Although cancer immunotherapy has made great strides in the clinic, it is still hindered by the tumor immunosuppressive microenvironment (TIME). The stimulator of interferon genes (STING) pathway which can modulate TIME effectively has emerged as a promising therapeutic recently. However, the delivery of most STING agonists, specifically cyclic dinucleotides (CDNs), is performed intratumorally due to their insufficient pharmacological properties, such as weak permeability across cell membranes and vulnerability to nuclease degradation. To expand the clinical applicability of CDNs, a novel pH-sensitive polycationic polymer-modified lipid nanoparticle (LNP-B) system was developed for intravenous delivery of CDNs. LNP-B significantly extended the circulation of CDNs and enhanced the accumulation of CDNs within the tumor, spleen, and tumor-draining lymph nodes compared with free CDNs thereby triggering the STING pathway of dendritic cells and repolarizing pro-tumor macrophages. These events subsequently gave rise to potent anti-tumor immune reactions and substantial inhibition of tumors in CT26 colon cancer-bearing mouse models. In addition, due to the acid-sensitive property of the polycationic polymer, the delivery system of LNP-B was more biocompatible and safer compared with lipid nanoparticles formulated with an indissociable cationic DOTAP (LNP-D). These findings suggest that LNP-B has great potential in the intravenous delivery of CDNs for tumor immunotherapy.

Cold tumors have a poor response to tumor immunotherapy due to low immune cell infiltration and the ability to evade immune attacks. Converting cold tumors into hot tumors can enhance the clinical effectiveness of anti-tumor immunotherapy. High-intensity focused ultrasound (HIFU) as a non-invasive treatment can damage tumors through mechanical effects, but there is a lack of research on its cytotoxic mechanisms at the cellular level and its role in inducing anti-immune responses. In this study, the role of HIFU in triggering tumor ferroptosis by disrupting the GSH/GSSG balance through mechanochemical action and the associated anti-tumor immune priming effect were investigated. The use of a nano-enhancer loaded with PFOB combined with HIFU could enhance ferroptosis in triple-negative breast cancer at a specific stage of tumor growth (UTGR = 0) while promoting the conversion of a cold tumor into a hot tumor, thereby improving the immune response. Overall, this provides valuable guidance for the clinical application of HIFU in tumor immunotherapy.

This report presents a case of a 9-year-old child with a complicated crown-root fracture of the maxillary central incisor, accompanied with a root fracture. The treatment strategy was minimally invasive, focusing on vital pulp preservation, root fracture recovery, and crown restoration. The fractured crown was reattached using resin short posts to enhance retention, resulting in aesthetic and functional restoration of the anterior teeth. A 2-year follow-up revealed favorable clinical and radiographic outcomes.
Humans ; Child ; Tooth Fractures/therapy* ; Tooth Root/injuries* ; Incisor/injuries* ; Tooth Crown/injuries* ; Post and Core Technique ; Dental Restoration, Permanent/methods* ; Maxilla

Objective:To analyze the expression of tumor-associated transmembrane protein 61 (TMEM61) in cholangiocarcinoma tissues and its influence on prognosis and immune infiltration, as well as the effect on the proliferation of cholangiocarcinoma cells.Methods:In the cholangiocarcinoma gene chip dataset (TCGA-CHOL), differentially expressed genes between cholangiocarcinoma tissues and normal bile duct tissues were screened, and the upregulated TMEM61 gene was selected for further analysis. Based on the TMEM61 expression, cholangiocarcinoma patients higher than the median value were classified as the high-expression group ( n=17), and those lower than the median value were classified as the low-expression group ( n=18). The Kaplan-Meier survival curve was plotted. Functional and pathway enrichment analyses were conducted on differentially expressed genes related to TMEM61, and the correlations between TMEM61 expression and immune cells and immune molecules were respectively analyzed. The expression level of TMEM61 in cholangiocarcinoma tissues was analyzed by immunohistochemistry; The effect of TMEM61 expression on the proliferation of cholangiocarcinoma cells was detected by Western blotting, CCK-8, clone formation assay, etc. Results:Compared with normal tissues, the expression of TMEM61 mRNA in cholangiocarcinoma tissues was significantly upregulated ( t=18.31, P<0.001). The overall survival rate of patients in the high-expression group of TMEM61 was significantly lower than that in the low-expression group, and the difference was statistically significant ( χ2=7.23, P=0.007). The differentially expressed genes related to TMEM61 were involved in cell proliferation, cell cycle and DNA replication, etc. Compared with normal tissues, regulatory T cells ( t=10.21, P<0.001) and M0-type macrophages ( t=5.89, P=0.008) were significantly increased in cholangiocarcinoma tissues. Plasma cells ( t=7.34, P=0.002), γδT cells ( t=9.87, P<0.001), and M2-type macrophages ( t=11.53, P<0.001) were significantly decreased in cholangiocarcinoma tissues. The expression of TMEM61 was correlated with neurociliary protein 1, tumor necrosis factor ligand superfamily member 15 and B7 homologous protein 3 (all P<0.05). The proportion of positive staining area of TMEM61 protein in normal tissues was (10.15±2.27) %, and that in cholangiocarcinoma tissues was (69.43±11.66) %. The difference was statistically significant ( t=14.97, P<0.001). Inhibition of TMEM61 expression led to a decrease in the number of cholangiocarcinoma cell clones and proliferation activity, and the differences were statistically significant (both P<0.01). Conclusion:The expression of TMEM61 is elevated in cholangiocarcinoma tissues and is associated with poor prognosis. The abnormally high expression of TMEM61 affects the infiltration of immune cells and promotes the proliferation of cholangiocarcinoma cells. TMEM61 is expected to become a potential biomarker for the prognosis assessment of cholangiocarcinoma.

Objective To investigate the risk factors and intervention measures for surgical site infec-tion following posterolateral approach lumbar fusion surgery.Methods A total of 1 078 patients who under-went posterolateral approach lumbar fusion surgery in the department of spine surgery from January 1,2022 to December 31,2023 were included.Patient related information was collected through the real-time nosocomi-al infection monitoring system,while medical visit information was obtained via the outpatient electronic med-ical record system.Multivariate logistic regression analysis was performed to identify risk factors for surgical site infection.Results Among the 1 078 patients,34 cases(3.15%)developed surgical site infections,while 1 044 cases did not.Multivariate logistic regression analysis revealed that age,smoking,hypertension,diabetes,concurrent hospital stay,operative time,duration of postoperative antimicrobial use after initial surgery,and total antimicrobial use duration were significant risk factors for surgical site infection(P＜0.05).Among the 34 infected patients,the duration of antimicrobial use varied significantly across different infection sites(P＜0.05),with the longest duration observed in patients with deep space infections.Conclusion Targeted surveil-lance of surgical site infections should be reinforced based on these risk factors.Perioperative infection control measures must be strictly implemented to improve the scientific,precise,and standardized management of sur-gical-related nosocomial infections.

BACKGROUND: Renal osteodystrophy (ROD) is a skeletal pathology associated with chronic kidney disease-mineral and bone disorder (CKD-MBD) that is characterized by aberrant bone mineralization and remodeling. ROD increases the risk of fracture and mortality in CKD patients. The underlying mechanisms of ROD remain elusive, partially due to the absence of an appropriate animal model. To address this gap, we established a stable mouse model of ROD using an optimized adenine-enriched diet and conducted exploratory analyses through ribonucleic acid sequencing (RNA-seq). METHODS: Eight-week-old male C57BL/6J mice were randomly allocated into three groups: control group ( n = 5), adenine and high-phosphate (HP) diet group ( n = 20), and the optimized adenine-containing diet group ( n = 20) for 12 weeks. We assessed the skeletal characteristics of model mice through blood biochemistry, microcomputed tomography (micro-CT), and bone histomorphometry. RNA-seq was utilized to profile gene expression changes of ROD. We elucidated the functions of differentially expressed genes (DEGs) using gene ontology (GO) analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, and gene set enrichment analysis (GSEA). DEGs were validated via quantitative real-time polymerase chain reaction (qRT-PCR). RESULTS: By the fifth week, adenine followed by an HP diet induced rapid weight loss and high mortality rates in the mouse group, precluding further model development. Mice with optimized adenine diet-induced ROD displayed significant abnormalities in serum creatinine and blood urea nitrogen levels, accompanied by pronounced hyperparathyroidism and hyperphosphatemia. The femur bone mineral density (BMD) of the model mice was lower than that of control mice, with substantial bone loss and cortical porosity. ROD mice exhibited substantial bone turnover with an increase in osteoblast and osteoclast markers. Transcriptomic profiling revealed 1907 genes with upregulated expression and 723 genes with downregulated expression in the femurs of ROD mice relative to those of control mice. Pathway analyses indicated significant enrichment of upregulated genes in the sphingolipid metabolism pathway. The significant upregulation of alkaline ceramidase 1 ( Acer1 ), alkaline ceramidase 2 ( Acer2 ), prosaposin-like 1 ( Psapl1 ), adenosine A1 receptor ( Adora1 ), and sphingosine-1-phosphate receptor 5 ( S1pr5 ) were successfully validated in mouse femurs by qRT-PCR. CONCLUSIONS Optimized adenine diet mouse model may be a valuable proxy for studying ROD. RNA-seq analysis revealed that the sphingolipid metabolism pathway is likely a key player in ROD pathogenesis, thereby providing new avenues for therapeutic intervention.
Animals ; Mice ; Chronic Kidney Disease-Mineral and Bone Disorder/genetics* ; Male ; Disease Models, Animal ; Mice, Inbred C57BL ; Sphingolipids/metabolism* ; Transcriptome/genetics* ; Signal Transduction/genetics* ; X-Ray Microtomography ; Adenine

Objective To investigate the temporal predictive value of soluble triggering receptor expressed on myeloid cells-1(sTREM-1),procalcitonin(PCT),and myeloid-related protein 8/14(MRP8/14)for secondary acute respiratory distress syndrome(ARDS)in patients with severe pulmonary tuberculosis.Methods A retro-spective cohort study was conducted among patients with severe pulmonary tuberculosis admitted between January 2021 and December 2024.Patients were randomly assigned in an 8∶2 ratio to a training set(n=148)and a valida-tion set(n=37).Serum sTREM-1,PCT,and MRP8/14 were extracted from the electronic medical record at three time points:on admission(day 0),day 3,and day 7.Multivariable logistic regression was used to identify risk factors,and predictive performance was evaluated using receiver operating characteristic(ROC)curves.Results A total of 185 patients were included.In the training set(n=148),27 developed ARDS and 121 did not;in the validation set(n=37),7 developed ARDS and 30 did not.In the training set,serum sTREM-1,PCT,and MRP8/14 levels showed significant temporal changes(P<0.05).At admission,day 3,and day 7,levels of sTREM-1,PCT,and MRP8/14 were higher in the ARDS group than in the non-ARDS group(all P<0.05).At each time point,sTREM-1,PCT,and MRP8/14 were independently associated with the development of ARDS(P<0.05).In the training set,the combination of sTREM-1,PCT,and MRP8/14 at admission yielded the largest area under the ROC curve[AUC=0.976;95%confidence interval(CI),0.952～1.000],with a sensitivity of 88.9%and a specificity of 98.3%.In the validation set,the same combination achieved an AUC of 0.957(95%CI,0.895～1.000),with a sensitivity of 100.0%and a specificity of 86.7%.Conclusion Dynamic changes in sTREM-1,PCT,and MRP8/14 provide temporal predictive value for ARDS in patients with severe pulmonary tuberculosis,and the combined assessment improves early warning accuracy.

Objective To investigate the effects of scribble planar cell polarity protein(SCRIB)on proliferation,apoptosis,and autophagy of glioblastoma(GBM)and elucidate its potential underlying mechanisms.Methods The expression level of SCRIB in GBM tissue was queried through the Biomarker Exploration of Solid Tumors(BEST)database.Lentivirus-mediated shRNA interference was employed to downregulate SCRIB expression in human glioblastoma cell lines U87 and U251,which were divided into negative control group(mock)and SCRIB shRNA interference groups(kd1 and kd2).SCRIB expression levels were detected using Western blotting(WB)and quantitative polymerase chain reaction(qPCR).EdU incorporation and cell apoptosis rates were detected by flow cytometry(FCM).CCK-8 assay was used to detect the proliferation vitality of U87 and U251 cells,and WB was used to detect the expression of proliferation-related proteins.Immunofluorescence(IF)staining was conducted to detect the expression of autophagy-related proteins LC3 and p62,followed by quantitative analysis across multiple fields.WB was also used to detect the expression levels of LC3,p62,and proteins in the JAK-STAT3 signaling pathway.Results Compared with that of normal tissues,SCRIB mRNA expression level was significantly upregulated in GBM tissues(P＜0.05).FCM results showed that EdU incorporation rates were significantly reduced(P＜0.001)while cell apoptosis rates were markedly increased(P＜0.001)in U87 and U251 cells with SCRIB knockdown.CCK-8 results indicated that compared with the mock group,the proliferation vitality of U87 and U251 cells in the SCRIB knockdown group was significantly downregulated(P＜0.001).IF staining showed that LC3 fluorescence aggregation was significantly enhanced(P＜0.001),while p62 fluorescence aggregation was significantly reduced(P＜0.001)in the SCRIB knockdown group.WB results showed that compared with the mock group,the protein expression levels of p27,LC3,p-JAK2 and p-STAT3 were upregulated,while C-Myc,Cyclin D1,MCM,PCNA and p62 were downregulated,with statistically significant differences(P＜0.05).Conclusion Downregulation of SCRIB may induce autophagy and apoptosis in glioblastoma cells by inhibiting the JAK-STAT3 signaling pathway,thereby suppressing cell proliferation.

Objective To evaluate answers to typical clinical questions related to neuro-ophthalmology generated by Artificial Intelligence(AI)Large Language Models(LLM)and to explore the performance of neuro-ophthalmology-related questions on LLM in a multidimensional manner using objective and expert assessment.Methods Multicenter,random-ized,cross-sectional pilot study.Thirty typical questions related to neuro-ophthalmology were selected based on four per-spectives:definition,etiology,clinical manifestations and signs,and treatment and prognosis,and were analyzed quantita-tively using Deepseek,Wenxin Yiyin 4.0,Doubao,and Kimi 1.5,which are four open-source LLMs in China,and quantita-tively analyzed with objective assessment;and quantitatively rated by three ophthalmologists using expert assessment for 120 answer texts.Three ophthalmology experts quantitatively scored the 120 answer texts.Three ophthalmologists quantita-tively scored the 120 answer texts.Level 3,5,and 4 Likert scales were developed according to the completeness,accura-cy,professionalism,relevance,and criticality of the question texts,respectively.The best-performing LLM was selected,and its performance was observed across the four types of questions.Additionally,three other experts assessed whether the best-performing one could be evaluated as a substitute for real-world doctor-patient communication.Results In the objective Chinese text reading difficulty analysis,the differences in total word count among the four LLMs were statistically significant(all P＜0.001).Of the four LLMs,Kimi 1.5 performed the best,with frequencies of 61％,29％,and 41％for the highest scores in completeness(3),accuracy and professionalism(5),and relevance and usefulness(4),respective-ly.Kimi 1.5 performed more consistently on the questions on the four areas of neuro-ophthalmologic disorders:definition,etiology,clinical manifestations and signs,treatment,and prognosis,with no between-group differences(P＞0.05).Con-clusion Chinese language LLMs have great potential in the clinical application of neuro-ophthalmology.Kimi 1.5 outper-forms other LLMs in terms of completeness,accuracy,professionalism,relevance,and usefulness,but it still cannot re-place real-world doctor-patient communication.There is a need to explore new diagnostic and therapeutic model of AI+physician in the future.