BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

BACKGROUND:Studies have confirmed that Pterocarya hupehensis skan total flavonoids(PHSTF)can improve the level of collagen-induced arthritis in rats,but there is still a lack of research on the regulation of Wnt/β-catenin signaling pathway in fibroblast-like synoviocytes and its effect on related cell functions.OBJECTIVE:To investigate the effect and mechanism of PHSTF on lipopolysaccharide-induced proliferation,migration and apoptosis of fibroblast-like synoviocytes based on the Wnt/β-catenin signaling pathwayMETHODS:Fibroblast-like synoviocytes were divided into control group,lipopolysaccharide group,lipopolysaccharide+low-,medium-,and high-dose PHSTF groups(10,20,and 40 μg/mL),lipopolysaccharide+Wnt pathway inhibitor DKK1 group,and lipopolysaccharide+Wnt pathway inhibitor DKK1+high-dose PHSTF group(40 μg/mL).The cell counting kit-8 method was used to detect the effect of PHSTF on the viability of fibroblast-like synoviocytes,and the final drug concentration and time were screened.Flow cytometry was used to detect the apoptosis of fibroblast-like synoviocytes.Cell scratch assay,EDU staining and cell cloning assay were used to detect the migration and proliferation of fibroblast-like synoviocytes.Western blot assay was used to detect the protein expression levels of Wnt3a,β-catenin,tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,Bax and Bcl-2 in fibroblast-like synoviocytes.RESULTS AND CONCLUSION:(1)Compared with the control group,the cell viability decreased significantly when the concentration of PHSTF was＞40 μg/mL(P＜0.01).Therefore,the drug concentration of≤40 μg/mL was selected for subsequent experiments.(2)Compared with the lipopolysaccharide group,the wound healing rate,cell clone formation rate and the number of EDU-positive cells in the low-,medium-and high-dose PHSTF groups were significantly reduced,while the apoptosis rate was significantly increased(P＜0.05-0.01).(3)Western blot results showed that compared with the lipopolysaccharide group,low-,medium-and high-dose PHSTF significantly inhibited cellular Wnt3a,β-catenin,cellular tumorigenic genes,matrix metalloproteinase 2,matrix metalloproteinase 9,and Bcl-2 protein expression,and promoted the expression of Bax protein(P＜0.01).(4)Compared with the DKK1 group,the combination of DKK1 and high-dose PHSTF significantly inhibited the protein expression of Wnt3a,β-catenin,matrix metalloproteinase 2,matrix metalloproteinase 9 and Bcl-2 protein expression and promoted the protein expression of Bax(P＜0.01).To conclude,PHSTF may inhibit the proliferation and migration of fibroblast-like synoviocytes and promote apoptosis by inhibiting the Wnt/β-catenin signaling pathway.

To compare the performance differences between the multidisciplinary team (MDT) model and the conventional diagnostic and treatment model for lung cancer, and to explore a high-quality development pathway for optimizing lung cancer diagnostic and treatment resources.

Objective:To analyze the prognostic influencing factors in patients with isocitrate dehydrogenase (IDH) wild-type glioma, and further evaluate the value of surgical resection in prognosis.Methods:The clinical data and molecular pathological information of 647 patients with IDH wild-type glioma were retrieved from the Chinese Glioma Genome Atlas (CGGA) database (from 2006 to 2019). The clinical characteristics were recorded, including gender, age, initial symptoms, preoperative Karnofsky performance status (KPS) score, tumor location, tumor laterality, extent of resection, O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation status and postoperative treatment. Kaplan-Meier survival curve was plotted to calculate overall survival (OS) and progression-free survival (PFS), and comparisons were performed using the log-rank test. Multivariate Cox regression analysis was used to identify the independent influencing factors of prognosis in patients with IDH wild-type glioma.Results:Among the 647 patients with IDH wild-type glioma, there were 120 cases of WHO grade Ⅱ, 115 cases of grade Ⅲ, and 412 cases of grade Ⅳ; the median OS was 20.3 months. There were statistical differences in age, initial symptoms, tumor location, preoperative KPS score, extent of resection, MGMT promoter methylation status and postoperative treatment among patients with different WHO grades (P<0.01), but there were no statistical difference in gender and tumor laterality (P>0.05). The median OS and PFS in patients with WHO grade Ⅱ were significantly longer than those in patients with WHO grade Ⅲ and WHO grade Ⅳ (169.9 months vs. 24.7 and 14.4 months, 138.8 months vs. 17.7 and 11.4 months), the indexes in patients with WHO grade Ⅲ were significantly longer than patients with WHO grade Ⅳ, and there were statistical differences (P<0.05). The median OS and PFS in patients with total resection were significantly longer than those in patients with subtotal resection and partial resection (37.2 months vs. 20.3 and 8.5 months, 29.5 months vs. 17.6 and 6.0 months), the indexes in patients with subtotal resection were significantly longer than patients with partial resection, and there were statistical differences (P<0.05). Among 153 patients with MGMT promoter methylation who received postoperative chemoradiotherapy, the median OS and PFS in patients with total resection (67 cases) were significantly longer than those in patients with subtotal resection (64 cases) and partial resection (22 cases): 28.8 months vs. 18.4 and 9.0 months, 25.6 months vs. 16.6 and 6.5 months), the indexes in patients with subtotal resection were significantly longer than patients with partial resection, and there were statistical differences (P<0.05). Among 202 patients with MGMT promoter non-methylation who received postoperative chemoradiotherapy, the median OS and PFS in patients with total resection (81 cases) were significantly longer than those in subtotal resection (81 cases) and partial resection (40 cases): 31.3 months vs. 21.9 and 14.0 months, 22.7 months vs. 16.8 and 9.7 months, and there were statistical differences (P<0.05), but there were no statistical difference in the indexes between patients with subtotal and patients with partial resection (P>0.05). Multivariate Cox regression analysis result showed that age, extent of resection, WHO grade (grade Ⅲ/grade Ⅳ) and postoperative treatment were independent OS and PFS influencing factors in patients with IDH wild-type glioma (OS: HR = 1.327, 3.295, 3.406/7.964 and 1.597; 95% CI 1.079 to 1.633, 2.627 to 4.132, 2.225 to 5.216/5.339 to 11.880 and 1.288 to 1.981; P<0.01. PFS: HR = 1.282, 2.655, 3.143/6.511 and 1.477; 95% CI 1.039 to 1.583, 2.123 to 3.322, 2.049 to 4.819/4.376 to 9.690 and 1.181 to 1.849; P<0.05 or <0.01).Conclusions:The age, extent of resection, WHO grade and postoperative treatment are independent prognosis influencing factors in patients with IDH wild-type glioma. Treatment strategies should fully consider the clinical and molecular pathological characteristics to achieve maximal safe tumor resection and optimal survival benefit.

Gallbladder cancer is a highly aggressive malignancy of the biliary system, often diagnosed at the advanced stage due to its insidious early symptoms, leading to poor overall progno-sis. In recent years, the rapid advancement of artificial intelligence (AI) technologies and their inte-gration into medicine have opened new avenues for the early diagnosis and precision treatment of gallbladder cancer. Currently, AI incorporating deep learning algorithm has significantly improved diagnostic sensitivity and specificity in ultrasound, computed tomography, and pathological analysis. However, clinical translation of AI models remains limited by challenges such as insufficient annota-ted data and limited model interpretability. Future research should focus on establishing multi-center data-sharing mechanisms, developing interpretability tools, and optimizing multimodal data integration strategies, thereby promoting the transformation of AI technologies from an auxiliary diagnostic tool to a core component of clinical decision-making.

Objective·To develop a graph neural network(GNN)-based auxiliary diagnostic model for gallbladder cancer on CT images,and validate its accuracy and feasibility.Methods·From January 2010 to November 2023,1 774 contrast-enhanced CT arterial-phase images were acquired from 887 patients with normal gallbladder,benign gallbladder disease,or gallbladder cancer at Xinhua Hospital and Renji Hospital,Shanghai Jiao Tong University School of Medicine.These images were randomly divided into training and testing sets at a 4∶1 ratio to develop a hybrid GNN-convolutional neural network(CNN)model,named VJK-GIN.The model constructed a pixel-level graph in which each pixel served as a node,and spatial adjacency defined the edges,enabling extraction of local texture features.In the model architecture design,VJK-GIN integrated a three-layer graph isomorphism network,augmented with virtual nodes and jump-knowledge connections;global pooling compressed node features into a graph-level representation,which was classified by a multi-layer perceptron head.Five-fold cross-validation was used to compare VJK-GIN with GNN baselines(GCN,GraphSAGE,GAT,and GIN)and CNN baselines(ViT,EfficientNetV2,and ConvNeXt)in terms of accuracy,precision,recall,F1-score,and area under the receiver operating characteristic curve(AUC).Results·The results of five-fold cross-validation showed that VJK-GIN achieved an F1-score of 0.799(95%CI 0.775?0.823),recall of 0.795(95%CI 0.773?0.817),precision of 0.799(95%CI 0.775?0.823),AUC of 0.812(95%CI 0.792?0.832),and accuracy of 0.773(95%CI 0.748?0.798),surpassing all competing models across every metric.Conclusion·The VJK-GIN model exhibits high stability and accuracy in identifying contrast-enhanced CT images of normal,benign,and malignant gallbladder conditions.

Gallbladder cancer is a highly aggressive malignancy of the biliary system, often diagnosed at the advanced stage due to its insidious early symptoms, leading to poor overall progno-sis. In recent years, the rapid advancement of artificial intelligence (AI) technologies and their inte-gration into medicine have opened new avenues for the early diagnosis and precision treatment of gallbladder cancer. Currently, AI incorporating deep learning algorithm has significantly improved diagnostic sensitivity and specificity in ultrasound, computed tomography, and pathological analysis. However, clinical translation of AI models remains limited by challenges such as insufficient annota-ted data and limited model interpretability. Future research should focus on establishing multi-center data-sharing mechanisms, developing interpretability tools, and optimizing multimodal data integration strategies, thereby promoting the transformation of AI technologies from an auxiliary diagnostic tool to a core component of clinical decision-making.

Objective·To develop a graph neural network(GNN)-based auxiliary diagnostic model for gallbladder cancer on CT images,and validate its accuracy and feasibility.Methods·From January 2010 to November 2023,1 774 contrast-enhanced CT arterial-phase images were acquired from 887 patients with normal gallbladder,benign gallbladder disease,or gallbladder cancer at Xinhua Hospital and Renji Hospital,Shanghai Jiao Tong University School of Medicine.These images were randomly divided into training and testing sets at a 4∶1 ratio to develop a hybrid GNN-convolutional neural network(CNN)model,named VJK-GIN.The model constructed a pixel-level graph in which each pixel served as a node,and spatial adjacency defined the edges,enabling extraction of local texture features.In the model architecture design,VJK-GIN integrated a three-layer graph isomorphism network,augmented with virtual nodes and jump-knowledge connections;global pooling compressed node features into a graph-level representation,which was classified by a multi-layer perceptron head.Five-fold cross-validation was used to compare VJK-GIN with GNN baselines(GCN,GraphSAGE,GAT,and GIN)and CNN baselines(ViT,EfficientNetV2,and ConvNeXt)in terms of accuracy,precision,recall,F1-score,and area under the receiver operating characteristic curve(AUC).Results·The results of five-fold cross-validation showed that VJK-GIN achieved an F1-score of 0.799(95%CI 0.775?0.823),recall of 0.795(95%CI 0.773?0.817),precision of 0.799(95%CI 0.775?0.823),AUC of 0.812(95%CI 0.792?0.832),and accuracy of 0.773(95%CI 0.748?0.798),surpassing all competing models across every metric.Conclusion·The VJK-GIN model exhibits high stability and accuracy in identifying contrast-enhanced CT images of normal,benign,and malignant gallbladder conditions.

Objective To develop an instrument for predicting postoperative constipation risks in patients with osteoporotic thoracolumbar fracture(OTLF)who have undergone percutaneous kyphoplasty(PKP).Methods A total of 858 OTLF patients who underwent PKP surgery between January 2020 and December 2024 were enrolled.The patients were randomly assigned to a training set(n=600)and a validation set(n=258)in a 7∶3 ratio.According to whether the patients had postoperative constipation,the training set was divided into a constipation group(n=205)and a non-constipation group(n=395),and the validation set was divided into a constipation group(n=90)and a non-constipation group(n=168).Logistic regression analysis was conducted to analyze the factors influencing postoperative constipation in OTLF patients after PKP,and a nomogram model was constructed accordingly.The receiver operating characteristic(ROC)curve and the calibration curve of the model were plotted,and the Hosmer-Lemeshow test for goodness of fit was performed.Results A total of 205 OTLF patients(34.17%)in the training set and 90 OTLF patients(34.88%)in the validation set experienced constipation after PKP.Univariate analysis revealed significant differences between the constipation and non-constipation groups in terms of operative time,postoperative water intake,time to first postoperative meal,postoperative bed rest time,the levels of Bifidobacterium,Lactobacillus,Enterococcus,and Enterobacter,the Nutrition Risk Screening 2002(NRS-2002)score,and the levels of sodium,potassium,and HbA1c(P<0.05).Least absolute shrinkage and selection operator(LASSO)regression was performed and operative time,time to first postoperative meal,the levels of Bifidobacterium,Lactobacillus,Enterococcus,and Enterobacter,the NRS-2002 score,and the levels of sodium,potassium,and HbA1c were identified as candidate predictors.Multivariate logistic analysis showed that the time to first postoperative meal,the levels of Bifidobacterium and Lactobacillus,the NRS-2002 score,and the levels of sodium and HbA1c were influencing factors of postoperative constipation in OTLF patients(P<0.05).The ROC curves showed that the area under the curve(AUC)of the training set was 0.842(95%CI:0.793-0.892),while that of the validation set was 0.860(95%CI:0.830-0.889).The calibration curves demonstrated good agreement between the prediction curve and the standard curve in both the training set and the validation set.Conclusion The time to the first postoperative meal,the NRS2002 score,and the levels of Bifidobacterium,Lactobacillus,sodium,and HbA1c are influencing factors of post-PKP constipation in OTLF patients.The nomogram model built based on these factors exhibited good predictive performance.

OBJECTIVE To establish an ultra performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS)method for determination of dexmedetomidine(DEX)in plasma of beagle dogs and evaluate the pharmacokinetics and sedation after nasal,buccal and sublingual mucosal admin-istration.METHODS A UPLC-MS/MS method was established and validated for dertermination of DEX in plasma of beagle dogs.DEX was administered to the nasal cavity,buccal and sublingual mucous membranes of beagle dogs,respectively.Blood samples were collected at different time points.The plasma concentration of DEX was measured by the established UPLC-MS/MS method.Pharmacokinetic parameters were fitted by Phoenix software and the sedative effect at different mucous membrane sites was evaluated in conjunction with behavioral and Ramsay scores.RESULTS The linearity of DEX was fine within the range of 0.05-100 μg·L-1(r>0.999),which was validated methodologically to meet the requirements of quantitative detection.The plasma concentration of the drug peaked the fastest with nasal administration.Tmax was 0.25 h,Cmax(4.43±1.19)μg·L-1,and the AUC0-6h was(8.92±2.07)μg·h·L-1,compared with 0.92 and 1 h,(2.87±0.69),(2.70±0.41)μg·L-1,and(7.99±1.77),(7.01±2.09)μg·h·L-1 with buccal and sublingual administration.Nasal administration had the fastest onset at 7 min,with a Ramsay score of 4,and sedation lasted for 36 min,compared with 33 and 35 min,and 38 and 37 min for buccal and sublingual administration.CONCLUSION The proposed method is sensitive,reliable and applicable to quantitative analysis of DEX in plasma of beagle dogs.Administration of DEX to the nasal cavity mucosa has a faster onset and a better sedative effect than to the buccal and sublingual mucosa.