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.

This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.

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.

This study aims to simultaneously detect three antibiotic resistance genes(blaNDM,mcr-1 and cfr).A triplex fluorescence quantitative PCR method was established.Plasmids,primers and probes were designed and optimized.The method could specifically detect blaNDM,mcr-1 and cfr,but not other antibiotic resistance genes.The R2 of the standard curves of the three antibiotic re-sistance genes were all greater than 0.999,and the coefficients of variation were all lower than 1％.The lowest detection limits of the plasmids were 1 × 102 copies/μL.This method was used to de-tect 800 bacterial samples.The results showed that 32 samples contained mcr-1 gene,40 samples contained blaNDM gene,2 samples contained cfr gene,8 samples contained both mcr-1 and blaNDM genes.There were no samples carrying three antibiotic resistance genes detected.The results indica-ted that the triplex fluorescence quantitative PCR method established in this experiment had the advantages of high sensitivity,specificity and stability.It was suitable for rapid detection of blaNDM,mcr-1 and cfr antibiotic resistance genes in clinical practice.It provided a convenient and quick method basis for the detection of antibiotic resistance genes.

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.

Immunoglobulin A nephropathy (IgAN) is the most common primary glomerular disease, and the "four-hit" theory represents its currently accepted pathogenic mechanism. Mucosal immunity triggered by infections in the respiratory tract, intestines, or other areas leads to antigen presentation, T cell stimulation, B cell maturation, and the production of IgA-producing plasma cells. The proteins B-lymphocyte stimulator (BLyS) and a proliferation-inducing ligand (APRIL) are involved in this process, and alternative complement and lectin pathway activation are also part of the pathogenic mechanism. Kidney Disease Improving Global Outcomes guidelines indicate that a specific effective treatment for IgAN is lacking, with renin–angiotensin–aldosterone system inhibitors being the primary therapy. Recent research shows that biological agents can significantly reduce proteinuria, stabilize the estimated glomerular filtration rate, and reverse some pathological changes, such as endocapillary proliferation and crescent formation. There are four main categories of biological agents used to treat IgA nephropathy, specifically anti-CD20 monoclonal antibodies, anti-BLyS or APRIL monoclonal antibodies, monoclonal antibodies targeting both BLyS and APRIL (telitacicept and atacicept), and monoclonal antibodies inhibiting complement system activation (narsoplimab and eculizumab). However, further research on the dosages, treatment duration, long-term efficacy, and safety of these biological agents is required.

BACKGROUND:The assessment of asymmetric gait quality plays a pivotal role in guiding rehabilitation training;however,the link between gait quality and kinematic-kinetic gait parameters remains ambiguous. OBJECTIVE:To formulate a machine-learning model for evaluating gait quality based on gait parameters,identify factors sensitive to gait quality from asymmetric gait parameters,investigate the relationship between gait indicators and gait quality,and provide guidance for asymmetric gait training and rehabilitation. METHODS:An asymmetric gait database was established through the creation of asymmetric conditions.Kinematic and kinetic data were collected from 8 young and 8 elderly subjects(all male,right dominant population)during gait tests.Gait quality for each test data set was assessed using symmetry indices,resulting in the creation of a gait parameter-gait quality dataset.Utilizing the Random Forest algorithm,a gait quality evaluation model was developed and key quality parameter factors were identified through differential analysis.This model was iteratively refined.The model's performance was evaluated through 10-fold cross-validation,and its effectiveness was verified using the cross-validation dataset. RESULTS AND CONCLUSION:(1)A gradient test was designed to categorize gait quality into optimal,suboptimal,intermediate,and poor groups,with 759,329,133,and 125 instances,respectively.(2)The application of the Random Forest algorithm in gait quality assessment was explored.A relationship model was established between gait indicators and gait quality,yielding a predictive model accuracy of 95.99%.(3)The 13 main parameters significantly influencing asymmetric gait quality were identified through the Random Forest model's feature importance ranking.(4)An analysis of gait quality sensitivity factors using the 13 important parameters led to the identification of five key sensitivity indexes.The Random Forest model utilizing these sensitivity factors achieved a predictive accuracy of 94.20%.

Immunoglobulin A nephropathy (IgAN) is the most common primary glomerular disease, and the "four-hit" theory represents its currently accepted pathogenic mechanism. Mucosal immunity triggered by infections in the respiratory tract, intestines, or other areas leads to antigen presentation, T cell stimulation, B cell maturation, and the production of IgA-producing plasma cells. The proteins B-lymphocyte stimulator (BLyS) and a proliferation-inducing ligand (APRIL) are involved in this process, and alternative complement and lectin pathway activation are also part of the pathogenic mechanism. Kidney Disease Improving Global Outcomes guidelines indicate that a specific effective treatment for IgAN is lacking, with renin–angiotensin–aldosterone system inhibitors being the primary therapy. Recent research shows that biological agents can significantly reduce proteinuria, stabilize the estimated glomerular filtration rate, and reverse some pathological changes, such as endocapillary proliferation and crescent formation. There are four main categories of biological agents used to treat IgA nephropathy, specifically anti-CD20 monoclonal antibodies, anti-BLyS or APRIL monoclonal antibodies, monoclonal antibodies targeting both BLyS and APRIL (telitacicept and atacicept), and monoclonal antibodies inhibiting complement system activation (narsoplimab and eculizumab). However, further research on the dosages, treatment duration, long-term efficacy, and safety of these biological agents is required.

Objective: To investigate the effect of lead exposure before pregnancy on bone lead mobilization and bone microstructure in pregnant rats, so as to provide the evidence for illustrating the potential mechanisms of bone lead mobilization during pregnancy. Methods: Twenty-six weaning female specific pathogen-free (SPF) rats of the Wistar strain were randomly divided into the exposure group and the control group. Rats in the exposure group were given 0.05% lead acetate solution for weeks, while animals in the control group were given 0.05% sodium acetate solution. Then, rats in both groups were given distilled water. Following removal of lead exposure for 4 weeks, female rats were co-caged with healthy males at the same age until pregnancy. The blood, femur and tibia specimens were collected from female rats on days 3 (GD3), 10 (GD10) and 17 (GD17) at pregnancy, and the blood and bone lead levels were measured using inductively coupled plasma mass spectrometry (ICP-MS). The unilateral rat femur was scanned using micro-computed tomography (micro CT), and the microstructure changes of cortical and trabecular bones were investigated. The structural and morphological changes of rat femur were observed using hematoxylin-eosin (HE) staining. Results: During the study period, satisfactory mental status and activity and good coat glossiness were observed in female rats in both groups, and there was no significant difference in the increase of rat body weight between groups. The blood lead level at GD17 and bone lead levels at GD3, GD10 and GD17 were significantly higher in rats in the exposure group than in the control group (P<0.05), and the trabecular bone lead level was significantly lower in rats in the exposure group at GD17 than at GD10 (P=0.015). The trabecular bone lead level correlated negatively with blood lead level (r=-0.578, P=0.049), and bone lead contributed 26.8% to blood lead. The bone mass, trabecular number, thickness and density of female rat trabecular bones all reduced in the exposure group at GD17, with an increase in trabecular space, and the proportion of trabecular areas reduced by 27.34% in the exposure group relative to the control group (t=2.851, P=0.046). Conclusions Lead exposure before pregnancy promotes the release of lead from trabecular bones into blood and affects bone microstructure in rats. There is bone lead mobilization during late pregnancy.