Purpose To investigate how IL-6-mediated monocytes/macrophages suppress the effector function of natural killer(NK)cells through the PD-L1/PD-1 axis in acute-on-chronic liver failure(ACLF).Methods HE stai-ning and Masson's trichrome staining were used to assess hepatic inflammatory infiltration and fibrosis.Immunofluores-cence staining was performed to detect IL-6 expression in hepatic macrophages.Flow cytometry was applied to evaluate the phenotypes and functions of monocytes/macrophages and NK cells.Recombinant IL-6 was used to treat peripheral blood mononuclear cells,and PD-L1 expression on monocytes was examined.Monocytes and NK cells were co-cul-tured,and after PD-1 blockade,NK cell effector function was evaluated.Results The ACLF patients,the degree of hepatic fibrosis increased with higher serum IL-6 level.Hepatic macrophages secreted more IL-6,and the circulating monocytes showed an increased percentage[(9.89±0.65)％],absolute count[(0.58±0.04)× 106/mL],prolif-erative capacity[(36.65±6.810)％],and IL-6 secretion[(16.97±2.387)％].In ACLF mice,hepatic inflamma-tory infiltration and fibrosis were aggravated,and IL-6+macrophages exhibited enhanced pro-inflammatory activity.IL-6 had mediated the upregulation of PD-L1 expression in monocytes,while NK cells showed increased PD-1 expression and impaired effector function.When monocytes and NK cells were co-cultured,PD-1 blockade restored,NK cell ef-fector functions.Conclusion In ACLF,elevated IL-6 levels were associated with hepatic fibrosis and disease severi-ty.IL-6 mediated monocytes-induced suppression of NK cell effector functions via the PD-L1/PD-1 axis,suggesting that IL-6 plays a critical role in ACLF-related immune dysregulation.

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*

The " no-accompany" service is an important measure to meet the patients′ needs for comprehensive, continuous, and high-quality diversified care and to enhance their medical experience. Starting in March 2024, the Second Affiliated Hospital of Zhejiang University School of Medicine launched a pilot " no-accompany" service in the breast surgery and dermatology wards. The hospital established a working team led by the nursing department and involving multiple administrative departments such as the medical affairs department, medical insurance office, and logistics management department. And the hospital has successfully advanced the implementation of the " no-accompany" ward program by establishing a multi-department collaborative management mechanism, standardizing the management of medical caregivers, developing a tiered fee scheme for the " no-accompany" service, and improving relevant service measures. The acceptance of the " no-accompany" service by patients increased from (3.93±0.83) in the second quarter of 2024 to (4.69±0.59) in the fourth quarter, and overall satisfaction rose from (4.18±0.73) to (4.50±0.54) (all P<0.001). The job competency of medical caregivers also improved significantly from (64.80±3.49) before starting work in March 2024 to (94.00±2.40) in the fourth quarter ( P<0.001). These findings provide a reference for the implementation of " no-accompany" services in public hospitals.

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.

Objective:To investigate the efficacy and safety of bridging thrombectomy and direct thrombectomy in patients with acute anterior circulation large core infarction.Methods:A retrospective cross-sectional study was performed; 131 patients with acute anterior circulation large core infarction with an onset time≤ 24 hours received endovascular treatment (EVT) at Department of Intervention Neurosurgery, Nanyang Central Hospital, Xinxiang Medical University, from June 2022 to June 2024 were chosen; they were divided into bridging thrombectomy group (accepted intravenous thrombolysis [IVT] before EVT, n=55) and direct thrombectomy group ( n=76). Statistical analysis was conducted to compare the differences in efficacy and safety between the two groups. Efficacy indicators included modified Rankin scale (mRS) score 90 days after thrombectomy, and good prognosis rate 90 days after thrombectomy (mRS scores of 0-2). Safety indicators included intracranial hemorrhage (ICH) incidence within 72 hours of thrombectomy, symptomatic intracranial hemorrhage (sICH) incidence within 72 hours of thrombectomy, mortality 90 days after thrombectomy, and ratio of patients receiving decompressive craniectomy after surgery. Results:No significant difference between the direct thrombectomy group and bridging thrombectomy group was noted in terms of mRS score 90 days after thrombectomy (3 [2, 6] vs. 3 [2, 6]), good prognosis rate 90 days after thrombectomy (38.2% vs. 25.5%), ICH incidence within 72 hours of thrombectomy (30.3% vs. 40.0%), sICH incidence within 72 hours of thrombectomy (21.1% vs. 21.8%), mortality rate within 90 days of thrombectomy (28.9% vs. 25.5%), and ratio of patients receiving decompressive craniectomy after surgery (9.2% vs. 9.1%, P>0.05). Core infarction volume at 100 mL was used as grouping standard, subgroup analysis showed that when the core infarction volume was 70-100 mL and >100 mL, respectively, no significant difference in sICH incidence was noted between the bridging thrombectomy group and direct thrombectomy group ( P>0.05). Conclusion:For patients with acute anterior circulation large core infarction with an onset time≤ 24 hours, bridging thrombectomy has similar efficacy and safety as direct thrombectomy.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

Purpose To explore the relationship between regulatory T cells(Treg cells)and natural killer(NK)cells with tumor occurrence and development in papillary thyroid carcinoma(PTC).Methods Hematoxylin and eosin(HE)staining was performed on pathological tissue sections from patients with nodular goiter and PTC to evaluate im-mune cell infiltration.Flow cytometry was used to detect the percentage and absolute count of Treg cells and NK cells in peripheral blood from healthy controls(HCs)and PTC patients.Treg cells phenotypes(ICOS、PD-1、Lag3)and their IL-10 secretion ability were analayzed,along with functional molecules(TNF-α、IFN-γ、Granzyme B、Perforin)ex-pressed by NK cells.A cytometric bead array was used to measure serum cytokine IL-10 concentration.Changes in Treg and NK cells and their correlations were analyzed.Results Immune cell infiltration in PTC tissues is higher than in nodular goiter tissue.Compared with healthy controls,PTC patients exhibited a significant increase in the percentage[(6.86±3.10)％]and absolute count[(4.97±2.82 × 106 cells/mL)]of circulating Treg cells,with an enhanced IL-10 secretion capacity[(7.18±3.57)％],which is related to lymph node metastasis and vascular invasion.Con-versely,NK cell percentage[(16.56±8.44)％]and absolute count[(3.29±3.12 × 106 cells/mL)]were reduced,with impaired effector functions.The NK-to-Treg cell ratio(2.49±1.60)significantly declined,and a negative corre-lation was observed between the percentage of the two groups cell types(P=0.04,r=-0.625).Serum IL-10 levels in PTC patients were elevated[(1.30±1.09)pg/mL],correlating with tumor size and lymph node metastasis.After treatment with 1,25-(OH)2-D3,PTC patients showed a reduction in the percentage[(7.01±2.13)％]and absolute count(4.27±1.62 × 106 cells/mL)of circulating CD4+CD25+CD127low Treg cells,and a decrease in serum cytokine IL-10 levels[(1.27±0.09)pg/mL].Circulating NK cells percentages[(20.6±6.72)％]and absolute counts[(5.63±2.93)× 106 cells/mL]partially recovered.Conclusion PTC patients exhibit a pronounced immunosup-pressive state.Dysregulated numbers and functions of Treg and NK cells are closely associated with the occurrence,de-velopment,and prognosis of PTC,highlighting their potential importance in tumor immunotherapy.

To evaluate the clinical application value of magnetic bead sorting technology (MBS) using the T-cell select kit combined with an automatic cell sorter for isolating peripheral blood mononuclear cells (PBMCs) to realize automated specimen pre-processing and process optimization in T-SPOT.TB assay. A cross-sectional study was conducted involving 300 patients recruited from the first affiliated hospital of Air Force Medical University from March 2023 to July 2024. Among them, there were 167 males and 133 females. The age range of the patients was 18 to 65 years, with a median age of 49.0 (33.3, 59.0) years and a mean age of (46.4±13.9) years. 4 ml of anticoagulated whole blood samples in duplicate were collected from each patient. One fresh sample (0-4 h) was processed immediately using density gradient centrifugation (DGC) for PBMC isolation, while the other was processed using MBS method at either 0-4 h or 34-54 h post-collection. The cycles for cell enrichment step of the automatic cell sorter were adjusted from the conventional 4 cycles to 2 cycles. Statistical analysis was performed using Cohen′s Kappa test to evaluate the concordance of T-SPOT.TB results across all processing groups. The results showed that for fresh samples (0-4 h), the T-SPOT.TB results for samples processed with 4-cycle and 2-cycle MBS enrichment steps demonstrated positive concordance rates of 91.7% and 93.1%, negative concordance rates of 100% for both, overall concordance rates of 98.0% and 97.9%, and Kappa values of 0.94 and 0.95, respectively, compared with the reference results for paired samples processed using DGC. Correspondingly, the results for samples processed with the 4-cycle and 2-cycle MBS methods at 34-54 h post-collection yielded positive concordance rates of 90.0% and 81.3%, negative concordance rates of 95.1% and 100%, overall concordance rates of 93.0% and 82.9%, and Kappa values of 0.86 and 0.43, respectively, compared with the reference results for fresh samples processed using DGC. In conclusion,for 0-4 h samples, the T-SPOT.TB results from both 4-cycle and 2-cycle MBS enrichment protocols were highly consistent with the reference results from conventional DGC methods. However, for 34-54 h stored samples, results from the 4-cycle MBS method rather than the 2-cycle protocol exhibited significantly superior concordance with the reference results. The MBS method using T-Cell Select kit achieves automated sample pre-processing for T-SPOT.TB assay, reduces hands-on time, and provides a viable alternative with reliable results for long-stored specimens.

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.

Purpose To investigate how IL-6-mediated monocytes/macrophages suppress the effector function of natural killer(NK)cells through the PD-L1/PD-1 axis in acute-on-chronic liver failure(ACLF).Methods HE stai-ning and Masson's trichrome staining were used to assess hepatic inflammatory infiltration and fibrosis.Immunofluores-cence staining was performed to detect IL-6 expression in hepatic macrophages.Flow cytometry was applied to evaluate the phenotypes and functions of monocytes/macrophages and NK cells.Recombinant IL-6 was used to treat peripheral blood mononuclear cells,and PD-L1 expression on monocytes was examined.Monocytes and NK cells were co-cul-tured,and after PD-1 blockade,NK cell effector function was evaluated.Results The ACLF patients,the degree of hepatic fibrosis increased with higher serum IL-6 level.Hepatic macrophages secreted more IL-6,and the circulating monocytes showed an increased percentage[(9.89±0.65)％],absolute count[(0.58±0.04)× 106/mL],prolif-erative capacity[(36.65±6.810)％],and IL-6 secretion[(16.97±2.387)％].In ACLF mice,hepatic inflamma-tory infiltration and fibrosis were aggravated,and IL-6+macrophages exhibited enhanced pro-inflammatory activity.IL-6 had mediated the upregulation of PD-L1 expression in monocytes,while NK cells showed increased PD-1 expression and impaired effector function.When monocytes and NK cells were co-cultured,PD-1 blockade restored,NK cell ef-fector functions.Conclusion In ACLF,elevated IL-6 levels were associated with hepatic fibrosis and disease severi-ty.IL-6 mediated monocytes-induced suppression of NK cell effector functions via the PD-L1/PD-1 axis,suggesting that IL-6 plays a critical role in ACLF-related immune dysregulation.