Objective To develop nanobodies with broad-spectrum reactivity,specificity,and high sensitivity that can be used for detecting multiple subtypes of influenza A virus,and to establish a nanobody-based enzyme-linked immunosorbent assay(ELISA)method.Methods Gene sequences of twelve nanobodies against influenza A virus were retrieved from the National Center for Biotechnology Information(NCBI)and nanobody databases.The nanoantibodies were prepared using molecular biological techniques including gene synthesis and recombinant expression.The binding activity,specificity,sensitivity,and affinity of these nanobodies were determined by ELISA screening and Gator affinity analysis.A double-antibody sandwich ELISA assay was established by combining the selected nanobody with a traditional mouse monoclonal antibody.Results Twelve nanobodies were expressed and purified.Two nanobodies capable of binding to multiple subtypes of influenza virus including H1,H3,H5,H7,and H9 were obtained and designated as VHH54 and KV108.Both nanobodies showed no cross-reactivity with other respiratory virus antigens.Furthermore,the KV108 nanobody exhibited the highest binding affinity,with a dissociation constant of 5.94×10-9mol/L for the influenza virus nucleoprotein(NP),and the lowest detection concentration for the NP antigen reached 0.00064 μg/mL.The double-antibody sandwich ELISA,using a combination of KV108 and a mouse monoclonal antibody,could sensitively detect the five common subtypes of influenza A virus(H1N1,H3N2,H5N1,H7N9,and H9N2).The lowest detection limit reached 110-403 PFU/mL,which was higher than that of the commercial colloidal gold kitfor influenza virus detection.Conclusion This study has identified a nanobody KV108,which is capable of binding to multiple subtypes of influenza virus,and established a nanobody-based ELISA method that can detect multiple subtypes of influenza A virus.This study can facilitate the development of nanobody-based influenza detection technologies.

Objective This study aims to investigate the selection intention of nursing interns and their influencing fac-tors,providing a scientific basis for nursing human resource management in hospitals and departments.Methods A convenience sampling method was used to conduct a questionnaire survey A questionnaire survey was conducted concerning the selection inten-tion on departments and influencing factors of intern nurses from several tertiary hospitals affiliated to three medical universities from July 2022 to December 2023.General data investigation and self-developed scales were employed,and descriptive analysis,chi-square test and other statistical methods were used to analyze the differences in department selection and related influencing factors among nursing interns.Results Analysis of 652 valid questionnaires revealed that surgery(65.80％)and operating room(38.84％)were the most preferred departments for nursing interns.The primary influencing factors included the working environment and equipment of the department(54.60％),as well as its cohesion and cultural atmosphere(51.53％).Pediat-rics(51.23％)and infectious medicine(48.77％)were found to be less favored by nursing interns.Their main concerns re-volved around work-related risks and population characteristics of patients(67.02％),along with work pressure and labor intensi-ty(65.49％).Epidemic factors have an impact on the attitudes and expectations of nursing interns towards their future practice(P＜0.01).Conclusion The choice of departments by nursing interns is influenced by various environmental,societal and personal factors;primarily based on departmental environment quality and team culture atmosphere considerations.During the in-ternship,nursing interns require more professional education guidance from both hospital administration and individual depart-ments in order to make informed career choices and plans.

Objective To explore the differential expression genes(DEGs)and related signaling pathways of the Qi deficiency and blood stasis(QDBS)and Yin deficiency and blood stasis(YDBS)syndromes in ischemic stroke(IS)at the molecular level,elucidating the potential mechanisms and biological basis of blood stasis syndrome in IS.Methods The mRNA expression profile dataset GSE100235 of the two syndromes of IS rats was downloaded from the GEO database,and the limma package of R language was used to screen DEGs.Subsequently,GO and KEGG analyses were performed using the DAVID database.The miRDB and miRWalk databases were used to predict the miRNAs of DEGs,and aa miRNA-mRNA regulatory network was constructed.A protein-protein interaction network was built using the STRING database,and the cytoHubba plug-in was utilized to identify the core target genes.The rat model of QDBS in IS was established by exhaustive swimming training combined with modified Longa's thread embolism,and the rat model of YDBS in IS was established by hydrocortisone injection and thread embolism.RT-qPCR was employed to validate the expression levels of core target genes in rat brain tissues of the two groups.Results A total of 47 DEGs were identified.These genes were mainly involved in biological processes such as neuronal apoptosis,inflammatory response,and adaptive immune response,as well as pathways related to lipid metabolism,atherosclerosis,and C-type lectin receptor signaling.The miRNA-mRNA interaction network consisted of 64 nodes and 55 edges.Five core target genes were obtained,including Ccl2,Selp,Timp1,Ccr1l1,and Fpr1.Conclusion There are significantly differentially expressed genes in QDBS and YDBS syndrome of IS rats.These genes are involved in a variety of biological processes and pathways,and are most closely related to lipid metabolism and atherosclerosis signaling pathways.

Objective This study aims to investigate the selection intention of nursing interns and their influencing fac-tors,providing a scientific basis for nursing human resource management in hospitals and departments.Methods A convenience sampling method was used to conduct a questionnaire survey A questionnaire survey was conducted concerning the selection inten-tion on departments and influencing factors of intern nurses from several tertiary hospitals affiliated to three medical universities from July 2022 to December 2023.General data investigation and self-developed scales were employed,and descriptive analysis,chi-square test and other statistical methods were used to analyze the differences in department selection and related influencing factors among nursing interns.Results Analysis of 652 valid questionnaires revealed that surgery(65.80％)and operating room(38.84％)were the most preferred departments for nursing interns.The primary influencing factors included the working environment and equipment of the department(54.60％),as well as its cohesion and cultural atmosphere(51.53％).Pediat-rics(51.23％)and infectious medicine(48.77％)were found to be less favored by nursing interns.Their main concerns re-volved around work-related risks and population characteristics of patients(67.02％),along with work pressure and labor intensi-ty(65.49％).Epidemic factors have an impact on the attitudes and expectations of nursing interns towards their future practice(P＜0.01).Conclusion The choice of departments by nursing interns is influenced by various environmental,societal and personal factors;primarily based on departmental environment quality and team culture atmosphere considerations.During the in-ternship,nursing interns require more professional education guidance from both hospital administration and individual depart-ments in order to make informed career choices and plans.

Objective To explore the differential expression genes(DEGs)and related signaling pathways of the Qi deficiency and blood stasis(QDBS)and Yin deficiency and blood stasis(YDBS)syndromes in ischemic stroke(IS)at the molecular level,elucidating the potential mechanisms and biological basis of blood stasis syndrome in IS.Methods The mRNA expression profile dataset GSE100235 of the two syndromes of IS rats was downloaded from the GEO database,and the limma package of R language was used to screen DEGs.Subsequently,GO and KEGG analyses were performed using the DAVID database.The miRDB and miRWalk databases were used to predict the miRNAs of DEGs,and aa miRNA-mRNA regulatory network was constructed.A protein-protein interaction network was built using the STRING database,and the cytoHubba plug-in was utilized to identify the core target genes.The rat model of QDBS in IS was established by exhaustive swimming training combined with modified Longa's thread embolism,and the rat model of YDBS in IS was established by hydrocortisone injection and thread embolism.RT-qPCR was employed to validate the expression levels of core target genes in rat brain tissues of the two groups.Results A total of 47 DEGs were identified.These genes were mainly involved in biological processes such as neuronal apoptosis,inflammatory response,and adaptive immune response,as well as pathways related to lipid metabolism,atherosclerosis,and C-type lectin receptor signaling.The miRNA-mRNA interaction network consisted of 64 nodes and 55 edges.Five core target genes were obtained,including Ccl2,Selp,Timp1,Ccr1l1,and Fpr1.Conclusion There are significantly differentially expressed genes in QDBS and YDBS syndrome of IS rats.These genes are involved in a variety of biological processes and pathways,and are most closely related to lipid metabolism and atherosclerosis signaling pathways.

Objective To identify stable reference genes for a comparison of the transcription levels of target host genes under viral infection in order to provide data for studies on interactions between the host and the influenza virus.Methods Reverse transcription quantitative real-time PCR(RT-qPCR)was performed to detect the relative expression levels of six candidate reference genes,including glyceraldehyde 3-phosphate dehydrogenase(GAPDH),β-actin,18S RNA,β2-microglobulin(B2M),ubiquitin-conjugating enzyme E2D2(UBE2D2),and ribosomal protein L37A(RPL37A)in classical cell models(A549 cells and THP-1 cells)under different conditions.The stability of the reference genes was evaluated using such methods as BestKeeper,GeNorm,NormFinder,and comparative A Ct method.Results The stability of reference genes varied depending on conditions.When such experimental factors as influenza virus infection and immune activation were taken into consideration,β-actin and GAPDH were identified as the most stable reference genes in A549 cells and THP-1 cells,followed by UBE2D2 and B2M.Conclusion The optimal reference genes in A549 cells and THP-1 cells under influenza virus infection or after being treated with interferons or LPS have been identified,which is of referential value for studying the mechanisms of viral infections.

As one of the flexible means of drug supervision， the “compassionate medicine use system” has become a new way to meet the drug accessibility of patients at present. The “compassionate medicine use system” in Britain， that is， Early Access to Medicine Scheme， intervenes in the drug development process of the applicant enterprises at an early stage through the cooperation of different government departments， which not only ensures patients obtain innovative drugs in time， but also accelerates the drug listing and payment and reimbursement process. At present， China has only made principled provisions on the “compassionate medicine use system”， but has not issued specific implementation rules. It is suggested that the access conditions of “compassionate medicine use”， the responsibilities and obligations of different subjects， the payment mechanism of expenses and how to promote the cooperation of relevant departments should be clearly defined as soon as possible in combination with the system experience of Britain， so as to form a standardized and operable “compassionate medicine use system” suitable for China’s national conditions.

The reform of medical insurance payment is a critical part of deepening the medical reform.It plays an important role in guiding the behavior of healthcare providers.The latest progress in the reform of health insurance payment in the US.is the imple-mentation of value-based payment models.The literature research method is applied to review the development of value-based pay-ment practices in the US.It finds the main value-based payment models in the United States are targeted hospitals and physicians as the incentive object,with the characteristics of the combination of reward and punishment,the combination of horizontal and ver-tical evaluation,the continuous improvement of evaluation system,and the full use of information means.On this basis,combined with the national conditions of China,it puts forward four suggestions on establishing effective incentive mechanism,formulating scientific evaluation system,insisting on continuous improvement mechanism and strengthening medical information construction for future reform of medical insurance.

The reform of medical insurance payment is a critical part of deepening the medical reform.It plays an important role in guiding the behavior of healthcare providers.The latest progress in the reform of health insurance payment in the US.is the imple-mentation of value-based payment models.The literature research method is applied to review the development of value-based pay-ment practices in the US.It finds the main value-based payment models in the United States are targeted hospitals and physicians as the incentive object,with the characteristics of the combination of reward and punishment,the combination of horizontal and ver-tical evaluation,the continuous improvement of evaluation system,and the full use of information means.On this basis,combined with the national conditions of China,it puts forward four suggestions on establishing effective incentive mechanism,formulating scientific evaluation system,insisting on continuous improvement mechanism and strengthening medical information construction for future reform of medical insurance.

The reform of medical insurance payment is a critical part of deepening the medical reform.It plays an important role in guiding the behavior of healthcare providers.The latest progress in the reform of health insurance payment in the US.is the imple-mentation of value-based payment models.The literature research method is applied to review the development of value-based pay-ment practices in the US.It finds the main value-based payment models in the United States are targeted hospitals and physicians as the incentive object,with the characteristics of the combination of reward and punishment,the combination of horizontal and ver-tical evaluation,the continuous improvement of evaluation system,and the full use of information means.On this basis,combined with the national conditions of China,it puts forward four suggestions on establishing effective incentive mechanism,formulating scientific evaluation system,insisting on continuous improvement mechanism and strengthening medical information construction for future reform of medical insurance.