ObjectiveTo investigate the change in the activity of hepatitis B virus （HBV）-specific CD8⁺ T cells after pegylated interferon-α-2b （PEG-IFN-α-2b） therapy in HBeAg-negative patients with chronic HBV infection. MethodsA total of 53 HBeAg-negative patients with chronic HBV infection who attended The First Affiliated Hospital of Xinxiang Medical University and Tangdu Hospital of Air Force Mdical University from April 2020 to June 2022 were enrolled and treated with PEG-IFN-α-2b （180 μg/week， subcutaneous injection） antiviral therapy. The study endpoint was HBsAg clearance （course of treatment<48 weeks） or 48 weeks （course of treatment≥48 weeks）. Peripheral blood mononuclear cells were isolated at baseline and study endpoint， and peripheral blood T cell counts were measured. Enzyme-linked immunospot assay was used to measure the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ. A total of 17 HLA-A^*02-restricted patients were selected， and CD8⁺ T cells were purified to establish direct- and indirect-contact co-culture systems for HBV-specific CD8⁺ T cells and HepG2.2.15 cells. The level of lactate dehydrogenase in supernatant was measured to calculate the mortality rate of HepG2.2.15 cells， and the levels of HBV DNA， cytotoxic molecules， and cytokines in supernatant were also measured. Flow cytometry was used to measure the expression of apoptosis ligands， and the cytotoxicity of HBV-specific CD8⁺ T cells was evaluated. The independent samples t-test or the paired t-test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test or the Wilcoxon rank-sum test was used for comparison of non-normally distributed continuous data between two groups. ResultsThe HBsAg clearance rate at study endpoint was 30.19% （16/53）. There were no significant differences in peripheral blood T cell counts （CD3⁺， CD4⁺， and CD8⁺ T cells） between baseline and study endpoint （P>0.05）. At study endpoint， there was a significant increase in the frequency of HBV-specific CD8⁺ T cells secreting perforin， granzyme B， and interferon-γ （U=177.50， t=11.90， U=186.50， all P<0.001）， and the patients with HBsAg clearance had a significantly higher frequency of such HBV-specific CD8⁺ T cells than those without HBsAg clearance （U=120.50， t=2.73， U=121.50， all P<0.01）. In the direct- and indirect-contact co-culture systems at study endpoint， HBV-specific CD8⁺ T cells induced a significant reduction in HBV DNA in the supernatant of HepG2.2.15 cells （all P<0.001） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （all P<0.05）； in the direct-contact co-culture system， HBV-specific CD8⁺ T cells induced significant increases in the mortality rate of HepG2.2.15 cells （13.62%±3.27% vs 11.39%±2.40%， t=2.27， P=0.030） and the secretion of perforin and granzyme B （t=72.50， U=52.50， both P<0.05）. In the direct- and indirect-contact co-culture systems， compared with HBV-specific CD8⁺ T cells from the patients without HBsAg clearance， the HBV-specific CD8⁺ T cells from patients with HBsAg clearance had a significantly greater reduction in HBV DNA （P<0.05） and significant increases in the secretion of interferon-γ and tumor necrosis factor-α （P<0.05）. ConclusionPEG-IFN-‍α‍-2b therapy can help to achieve a relatively high HBsAg clearance rate in HBeAg-negative patients with chronic HBV infection， and the activity of HBV-specific CD8⁺ T cells is significantly enhanced， which is closely associated with HBsAg clearance.

OBJECTIVE To analyze and identify the metabolites of pirtobrutinib （PTN） in rats， and clarify the possible metabolic pathways of PTN in rats. METHODS Six rats were intragastrically administered with 10 mg/kg PTN suspension. Blood samples were collected from the rats 30 minutes before administration and at 0.25， 0.5， 1， 2， 4， 6， 8， 12， 24 hours after administration. Urine and feces samples were collected 12 hours before administration and 24 hours after administration. UHPLC- Orbitrap Exploris 240 system combined with Compound Discoverer 3.0 and Xcalibur 2.0 software were adopted for structural identification and metabolic pathway analysis of PTN metabolites in rat plasma， urine， and feces. RESULTS A total of 29 PTN metabolites were identified， including 17， 19 and 22 metabolites in plasma， urine and feces， respectively. The metabolic pathways of PTN mainly included oxidation， sulfation， glucuronidation， etc.， and its metabolites were mostly combination products of two or more different metabolic forms. In detail， a total of 26 metabolites were associated with phase Ⅰ metabolic reactions （14 oxidation metabolites， 9 reduction/dehydrogenation metabolites， 8 demethylation metabolites， and 5 hydrolysis metabolites）. Meanwhile， a total of 20 products were involved in phase Ⅱ metabolites （14 sulfation metabolites and 8 glucuronic acid binding metabolites）. CONCLUSIONS PTN exhibits a diverse range of metabolites in rat fecal samples， with the primary metabolic pathways being oxidation， sulfation， glucuronidation， and others.

Objective:To utilize single-nucleus RNA sequencing(snRNA-seq) technology to investigate the primary cell subpopulations in human limb venous malformations (VMs) tissue and the role of the key gene RhoJ.Methods:Surgical resection specimens of VMs tissues and surrounding normal vein tissues were collected from 100 clinically diagnosed and screened patients with limb VMs at the Department of Hemangioma Surgery of the Third Affiliated Hospital of Zhengzhou University from January 2021 to December 2023. (1) Transcriptome analysis: Three patient samples were randomly selected for snRNA-seq studies, with the surgically removed VMs tissue serving as the experimental group and the surrounding normal vein tissue as the control group. A gene expression matrix for cell nuclei was established, followed by data quality control, dimensionality reduction, clustering, and cell type annotation. Cell-to-cell communication analysis was performed using the R language CellChat package to identify dominant cell subpopulations. The FindMarkers function was utilized to screen for differentially expressed genes (DEGs) between the dominant cell subpopulations of the experimental and control groups, and functional enrichment analysis was conducted. (2) Tissue experiments: An additional 35 patient samples from both the experimental and control groups were randomly selected. The mRNA and protein expression levels of the RhoJ gene were measured using real-time quantitative PCR (RT-qPCR) and Western blotting, respectively. (3) Validation experiments with human umbilical vein endothelial cells (HUVECs): HUVECs were transfected with pcDNA3.1-NC (blank control) and pcDNA3.1-RhoJ (plasmid expression vector carrying the RhoJ gene), respectively. The biological behavior differences between the two groups of cells were examined using the CCK-8 cell proliferation assay, Transwell invasion assay, and Matrigel angiogenesis assay. Measurement data conforming to a normal distribution were expressed as Mean±SD, and comparisons between the two groups were performed using an independent samples t-test. Results:Through CellChat intercellular communication analysis, it was discovered that endothelial cells were the predominant cell subpopulation in both the experimental and control groups, exhibiting strong communication links with other cell subpopulations. In the analysis of DEGs, it was found that the RhoJ gene in endothelial cells was significantly involved in the biological processes of angiogenesis and regulation. In tissue experiments, RT-qPCR and Western bloting results indicated that the relative expression levels of RhoJ mRNA (4.48±1.29 vs. 1.01±0.17) and protein (1.22±0.03 vs. 0.51±0.20) in the experimental group were significantly higher than those in the control group, with statistically significant differences ( P<0.01 for both). The results of the HUVECs validation experiment showed that the cell proliferation, invasion, and angiogenesis abilities of the pcDNA3.1-RhoJ group were significantly enhanced compared to the pcDNA3.1-NC group. Conclusion:Endothelial cells represent the dominant cell subpopulation during the occurrence and locally invasive progression of VMs, playing a crucial role in this process. The RhoJ gene is significant in regulating the biological behavior of VMs endothelial cells.

Objective To investigate the current status and barriers to implementing non-accompanied care services in medical institutions in Zhejiang Province,and to provide a basis for the improvement of standardized management of such services.Methods A self-designed questionnaire was conducted from September to October 2024 among all secondary and tertiary medical institutions in Zhejiang Province to assess the implementation status and barriers to non-accompanied care services.Results A total of 397 questionnaires were distributed,with 389 valid responses,yielding a valid response rate of 97.98％.Non-accompanied care services were implemented in 118 institutions(30.33％).Among these,90 institutions(76.27％)had established management systems for non-accompanied wards;71 institutions(60.17％)had a medical nursing assistant-to-bed ratio lower than 1∶5;41 institu-tions(34.75％)provided tiered training for medical nursing assistants;93 institutions(78.81％)required patients to bear the full cost of the service.Compared with secondary medical institutions,tertiary medical institutions have more complete management system for non-accompanied care services.The main obstacles hindering the development of non-accompanied care services include an imperfect management system for non-accompanied wards,a shortage of medical nursing assistants,a lack of standardized training for such assistants,inconsistent charging standards,and low acceptance among patients and their families.Conclusion The promotion of non-accompanied care services in medical institutions in Zhejiang Province has achieved initial success.However,challenges persist,including incomplete management systems,uneven development across hospital tiers,and imperfect charging mechanisms.It is recommended that relevant authorities strengthen policy support,enhance standardized training for healthcare nursing assistants,refine cost-sharing mechanisms,and improve the quality and sustainability of non-accompanied care services through multi-party collaboration.

In this study, araucarene diterpenes, characterized by a pimarene skeleton with a variably oxidized side chain at C-13, were investigated. A total of 16 araucarene diterpenoids and their derivatives were isolated from the woods of Agathis dammara, including 11 previously unreported compounds: dammaradione (1), dammarones D-G (2, 5, 14, 15), dammaric acids B-F (8-12), and dammarol (16). The structures of these new compounds were elucidated using high-resolution electrospray ionization mass spectroscopy (HR-ESI-MS) and one-dimensional/two-dimensional (1D/2D) nuclear magnetic resonance (NMR), while their absolute configurations were determined through the electronic circular dichroism (ECD) exciton chirality method and Snatzke's method. The hypoglycemic activity of all isolated compounds was evaluated using a transgenic zebrafish model, and a structure-activity relationship (SAR) analysis was conducted. Araucarone (3) and dammaric acid C (9), serving as representative compounds, demonstrated significant hypoglycemic effects on zebrafish. The primary mechanism involves the promotion of pancreatic β cell regeneration and glucose uptake. Specifically, these compounds enhance the differentiation of pancreatic endocrine precursor cells (PEP cells) into β cells in zebrafish.
Zebrafish ; Animals ; Diterpenes/isolation & purification* ; Insulin-Secreting Cells/cytology* ; Glucose/metabolism* ; Hypoglycemic Agents/isolation & purification* ; Molecular Structure ; Structure-Activity Relationship ; Plant Extracts/pharmacology* ; Regeneration/drug effects*

Objective To explore the epidemiological characteristics and differences in antimicrobial resistance be-tween catheter-associated urinary tract infection(CAUTI)and non-CAUTI of healthcare-associated infection(HAI),and provide scientific basis for precise clinical prevention and control.Methods Based on the regional HAI surveillance platform in Suzhou City,urinary tract infection(UTI)surveillance data reported by 61 member units from January 2020 to December 2024 were analyzed retrospectively.Pathogen distribution,detection rate of multi-drug-resistant organisms(MDROs),and antimicrobial resistance spectrum characteristics of patients in the CAUTI group and non-CAUTI group were compared.Results The incidence of CAUTI in patients in CAUTI group was 0.99‰,the incidence of healthcare-associated UTI in patients in non-CAUTI group was 0.14％.There was statis-tically significant difference in the distribution of UTI pathogens between the two groups(P＜0.05).The patho-gens of the CAUTI group were mainly Gram-negative bacteria(56.1％),with high proportions of Escherichia coli(19.6％)and Klebsiella pneumoniae(15.0％).In the non-CAUTI group,the proportion of Gram-negative bacteria was higher(64.7％).Antimicrobial susceptibility testing results showed that the resistance rates of Escherichia co-li to tobramycin,cephalosporins,and carbapenems in the CAUTI group were all higher than those in the non-CAU-TI group(all P＜0.05).Except for tigecycline,the resistance rates of Klebsiella pneumoniae to other antimicrobial agents in the CAUTI group were all significantly different from the non-CAUTI group(all P＜0.05).The resis-tance rates of Acinetobacterbaumannii to ticarcillin/clavulanic acid,quinolones,most cephalosporins,carbapenems,and aminoglycosides in the CAUTI group were higher than those of the non-CAUTI group(all P＜0.05).The de-tection rates of MDROs were higher in the CAUTI group,especially that of carbapenem-resistant Klebsiella pneu-moniae,accounting for 57.8％.Conclusion There are significant differences in pathogen distribution and antimi-crobial resistance of UTI between the CAUTI group and the non-CAUTI group.It is necessary to establish a re-gional antimicrobial resistance surveillance system for pathogens in UTI,and provide basis for the rational use of an-timicrobial agents in clinical practice.

Objective To examine the association between lymphocyte subsets and renal clinicopathological characteristics as well as prognosis in patients with IgA nephropathy(IgAN).Methods The retrospective analysis included general clinical data and pathological examination results of IgAN patients diagnosed by renal biopsy at the Fourth Hospital of Hebei Medical University from January 2018 to January 2022.Correlation tests were conducted to examine the relationship between lymphocyte subsets and other significant clinicopathological parameters.The optimal cut-off value of CD4+T determined using the Youden index,and patients were grouped accordingly.Kaplan-Meier survival curves and Cox regression analyses were employed to compare the low and high CD4+T lymphocyte groups among IgAN patients,identifying factors influencing renal function progression.The endpoint event was defined as a decrease in estimated glomerular filtration rate(eGFR)of≥30%from baseline,progression to end-stage renal disease(ESRD)[eGFR<15 mL/(min·1.73 m2)or initiation of renal replacement therapy],or all-cause mortality.Results Low CD4+T lymphocytes were significantly positively correlated with blood IgA levels and the proportion of glomerular crescents in IgAN patients(all P<0.05).This study included a total of 53 IgAN patients,divided into two groups based on CD4+T lymphocyte counts:20 patients in the low CD4+T lymphocyte group and 33 patients in the high CD4+T lymphocyte group.In the low CD4+T lymphocyte group,there was a higher proportion of males and a lower proportion of glomerular crescents(P<0.05).Kaplan-Meier survival analysis revealed that patients with low CD4+lymphocytes had a significantly lower cumulative renal survival rate(Log-Rank test χ2=4.188,P=0.041).Cox regression analysis indicated that low CD4+lymphocytes were an independent risk factor for the progression of renal function decline in IgAN patients(HR=2.614,95%CI:1.006～6.788,P=0.048).Conclusions Patients with higher levels of CD4+T lymphocytes exhibit a lower risk of adverse renal outcomes.In contrast,patients with IgA nephropathy and low CD4+T lymphocyte counts tend to have poorer renal survival rates.

OBJECTIVE To retrospectively investigate the incidence of surgical site infection(SSI),distribution and drug resistance of pathogens in 53 hospitals of Suzhou area from 2020 to 2023 so as to provide strategies for pre-vention and control of the hospital-associated infections in Suzhou area.METHODS The data regarding the SSI,distribution and drug resistance of pathogens were statistically analyzed through Suzhou City Regional Hospital In-fection Surveillance Platform.RESULTS The incidence of SSI was 0.24％among the 53 hospitals from 2020 to 2023.Staphylococcus aureus,Enterococcus faecalis,Staphylococcus epidermidis,Enterococcus faecium and Staphylococcus haemolyticus ranked the top 5 species of gram-positive bacteria;Escherichia coli,Klebsiella pneumoniae,Pseudomonas aeruginosa,Enterobacter cloacae and Acinetobacter baumannii ranked the top 5 spe-cies of gram-negative bacteria.There were significant differences in the distributions of S.aureus,K.pneumoniae and A.baumannii in the superficial,deep and organ cavities(P＜0.05).The isolation rate of carbapenem-resistant Acinetobacter baumannii(CRAB)was 69.23％,and the isolation rate of methicillin-resistant Staphylococcus au-reus(MRSA)was 42.14％.The drug resistance rate of the S.aureus strains to oxacillin reached up to 40.24％;the drug resistance rates of the A.baumannii to imipenem and meropenem reached up 63.00％and 65.80％,re-spectively.The drug resistance rates of the K.pneumoniae strains to imipenem and meropenem approximated to 30％,and the drug resistance rate to tigecycline was 19.84％.CONCLUSIONS The total incidence of SSI shows a downward trend in the 53 hospitals of Suzhou area.The S.aureus is the major species of pathogens,and some of the pathogens show high drug resistance rates.

ln the context of the high-quality development of public hospitals,research management is transitioning from a results-oriented approach to a full-process management model,necessitating the exploration of a governance system that integrates topic guidance,process support,and outcome transformation.Based on the Knowledge-to-Action and SECl model,it analyzes the construction logic and application effectiveness of the nursing research rounds mechanism at a teritary grade A hospital in Hainan Province.Through a"three-tier stratification+dual-track collaboration+full-process closed-loop"design,the mechanism bridges the chain from"clinical problem identification to research path optimization to outcome transformation",significantly enhancing research participation,research plan standardization,and research output.The findings validate the practical value of the research rounds mechanism in promoting knowledge transformation,capability enhancement,and organizational knowledge retention.

Objective:To develop a novel fibroblast activation protein (FAP) cyclic peptide imaging agent, Al 18F-FAP-NOX, evaluate its in vitro and in vivo properties, and explore its feasibility of PET/CT imaging in tumors with FAP positive expression. Methods:Al 18F-FAP-NOX was manually synthesized. The in vitro stability of Al 18F-FAP-NOX was determined using radio high performance liquid chromatography (HPLC). The lipid water partition coefficient log P, in vitro cell uptake experiments, microPET/CT imaging and biodistribution in 293T-FAP tumor-bearing mice were conducted to preliminarily evaluate the pharmacokinetics and biological efficacy of Al 18F-FAP-NOX. Afterwards, a patient (male, 65 years old) with lung cancer underwent Al 18F-FAP-NOX PET/CT imaging. Results:Al 18F-FAP-NOX was successfully synthesized with a yield of (26.28±2.31)% without attenuation correction ( n=4), and the radiochemical purity was more than 95%. Al 18F-FAP-NOX exhibited good stability and hydrophilicity (log P=-3.02±0.08, n=5). In cell assays, the uptake of Al 18F-FAP-NOX in HT1080-FAP cells reached the plateau phase at 15 min ((7.31±0.53) percentage activity of injection dose per million cells (%ID/mio cells)), exhibiting high cellular uptake. The uptake of Al 18F-FAP-NOX could be significantly inhibited by 1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid (DOTA)-FAP-2286. The microPET/CT results of 293T-FAP tumor-bearing mice in vivo showed that Al 18F-FAP-NOX was highly uptaken in FAP-positive tumor tissues (60 min: (12.47±1.66) percentage activity of injection dose per gram of tissue (%ID/g)), while the uptake was very low in FAP-negative tumors. The biodistribution results were similar to the microPET/CT imaging results of tumor-bearing mice. The human clinical imaging showed an abnormal increase in Al 18F-FAP-NOX uptake (SUV max 5.5) of the lung cancer lesions. Conclusions:A novel cyclic peptide radiopharmaceutical, Al 18F-FAP-NOX, demonstrates good stability and hydrophilicity. It can be quickly distributed to tumor tissue in vivo. The human clinical PET/CT imaging shows certain diagnostic ability of Al 18F-FAP-NOX for lung cancer lesions. It is a promising cyclic peptide agent for PET imaging.