ObjectiveThe malaria parasites remodel the host erythrocyte structure by exporting parasite proteins that interact with the membrane skeleton proteins of red blood cells (RBCs), facilitating their intracellular survival and pathogenicity. Skeleton-binding protein 1 (SBP1) is a conserved exported protein across Plasmodium species. In Plasmodium falciparum, SBP1 has been reported to interact with erythrocyte membrane skeleton proteins 4.1R and spectrin, while its contribution to erythrocyte remodeling and parasite virulence in Plasmodium berghei (Pb) remains unclear. This study aims to determine whether PbSBP1 associates with the host cytoskeletal protein 4.1R and to investigate its role in the remodeling of host RBCs and the pathogenicity of Plasmodium berghei. MethodsIn Plasmodium berghei, the relationship between PbSBP1 and the erythrocyte cytoskeletal protein 4.1R was examined using co-immunoprecipitation. A Pbsbp1 gene knockout mutant of Plasmodium berghei (Pbsbp1∆) was generated based on the principle of double crossover homologous recombination. The deformability of erythrocytes infected with Pbsbp1∆ parasites was assessed using microfluidic methods. Microchannels with an array of cylindrical pillars were used to detect modifications in infected RBC deformability. The infected RBCs were squashed between the rows and recovered between the columns and the transit velocity (μm/s) of infected RBCs travelling through the microchannel was recorded. The component of the erythrocyte membrane skeleton junctional complex, tropomodulin (TMOD), was fluorescently labeled, and the cytoskeletal network of infected erythrocytes was imaged using super-resolution stochastic optical reconstruction microscopy (STORM) to analyze ultrastructural changes in the cytoskeleton of wild-type (WT) and Pbsbp1∆-infected erythrocytes. Actin-based junctional complexes were displayed as individual clusters by the labeled TMOD in the STORM images, and the cluster densities and distances between adjacent clusters of infected RBCs were calculated. Additionally, rodent malaria models (BALB/c mice) and experimental cerebral malaria models (C57BL/6 mice) were employed to monitor the growth of Pbsbp1∆ and WT parasites during the intraerythrocytic stage and their capacity to induce cerebral malaria in mice. ResultsPbSBP1 may participate in the remodeling of infected erythrocytes through direct or indirect interaction with the erythrocyte cytoskeletal protein 4.1R. Microfluidic assays revealed that the deformability of erythrocytes infected with Pbsbp1∆ parasites was significantly enhanced compared to those infected with WT parasites. STORM imaging further demonstrated that the ultrastructure of the erythrocyte cytoskeleton in Pbsbp1∆-infected cells was altered relative to that in WT-infected erythrocytes. The distances between nearest neighbors of clusters had a tendency to increase while the cluster densities were decreased in Pbsbp1∆-infected RBCs compared to WT-infected RBCs. Subsequent phenotypic analysis indicated that the growth rate of Pbsbp1∆ parasites during the intraerythrocytic stage was significantly slower than that of WT parasites, and their ability to induce cerebral malaria in mice was also attenuated. These findings suggest that PbSBP1 is involved in the remodeling of the erythrocyte membrane skeleton, likely through its direct or indirect interaction with protein 4.1R, thereby regulating the deformability of infected erythrocytes and influencing the pathogenicity of the blood-stage parasites. ConclusionThis study establishes a role for PbSBP1 in host erythrocyte remodeling and parasite virulence, providing new research strategies for the prevention and treatment of malaria.

Stroke is one of the leading causes of disability and mortality worldwide. Research into its mechanisms and the development of therapeutic strategies heavily rely on animal models that accurately replicate the pathological features of human disease. An ideal animal model for stroke should not only reproduce the neurological deficits and pathological changes observed in clinical patients but also demonstrate good reproducibility and translational value. This review focuses on the preparation and evaluation methods of ischemic stroke animal models. Firstly, it elaborates on the selection criteria, advantages, and disadvantages of experimental animals, including rodents (rats, mice) and non-rodents (non-human primates, miniature pigs, rabbits, zebrafish). Secondly, it provides a detailed overview of the modeling principles, key procedures, and application scopes for ischemic stroke models and hemorrhagic stroke models. Furthermore, the review summarizes advances in the applications of emerging technologies—including gene editing [e.g., clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) gene editing], multimodal imaging (e.g., two-photon microscopy, photoacoustic imaging), artificial intelligence, optogenetics, 3D bioprinting, organoid models, and multi-omics–in model optimization, precise assessment, and mechanistic investigation. Finally, based on a systematic analysis of relevant domestic and international literature from 2019 to 2024, this review discusses model selection strategies based on research objectives, a multidimensional evaluation system encompassing behavioral, imaging, and molecular pathological assessments, and envisions future directions involving technological integration to achieve model precision and individualization. This article aims to provide a comprehensive methodological reference to help researchers select appropriate animal models of stroke according to specific scientific questions.

Objective To construct and evaluate a prognostic model for clear cell renal cell carcinoma(ccRCC)based on preoperative lipid ratios, cM0(i+)staging, and other clinical characteristics, so as to provide a precise tool for clinical prognosis assessment. Methods A retrospective analysis was conducted on the clinical data of 215 ccRCC patients treated in our hospital during May 2014 and May 2023. Lipid ratios were calculated using preoperative lipid data, and patients were divided into cM0(i+)stage or cM0 stage according to postoperative circulating tumor cells(CTCs)test results. The optimal lipid ratio and cutoff value were selected using receiver operating characteristic(ROC)curves and the X-tile method. Patients were subdivided into three groups based on the level of the low-density lipoprotein cholesterol to non-low-density lipoprotein cholesterol ratio(LNLR)and cM0(i+)staging:Group A [LNLR＞1.68 and diagnosed with cM0(i+)], Group B [LNLR＞1.68 or diagnosed with cM0(i+)], and Group C(LNLR ≤1.68 and diagnosed with cM0). Kaplan-Meier survival analysis was used to plot survival curves for patients in different groups. The log-rank test was employed to compare differences in recurrence-free survival(RFS)among the subgroups. Multivariate Cox regression analysis was conducted to identify the independent risk factors influencing RFS, and a nomogram prediction model was constructed based on these results. The predictive performance of the model was validated using ROC curves, calibration curves, and decision curves. Results ROC curves were plotted for lipid markers, and LNLR was identified as the most predictive for RFS. Its optimal cutoff point was 1. 68. Patients in Group A experienced a significantly shorter postoperative RFS. Multivariate Cox regression analysis identified preoperative LNLR, cM0(i+)staging, pathological grade, and stage as independent risk factors for RFS. A nomogram model was constructed based on these risk factors. The area under the ROC curve(AUC)for 1-, 3-, and 5-year RFS was 0.896(95% CI:0.8121-0.9627), 0.890(95%CI:0.7879-0.9641), and 0.870(95%CI:0.7697-0.9526), indicating good discriminatory ability and predictive performance. Calibration plots demonstrated good agreement between predicted and actual outcomes. Clinical decision curve analysis showed high clinical net benefit. Conclusion Preoperative LNLR level is an independent risk factor for RFS in ccRCC patients. The prognostic prediction model based on LNLR, cM0(i+)staging, patient pathological grade, and staging demonstrates good predictive performance for RFS and holds potential clinical application value.

Objective To address the need for rapid review of automated organ-at-risk (OAR) contouring for head-and-neck cancer in online adaptive radiotherapy, this study used dosimetric indices as the reference standard to evaluate the suitability of geometric similarity metrics and to develop a rapid screening model that balances the risk of missed errors with review efficiency. Methods This retrospective study included 29 patients with head-and-neck radiotherapy, yielding 243 pairs of OAR contours (auto-generated vs. manual). Geometric similarity metrics, including the Dice similarity coefficient (Dice), the 95th percentile Hausdorff distance (HD95), and the maximum Hausdorff distance (HD100), were computed between the two contour sets. Using the dose distributions from clinical treatment plans, dosimetric differences between the two contour sets were calculated for key indices, including mean dose (D_mean) and maximum dose (D_max). Correlations between geometric metrics and dosimetric differences were analyzed. Dosimetric discrepancy events were defined as Y_3Gy using a threshold of 3 Gy. Within a univariable logistic regression triage framework, each geometric metric was used as an independent variable to estimate the probability of Y_3Gy. Discriminative performance was assessed using receiver operating characteristic (ROC) curves and precision-recall (PR) curves, with the area under the curve (AUC) including ROC-AUC and PR-AUC. Practical operating thresholds were determined via threshold sweeping. Results Geometric similarity metrics showed weak-to-moderate correlations with dosimetric differences; compared with maximum dose, correlations with mean dose differences were more consistent. Among the evaluated metrics, HD95 achieved the best classification performance for Y_3Gy. Threshold sweeping suggested that an HD95 threshold in the range of 4-9 mm can balance the risk of missed discrepancies with the efficiency of automated contour quality assurance (QA). Conclusion Several geometric similarity metrics demonstrated only weak-to-moderate associations with dosimetric differences. For head-and-neck OAR contour QA in this study, HD95 provided the best discriminative performance and can support rapid triage of automated contours, with a practical operating threshold range of 4-9 mm, potentially improving the efficiency of adaptive radiotherapy workflows.

Objective To address the need for rapid review of automated organ-at-risk (OAR) contouring for head-and-neck cancer in online adaptive radiotherapy, this study used dosimetric indices as the reference standard to evaluate the suitability of geometric similarity metrics and to develop a rapid screening model that balances the risk of missed errors with review efficiency. Methods This retrospective study included 29 patients with head-and-neck radiotherapy, yielding 243 pairs of OAR contours (auto-generated vs. manual). Geometric similarity metrics, including the Dice similarity coefficient (Dice), the 95th percentile Hausdorff distance (HD95), and the maximum Hausdorff distance (HD100), were computed between the two contour sets. Using the dose distributions from clinical treatment plans, dosimetric differences between the two contour sets were calculated for key indices, including mean dose (D_mean) and maximum dose (D_max). Correlations between geometric metrics and dosimetric differences were analyzed. Dosimetric discrepancy events were defined as Y_3Gy using a threshold of 3 Gy. Within a univariable logistic regression triage framework, each geometric metric was used as an independent variable to estimate the probability of Y_3Gy. Discriminative performance was assessed using receiver operating characteristic (ROC) curves and precision-recall (PR) curves, with the area under the curve (AUC) including ROC-AUC and PR-AUC. Practical operating thresholds were determined via threshold sweeping. Results Geometric similarity metrics showed weak-to-moderate correlations with dosimetric differences; compared with maximum dose, correlations with mean dose differences were more consistent. Among the evaluated metrics, HD95 achieved the best classification performance for Y_3Gy. Threshold sweeping suggested that an HD95 threshold in the range of 4-9 mm can balance the risk of missed discrepancies with the efficiency of automated contour quality assurance (QA). Conclusion Several geometric similarity metrics demonstrated only weak-to-moderate associations with dosimetric differences. For head-and-neck OAR contour QA in this study, HD95 provided the best discriminative performance and can support rapid triage of automated contours, with a practical operating threshold range of 4-9 mm, potentially improving the efficiency of adaptive radiotherapy workflows.

Objective To address the need for rapid review of automated organ-at-risk (OAR) contouring for head-and-neck cancer in online adaptive radiotherapy, this study used dosimetric indices as the reference standard to evaluate the suitability of geometric similarity metrics and to develop a rapid screening model that balances the risk of missed errors with review efficiency. Methods This retrospective study included 29 patients with head-and-neck radiotherapy, yielding 243 pairs of OAR contours (auto-generated vs. manual). Geometric similarity metrics, including the Dice similarity coefficient (Dice), the 95th percentile Hausdorff distance (HD95), and the maximum Hausdorff distance (HD100), were computed between the two contour sets. Using the dose distributions from clinical treatment plans, dosimetric differences between the two contour sets were calculated for key indices, including mean dose (D_mean) and maximum dose (D_max). Correlations between geometric metrics and dosimetric differences were analyzed. Dosimetric discrepancy events were defined as Y_3Gy using a threshold of 3 Gy. Within a univariable logistic regression triage framework, each geometric metric was used as an independent variable to estimate the probability of Y_3Gy. Discriminative performance was assessed using receiver operating characteristic (ROC) curves and precision-recall (PR) curves, with the area under the curve (AUC) including ROC-AUC and PR-AUC. Practical operating thresholds were determined via threshold sweeping. Results Geometric similarity metrics showed weak-to-moderate correlations with dosimetric differences; compared with maximum dose, correlations with mean dose differences were more consistent. Among the evaluated metrics, HD95 achieved the best classification performance for Y_3Gy. Threshold sweeping suggested that an HD95 threshold in the range of 4-9 mm can balance the risk of missed discrepancies with the efficiency of automated contour quality assurance (QA). Conclusion Several geometric similarity metrics demonstrated only weak-to-moderate associations with dosimetric differences. For head-and-neck OAR contour QA in this study, HD95 provided the best discriminative performance and can support rapid triage of automated contours, with a practical operating threshold range of 4-9 mm, potentially improving the efficiency of adaptive radiotherapy workflows.

Background Lung tissue fibrosis caused by exposure to free silica (SiO₂) dust is the core pathological feature of silicosis. Lipocalin-2 (LCN2), as a secreted glycoprotein, plays an important role in various inflammatory diseases, but its specific role in silicosis-induced inflammatory injury remains unclear. Objective To identify differentially expressed genes (DEGs) and enriched pathways in the lung tissues of rats with silicosis, and to explore the regulatory mechanism of LCN2 in silicosis inflammation. Methods Twenty specific pathogen-free male SD rats were randomly divided into a control group and a silicosis model group (n=10 per group). The silicosis model was established via a single intratracheal instillation of silicon dioxide (SiO₂) suspension (50 mg), while the control group received an equal volume of normal saline. In vitro, NR8383 cells were divided into a control group and a SiO₂ treatment group. Hematoxylin-eosin (HE) staining and Van Gieson (VG) staining were used to evaluate alveolar structure damage, inflammatory infiltration, and collagen deposition in the lung tissues. Transcriptomic sequencing was applied to screen for DEGs and enriched signaling pathways. Western blot was used to detect the protein expression of interleukin-6 (IL-6), E-cadherin (ECA), cluster of differentiation 36 (CD36), liver X receptor (LXR), ATP-binding cassette transporter A1 (ABCA1), and LCN2 in both rat lung tissues and NR8383 cells. Immunohistochemistry was used to further assess LCN2 expression and localization. Additionally, NR8383 cells were treated with the LCN2 inhibitor ZINC00640089 and assigned to four groups: control, ZINC00640089-only, SiO₂, and SiO₂+ZINC00640089. Western blot was used to evaluate the expression of LCN2, IL-6, and interleukin-1β (IL-1β). Results HE and VG staining revealed disorganized lung tissue structure, damaged alveolar walls, nodule formation, and increased collagen deposition in the silicosis model group compared to the controls. Transcriptomic analysis identified 2723 DEGs in the lung tissues of silicosis rats (312 down-regulated and 2411 up-regulated) with LCN2 expression being significantly up-regulated. These DEGs were primarily enriched in pathways related to the innate immune response, extracellular matrix (ECM)-receptor interaction, and inflammatory response. Immunohistochemical staining confirmed elevated LCN2 levels in the lung tissues of the model rats. Western blot demonstrated that the protein levels of IL-6, CD36, and LCN2 were significantly increased (P<0.05), whereas ABCA1 and ECA were significantly decreased in the silicosis group (P<0.05). Similarly, in SiO₂-treated NR8383 cells, IL-6, CD36, and LCN2 expressions significantly increased (P<0.05), while ABCA1 and LXR significantly decreased (P<0.05). Compared with the SiO₂ group, treatment with ZINC00640089 significantly reduced the expression levels of LCN2, IL-6, and IL-1β (P<0.05). Conclusion LCN2 is significantly upregulated in both the lung tissues of silicosis rats and SiO₂-treated NR8383 cells. Inhibiting LCN2 expression effectively reduces the SiO₂-induced inflammatory response, indicating that LCN2 serves as a potential therapeutic target for silicosis.

The recombinant human thyroid-stimulating hormone (rhTSH) can be used for differentiated thyroid cancer (DTC) preparation before ¹³¹I therapy, and follow-up. Since its approval by the American Food and Drug Administration (FDA) in 1998, rhTSH has been clinically used in European countries, the United States, and other countries for nearly 30 years. In April 2024, China’s first independently developed rhTSH agent was approved. Based on clinical experience and evidence from domestic and international studies, an expert consensus on the application of rhTSH has been developed following thorough consultations among experts. The consensus offers evidence-based recommendations to guide the appropriate use of rhTSH in clinical practice.

Object:To explore the feasibility of using artificial intelligence for quality control of echocardiographic images.Methods:Retrospectively，5 000 two-dimensional echocardiographic video images within the period from 2021 to 2023 were randomly retrieved from the echocardiography database of Nanjing Drum Tower Hospital，Affiliated Hospital of Medical School，Nanjing University. Among these selected images，1 559 of them were apical views. The physician team formulated the scoring rules，which specifically included four scoring criteria：gain，scaling ratio，cardiac axis angle，and structure. Subsequently，the data were labeled with view classification and image quality scores. The labeled data were further partitioned into the training set（ n = 643），the validation set（ n = 276），and the test set（ n = 640）. The training and validation sets were utilized for constructing the models for view classification and quality assessment，while the test set was employed to verify the models' effectiveness. The view classification module was implemented using the SlowFast model，and the quality assessment module involved algorithms such as ResNet，Video Swin Transformer，SSD，and U-Net. Results:The average accuracy，precision，recall rate and F1 score of the classification model in identifying each apical view were 0.987 1，0.983 0，0.987 1 and 0.984 9 respectively，and the inference time was（333.4 ± 105.4）ms. The average accuracies of the quality assessment module in terms of gain，scaling ratio，cardiac axis angle and display of main structures were 0.915 1，0.928 2，0.938 7 and 0.965 6 respectively，and the overall scoring accuracy was 0.912 7.Conclusions:The echocardiogram quality control system developed in this research can effectively classify and evaluate the quality of two-dimensional images of the apical views in echocardiograms. Moreover，it guarantees the objectivity，timeliness and high-efficiency of quality control，which has reference value for the establishment of the echocardiogram quality control system.

Objective:To investigate the association between serum 25-hydroxyvitamin D [25(OH)D] levels and 24-h urinary calcium excretion (24-h UCaE) and hypercalciuria in Chinese adults.Methods:This cross-sectional study was conducted from March 2022 to March 2023 in nine cities in China and included 1 239 residents. Demographic characteristics were collected through questionnaires and physical examinations, fasting blood samples were assessed for bone metabolism indicators, and 24-h urine samples were used to determine the 24-h UCaE. Multiple linear regression analysis was used to explore the relationship between serum 25(OH)D and 24-h UCaE and bone metabolism indexes. The relationship between serum 25(OH)D and hypercalciuria was analyzed using a multiple logistic regression model combined with restricted cubic spline modeling.Results:The mean participant age was (47.9±18.1) years, of which 453 (36.6%) were male. The percentages of vitamin D sufficiency, insufficiency, and deficiency were 7.6% (94/1 239), 29.0% (359/1 239), and 63.4% (786/1 239), respectively. The multiple linear regression model showed that after adjusting for the covariates the 24-h UCaE gradually increased with higher levels of 25(OH)D ( P overall <0.001, P nonlinear <0.001). The logistic regression analysis revealed that compared with the vitamin D deficient group, the OR for the prevalence of hypercalciuria in the vitamin D sufficient and vitamin D insufficient groups were 3.290 (95% CI 1.745 to 6.202) and 3.742 (95% CI 2.458 to 5.697), respectively. The results of the restricted cubic spline modeling showed a positive nonlinear relationship between 25(OH)D and the prevalence of hypercalciuria ( P overall <0.001, P nonlinear <0.001). The prevalence of hypercalciuria increased when 25(OH)D was >17.00 μg/L and peaked at 26.71 μg/L, after which there was a decreasing trend in the prevalence of hypercalciuria with increasing 25(OH)D. Conclusion:Associations between serum 25(OH)D levels and urinary calcium excretion and the prevalence of hypercalciuria were observed in the Chinese adult population.