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.

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.

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.

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.

BACKGROUND:Diabetic ulcers are a common complication of diabetes mellitus,which is manifested as foot ulcers complicated with infection,long treatment cycle,high disability rate and mortality rate,and brings a heavy burden to patients and social care. OBJECTIVE:To review the mechanism of action and the latest treatment progress of traditional Chinese medicine(TCM)in the treatment of diabetic ulcers,and to provide a basis for further theoretical research and clinical application. METHODS:CNKI,WanFang Database and PubMed database were searched for relevant literature using the keywords of"diabetic ulcer,medicinal herb,inflammation,interleukin-1β,interleukin-6,tumor necrosis factor,hypersensitive C-reactive protein,γ-interferon,interleukin-4,interleukin-10"in Chinese and English,respectively.The relevant literature in recent years was searched,and finally 75 articles were included for review. RESULTS AND CONCLUSION:The high glucose environment of the body will increase the level of pro-inflammatory cytokines,so that diabetic ulcer wounds are in a state of chronic inflammatory response for a long time,and difficult to heal or even not heal.TCM has summed up a lot of experience in the long-term struggle with diabetic ulcer.At present,TCM divides diabetic ulcers into four syndrome types:dampness and heat poison syndrome,blood and blood stasis obstruction pattern,heat poison injury Yin pattern,and Qi and blood deficiency syndrome,as well as representative prescriptions for treatment.According to their clinical characteristics,diabetic ulcers can be also divided into three stages:primary,middle and late stages.Different treatment methods are proposed:"clear method,""warm and clear combined use"and"maintenance method."Under the guidance of dialectical typing and staging of TCM,TCM monomers,extracts and compounds inhibit the inflammatory response and promote the healing of diabetic ulcers by down-regulating the expression of pro-inflammatory factors and/or up-regulating the expression of anti-inflammatory factors.Compared with modern medicine,TCM has significant advantages in the treatment of diabetic ulcers.There are many TCM monomers,extracts and compounds for the treatment of diabetic ulcers,such as angelica,curcumin,improved Chonghe ointment,Sanhuang blood exhaustion prescription and sore-ulcer I.formula,etc.It has been found that TCM for the treatment of diabetic ulcers is mainly heat-clearing and detoxifying,invigorating blood circulation and removing blood stasis,and amassing sores and muscle-building drugs,and the frequency of use,treatment scope and therapeutic effect of TCM compounds are obviously better than those of TCM monomers and extracts.Among them,the most commonly used are the Sanhuang blood exhaustion prescription and the sore-ulcer I as well as prescription for the treatment of damp heat toxicity syndrome and Zizhu ointment for the treatment of non-ischemic diabetic ulcers.However,there are also some shortcomings in the treatment of diabetic ulcers with TCM.First,there are few clinical syndrome studies on diabetic ulcers.Secondly,there are a wide variety of TCM monomers,extracts and compounds for the treatment of diabetic ulcers,and the relevant research is insufficiently in-depth.Finally,the research on the mechanism underlying TCM treatment of diabetic ulcers is still in the preliminary exploration stage,and the mechanism of action still needs to be further explored.In the future,it is necessary to strengthen the research on the pharmacology of TCM and the clinical syndrome of diabetic ulcers,analyze the potential targets and related signaling pathways of TCM in the treatment of diabetic ulcers,give full play to the therapeutic advantages of TCM with multiple targets,multiple pathways,multiple levels and multiple systems,and develop TCM with significant efficacy,active ingredients and clear targets.

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.

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.