Senecavirus A (SVA) is a major viral pathogen causing disease in pigs, and effective monitoring of SVA infection is critical for disease control. In this study, we aimed to develop a reliable ELISA method for rapidly detecting neutralizing antibodies against SVA. We used HEK293F cells to express an SVA-specific porcine Fab antibody and verified the biological activity of the Fab antibody by indirect ELISA, immunofluorescence assay, virus neutralization test, and Western blotting. The Fab antibody was biotinylated and used as a competitive antibody to establish a competitive ELISA (C-ELISA) for detecting neutralizing antibodies against SVA. We then evaluated the C-ELISA in terms of sensitivity, specificity, repeatability, and result agreement rate with the VNT. The results showed that we successfully prepared an SVA-specific porcine Fab antibody, which showed high affinity for SVA. We named this antibody 1M33Fab and designated it as Bio-1M33Fab after biotin labeling. The assay conditions were optimized as follows: the coating concentration of SVA particles being 1 μg/mL, the working concentration of Bio-1M33Fab being 0.5 μg/mL, the optimal serum dilution of 1:10, and the optimal dilution of enzyme-labeled avidin being 1:30 000. At a percent inhibition (PI) of 47%, the assay demonstrated the highest sensitivity (96.88%) and specificity (100%), with no cross-reactivity observed with the positive sera of major porcine viral diseases. The intra-assay coefficient of variation ranged from 1.12% to 7.34%, while the inter-assay coefficient of variation ranged from 1.10% to 8.97%, indicating good repeatability. In the detection of 224 clinical pig serum samples, C-ELISA and VNT showed a result agreement rate of 93.75%. In conclusion, we successfully develop a C-ELISA method for detecting neutralizing antibodies against SVA by using a porcine-derived Fab antibody, which lays a foundation for the development of detection kits.
Animals ; Swine ; Antibodies, Neutralizing/immunology* ; Enzyme-Linked Immunosorbent Assay/methods* ; Immunoglobulin Fab Fragments/immunology* ; Antibodies, Viral/immunology* ; Picornaviridae/immunology* ; Humans ; HEK293 Cells ; Swine Diseases/diagnosis* ; Picornaviridae Infections/diagnosis*

This study started from the connotation of public welfare and, from the perspective of equal rights and responsibilities, interpreted the public welfare responsibilities and entitlements of hospitals. It developed corresponding quantitative measurement tools and constructed a Public Welfare Rights and Responsibilities Index. Public welfare responsibilities were reflected in the provision of equitable, accessible, appropriate, and reasonable basic medical and public health services. These responsibilities were quantified using the market price of basic medical services, the average cost per case-mix index unit, and public service expenditures. Entitlements were reflected in financial subsidies, tax exemptions, and land policy support. Based on this framework, the study applies the Public Welfare Rights and Responsibilities Index to evaluate 147 public hospitals in City S from 2019 to 2021. The results show that the overall level of public welfare among public hospitals in City S was relatively high (with an average index of 2.39), but showed a slight downward trend. Differences were observed across hospital grades and types, with secondary-level hospitals and general hospitals demonstrating relatively higher levels of public welfare. This study could provide a practical and quantifiable method for measuring hospital public welfare within an equal rights and responsibilities framework, offering a novel analytical tool and empirical evidence to support policy formulation and performance evaluation.

Objective To compare and find an optimal model for predicting the risk of DKD occurrence in patients with type 2 diabetes mellitus(T2DM).Methods A total of 2005 patients with T2DM were enrolled in this study from The Second Hospital of Shijiazhuang City during December 2017 to December 2022.All the subjects were divided into a training set(n=1403)and a validation set(n=602)according to the ratio of 3∶1 by simple random sampling.With the occurrence of DKD as the outcome variablein the training set,important feature variables were screened by LASSO regression.Six different machine learning models were established according to the feature variables,thenthe optimal model was determined by comparison,and anonlinerisk predictor for DKD occurrence was constructed in patients with T2DM.Results Taking the occurrence of DKD as the outcome variable in the training set,the results of LASSO regression analysis showed that the optimal value of the model was 10-fold cross validation lambda.1se=0.01662473,and 15 characteristic variables with nonzero coefficient were screened out to be related to the occurrence of DKD.The data included sex,age,family history of DM,DM duration,LDL-C,HbA1c,WBC,PDW,Scr,urine α1-microglobulin,urine β2-microglobulin,urine microalbumin,hypertension,hypokalemia,and DR.In the training set and validation set,the prediction performance of XGBoost model was better than that of other models(AUC=0.872,0.893,95%CI 0.853～0.891,0.865～0.921),the sensitivity was 0.779,0.863,and the specificity was 0.721,0.758,respectively.The F1 scores were 0.774 and 0.787.DCA analysis showed that the XGBoost model had a greater net benefit and threshold probability.According to the XGBoost model,the online predictor of DKD risk in T2DM patients was laid out,and two patients were selected for application,the results showed that the predictive value of the model was 0.185 in non-DKD patients,and the predictive value was 0.510 in DKD patients.Conclusions The XGBoost model is the best model for predicting the occurrence of DKD in T2DM patients,and an online predictor was successfully built.

Objective To summarize the outcomes of a pricing reform pilot program in a tertiary hospital in Ningxia and offer relevant suggestions.Methods The key measures used in the hospital for adjusting and optimizing the medical revenue structure were summarized.Trend and ratio analysis was employed to compare the economic performance indicators before and af-ter the pilot reform.Results During the pilot period,key performance indicators for national tertiary public hospitals demonstra-ted steady improvements.Meanwhile,the annual growth rate of medical revenue was 4.7 percentage points lower than in 2019,with medical service revenue increased by 5.6 percentage points.Moreover,outpatient and inpatient costs per visit were lower than in same-level,same-type hospitals in the region and in provincial-level general hospitals nationwide.Unreasonable growth in total medical costs was effectively curbed,and the pricing ratio of medical services became more rational.Annual management re-duced drug,consumable,examination and testing revenue by 200 million yuan,while employee average wage income rose by 10.95％from 2019 and employee satisfaction reached 85.2％,ensuring the hospital's healthy and sustainable development.Conclusion The pricing reform pilot project implemented in Ninxia provides a reference for public hospitals to adjust and opti-mize their medical revenue structure.

Immune thrombocytopenia(ITP)is an autoimmune disease characterized primarily by bleeding and fatigue.ITP-related fatigue can substantially affect health-related quality of life and has been recognized as a treatment indication in the latest guidelines.However,its pathogenesis remains unclear,and targeted therapies are lacking.This paper explores the theoretical foundation of the"excessive fire consuming healthy qi"theory,linking two key conceptual pairs:"excessive fire"with inflammation and"healthy qi consumption"with mitochondrial dysfunction.Chronic inflammation leads to continuous"qi consumption"and damage to mitochondrial structure and function,ultimately resulting in symptoms of qi deficiency such as tiredness and fatigue.The imbalance between qi and fire,leading to imbalance of yin and yang,is the primary pathogenesis of ITP-related fatigue,with the disease mainly affecting the spleen.Considering the etiology and pathogenesis of ITP,and relative equilibrium of yin-yang,the therapeutic approach of invigorating spleen and benefiting qi to promote the production of blood serves as a core treatment strategy.Previous research has demonstrated the clinical efficacy of Jianpi Yiqi Shexue Formula,consisting of milkvetch root,tangshen,Indian bread,largehead atractylodes rhizome,India madder root,and ass hide glue in treating patients with ITP-related fatigue.Designed to invigorate the spleen and regulate qi and blood,this formula aligns with the pathogenesis of ITP-related fatigue characterized by deficiency of spleen qi and stomach qi.The formula also modulates the expression levels of key proteins and genes involved in mitochondrial energy metabolism and alleviates the immune dysfunction.These findings highlight the potential for further research into the mechanism of ITP-related fatigue,particularly by using inflammation-mediated mitochondrial energy metabolism as a promising entry point.

Objective To compare the benefits and drawbacks of primary patch expansion versus pericardial tube right ventricular-pulmonary artery connection in patients diagnosed with pulmonary atresia with ventricular septal defect (PA/VSD). Methods A retrospective study was conducted on patients diagnosed with PA/VSD who underwent primary right ventricular-pulmonary artery connection surgery at our center between 2010 and 2020. Patients were categorized into two groups based on the type of right ventricular-pulmonary artery connection: a pericardial tube group and a patch expansion group. Clinical data and imaging findings were compared between the two groups. Results A total of 51 patients were included in the study, comprising 31 males and 20 females, with a median age of 12.57 (4.57, 49.67) months. The pericardial tube group included 19 patients with a median age of 17.17 (7.33, 49.67) months, while the patch expansion group consisted of 32 patients with a median age of 8.58 (3.57, 52.72) months. In both groups, the diameter of pulmonary artery, McGoon index, and Nakata index significantly increased after treatment (P<0.001). However, the pericardial tube group exhibited a longer extracorporeal circulation time (P<0.001). The reoperation rate was notably high, with 74.51% of patients requiring further surgical intervention, including 26 (81.25%) patients in the patch expansion group and 12 (63.16%) patients in the pericardial tube group. No statistical differences were observed in long-term cure rates or mortality between the two groups (P＞0.005). Conclusion In patients with PA/VSD, both patch expansion and pericardial tube right ventricular-pulmonary artery connection serve as effective initial palliative treatment strategies that promote pulmonary vessel development and provide a favorable foundation for subsequent radical operations. However, compared to the pericardial tube approach, the patch expansion technique is simpler to perform and preserves some intrinsic potential for pulmonary artery development, making it the preferred procedure.

AIM: To conduct a CiteSpace-based visualization analysis of the literature on traditional Chinese medicine(TCM)treatment of diabetic retinopathy(DR)from January 2014 to January 2024.METHODS: The literatures were retrieved from the China National Knowledge Infrastructure(CNKI)database. Co-occurrence, clustering, and emergence analyses of annual publications, authors, research institutions, and keywords were performed using Note Express and CiteSpace software to generate knowledge graph.RESULTS: A total of 643 Chinese-language articles were included. The annual publication trend was characterized by fluctuations. The primary research institutions were Liaoning University of Traditional Chinese Medical and Eye Hospital China Academy of Chinese Medical Sciences. Twenty high-frequency keywords were identified, including diabetes, TCM, and review, along with emergent terms such as Qiming granules and data mining. The research focus in 2014 was on exploring therapeutic methods and the efficacy of proprietary Chinese medicines. From 2016 to 2018, the research perspective expanded to include symptoms and multi-factorial analysis. Between 2018 and 2020, there was an in-depth investigation into evidence types and disease stages. After 2019, the emphasis shifted towards TCM-specific therapies and patient-centered concepts. From 2022 to 2024, innovation in research methods and the theory of TCM collaterals theory emerged as new hotspots.CONCLUSION: The research on TCM treatment for DR is still in an ascending phase. The primary research hotspot currently centers on Qiming granules. Future trends are likely to focus on the efficacy and mechanisms of action. The combined application of different therapies and long-term follow-up experimental research could provide a more robust basis for evidence-based medicine, unearth the potential of TCM, and promote its development in the prevention and treatment of DR.

Objective: To establish a method for determining the molecular weight and molecular weight distribution of Poly(Lactide-co-Glycolide Acid) (PLGA) using Size Exclusion Chromatography-Refractive Index-Multiangle Laser Light Scattering (SEC-RI-MALLS) and Size Exclusion Chromatography-Refractive Index (SEC-RID), and to compare the results obtained from these two methods.
Methods: For SEC-RI-MALLS, tetrahydrofuran was used as the mobile phase, Shodex GPC KF-803L was employed as the chromatographic column with a flow rate of 1 mL·min^-1, column temperature at 30 ℃, and an injection volume of 100 μL. For SEC-RID, tetrahydrofuran was also used as the mobile phase, Agilent PLgel 5 μm MIXD-D was used as the chromatographic column with a flow rate of 1 mL·min^-1, column temperature at 30 ℃, differential detector temperature at 35 ℃, and an injection volume of 20 μL. The molecular weight and molecular weight distribution were calculated using Agilent’s GPC software. The newly established methods were validated methodologically, and the molecular weight and molecular weight distribution of 13 batches of samples were determined.
Results: The precision, accuracy, stability, and repeatability tests for SEC-RI-MALLS showed RSD values of 1.35%, 1.58%, 1.53%, and 1.26%, respectively. The SEC-RID method exhibited good linearity (r=0.999 9), with RSD values for precision, accuracy, stability, and repeatability tests (n=6) of 2.05%, 1.62%, 1.30%, and 2.97%, respectively. The results obtained from SEC-RI-MALLS were lower than those from SEC-RID, and the molecular weight distribution coefficient was smaller, but the results from the paired T-test performed with the value measured by SEC-RID method and the value measured by SEC-RI-MALLS method multiplied a conversion coefficient of 1.5 showed no significant difference between the two methods.
Conclusion: Both methods are stable and reliable, and can be used for the determination of PLGA molecular weight and molecular weight distribution based on the specific situations.

Objective:To investigate the role of heme oxygenase-1(HO-1)regulating autophagy and its impact on the intracel-lular growth of Mycobacterium abscessus(M.abs).Methods:The standard strain of M.abs ATCC19977 was used to stimulate mouse RAW264.7 macrophages with a specified multiple of infection(MOI)for a specified time.Western blot analysis was performed to detect the expression levels of HO-1 protein and autophagy related proteins Atg5,LC3Ⅱ and p62.Macrophages were pretreated with HO-1 specific inducer CoPP and enzyme activity inhibitor SnPP for 12 h,and incubated with M.abs for 2 h.Amikacin removed extra-cellular bacteria and continued to culture until the specified time,and cell activity was detected by CCK-8 method.Western blot and LysoTracker Red were used to detect the regulatory relationship between HO-1 protein and autophagy related proteins.Bacterial survival and TNF-α secretion levels were detected by colony count and ELISA.Results:Compared with the control group,M.abs could induce increased expression of HO-1,Atg5,LC3Ⅱ and p62 proteins.Over-expression and inhibition of HO-1 were found to effectively regu-late the expressions of Atg5,LC3Ⅱ and p62 induced by M.abs.LysoTracker Red,colony count,and ELISA results demonstrated that SnPP promoted the increasement of intracellular lysosomes,significantly inhibited M.abs intracellular proliferation,and reduced TNF-α secretion.Conclusion:Inhibition of HO-1 can enhance the autophagy flow induced by M.abs and reduce the intracellular growth of M.abs,which provides scientific basis for the development of targeted drugs targeting HO-1.