OBJECTIVETo investigate the inhibitory effects of intervention of the tumor necrosis factor-alpha (TNFa)/nuclear factor-kappa B (NF-kappaB) signaling pathway activation on hepatoma cell proliferation and to explore its mechanism.

METHODSA rodent hepatoma model was established by feeding N-2-fluorenylacetamide (2-N-FAA) to male Sprague-Dawley rats. Human subjects with various liver diseases were enrolled in the study, and serum and peripheral blood nuclear cells were collected for analysis. HepG2 cells were cultured in vitro and treated with anti-TNFa (monoclonal antibody, mAb) to down-regulate its expression or transfected with siRNA targeting the p65 subunit of NF-kappaB to inhibit its activation. The liver cell line L02 was used as a control. Changes in protein and gene expression levels of NF-kappaB and TNFa were analyzed by Western blotting or enzyme-linked immunosorbent assay and real-time PCR, respectively. Changes in the cell cycle or apoptosis were evaluated by flow cytometry or Annexin-V/PI double-labeling assay, respectively.

RESULTSTNFa and NF-kappaB expression showed increasing trends during the malignant transformation of rat hepatocytes, and the differential expression patterns showed association with histopathological alterations in the hepatocytes. Following treatment with the TNFa mAb, the HepG2 cells showed a higher percentage of apoptotic cells than the untreated control cells (21.45% +/- 4.07% vs. 5.63% +/- 0.93%, q =10.07, P less than 0.01).There was a significant difference in the rate of cells in the G0/G1 phase in the p65-siRNA transfected cells (66.23% +/- 1.29% vs. untreated control cells: 59.00% +/- 1.02%, q =10.98, P less than 0.01). The decreased expression of TNFa and NF-kappaB in cell culture supernatants was positively correlated with the dose of treatment (r =0.89, P less than 0.01), with the most robust decreases being achieved with the highest concentrations ( P less than 0.01). NF-kappaB expression was significantly higher in the HepG2 cells than in the L02 cells, and transfection of p65-siRNA reduced the mRNA (93%) and protein (62%) levels and increased the cell apoptosis index (to 85%).

CONCLUSIONProliferation of hepatoma cells may be significantly inhibited by intervening in the activation of the TNFa/NF-kappaB signaling pathway, which promotes cell apoptosis and blocks cell cycling.

Adult ; Aged ; Animals ; Apoptosis ; Carcinoma, Hepatocellular ; pathology ; Cell Proliferation ; Female ; Hep G2 Cells ; Hepatocytes ; metabolism ; Humans ; Liver Neoplasms ; pathology ; Male ; Middle Aged ; NF-kappa B ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; Transfection ; Tumor Necrosis Factor-alpha ; metabolism

Objective:To investigate the relationship between hyperuricemia and outcome in patients with acute ischemic stroke.Methods:Patients with acute ischemic stroke admitted to Department of Neurology, Putuo Hospital, Shanghai University of Tranditional Chinese Medicine between January 2020 and September 2020 were enrolled retrospectively. The modified Rankin Scale (mRS) score was used to evaluate the clinical outcome 3 months after the onset. ≤2 was considered as good outcome, and >2 was considered as poor outcome. The demography and baseline characteristics were compared between the good outcome group and the poor outcome group. Multivariate logistic regression analysis was used to determine the independent influencing factors of the outcome. Results:A total of 210 patients were included, their age was 69.87±62.62 years. There were 125 males (59.52%) and 85 females (40.48%). The baseline median National Institutes of Health Stroke Scale (NIHSS) score was 4. The serum uric acid level in 169 patients (80.48%) was normal and 41 (19.52%) had hyperuricemia; 120 patients (57.14%) had a good outcome, and 90 (42.86%) had a poor outcome. Blood glucose level, serum uric acid level, baseline NIHSS score and the proportions of diabetes mellitus, history of stroke or transient ischemic attack, hyperuricemia in the poor outcome group were significantly higher than those in the good outcome group (all P<0.05). Multivariate logistic regression analysis showed that diabetes mellitus (odds ratio [ OR] 2.735, 95% confidence interval [ CI] 1.461-5.121; P=0.002), hyperuricemia ( OR 2.400, 95% CI 1.102-5.228; P=0.027), and higher baseline NIHSS score ( OR 1.233, 95% CI 1.118-1.360; P<0.001) were the independent risk factors for poor outcome in patient with acute ischemic stroke. Conclusion:Hyperuricemia is an independent risk factor for poor outcome in patients with acute ischemic stroke.

Objective:To study the risk factors of lymph node metastases in patients with intrahepatic cholangiocarcinoma (ICC) and to establish a risk prediction model of lymph node metastases in ICC.Methods:The clinicopathological data of 587 ICC patients who underwent radical hepatectomy and lymph node dissection at Third Affiliated Hospital of Naval Medical University (Shanghai Eastern Hepatobiliary Surgery Hospital) from January 2007 to December 2011 were retrospectively analyzed. There were 395 males and 192 females with ages which ranged from 20 to 82 (54.7±10.8) years. Independent risk factors of lymph node metastases were studied using univariate and multivariate logistic regression analysis, and a risk prediction model was established. Receiver operating characteristic (ROC) curve was used to evaluate the accuracy of this model.Results:Of 587 patients, 158 (26.9%) had lymph node metastases. Multivariate logistic regression analysis showed that platelet count >300×10 9/L ( OR=1.985, 95% CI: 1.030-3.824, P=0.041), carbohydrate antigen 19-9 >37 U/ml ( OR=2.978, 95% CI: 1.994-4.448, P<0.001), tumor situated in left hemiliver ( OR=1.579, 95% CI: 1.065-2.341, P=0.023), multiple tumors ( OR=1.846, 95% CI: 1.225-2.783, P=0.003), and absence of cirrhosis ( OR=2.125, 95% CI: 1.192-3.783, P=0.011) were independent risk factors for lymph node metastases in ICC. The area under the ROC curve was 0.714, with a cutoff value of 0.215, and the sensitivity and specificity being 75.9% and 58.3%, respectively. Conclusions:The risk prediction model of ICC lymph node metastases was established using readily available clinical data obtained before operation. This model has good predictive values and can provide a reference for treatment decision on patients with ICC.