Renal lymphatic vessel density is clinically relevant in kidney disease but is still assessed by slow, subjective visual estimation. We evaluated a weakly supervised, attention-based multiple-instance learning framework for automated detection and quantification of renal lymphatic vessel density on D2-40-stained whole-slide images (WSIs). Methods: Two independent internal datasets from Tongji Hospital were collected, including 198 cases of chronic kidney disease (CKD) and 50 cases of hypertensive nephropathy (HTN). All biopsies were immunohistochemically stained for D2-40 and digitized as WSIs. Pathologists provided only slide-level labels (D2-40 high vs. D2-40 low). Tissue regions were automatically segmented, tiled into patches, and encoded using a pretrained convolutional neural network. Patch embeddings were then analyzed with a clustering-constrained attention multiple-instance learning (CLAM) model. Unlike conventional multiple-instance learning (MIL) methods that only weight instances, CLAM jointly performs attention-based instance selection and instance-level clustering to distinguish positive from negative evidence within each slide, yielding more discriminative slide-level features and interpretable attention maps. Performance was compared with a classic MIL model trained on the same features. Results: CLAM achieved area under the receiver operating characteristic curves of 0.942 and 0.858 on the CKD and HTN datasets, respectively, outperforming classic MIL (0.866 and 0.801). Attention maps highlighted lymphatic-rich regions consistent with renal pathologists’ assessments. Conclusions: This clustering-constrained, attention-based weakly supervised framework enables fully automated, reproducible quantification of renal lymphatic vessel density from WSIs, providing renal pathologists with rapid visual and numerical support for diagnosis and risk stratification in CKD and HTN.

Background/Aims: Dysregulated cholesterol metabolism is a hallmark of hepatocellular carcinoma (HCC) that drives tumor initiation and progression. However, clinical targeting of cholesterol metabolism has yielded limited benefits due to stringent feedback in tumor cells. Identifying a central mediator capable of restoring cholesterol homeostasis within the cell’s intrinsically fine-tuned regulatory framework is urgently needed. Methods: We integrated a proteomic dataset from patients with cholesterol-dysregulated HCC into a global cholesterol metabolic regulatory network to identify potential therapeutic targets for disrupted cholesterol homeostasis. The prognostic significance of the candidate targets was further validated in an independent cohort through immunohistochemistry. Functional and mechanistic studies were conducted in vitro using HCC cell lines and in vivo using mouse models. The pharmacological efficacy of the candidate agent was evaluated in both subcutaneous and orthotopic HCC mouse models. Results: ER lipid raft-associated 1 (ERLIN1), a pivotal regulator of cholesterol metabolism reprogramming, was identified as an independent favorable prognostic indicator in HCC. ERLIN1 constrains HCC progression both in vitro and in vivo by stabilizing the INSIG1–SCAP–SREBP2 axis and maintaining the metabolic balance of intracellular cholesterol. Under hypoxia, impaired factor-inhibiting hypoxia-1-dependent hydroxylation of ASB11 at asparagine residues 90 and 92 enhances ASB11-mediated ERLIN1 degradation. Pharmacological targeting of this axis using zoledronic acid (ZoA) attenuated HCC progression by weakening the ASB11–ERLIN1 interaction and restoring cholesterol homeostasis. Conclusions ERLIN1 represents a druggable metabolic vulnerability in cholesterol-dysregulated HCC. Targeting the ASB11–ERLIN1 axis with the clinically approved ZoA reestablishes cholesterol homeostasis and offers a promising therapeutic strategy to overcome the current limitations of cholesterol-targeted HCC therapies.

Purpose: The aim of this study was to investigate the feasibility of an intelligent handheld ultrasound (US) device for assisting non-expert general practitioners (GPs) in detecting carotid plaques (CPs) in community populations. Methods: This prospective parallel controlled trial recruited 111 consecutive community residents. All of them underwent examinations by non-expert GPs and specialist doctors using handheld US devices (setting A, setting B, and setting C). The results of setting C with specialist doctors were considered the gold standard. Carotid intima-media thickness (CIMT) and the features of CPs were measured and recorded. The diagnostic performance of GPs in distinguishing CPs was evaluated using a receiver operating characteristic curve. Inter-observer agreement was compared using the intragroup correlation coefficient (ICC). Questionnaires were completed to evaluate clinical benefits. Results: Among the 111 community residents, 80, 96, and 112 CPs were detected in settings A, B, and C, respectively. Setting B exhibited better diagnostic performance than setting A for detecting CPs (area under the curve, 0.856 vs. 0.749; P<0.01). Setting B had better consistency with setting C than setting A in CIMT measurement and the assessment of CPs (ICC, 0.731 to 0.923). Moreover, measurements in setting B required less time than the other two settings (44.59 seconds vs. 108.87 seconds vs. 126.13 seconds, both P<0.01). Conclusion Using an intelligent handheld US device, GPs can perform CP screening and achieve a diagnostic capability comparable to that of specialist doctors.

Objective:To observe the role of impulsivity in cognitive impairment in manic episodes of bipolar Ⅰ disorder(BPDTI)patients.Methods:180 cases of patients with BPDTI manic episodes admitted to our hospital from February 2024 to December 2024 were selected as the manic group,and another 180 cases of volunteers who were psychologically tested as normal during the same period were selected as the healthy group.The impulse Barratt-11 scale(BIS-11)and the MATRICS Consensus Cognitive Test(MCCB)were used to investigate and evaluate impulsivity and cognitive function,and correlation analysis and risk factor analysis were conducted.Results:In the BIS-11 scale,there was no statistically significant difference between the two groups in the unplanned factor(P＞0.05),but there were significant differences compared between the two groups in cognitive,motor factors,and total score(P＜0.05).The MCCB scores of the manic group in information processing speed,working memory,word learning,visual learning,social cognition,reasoning and problem solving,and attention vigilance were significantly lower than those of the healthy group(P＜0.05).The relationship between BIS-11 total score and cognitive impulsivity factor and MCCB total score and word learning showed there were negative correlation(P＜0.05);The exercise factor in the BIS-11 scale were negatively correlated with the MCCB total score and working memory factor(P＜0.05);The unplanned factor in the BIS-11 scale showed there were negative correlation with the MCCB total score,word learning(P＜0.05),and information processing speed.BIS-11,the number of years of education was an influencing factor for BPDTI mania(P＜0.05).Conclusion:BPDTI mania patients had high impulsivity and cognitive impairment,high impulsivity and number of years of education were influencing factors for mania,high impulsivity affected cognitive function through corresponding brain regions,aggravating the condition.

Pancreatic cancer is highly aggressive and often diagnosed at an advanced stage,leaving most patients ineligible for radical resection.This study retrospectively analyzed four patients with locally advanced or advanced pancreatic cancer to evaluate the clinical efficacy and safety of high-intensity focused ultrasound(HIFU)ablation combined with chemotherapy as a neoadjuvant and conversion therapy.All cases were reviewed and individualized treatment plans were formulated through a multidisciplinary team evaluation.All patients received HIFU plus gemcitabine and nab-paclitaxel chemotherapy,with assessments of tumor volume,vascular involvement,surgical conversion,symptom relief,and adverse events.Three patients achieved marked tumor shrinkage and reduction of vascular invasion,enabling successful R0 resection without recurrence during follow-up.The remaining patient achieved disease stability,significant pain relief,and maintained good quality of life under repeated HIFU therapy.All treatments were well tolerated,and no severe adverse reactions occurred.The combination of HIFU and chemotherapy demonstrated synergistic local and systemic effects,effectively achieving tumor downstaging,improving resectability,and alleviating symptoms.As a safe,noninvasive,and repeatable therapeutic approach,this strategy offers a promising option for patients with advanced pancreatic cancer.Further large-scale prospective studies are warranted to validate its long-term efficacy and elucidate underlying mechanisms.

Objective:To explore the effect of Lactobacillus reuteri on testicular injury in mice exposed to neonicotinoid insec-ticides(NNI).Methods:Fifteen C57BL/6 male mice were randomly divided into control group(CTRI.group),exposure group(NNI group)and Lactobacillus intervention group(NNI-L group).The mice in CTRL group were given 0.02ml/g of 0.5％carboxym-ethyl cellulose sodium solution by gavage for 14 days.The mice in NNI group were given 0.02 ml/g of NNI mixture by gavage for 14 days.The mice in NNI-L group were given 0.02 ml/g of NNI mixture by gavage and 5 × 108cfu/ml of Lactobacillus reuteri powder so-lution for 14 days.Then,the histomorphology and function of testicle were evaluated by hematoxylin-eosin staining,immunofluores-cence staining and RNA sequencing.Results:Compared with CTRL group,the thickness of testicular seminiferous epithelium in the NNI group was significantly thinner.And the decline in the number of spermatogenic cells and sperm was observed.And the expression of spermatogonial stem cell marker UCHL1 was down-regulated which was significantly improved in NNI-L group compared with the NNI group.The abnormal expressions of hormone and sperm methylation related genes in testis of NNI group were detected by RNA sequen-cing,with significant down-regulation being found in NPFF and IGF2.While the expression of HSD3B8 was significantly up-regulated.The abnormal expression of these genes could be significantly improved after oral administration of Lactobacillus reuteri.Conclusion:Testicular spermatogenesis and endocrine function can be damaged by NNI exposure.And oral administration of Lactoba-cillus reuteri protects testis from the adverse effects of NNI toxicity.

Objective:To study the relationship between microRNA (miRNA, miR)-675-3p, miR-675-5p, miR-29b-3p, miR-let-7b-3p and fluoride induced articular cartilage injury in rats.Methods:Using the factorial design, thirty 3-week-old specific pathogen free grade male Wistar rats (weighted 125 - 150 g) were selected and randomly divided into a control group, a 25 mg/L fluoride group, and a 50 mg/L fluoride group using a random number table method, with 10 rats in each group. The control group drank distilled water, while the fluoride exposure groups drank distilled water with fluoride ion concentrations of 25 and 50 mg/L, respectively. Five rats were euthanized in each group at 3 and 6 months of feeding, respectively. Visual observation was used to observe the occurrence of dental fluorosis in rats, and fluoride ion selective electrode method was used to detect the fluoride level in blood, urine, and cartilage. Hematoxylin-eosin staining and safranin O-fast green staining were used to observe the pathological changes of articular cartilage, and Mankin score was used to evaluate the grading of cartilage injury. Real-time fluorescence quantitative PCR was used to detect the expression levels of miR-675-3p, miR-675-5p, miR-29b-3p, and miR-let-7b-3p in cartilage.Results:After 3 and 6 months of fluoride exposure, no dental fluorosis was observed in the control group, while rats in the 25 and 50 mg/L fluoride groups showed varying degrees of dental fluorosis. There were statistically significant differences in the levels of blood fluoride (mg/L: 0.11 ± 0.04, 0.57 ± 0.32, 0.29 ± 0.06, 0.07 ± 0.01, 0.31 ± 0.05, 0.38 ± 0.06), urine fluoride (mg/L: 1.81 ± 0.58, 13.18 ± 2.29, 66.11 ± 20.74, 2.35 ± 1.08, 14.79 ± 3.87, 28.32 ± 4.79), and cartilage fluoride (mg/kg: 341.83 ± 44.07, 612.99 ± 174.72, 991.26 ± 227.32, 338.29 ± 72.53, 957.09 ± 195.86, 1 535.53 ± 89.01) among in rats the control group, 25 mg/L fluoride group, and 50 mg/L fluoride group ( F = 7.76, 42.78, 40.54, 23.10, 18.96, 80.81, P < 0.05). In the 50 mg/L fluoride group, there were statistically significant differences in the levels of urine fluoride and cartilage fluoride of rats exposed for different times ( t = 4.45, - 3.80, P < 0.05). The Mankin score grading for cartilage injury showed that at 3 months of fluoride exposure, there were 4, 0, and 0 rats with normal cartilage in the control group, 25 mg/L fluoride group, and 50 mg/L fluoride group, 1, 4, and 1 rats with mild injury, and 0, 1, and 4 rats with moderate injury, respectively. At 6 months of fluoride exposure, there were 4, 0, and 0 rats with normal cartilage in the control group, 25 mg/L fluoride group, and 50 mg/L fluoride group, 1, 3, and 0 rats with mild injury, 0, 1, and 3 rats with moderate injury, and 0, 1, and 2 rats with severe injury, respectively. Real-time fluorescence quantitative PCR results showed that fluoride exposure dose had individual effects on the expression of miR-675-3p, miR-675-5p, miR-29b-3p, and miR-let-7b-3p in cartilage ( F = 8.68, 7.97, 9.34, 10.14, P < 0.05). There was no individual effect of fluoride exposure time on the expression of miR-675-3p, miR-675-5p, miR-29b-3p, and miR-let-7b-3p in cartilage ( F = 0.00, 0.15, 0.63, 0.53, P > 0.05). However, there was no interaction effect between fluoride exposure time and dose on the above-mentioned miRNA ( F = 0.68, 0.05, 0.22, 0.24, P > 0.05). The correlation analysis results showed that miR-675-3p and miR-675-5p in cartilage were negatively correlated with blood fluoride, urine fluoride, and cartilage fluoride ( r = - 0.37, - 0.42, - 0.56, - 0.53, - 0.57, - 0.53, P < 0.05), while miR-29b-3p and miR-let-7b-3p were positively correlated with urine fluoride and cartilage fluoride ( r = 0.58, 0.40, 0.48, 0.47, P < 0.05). The results of ordered logistic regression analysis showed that miR-675-3p, miR-675-5p, miR-29b-3p, and miR-let-7b-3p were influencing factors of dental fluorosis grading ( OR = 0.13, 0.04, 1.55, 2.58, P < 0.05) and Mankin score grading ( OR = 0.04, 0.06, 1.41, 1.58, P < 0.05). Conclusion:MiR-675-3p, miR-675-5p, miR-29b-3p, and miR-let-7b-3p may be involved in the process of fluoride induced articular cartilage injury.

Aim To investigate the antidepressant mechanism of hyperforin via the utilization of single-cell sequencing technology.Methods C57BL/6 mice were randomly divided into the control group,depres-sion model group,and hyperforin intervention group.The chronic unpredictable mild stress(CUMS)model was induced and drug interventions were administered for 28 d.Behavioral experiments were conducted to as-sess depressive symptoms,and hippocampal tissue was collected for single-cell RNA sequencing.Key cell populations and differentially expressed genes across groups were identified,followed by PPI network,GO,and KEGG enrichment analysis.Results Behavioral experiments indicated that CUMS successfully induced depressive symptoms in mice,while hyperforin im-proved depressive behavior.In the depression model group,the proportion of brain perivascular macrophages(PVM)increased,and this proportion decreased after hyperforin intervention,approaching the level seen in the control group.The top 20 common differentially ex-pressed genes in the PVM subpopulation were Saa3,Hbb-bs and Ccl24.PPI network analysis identified core targets,including Ccl2,Dhx9,C3,Msr1,Cxcl2 and Cx3cr1.KEGG enrichment analysis revealed pathways related to chemokines,phagosome formation,and inosi-tol phosphate metabolism.Conclusion The antide-pressant mechanism of hyperforin may be related to the regulation of Ccl24 and its related chemokine signaling pathway by PVM.

Objective In view of the potential threat of adverse drug reaction(ADR)to patients'health and the limitations of traditional monitoring methods,this study investigates the construction method of an ADR knowledge graph based on a knowledge-driven technology path to promote the intelligent upgrading of pharmacovigilance field.Methods In this study,an adverse drug reaction knowledge base was established,and the data within the knowledge base were preprocessed.The BERT-CRF model was utilized to identify and extract the relevant entities,relationships and attribute information,and knowledge alignment was carried out by combining the rule method and statistical method.The knowledge fusion data was stored in the Neo4j graph database to construct the knowledge graph.Results Based on the established knowledge base,this study explored the method of constructing knowledge graph of adverse drug reaction by knowledge-driven technology,laying the groundwork for data mining and the development of intelligent auxiliary system tools based on knowledge graph.Conclusions The study on the construction method of an ADR knowledge graph based on a knowledge-driven technology path provides a new perspective and technical path for the intelligent upgrading of pharmacovigilance field.It holds important theoretical and practical significance for improving drug safety,protecting patients'health,and promoting the healthy development of pharmaceutical industry.

Objective To investigate the biomechanical characteristics of Salto Talaris tibial components with different heights at the bone-prosthesis interface during different gait support phases after total ankle replacement.Methods An ankle joint model was reconstructed using a weight-bearing CT from a 61-year-old female patient with ankle arthritis,and Salto Talaris tibial components with different heights(5,7,9,11 mm)were modelled to simulate the loading of the tibial-prosthesis during four gait support phases,and to analyse the micromotion and stresses at the bone-prosthesis interface.Results The 11 mm and 9 mm models had a poorer prosthesis stability,with the peak micromotion exceeding 50 μm and the peak internal tibial stresses of 30.75 MPa and 29.86 MPa,respectively,which exceeded the yield stress of the cancellous bone.The tibial stresses of the 7 mm and 5 mm models were within reasonable ranges and the average peak micromotions were only 42.66 μm and 40.32 μm.In contrast,the initial stability of the 5 mm model prosthesis was the best.Conclusions For total ankle replacement with Salto prosthesis,the height of the tibial component should be chosen appropriately,and the optimal height was about 5 mm.Excessive flexion and extension activities of the ankle joint should be avoided to maintain the stability of the prosthesis after surgery.This study provides a theoretical basis for the improvement of the structural parameters of the Salto prosthesis,which is valuable for the selection of clinical surgical prostheses and helps to improve the results of total ankle replacement.