Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

ObjectiveTo investigate whether co-activated mesenchymal stem cells（MSCs） exert therapeutic effects against schistosomiasis by modulating macrophage polarization. MethodsTwenty adult male Balb/c mice were randomly divided into four groups： uninfected， infected， MSC-treated， and MSC^TLR4+IFN-γ-treated groups. The Schistosoma japonicum infection model was established via abdominal patch method with cercariae. At week 5 post-infection， praziquantel was administered orally for antiparasitic treatment. At week 6， mice received either MSCs treatments （with or without pre-activation） or no treatment. Body weight changes were monitored weekly. Hepatic pathological alterations were evaluated via HE and Masson staining. RT-qPCR was used to assess α-SMA and collagen （Col-I， Col-Ⅲ） mRNA levels to quantify fibrosis. The mRNA levels of hepatic inflammatory cytokines and matrix metalloproteinases（MMP） were analyzed to explore fibrotic mechanisms. The expressions of i-Nos and Arg-1 in liver tissues were detected by RT-qPCR， and the ratio of M1 or M2 macrophages was detected by immunofluorescence staining， aiming to analyze the correlation between MSCs treatment and macrophage polarization. An in vitro co-culture system validated direct MSC-macrophage interactions. ResultsCompared with the infected group， the MSC^TLR4+IFN-γ group exhibited increased body weight gain （P< 0.01）， reduced hepatic granulomatous lesion area （P< 0.001）， and decreased α-SMA， Col-I， and Col-Ⅲ mRNA levels （P< 0.01）. Additionally， the MSC^TLR4+IFN-γ group showed reduced TNF-α and IL-1β expression （P< 0.05）， as well as elevated MMP2， Mmp9， and MMP13 levels （P< 0.01）. The MSC^TLR4+IFN-γ group showed higher expression of M2 marker Arg-1 mRNA compared with the infection group （P < 0.001） ， while the expression of M1 marker i-Nos decreased （P< 0.05）. Immunofluorescence confirmed a lower i-Nos+ cell ratio （P< 0.05） and higher F4/80⁺CD206⁺ cell ratio （P< 0.000 1） in the MSC^TLR4+IFN-γ group compared with the infection group. In vitro co-culture experiments further demonstrated that MSC^TLR4+IFN-γ promoted Arg-1 expression， suppressed pro-inflammatory cytokine i-Nos and TNF-α levels， consistent with ELISA results. ConclusionsThis study reveals that TLR4 and IFN-γ co-activated MSCs alleviate Schistosoma japonicum-induced hepatic fibrosis， potentially through modulating macrophage polarization toward the M2 phenotype. This mechanism may suppress inflammation and enhance extracellular matrix degradation， providing a therapeutic strategy for schistosomiasis-associated liver fibrosis.

Objective To construct a competency-based and functioning-oriented training curriculum system for inclusive education teachers in primary and secondary schools based on World Health Organization International Classification of Functioning,Disability and Health(ICF)and rehabilitation competency framework(RCF).Methods Based on the policy documents of inclusive education from United Nations Educational,Scientific and Cultur-al Organization and the European Union,and aligning with China's national policies such as the 14th Five-Year Plan for Special Education Development and Improvement,this study systematically analyzed the core competen-cies required for primary and secondary inclusive education teachers using ICF and RCF,and developed a compe-tency-based and functioning-oriented training curriculum system.Results The core competencies for primary and secondary school teachers encompassed four dimensions under the con-text of inclusive education.In the dimension of professional identity and ethics,teachers needed to establish in-clusive education values and be able to use ICF to comprehensively understand students'functioning perfor-mance and developmental needs.In the dimension of inclusive teaching and support practices,it was emphasized the ability for inclusive instructional design based on functioning assessment,including applying universal design for learning,creating barrier-free learning environments and implementing multiple assessments.In the dimen-sion of collaboration and communication,it was needed to build home-school-community support networks and possess communication skills for interprofessional teamwork.In the dimension of continuous reflection and pro-fessional development,it was focused on teachers'continuous learning and professional growth.A curriculum system comprising four major modules including concepts and foundations,core skills for teaching and support,collaboration and expansion,and practical reflection and research were constructed based on this competency model.Conclusion Based on ICF and RCF,a competency-based and functioning-oriented training curriculum system has been established,providing a theoretical framework and practical pathway for improving the quality of inclusive edu-cation teachers in China.

Objective To construct a competency-based and functioning-oriented training curriculum system for inclusive education teachers in primary and secondary schools based on World Health Organization International Classification of Functioning,Disability and Health(ICF)and rehabilitation competency framework(RCF).Methods Based on the policy documents of inclusive education from United Nations Educational,Scientific and Cultur-al Organization and the European Union,and aligning with China's national policies such as the 14th Five-Year Plan for Special Education Development and Improvement,this study systematically analyzed the core competen-cies required for primary and secondary inclusive education teachers using ICF and RCF,and developed a compe-tency-based and functioning-oriented training curriculum system.Results The core competencies for primary and secondary school teachers encompassed four dimensions under the con-text of inclusive education.In the dimension of professional identity and ethics,teachers needed to establish in-clusive education values and be able to use ICF to comprehensively understand students'functioning perfor-mance and developmental needs.In the dimension of inclusive teaching and support practices,it was emphasized the ability for inclusive instructional design based on functioning assessment,including applying universal design for learning,creating barrier-free learning environments and implementing multiple assessments.In the dimen-sion of collaboration and communication,it was needed to build home-school-community support networks and possess communication skills for interprofessional teamwork.In the dimension of continuous reflection and pro-fessional development,it was focused on teachers'continuous learning and professional growth.A curriculum system comprising four major modules including concepts and foundations,core skills for teaching and support,collaboration and expansion,and practical reflection and research were constructed based on this competency model.Conclusion Based on ICF and RCF,a competency-based and functioning-oriented training curriculum system has been established,providing a theoretical framework and practical pathway for improving the quality of inclusive edu-cation teachers in China.

Natural killer cells(NK)are important innate immune cells that do not require prior antigen exposure and can directly recognize and attack virus-infected cells and tumor cells.The activation and effector functions of NK cells are regulated by a balance of signals delivered through their surface activating receptors and inhibitory re-ceptors,which bind to ligands on target cells to achieve cytotoxicity via"induced self"and"missing self"recogni-tion models.The killing mechanisms of NK cells primarily include release of cytotoxic granules such as perforin and granzymes to induce target cell lysis,death receptor-mediated apoptosis,secretion of various cytokines,chemokines and growth factors to coordinate with other immune cells in killing tumor cells,thereby generating secondary im-mune responses and antibody-dependent cellular cytotoxicity(ADCC).

Objective To explore the optimal grouping scheme of diagnosis related groups (DRG) for acquired immunodeficiency syndrome(AIDS) patients and develop cost criteria to provide a reference for implementing DRG payment reform in this region. Methods Information on the first pages of 1987 cases of AIDS patients from Nantong AIDS designated hospitals between 2018 and 2022 was collected, the influencing factors of hospitalization costs were analyzed by applying univariate and multiple linear regression and screening out the classification nodes, and the DRG grouping scheme was constructed by using a decision tree model. Results Complications or comorbidities, number of other diagnoses, and case type were used as classification nodes to form a total of 6 case combinations and corresponding hospitalization cost criteria. The difference in hospitalization cost between groups was statistically significant (P<0. 001), the reduction in variation(RIV) value was 51. 0％, and the coefficient of variation(CV) value of each group was less than 1 (0. 33～0. 63), with good inter-group heterogeneity and intra-group homogeneity. Conclusion The grouping scheme constructed based on the decision tree model is more reasonable, and the standard cost can objectively reflect the actual level of medical consumption of patients, providing a reference for improving DRG grouping and cost payment for AIDS patients in the region.

Objective:To establish a method for detecting serum non-ceruloplasmin-bound copper (NCC) by immunoprecipitation-inductively coupled plasma mass spectrometry (IP-ICP-MS) and evaluate its clinical application.Methods:Methodological evaluation research. Immunoprecipitation was first used to separate serum ceruloplasmin, followed by detection of serum NCC levels using ICP-MS. Two levels of quality control serum were reconstituted to determine the limit of detection (LOD), limit of quantitation (LOQ), accuracy, and precision of the self-developed method. The serum samples of 131 healthy individuals (healthy group) and 69 first-time diagnosed Wilson′s disease (WD) patients(WD group) from November 2023 to June 2024 in the Affiliated Hospital of Institute of Neurology, Anhui University of Traditional Chinese Medicine were collected. Using self-developed method detected the serum NCC levels, established the healthy reference intervals, and NCC levels between the two groups were compared using the Mann-Whitney U test. Results:The calibration curve exhibited excellent lineary across the concentration range of 1.562 to 300.000 μg/L, as demonstrated by coefficient of determination ( R2) of 0.999. The self-developed NCC method exhibited that the LOD was 0.99 μg/L, the LOQ was 3.29 μg/L, the accuracy (spike and recovery experience) was 87.67%?106.27%, and the intra-batch and inter-batch imprecision expressed as coefficient of variation ( CV) was 2.8%?7.3%, and the healthy reference range was 34.31-71.79 μg/L. The serum NCC levels in the WD group were 102.39 (74.38, 144.04) μg/L, which was significantly higher than those in healthy group (51.45±10.34) μg/L ( Z=?7.967, P<0.01). Conclusions:The established IP-ICP-MS method for detecting serum NCC meets the required analytical performance criteria. It is simply operative, highly sensitive, and provides accurate and reliable results, which could be used for clinical detection.

Objective To explore the optimal grouping scheme of diagnosis related groups (DRG) for acquired immunodeficiency syndrome(AIDS) patients and develop cost criteria to provide a reference for implementing DRG payment reform in this region. Methods Information on the first pages of 1987 cases of AIDS patients from Nantong AIDS designated hospitals between 2018 and 2022 was collected, the influencing factors of hospitalization costs were analyzed by applying univariate and multiple linear regression and screening out the classification nodes, and the DRG grouping scheme was constructed by using a decision tree model. Results Complications or comorbidities, number of other diagnoses, and case type were used as classification nodes to form a total of 6 case combinations and corresponding hospitalization cost criteria. The difference in hospitalization cost between groups was statistically significant (P<0. 001), the reduction in variation(RIV) value was 51. 0％, and the coefficient of variation(CV) value of each group was less than 1 (0. 33～0. 63), with good inter-group heterogeneity and intra-group homogeneity. Conclusion The grouping scheme constructed based on the decision tree model is more reasonable, and the standard cost can objectively reflect the actual level of medical consumption of patients, providing a reference for improving DRG grouping and cost payment for AIDS patients in the region.

The management of antibiotic-resistant, bacterial biofilm infections in skin wounds poses an increasingly challenging clinical scenario. Pseudomonas aeruginosa infection is difficult to eradicate because of biofilm formation and antibiotic resistance. In this study, we identified a new benzothiazole derivative compound, SN12 (IC₅₀ = 43.3 nmol/L), demonstrating remarkable biofilm inhibition at nanomolar concentrations in vitro. In further activity assays and mechanistic studies, we formulated an unconventional strategy for combating P. aeruginosa-derived infections by targeting the two-component (Gac/Rsm) system. Furthermore, SN12 slowed the development of ciprofloxacin and tobramycin resistance. By using murine skin wound infection models, we observed that SN12 significantly augmented the antibacterial effects of three widely used antibiotics-tobramycin (100-fold), vancomycin (200-fold), and ciprofloxacin (1000-fold)-compared with single-dose antibiotic treatments for P. aeruginosa infection in vivo. The findings of this study suggest the potential of SN12 as a promising antibacterial synergist, highlighting the effectiveness of targeting the two-component system in treating challenging bacterial biofilm infections in humans.

ObjectiveTo evaluate the clinical efficacy and safety of Tongluo Mingmu capsules in the treatment of diabetic retinopathy with blood stasis， collateral obstruction， and Qi and Yin deficiency syndrome. MethodA randomized， double-blind， positive-control， and multi-center clinical trial design method was used. 416 patients with diabetic retinopathy with blood stasis， collateral obstruction， and Qi and Yin deficiency syndrome in four test centers were included （the ratio of the treatment group to the control group was 3∶1）. On the basis of standardized hypoglycemic treatment， the treatment group was given both four Tongluo Mingmu capsules and two Calcium Dobesilate capsule agents three times a day， while the control group were given both two Calcium Dobesilate capsules and four Tongluo Mingmu capsule agents three times a day. The course of treatment was 12 weeks. The curative effect of Tongluo Mingmu capsules was evaluated by comparing the comprehensive curative effect of diabetic retinopathy， traditional Chinese medicine（TCM） syndrome score， corrected visual acuity， fundus changes， fundus fluorescence angiography， and other curative effect indexes before and after treatment in the two groups. At the same time， general examination， laboratory examination， and adverse events were performed to evaluate the safety of the drug. ResultThe baseline demographic data and disease characteristics of the treatment group and the control group were balanced and comparable， with the difference not statistically significant. After 12 weeks of treatment， the total effective rate of the comprehensive curative effect of diabetic retinopathy in the treatment group （61.0%， 189/310） was better than that in the control group （44.1%， 45/102）， and the difference was statistically significant （χ²=8.880， P<0.01）. The total effective rate of TCM syndromes in the treatment group （88.4%， 259/293） was better than that in the control group （69.9%， 65/93）， and the difference was statistically significant （χ²=17.927， P<0.01）. The disappearance rate of dry eyes （χ²=8.305）， dull complexion （χ²=4.053）， lassitude （χ²=10.267）， shortness of breath （χ²=8.494）， and dry stool （χ²=8.657） in the treatment group was higher than that in the control group， and the difference between the groups was statistically significant （P<0.05， P<0.01）. In terms of improving corrected visual acuity （χ²=8.382）， fundus changes （χ²=6.026） ， the treatment group was significantly better than the control group （P<0.05）. During the trial， the incidence of adverse events in the treatment group and the control group was 1.3% and 2.9%， respectively. There was no significant difference between the two groups. In addition， there were no serious adverse events and adverse events leading to withdrawal in both groups. ConclusionTongluo Mingmu capsules can improve the comprehensive curative effect of diabetic retinopathy and enhance the efficacy of TCM syndromes， visual acuity， fundus changes， and fundus fluorescein angiography， with great safety. Therefore， it can provide a new alternative therapeutic drug for patients with diabetic retinopathy.