significant differences ( t=16.62, 5.93, 11.68, P<0.05). Western blot analysis confirmed that the relative protein expression levels of these three genes in the experimental group (0.35±0.04, 0.36±0.09, 0.31±0.04) were significantly higher than those in the control group (0.19±0.01, 0.13±0.02, 0.14±0.04), with statistically significant differences ( t=7.05, 4.61, 5.93, P<0.05). Conclusion:EGFL7, FLT1, and TEK in EC may play crucial roles in the occurrence and invasion of VMs.
10.Investigation on the status and barriers of non-accompanied care services in medical institutions in Zhejiang Province
Qiaomin TANG ; Jianping SONG ; Xiuqin FENG ; Leiwen TANG ; Yuping ZHANG ; Xiangying BAO ; Haiyan ZHENG ; Sumin MA ; Meijuan LAN
Chinese Journal of Nursing 2025;60(20):2507-2513
Objective To investigate the current status and barriers to implementing non-accompanied care services in medical institutions in Zhejiang Province,and to provide a basis for the improvement of standardized management of such services.Methods A self-designed questionnaire was conducted from September to October 2024 among all secondary and tertiary medical institutions in Zhejiang Province to assess the implementation status and barriers to non-accompanied care services.Results A total of 397 questionnaires were distributed,with 389 valid responses,yielding a valid response rate of 97.98%.Non-accompanied care services were implemented in 118 institutions(30.33%).Among these,90 institutions(76.27%)had established management systems for non-accompanied wards;71 institutions(60.17%)had a medical nursing assistant-to-bed ratio lower than 1∶5;41 institu-tions(34.75%)provided tiered training for medical nursing assistants;93 institutions(78.81%)required patients to bear the full cost of the service.Compared with secondary medical institutions,tertiary medical institutions have more complete management system for non-accompanied care services.The main obstacles hindering the development of non-accompanied care services include an imperfect management system for non-accompanied wards,a shortage of medical nursing assistants,a lack of standardized training for such assistants,inconsistent charging standards,and low acceptance among patients and their families.Conclusion The promotion of non-accompanied care services in medical institutions in Zhejiang Province has achieved initial success.However,challenges persist,including incomplete management systems,uneven development across hospital tiers,and imperfect charging mechanisms.It is recommended that relevant authorities strengthen policy support,enhance standardized training for healthcare nursing assistants,refine cost-sharing mechanisms,and improve the quality and sustainability of non-accompanied care services through multi-party collaboration.

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