ObjectiveTo obtain a construct chimeric antigen receptor macrophages （CAR-M） targeting disialoganglioside GD2 （GD2） and harboring the intracellular domain of macrophage-specific activating signaling adaptor protein DNAX-activating protein 12 （DAP12）. MethodsA CAR molecule targeting GD2 with the DAP12 intracellular domain as the signaling module ［Anti-GD2 CAR （DAP12）］ was designed， synthesized， and cloned into an adenoviral vector. This virus was used to infect PMA-differentiated THP-1 macrophages to construct green fluorescent protein （GFP）-labeled Anti-GD2 CAR THP-1 cells. CAR expression was confirmed using live-cell imaging. In vitro co-culture assays were performed to evaluate the phagocytic function of empty， Anti-GD2 CAR-M （CD3ζ）， and Anti-GD2 CAR-M （DAP12） groups against glioma cells with high （T98G） and low （U251） GD2 expression. ResultsThe CAR adenovirus efficiently transduced THP-1 macrophages， resulting in successful generation of Anti-GD2 CAR-M. Flow cytometry analysis following co-culture with glioma cells showed that compared to the empty group， the Anti-GD2 CAR-M （CD3ζ） group exhibited significantly enhanced phagocytosis （P<0.05）. Furthermore， compared to the Anti-GD2 CAR-M （CD3ζ） group， the Anti-GD2 CAR-M （DAP12） group demonstrated a further increase in phagocytic capacity specifically against GD2-high T98G cells （P<0.05）. ConclusionAnti-GD2 CAR-M （DAP12） cells are successfully established by transducing PMA-differentiated THP-1-M with recombinant Anti-GD2 CAR （DAP12） adenovirus. The engineered CAR-M can specifically target phagocytose glioma cells with high GD2 expression.

ObjectiveTo obtain stably transfected chimeric antigen receptor macrophages （CAR-M） targeting disialosylganglioside （GD2） based on the constitutive expression of myeloid differentiation factor 88 （MYD88）. MethodsAn Anti-GD2 CAR adenoviral vector incorporating the CD3ζ （cluster of differentiation 3 zeta chain）-MYD88 fusion construct was designed and constructed. THP-1-derived macrophages （THP-1M）， induced by phorbol 12-myristate 13-acetate （PMA） differentiation， were infected with this adenovirus to generate Anti-GD2 CAR THP-1 cells bearing a green fluorescent protein （GFP） tag. CAR expression was monitored using a live-cell imaging system. The phagocytic function of Empty CAR-M， Anti-GD2 CAR-M（CD3ζ）， and Anti-GD2 CAR-M（CD3ζ-MYD88） groups against glioma cells was evaluated in vitro through co-culture experiments， and their polarization status was also analyzed. ResultsCAR adenovirus successfully transfected THP-1 to obtain Anti-GD2 CAR-M. After co-culture with glioma cells， flow cytometry results showed that compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88） group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）. Compared with the Empty CAR-M group and the Anti-GD2 CAR-M（CD3ζ） group， the Anti-GD2 CAR-M（CD3ζ-MYD88）CAR-M group exhibited enhanced phagocytosis against T98G cells with high GD2 expression （P<0.05） and also showed upregulation of CD86 （P<0.05）， and the expression of inflammatory cytokines interleukin （IL）-6 and IL-12 was increased in this group （P<0.05）. ConclusionThis study successfully constructed Anti-GD2 CAR-M cells and verified their phagocytic activity， demonstrating that constitutive expression of MYD88 can promote inflammatory polarization of CAR-M. This provides a potential structural modification approach for the development of pro-inflammatory CAR-M.

Methods: Motor-evoked potentials (MEPs) and somatosensory-evoked potentials (SEPs) were recorded in 48 patients with TOLF (hybrid 20 vs. en bloc 28) during surgery. Patients were categorized based on MEP/SEP improvement, deterioration, or no change, and MEP/SEP improvement rates were measured in the improvement group. Furthermore, all patients were assessed using the Ashworth and modified Japanese Orthopedic Association scores. Results: The incidences of both MEP/SEP improvement (21.4% vs. 25.0%, p=0.772) and deterioration (21.4% vs. 20.0%, p=0.904) were similar between the en bloc and hybrid laminectomy groups, and no difference in preoperative and postoperative clinical assessments was observed between the two groups (p>0.05). In four patients (4/28, 14.3%) undergoing en bloc laminectomy, MEP amplitudes initially increased after OLF removal but gradually decreased. This delayed MEP reduction did not occur in the hybrid laminectomy group. Furthermore, more patients undergoing en bloc laminectomy had CFL than those undergoing hybrid laminectomy (46.4% vs. 15.0%, p=0.023). In the improvement group, the hybrid laminectomy group exhibited higher MEP improvement rates in the bilateral abductor hallucis than the en bloc laminectomy group (left side: 213.4%±35.9% vs. 152.5%±41.0%, p=0.028; right side: 201.2%±32.0% vs. 145.2%±46.3%, p=0.043). Conclusions Compared with en bloc laminectomy, hybrid laminectomy may be a safe and effective method for treating multilevel TOLF, potentially reducing intraoperative spinal cord irritation and CFL and causing relatively better functional recovery.

Methods: Motor-evoked potentials (MEPs) and somatosensory-evoked potentials (SEPs) were recorded in 48 patients with TOLF (hybrid 20 vs. en bloc 28) during surgery. Patients were categorized based on MEP/SEP improvement, deterioration, or no change, and MEP/SEP improvement rates were measured in the improvement group. Furthermore, all patients were assessed using the Ashworth and modified Japanese Orthopedic Association scores. Results: The incidences of both MEP/SEP improvement (21.4% vs. 25.0%, p=0.772) and deterioration (21.4% vs. 20.0%, p=0.904) were similar between the en bloc and hybrid laminectomy groups, and no difference in preoperative and postoperative clinical assessments was observed between the two groups (p>0.05). In four patients (4/28, 14.3%) undergoing en bloc laminectomy, MEP amplitudes initially increased after OLF removal but gradually decreased. This delayed MEP reduction did not occur in the hybrid laminectomy group. Furthermore, more patients undergoing en bloc laminectomy had CFL than those undergoing hybrid laminectomy (46.4% vs. 15.0%, p=0.023). In the improvement group, the hybrid laminectomy group exhibited higher MEP improvement rates in the bilateral abductor hallucis than the en bloc laminectomy group (left side: 213.4%±35.9% vs. 152.5%±41.0%, p=0.028; right side: 201.2%±32.0% vs. 145.2%±46.3%, p=0.043). Conclusions Compared with en bloc laminectomy, hybrid laminectomy may be a safe and effective method for treating multilevel TOLF, potentially reducing intraoperative spinal cord irritation and CFL and causing relatively better functional recovery.

Methods: Motor-evoked potentials (MEPs) and somatosensory-evoked potentials (SEPs) were recorded in 48 patients with TOLF (hybrid 20 vs. en bloc 28) during surgery. Patients were categorized based on MEP/SEP improvement, deterioration, or no change, and MEP/SEP improvement rates were measured in the improvement group. Furthermore, all patients were assessed using the Ashworth and modified Japanese Orthopedic Association scores. Results: The incidences of both MEP/SEP improvement (21.4% vs. 25.0%, p=0.772) and deterioration (21.4% vs. 20.0%, p=0.904) were similar between the en bloc and hybrid laminectomy groups, and no difference in preoperative and postoperative clinical assessments was observed between the two groups (p>0.05). In four patients (4/28, 14.3%) undergoing en bloc laminectomy, MEP amplitudes initially increased after OLF removal but gradually decreased. This delayed MEP reduction did not occur in the hybrid laminectomy group. Furthermore, more patients undergoing en bloc laminectomy had CFL than those undergoing hybrid laminectomy (46.4% vs. 15.0%, p=0.023). In the improvement group, the hybrid laminectomy group exhibited higher MEP improvement rates in the bilateral abductor hallucis than the en bloc laminectomy group (left side: 213.4%±35.9% vs. 152.5%±41.0%, p=0.028; right side: 201.2%±32.0% vs. 145.2%±46.3%, p=0.043). Conclusions Compared with en bloc laminectomy, hybrid laminectomy may be a safe and effective method for treating multilevel TOLF, potentially reducing intraoperative spinal cord irritation and CFL and causing relatively better functional recovery.

Methods: Preoperatively, all 115 patients underwent BMD assessments using both CT-derived HUs and dual-energy X-ray absorptiometry and were administered multiple questionnaires, including the Pain Catastrophizing Scale (PCS), Beck Anxiety Inventory (BAI), and Beck Depression Index (BDI). Both pain intensity and pain-related disability were assessed before and after lumbar fusion surgery. Results: Postoperatively, 14 patients (14/115, 12.2%) experienced FBSS. Multivariate logistic regression was used to examine all preoperative covariates with significant differences between the patients with and without FBSS. The numeric rating pain scale score at rest, BAI score, PCS score, and HU value were found to be independently associated with FBSS (p<0.05). Conclusions This study revealed that preoperative BMD, as quantified by CT-derived HU values, may be associated with FBSS. Preoperative assessments of CT-derived HU values might provide additional details for identifying patients susceptible to FBSS, which could help prevent this complication.

Objective To investigate the common molecular features of immunothrombosis, thus enhancing the comprehension of thrombosis triggered by immune and inflammatory responses and offering crucial insights for identifying potential diagnostic and therapeutic targets. Methods Differential gene expression analysis and functional enrichment analysis were conducted on datasets of systemic lupus erythematosus (SLE) and venous thromboembolism (VTE). The intersection of differentially expressed genes in SLE and VTE with those of neutrophil extracellular traps (NET) yielded cross-talk genes (CG) for SLE-NET and VTE-NET interaction. Further analysis included functional enrichment and protein-protein interaction (PPI) network assessments of these CG to identify hub genes. Venn diagrams and receiver operating characteristic (ROC) curve analysis were employed to pinpoint the most effective shared diagnostic CG, which were validated using a graft-versus-host disease (GVHD) dataset. Results Differential expression genes in SLE and VTE were associated with distinct biological processes, whereas SLE-NET-CG and VTE-NET-CG were implicated in pathways related to leukocyte migration, inflammatory response, and immune response. Through PPI network analysis, several hub genes were identified, with matrix metalloproteinase 9 (MMP9) and S100 calcium-binding protein A12 (S100A12) emerging as the best shared diagnostic CG for SLE (AUC: 0.936 and 0.832) and VTE (AUC: 0.719 and 0.759). Notably, MMP9 exhibited good diagnostic performance in the GVHD dataset (AUC: 0.696). Conclusion This study unveils the common molecular features of SLE, VTE, and NET, emphasizing MMP9 and S100A12 as the optimal shared diagnostic CG, thus providing valuable evidence for the diagnosis and therapeutic strategies related to immunothrombosis. Additionally, the expression of MMP9 in GVHD highlights its critical role in the risk of VTE associated with immune system disorders.
Humans ; Computational Biology/methods* ; Lupus Erythematosus, Systemic/immunology* ; Protein Interaction Maps/genetics* ; Venous Thromboembolism/therapy* ; Matrix Metalloproteinase 9/genetics* ; Extracellular Traps/metabolism* ; Gene Regulatory Networks ; Thrombosis/immunology* ; Graft vs Host Disease/genetics* ; Gene Expression Profiling

Objective To systematically review and quantify the content and structure of the non-accompanied wards policy texts at the provincial level in China from the perspective of policy tools,providing references and insights for optimizing and implementing future policies.Methods Using the policy analysis tools as framework,it applies content ana lysis to construct a two-dimensional analytical framework with the X-dimension and Y-dimension.A total of 19 policy texts related to non-accompanied ward issued by provincial governments in China from January 2000 to August 2024 were coded and analyzed.Results A total of 141 entries were coded.In the X-dimension,supply-oriented,demand-oriented,and environment-oriented policy tools accounted for 21.28%,19.86%,and 58.86%,respectively,indicating a greater reliance on environmental-oriented policy tools;In the Y-dimension,policies from the"12th Five-Year Plan"(16.31%),"13th Five-Year Plan"(14.89%),and"14th Five-Year Plan"(68.80%)phases were analyzed.The number of policies in the"14th Five-Year Plan"phase was the highest among the three periods.Conclusion The policy framework for non-accompanied wards in China is still in its initial exploration phase and has room for improvement.It is recommended that future policies optimize the use of policy tools,strengthen coordination among them,and support the development of non-accompanied wards.

Objective:To design and develop a cone-beam CT imaging system for small animals with continuously adjustable spatial resolution.Methods:The imaging system used an X-ray source with a focal spot size of 30 μm and a flat panel detector with a pixel size of 100 μm. On this premise, a " stepping-focusing-rotating" image acquisition mode was proposed, in which the " focusing" and " stepping" systems were sequentially embedded in the " rotating" system. In this acquisition mode, the X-ray source and flat panel detector were relatively stationary to form the " focusing" system. When the " stepping" system accurately transported the object to the scanning position, the " focusing" system could achieve adjustable spatial resolution by making linear motion around the object to be scanned according to different experimental requirements. Finally the " rotating" system achieve high-quality imaging.Results:The variable spatial resolution of small animal CBCT ranges from 35.7 μm to 71.4 μm, and the FOV ranges from 39.6 mm to 108.0 mm. The conversion time for the limit spatial resolution is 19.125 s, which allowed accurate 3D reconstruction of normal mice at different resolutions with high reproducibility.Conclusions:A cone-beam CT suitable for small animals has been developed, whose spatial resolution and FOV can be adjusted arbitrarily within a certain range, which can meet the different imaging requirements in rodent experiments.

Objective:To design and develop a cone-beam CT imaging system for small animals with continuously adjustable spatial resolution.Methods:The imaging system used an X-ray source with a focal spot size of 30 μm and a flat panel detector with a pixel size of 100 μm. On this premise, a " stepping-focusing-rotating" image acquisition mode was proposed, in which the " focusing" and " stepping" systems were sequentially embedded in the " rotating" system. In this acquisition mode, the X-ray source and flat panel detector were relatively stationary to form the " focusing" system. When the " stepping" system accurately transported the object to the scanning position, the " focusing" system could achieve adjustable spatial resolution by making linear motion around the object to be scanned according to different experimental requirements. Finally the " rotating" system achieve high-quality imaging.Results:The variable spatial resolution of small animal CBCT ranges from 35.7 μm to 71.4 μm, and the FOV ranges from 39.6 mm to 108.0 mm. The conversion time for the limit spatial resolution is 19.125 s, which allowed accurate 3D reconstruction of normal mice at different resolutions with high reproducibility.Conclusions:A cone-beam CT suitable for small animals has been developed, whose spatial resolution and FOV can be adjusted arbitrarily within a certain range, which can meet the different imaging requirements in rodent experiments.