Histological staining is the basis of pathological analysis,but the traditional staining method relies on chemical reagents,which not only consumes a lot of resources,but also causes harm to the environment and human health.In recent years,with the rapid development of deep learning technology,virtual staining technology,as a new method,is expected to effectively replace and supplement the traditional histological staining methods.It uses neural networks to analyze unstained tissue images,generate digital images that are highly similar to chemical staining effects,and even realize the mutual conversion between different staining modes,reducing the laboratory's dependence on chemical reagents and providing sustainable research programs.In this paper,the basic principles of virtual staining and its potential applications in histopathology are introduced in detail,and the current challenges and future research directions are discussed.

Background and Aims:As a core data source in the development of medical informatization,the front page of inpatient medical records plays a crucial role in determining the usability of research data and the scientific quality of clinical decision-making.Given the relatively standardized diagnostic and treatment protocols for thyroid malignancies,this disease serves as a representative model for evaluating the data quality of medical record front sheets.This study aimed to systematically assess the accuracy of diagnostic and surgical procedure coding for thyroid malignancies on the medical record front page,identify existing problems,and explore strategies for improvement and the potential for future application in clinical research.Methods:A total of 3 361 patients who underwent initial surgical treatment and were pathologically confirmed with thyroid cancer at the Department of Thyroid Surgery,the First Affiliated Hospital of Kunming Medical University from January 2018 to December 2020 were retrospectively included.Postoperative pathology reports and surgical records were used as reference standards to evaluate the accuracy of primary and secondary diagnosis codes,as well as primary and secondary surgical procedure codes on the medical record front page.Results:The accuracy rate of primary diagnosis coding reached 98.8%.However,among 1 368 patients with cervical lymph node metastasis,only 31.2%had correctly recorded secondary diagnosis codes and metastatic sites,with an omission rate as high as 67.8%.The overall accuracy rate for primary surgical procedure coding was 90.4%,with thyroid lobectomy(TL)and total thyroidectomy(TT)coding accuracies of 85.5%and 96.0%,respectively.Among the coding errors,70.0%involved TL procedures being incorrectly coded as TT.Secondary surgical procedure codes—mainly for lymph node dissection—were highly inconsistent,with up to 14 different coding types,reflecting a lack of standardization and unified input criteria.Notably,the error rate for primary surgical coding in 2020 increased significantly compared with the previous two years,suggesting subjectivity and variability in manual data entry.Conclusion:This study highlights major accuracy deficiencies in surgical procedure coding for thyroid malignancies on the medical record front page,which may compromise the scientific validity of real-world data.Going forward,leveraging artificial intelligence technologies to support structured documentation and automated coding,alongside establishing a proactive quality control system through multidisciplinary collaboration,may significantly improve data accuracy.These efforts are essential for strengthening data foundations for precision treatment,performance evaluation,insurance payment,and clinical research on thyroid cancer.

Cardiovascular diseases that develop from hypercholesterolemia-induced atherosclerosis have emerged as a significant threat to human health.Recently,probiotics exhibiting cholesterol-lowering properties have emerged as a prominent area of research.Numerous studies have demonstrated that Lactobacillus reuteri can effectively reduce endogenous cholesterol synthesis,regulate cholesterol transport,and promote cholesterol degradation by modulating the expression of key genes,such as sterol-regulatory element binding protein 2,3-hydroxy-3-methyl-glutaryl coenzyme A reductase,and cholesterol 7 alpha-hydroxylase,in both the liver and intestinal epithelial cells of the host.This leads to a notable decrease in total cholesterol and low-density lipoprotein cholesterol levels in the host serum.The present paper offers a comprehensive overview of the underlying mechanisms responsible for the cholesterol-lowering effects exerted by L.reuteri,aiming to provide valuable insights into the treatment of hypercholesterolemia and the development of probiotics with cholesterol-lowering properties.

OBJECTIVE: To observe the protective effect of Fisetin on sepsis-associated brain injury and explore its possible mechanism from the perspective of ferroptosis. METHODS: Sprague-Dawley (SD) rats (6-8-week-old male) were randomly divided into three groups: sham operation group (Sham group), colonic ligation and puncture (CLP) induced sepsis model group (CLP group) and Fisetin preprocessing group (CLP+Fisetin group), with 18 rats in each group (12 for observing survival rate and 6 for indicator testing). The CLP+Fisetin group was given Fisetin solution 50 mg×kg^-1×d^-1 by gavage continuously for 5 days before CLP, with dimethyl sulfoxide (DMSO) as the solute, while Sham group and CLP group were given the same dose of DMSO. The model was established at 2 hours after the last gavage. The general condition of each group of rats were observed, and the 10-day mortality were record. The behavioral testing (new object recognition experiment, elevated cross maze experiment) were performed after 7 days of modeling. After 24 hours of modeling, nerve reflex scoring was performed, and then the rats were euthanized and brain tissue was collected. The pathological changes of brain tissue were observed under a microscope by hematoxylin-eosin (HE) staining, the deposition of iron ion in brain tissue was observed by Prussian blue staining. The content of iron in brain tissue was determined by tissue iron kit, and the content of malondialdehyde (MDA) in brain tissue was determined by colorimetry. The expressions of tumor necrosis factor-α (TNF-α), neuron damage marker S100β, nuclear factor E2-related factor 2 (Nrf2), heme oxygenases-1 (HO-1) and glutathione peroxidase 4 (GPX4) were detected by Western blotting. RESULTS: On day 10 post-operation, 12, 3, and 7 animals survived in the Sham group, CLP group, and CLP+Fisetin group, respectively. Compared with the Sham group, rats in the CLP group showed significantly decreased nerve reflex score, new object discrimination index and open arm dwell time. HE staining showed arranged disorderly of neuronal cells, cytoplasm deep staining, nuclear condensation, unclear structures, neuron loss, and significant inflammation in the hippocampus in the hippocampus. Prussian blue staining showed iron ion deposition in the brain tissue. The contents of iron and MDA in brain tissue were elevated, and the expressions of TNF-α and S100β were up-regulated, while the expressions of Nrf2, HO-1, and GPX4 were down-regulated. Compared with the CLP group, the CLP+Fisetin group showed significantly increased neurological reflex score (7.33±1.15 vs. 4.67±1.53), improved new object discrimination index (0.44±0.02 vs. 0.32±0.04), and longer open arm dwell time (minutes: 78.33±9.29 vs. 41.15±9.64). Neuronal cells in the hippocampus were more organized, with less cytoplasmic staining, nuclear condensation, reduced neuronal loss, and fewer inflammatory cells. Iron ion deposition was reduced, and the contents of iron ions and MDA in brain tissue were decreased [iron ion (μg/g): 151.27±14.90 vs. 224.69±17.64, MDA (μmol/g): 470.0±44.3 vs. 709.3±65.4]. The expressions of TNF-α and S100β were significantly decreased (TNF-α/GAPDH: 0.651±0.060 vs. 0.896±0.022, S100β/GAPDH: 0.685±0.032 vs. 0.902±0.014), while the expressions of Nrf2, HO-1, and GPX4 were significantly increased (Nrf2/GAPDH: 0.708±0.108 vs. 0.316±0.112, HO-1/GAPDH: 0.694±0.022 vs. 0.538±0.024, GPX4/GAPDH: 0.620±0.170 vs. 0.317±0.039). All differences were statistically significant (all P < 0.05). CONCLUSION Fisetin pretreatment can inhibit ferroptosis and reduce sepsis-associated brain injury by Nrf2/HO-1/GPX4 pathway.
Animals ; Ferroptosis/drug effects* ; Rats, Sprague-Dawley ; NF-E2-Related Factor 2/metabolism* ; Sepsis/complications* ; Male ; Rats ; Phospholipid Hydroperoxide Glutathione Peroxidase ; Neurons/drug effects* ; Signal Transduction ; Brain Injuries/metabolism* ; Flavonols ; Flavonoids/pharmacology* ; Heme Oxygenase-1/metabolism* ; Heme Oxygenase (Decyclizing)

Purpose To investigate the relationship between the expression of POSTN and NECTIN-3 in urothelial carcinoma(UC)and its clinicopathologic features and prognosis.Methods Clinical data of 115 UC patients were col-lected.EnVision two-step method was used for immunohistochemical analysis to determine the expression of POSTN and NECTIN-3 in UC and their correlation with clinicopathological features and prognosis.Transcriptomic data from TCGA database were used to analyze the correlation between the expression of POSTN and NECTIN-3 in bladder cancer and the pathological stage and prognosis of bladder cancer.qPCR and Western blot were used to detect the expression levels of POSTN and NECTIN-3 in tumor tissues and adjacent tissues of patients.Results TCGA database analysis showed that the expression level of NECTIN-3 in urinary bladder urothelial carcinoma(UBUC)was significantly lower than that in paracancer tissues,and POSTN expression was positively correlated with pathological stage.Prognostic a-nalysis showed that POSTN expression was negatively correlated with the overall survival of UBUC(P＞0.05),and NECTIN-3 expression was negatively correlated with disease-free survival of UBUC(P＞0.05).The experimental re-suits showed that patients with positive POSTN expression were more prone to perineural invasion[25 cases(86.2％)vs 4 cases(13.8％),P=0.019],vascular invasion[36 cases(83.7％)vs 7 cases(16.3％),P=0.007],and lymph node metastasis[24 cases(88.9％)vs 3 cases(11.1％),P=0.033].Additionally,the positive expression rate of POSTN in UBUC was significantly higher than that in upper tract urothelial carcinoma(UTUC)(75.0％vs 54.3％,P=0.028).Higher expression levels of POSTN and NECTIN-3 were associated with shorter overall survival,but the difference was not statistically significant(P＞0.05).Conclusion There is no significant correlation between NECTIN-3 and the clinicopathological features and prognosis of urothelial carcinoma,while the expression of POSTN is correlated with the invasive clinicopathological features of UC,which has certain suggestive significance for clinical stage and prognosis.

Introduction: Early identification of patients at risk for severe multisystem inflammatory syndrome in children (MIS-C) is essential for favourable clinical outcomes. This study aims to identify the clinical characteristics, factors and outcomes associated with severe MIS-C. Materials and methods: In this retrospective cohort study involving 14 major hospitals in Malaysia, children <15 years who met the United States Centres for Disease Control and Prevention case definition for MIS-C were included. Severe MIS-C was defined as children who required inotropic support, ventilatory support (invasive or non-invasive ventilation), or left ventricular ejection fraction of <55%. The factors investigated for severe MIS-C were demographic characteristics, the presence of comorbidities, clinical characteristics, and laboratory measures. Multivariable logistic regression was used to compute the adjusted odds ratio (aORs) of factors associated with severe MIS-C. Results: Among the 155 patients, 91 (58.7%) presented with severe MIS-C. Severe MIS-C was more likely in patients aged ≥5 years old (aOR 2.13, 95% confidence interval [CI] 1.08-4.21), with dehydration (aOR 3.80, 95% CI 1.53-9.45), lethargy (aOR 2.02, 95% CI 0.97-4.18), tachycardia (aOR 8.33, 95% CI 3.27-21.22), albumin <30g/L (aOR 3.36, 95% CI 1.58-7.13), creatine kinase >200U/L (aOR 3.68, 95% CI 1.57-8.64), D-dimer >3.0µg/mL (aOR 2.11, 95% CI 1.08-4.13), ferritin >500ng/mL (aOR 3.77, 95% CI 1.88-7.55), prothrombin time >12.7 seconds (aOR 3.22, 95% CI 1.61-6.43), and urea >6mmol/L (aOR 5.09, 95% CI 2.04-12.71). Conclusion: Identification of these associated factors of severity in MIS-C could aid in early recognition and prompt escalation of care, leading to better outcomes.

Surgery is the most important and effective way to treat thyroid cancer. How to reduce surgical complications while completely treating tumor has always been the focus and difficulty of thyroid surgery. Carbon nanoparticles suspension injection is the third generation lymph node tracer, with strong staining specificity, rapid development, long staining time, simple use, harmless to human body and other characteristics, showing unique clinical value in lymph node tracing, parathyroid identification and protection. However, the application of carbon nanoparticles in thyroid surgery is still controversial. After literature review, the authors address the clinical application of carbon nanoparticles in thyroid cancer surgery.

Objective:The purpose of this study is to explore the mediating effect of 25-hydroxyvitamin D3[25(OH)D3]on the relationship between psychological status and cognitive function in elderly obese patients.Methods:130 elderly obese patients who visited Hongdu Traditional Chinese Medicine Hospital in Nanchang from January 2023 to June 2024 were selected,they were divided into cognitive impairment group(n=27)and cognitive normal group(n=103)based on the Mini Mental State Examination(MMSE)score.General information were prospectively collected,their psychological status used Hamilton Depression Rating Scale(HAMD)and Hamilton Anxiety Rating Scale(HAMA)to evaluated,25(OH)D3 levels were detected by enzyme-linked immunosorbent assay.General information,psychological status scores,and 25(OH)D3 levels were compare between two groups.Analysis of the correlation between 25(OH)D3 and psychological status and cognitive function by Pearson method.Analysis of the mediating effect of 25-hydroxyvitamin D3[25(OH)D3]on the relationship between psychological status and cognitive function in elderly obese patients.Results:Compared with cognitive normal group,cognitive impairment group had higher age and lower years of education(P＜0.05).HAMA and HAMD scores in the cognitive impairment group were significantly higher than those in the cognitive normal group(P＜0.05),and the 25(OH)D3 level was significantly lower than that in the cognitive normal group(P＜0.05).Pearson correlation analysis showed that,25(OH)D3 was negatively correlated with HAMA score and HAMD score,and positively correlated with MMSE score(P＜0.05);HAMA score,HAMD score,and MMSE score were all negatively correlated(P＜0.05).The structural equation model shows good fit[x2/df=1.87,root mean square error approximate=0.062,comparative fit index=0.945,Tucker-Lewis index=0.931].The direct effect of psychological status(HAMA+HAMD)on cognitive function was-0.325(95％confidence Interval(CI):-0.454～-0.216,P＜0.05),and the mediating effect through 25(OH)D3 was-0.182(95％CI:-0.271～-0.112,P＜0.05),accounting for 36.00％of the total effect.Conclusion:In elderly obese patients,25(OH)D3 is positively correlated with cognitive function and negatively correlated with psychological status,moreover,25(OH)D3 plays a significant mediating role in the relationship between psychological status and cognitive function,and is an important mediating factor affecting the relationship between the two.

Surgery is the most important and effective way to treat thyroid cancer. How to reduce surgical complications while completely treating tumor has always been the focus and difficulty of thyroid surgery. Carbon nanoparticles suspension injection is the third generation lymph node tracer, with strong staining specificity, rapid development, long staining time, simple use, harmless to human body and other characteristics, showing unique clinical value in lymph node tracing, parathyroid identification and protection. However, the application of carbon nanoparticles in thyroid surgery is still controversial. After literature review, the authors address the clinical application of carbon nanoparticles in thyroid cancer surgery.