This study aims to establish an antibody detection method for porcine deltacoronavirus (PDCoV). The recombinant proteins PDCoV-N1 and PDCoV-N2 were expressed via the prokaryotic plasmid pColdII harboring the N gene sequence of the PDCoV strain CH/XJYN/2016. The reactivity and specificity of PDCoV-N1 and PDCoV-N2 with anti-PEDV sera were analyzed after the recombinant proteins were analyzed by SDS-PAGE and purified by the Ni-NTA Superflow Cartridge. Meanwhile, Western blotting and indirect immunofluorescence assay were carried out separately to validate the recombinant proteins PDCoV-N1 and PDCoV-N2. Finally, we established an indirect ELISA method based on the recombinant protein PDCoV-N2 after optimizing the conditions and tested the sensitivity, specificity, and reproducibility of the method. Then, the established method was employed to examine 102 clinical serum samples. The recombinant protein PDCoV-N2 showed low cross-reactivity with anti-PEDV sera. The optimal conditions of the indirect ELISA method based on PDCoV-N2 were as follows: the antigen coating concentration of 1.25 μg/mL and coating at 37 ℃ for 1 h; blocking by BSA overnight at 4 ℃; serum sample dilution at 1:50 and incubation at 37 ℃ for 1 h; secondary antibody dilution at 1:80 000 and incubation at 37 ℃ for 1 h; color development with TMB chromogenic solution at 37 ℃ for 10 min. The S/P value ≥ 0.45, ≤0.38, and between 0.45 and 0.38 indicated that the test sample was positive, negative, and suspicious, respectively. The testing results of the antisera against porcine epidemic diarrhea virus (PEDV), porcine circovirus 2 (PCV2), transmissible gastroenteritis virus (TGEV), foot-and-mouth disease virus (FMDV), and African swine fever virus (ASFV) showed that the S/P values were all less than 0.38. The testing results of the 800-fold diluted anti-PDCoV sera were still positive. The results of the inter- and intra-batch tests showed that the coefficients of variation of this method were less than 10%. Clinical serum sample test results showed the coincidence rate between this method and neutralization test was 94.12%. In this study, an ELISA method for the detection of anti-PDCoV antibodies was successfully established based on the truncated N protein of PDCoV. This method is sensitive, specific, stable, and reproducible, serving as a new method for the clinical diagnosis of PDCoV.
Animals ; Enzyme-Linked Immunosorbent Assay/methods* ; Swine ; Antibodies, Viral/blood* ; Recombinant Proteins/genetics* ; Deltacoronavirus/isolation & purification* ; Coronavirus Infections/virology* ; Swine Diseases/diagnosis* ; Coronavirus Nucleocapsid Proteins ; Sensitivity and Specificity

Objective To investigate the effect and mechanism of methyltransferase-like 3(METTL3)on the pro-liferation,migration,and secretion of inflammatory factors by synovial fibroblasts from rheumatoid arthritis(RA).Methods The expression of METTL3 in synovial tissue(SF)from 25 patients with rheumatoid arthritis and 25 pa-tients with osteoarthritis was detected by RT-qPCR and immunohistochemistry,respectively.The concentration of RNA m6A was detected by ELISA.RA synovial fibroblasts were isolated and cultured,and divided into NC(nor-mal control)group,hi-METTL3(overexpression of METTL3)group,si-METTL3(knock-down METTL3)group,and STM2457(METTL3 specific inhibitor)intervention group.Cell proliferation was detected by CCK-8 method.Apoptosis was detected by flow cytometry.And the concentrations of interleukin-6(IL-6),interleukin-17A(IL-17A),receptor activator of nuclear factor-kappa B ligand(RANKL),and osteoprotegerin(OPG)in the superna-tant of cell culture were detected by ELISA.Results Compared with synovial tissue of osteoarthritis,the expres-sion of mRNA m6A and METTL3 in synovial tissue of RA significantly increased(P<0.05).After overexpression of METTL3,the expression of m6A in synovial fibroblasts increased.The proliferation and migration abilities of SF in hi-METTL3 group were significantly improved,and their apoptosis did not change significantly.The secretion of cytokines IL-6 and RANKL of SF in hi-METTL3 group significantly increased,while the OPG significantly de-creased(P<0.05).After interfering with METTL3 expression,the expression of m6A in synovial fibroblasts de-creased.Cell proliferation and migration of SF in siMETTL3 group significantly decreased.The secretion of cyto-kines IL-6 and RANKL significantly decreased,and OPG significantly increased(P<0.05).After intervention with METTL3 inhibitor STM2457,the proliferation and migration of synovial fibroblasts were significantly reduced,and the secretion of cytokines IL-6 and RANKL significantly reduced,and OPG significantly increased(P<0.05).There was no significant difference in the expression of IL-17A among each group.Conclusion METTL3 may promote the proliferation and migration of RA synovial fibroblasts,enhance the expression of IL-6 and RANKL,and inhibit the expression of OPG through RNA m6A methylation modification.

It collected relative policy documents systematically and analyzed the development process and policy objective evolution of Chinese basic medical insurance system.Based on this,it determined the performance assessment objectives of the designated medical institutions of basic medical insurance and constructed the assessment framework guided by the goal realization.Then,from the perspective of contract management and performance management,it determined the assessment elements of contract enforcement assessment system of designated medical institutions and the logical relationship between the elements,and to construct a conceptual model,which provides a reference for guiding the research of the contract enforcement assessment of the designated medical institutions of basic medical insurance.

Feces and intestinal contents of pigs suspected with porcine epidemic diarrhea virus were collected from a farm in Minqin County,Gansu Province,China.After the suspected positive sam-ples were detected by RT-PCR,Vero cells were used to isolate and culture them in vitro.The suc-cessfully isolated virus was identified in the laboratory,and its whole genome sequence was ana-lyzed for genetic evolution.The pathogenicity was evaluated by animal regression test.The results showed that typical syncytial lesions could be observed when the PEDV-positive treatment solu-tion was inoculated with Vero cells in the 4th generation,and the virus titer in the 6th generation reached 10-4 75TCID50/mL.PEDV-like virions with a diameter of about 100 nm and a round shape with obvious capsular membranes and spikes were observed by electron microscopy.Whole genome sequencing analysis showed that the total length of this strain was 28 085 bp,which was far from the G1 subtype represented by the classical strain CV777(96.6％),and had a high homology with the G2b strains BC-2011-1,IA1,USA/Colorado/2013 and WELL(98.6％).This indicated that the strain belonged to the G2b epidemic strain.The animal regression test showed that the 5-day-old piglets developed vomiting,acute watery diarrhea,emaciation and mental depression within 12 h after the attack,and the symptoms worsened and died within 24 h.After autopsy,the infected piglets could be observed with stomach swelling,high intestinal heave,thin and transparent intesti-nal wall,and undigested milk clots inside.In summary,a PEDV G2b epidemic strain was success-fully isolated and identified in this study,and its whole genome sequence and pathogenicity were analyzed,providing research materials for future studies on PEDV gene function,pathogenic mech-anism and vaccine development.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Reasonable disciplinary planning is an important prerequisite for the high-quality development of multi-campus hospitals. In order to promote the rational layout and homogeneous management of multi-campus hospitals, a large public hospital explored multi-campus discipline planning from 2021 to 2022 based on preliminary research, following the principles of management integration, layout differentiation, resource optimization, maximum efficiency, and high-quality development, as well as according to the functional positioning of each hospital(headquarters, south, west, north, and infectious disease hospital), the needs of surrounding residents, and resource allocation. This practice had improved the development of advantageous specialties of sub-campus, promoted the innovation of technologies, and improved the overall medical service capacity of the hospital. The number of outpatients/emergency patients in the hospital had increased from 5.013 million in 2020 to 6.7 million in 2023, and the average length of stay had been shortened from 7.76 days to 6.17 days. It had initially achieved a disciplinary development strategy of " grouping, differentiation, and integration", which could provide reference and guidance for other public hospitals to promote the discipline development and construction for multi-campus hospitals.

To screen and identify the key host proteins interacting with the non-structural protein 15 (Nsp15) of porcine epidemic diarrhea virus (PEDV). The IP/pull-down assay and mass spectrometry were employed to screen and identify the host proteins interacting with Nsp15. The interaction between the host protein and Nsp15 was studied by co-immunoprecipitation and laser scanning confocal microscopy. Finally, Western blotting and RT-qPCR were employed to examine the interaction between SLC25a3 and PEDV. The recombinant eukaryotic expression vector pcDNA3.1(+)-Flag-Nsp15 was successfully constructed, and the host protein SLC25a3 interacting with PEDV Nsp15 was screened out. An interaction existed between SLC25a3 and Nsp15, and SLC25a3 significantly inhibited PEDV replication in a dose-dependent manner. SLC25a3 inhibits PEDV replication. The results of this study provide a basis for deciphering the role and mechanism of SLC25a3 in the host immune response to PEDV infection.
Porcine epidemic diarrhea virus/genetics* ; Viral Nonstructural Proteins/metabolism* ; Animals ; Swine ; Virus Replication ; Coronavirus Infections/veterinary* ; Swine Diseases/metabolism*

ObjectiveTo establish a multi index fusion hand grip fatigue prediction model to evaluate the power-assisted effect of the glove exoskeleton prototype for extravehicular clothing. MethodsBP neural network algorithm was used to establish a hand fatigue prediction model. The related factors of hand fatigue were determined with isometric grasping fatigue experiment, and the input variables of BP neural network were determined as cylinder diameter, grasping force, grasping duration and root mean square of electromyography. The fatigue data corresponding to variables of each group were obtained through experiments and subjective fatigue measurement scales, and a fatigue evaluation model based on multi-source fusion of BP neural network algorithm was established. The relationship model between fatigue and assistance effect was established, and the assistance effect of the exoskeleton prototype was evaluated through the degree of fatigue relief. ResultsThe correlation coefficient was 0.974 between the predicted results of the model and the target value. Moreover, it effectively predicted the assistance effect of different prototypes. ConclusionThe BP neural network model established by combining the grasping strength, grasping object parameters and human electromyography can predict hand fatigue, which can be used to evaluate the assistance effect of glove exoskeleton and other hand aids.

Objective:In order to explore the impact of corona virus disease 2019(COVID-19)on the hospitalization of children with bronchiolitis and to improve clinicians′ understanding of the characteristics of bronchiolitis during the COVID-19 epidemic.Methods:This was a multicenter clinical study, and the data have been collected from 23 children′s medical centers in China.All the clinical data were retrospectively collected from children with bronchiolitis who were hospitalized at each study center from January 1, 2019 to December 31, 2021.The results included gender, age at hospitalization, length of stay, respiratory syncytial virus(RSV) test results, severity rating, ICU treatment, and the total number of children hospitalized with respiratory tract infection during the same period.The clinical data of children with bronchiolitis in 2019 before COVID-19 epidemic and in 2020、2021 during COVID-19 epidemic were statistically analyzed and compared.Results:According to a summary of data provided by 23 children′s medical centers, there were 4 909 cases of bronchiolitis in 2019, 2 654 cases in 2020, and 3 500 cases in 2021.Compared with 2019, the number of bronchiolitis cases decreased by 45.94% in 2020 and 28.70% in 2021.In 2019, 2020 and 2021, there were no significant differences in gender ratio, age, and duration of hospitalization.Compared with 2019, the ratio of bronchiolitis to the total number of hospitalizations for respiratory tract infection decreased significantly in 2020 and 2021( χ2=12.762, P<0.05; χ2=84.845, P<0.05).The proportion of moderate to severe bronchiolitis cases in both 2020 and 2021 was lower than that in 2019, and the difference was statistically significant ( χ2=4.054, P<0.05; χ2=8.109, P<0.05).There was no statistically significant difference in the proportion of bronchiolitis cases requiring ICU treatment between 2019, 2020, and 2021 ( χ2=1.914, P>0.05).In 2019, a total of 52.60%(2 582/4 909) of children with bronchiolitis underwent RSV pathogen testing, and among them, there were 708 cases with RSV positive, accounting for 28.00%.In 2020, 54.14%(1 437/2 654) of children with bronchiolitis underwent RSV pathogen testing, and there were 403 cases with RSV positive, accounting for 28.04%.In 2021, 66.80%(2 238/3 500) of children with bronchiolitis underwent RSV pathogen testing, and there were 935 cases with RSV positive, accounting for 41.78%.Compared with 2019 and 2020, the RSV positive rate in 2021 showed a significant increase( χ2=99.673, P<0.05; χ2=71.292, P<0.05). Conclusion:During the COVID-19 epidemic, the implementation of epidemic prevention and control measures reduced the hospitalization rate and severity of bronchiolitis, but did not reduce the positive rate of RSV detection.

Twenty general practice residents who received training in Tongren Hospital from 2018 to 2020 were enrolled. The residents undertook rotation in chronic disease care clinic of the general medicine department, and a multi-disciplinary integrated teaching mode was applied. The theoretical knowledge test, skill assessment and self-evaluation were performed before and after rotation. The 360-degree evaluation was carried out when leaving the department (study group), the results were compared with that of internal medicine (control group) in the same period. The teaching effect was comprehensively evaluated. The scores of theoretical knowledge and skill assessment after rotation were significantly higher than those before rotation ( P<0.01). The self-evaluated ability was significantly higher than that at admission, especially in general practice thinking (7.85±0.88), doctor-patient communication (7.95±0.69), health education (7.80±0.70) and disease management (8.20±0.62) ( P<0.01). The 360-degree evaluation showed that the study group was better than the control group in interpersonal communication skills [(4.75±0.44) vs. (4.41±0.50)], system-based work ability[(4.75±0.44) vs. (4.31±0.47)], practice-based learning and improvement [(4.80±0.41) vs. (4.33±0.48)], patient care [(4.75±0.44) vs. (4.28±0.46)] and training mode satisfaction [(4.85±0.37) vs. (4.38±0.49)] (all P<0.05). The study indicates that the multi-disciplinary integrated teaching mode applied for rotation in the chronic disease care clinic of the general medicine department can improve the teaching effect and raise the teaching quality for general practice residency training.