Objective To investigate whether cisplatin induces tumor necrosis factor-α(TNF-α)secretion in human head and neck squamous cell carcinoma(HNSCC)cells to trigger RIP1/RIP3/MLKL-dependent necroptosis of the cells.Methods HNSCC cell lines HN4 and SCC4 treated with cisplatin(CDDP)or the combined treatment with CDDP and z-VAD-fmk(a caspase inhibitor)or Nec-1(a necroptosis inhibitor)for 24 h were examined for changes in cell viability using CCK8 assay and expressions of caspase-8 and necroptosis pathway proteins(RIP1/RIP3/MLKL)using Western blotting.The changes in migration of the cells were assessed with cell scratch assay,and the expressions of epithelial-mesenchymal transition(EMT)marker proteins N-cadherin,vimentin,and E-cadherin as well as the expressions of NF-κB(p65)and TNF-α were detected with Western blotting.Results The IC50 of cisplatin was 10 μg/mL in HN4 cells and 15 μg/mL in SCC4 cells.Cisplatin treatment significantly decreased the expressions of caspase-8,N-cadherin and vimentin and increased the expressions of E-cadherin,the necroptosis pathway proteins(RIP1/RIP3/MLKL),TNF-α,and NF-κB(p65),and these changes were obviously inhibited by treatment with Nec-1.Cisplatin stimulation also significantly lowered migration of the cells,and this inhibitory effect was strongly attenuated by Nec-1 treatment.Conclusion Cisplatin activates nuclear factor-κB signaling in HNSCCs to promote TNF-α autocrine and induce RIP1/RIP3/MLKL-dependent necroptosis,thus leading to inhibition of cell proliferation.

Objective To investigate whether cisplatin induces tumor necrosis factor-α(TNF-α)secretion in human head and neck squamous cell carcinoma(HNSCC)cells to trigger RIP1/RIP3/MLKL-dependent necroptosis of the cells.Methods HNSCC cell lines HN4 and SCC4 treated with cisplatin(CDDP)or the combined treatment with CDDP and z-VAD-fmk(a caspase inhibitor)or Nec-1(a necroptosis inhibitor)for 24 h were examined for changes in cell viability using CCK8 assay and expressions of caspase-8 and necroptosis pathway proteins(RIP1/RIP3/MLKL)using Western blotting.The changes in migration of the cells were assessed with cell scratch assay,and the expressions of epithelial-mesenchymal transition(EMT)marker proteins N-cadherin,vimentin,and E-cadherin as well as the expressions of NF-κB(p65)and TNF-α were detected with Western blotting.Results The IC50 of cisplatin was 10 μg/mL in HN4 cells and 15 μg/mL in SCC4 cells.Cisplatin treatment significantly decreased the expressions of caspase-8,N-cadherin and vimentin and increased the expressions of E-cadherin,the necroptosis pathway proteins(RIP1/RIP3/MLKL),TNF-α,and NF-κB(p65),and these changes were obviously inhibited by treatment with Nec-1.Cisplatin stimulation also significantly lowered migration of the cells,and this inhibitory effect was strongly attenuated by Nec-1 treatment.Conclusion Cisplatin activates nuclear factor-κB signaling in HNSCCs to promote TNF-α autocrine and induce RIP1/RIP3/MLKL-dependent necroptosis,thus leading to inhibition of cell proliferation.

Objective:To investigate the potential risk factors of the tumor abnormal protein (TAP) in papillary thyroid carcinoma (PTC) and analyze the clinical significance of TAP.Methods:The clinical data of 792 patients who underwent thyroid surgery in the thyroid surgery department of the First Affiliated Hospital of Zhengzhou University from Jun. 2018 to Jun. 2019 was collected, of whom 564 were PTC patients and 228 were benign thyroid tumor patients. Patients were divided into two groups by postoperative pathology: PTC group and thyroid benign tumor group. The potential risk factors the TAP in PTC were analyzed by univariate and multivariate Logistic regression analysis, and then the diagnostic value of TAP for PTC was assessed by ROC curve analysis.Results:Multivariate analysis showed that tumor maximum diameter (≥1.0 cm) ( OR:1.555; 95% CI: 1.031~2.344, P=0.035) and multifocal tumor ( OR:1.789; 95% CI: 1.098~2.916, P=0.019) were risk factors for elevation of TAP. Gender, age, body weight, extracapsular extension of cancer, invasive growth of cancer, Hashimoto's thyroiditis, central lymph node metastasis, lateral lymph node metastasis, and BrafV600E mutation were not associated with TAP ( P>0.05) . TAP in patients with PTC was significantly higher than that in patients with benign thyroid tumors[ (122.36±49.37) μm 2 vs (105.04±40.61) μm 2, P<0.001]. The sensitivity and specificity of TAP in diagnosing PTC were 78.90% and 40.35%, respectively. Conclusions:Tumor maximum diameter (≥1.0 cm) and multifocal tumor in PTC were risk factors for elevation of TAP. TAP has certain clinical value in differentiating PTC and thyroid benign tumor, which could be used as an auxiliary indicator.

Objective:To evaluate the potential factors influencing the parathyroid autofluorescence intensity of near-infrared fluorescent (NIRF) and further value of NIRF in identifying the parathyroid during surgery.Methods:The clinical data of 51 patients who underwent thyroid or parathyroid surgery in the Department of Thyroid Surgery, the First Affiliated Hospital of Zhengzhou University from April to June 2019 were retrospectively analyzed, including 16 males and 35 females, aged 18 to 74 years.The fluorescence intensity (FI) of the parathyroid glands, thyroid glands and background, and the number of parathyroid glands detected by NIRF and white light were measured. Variance analysis, two independent samples t test and Spearman rank correlation analysis were used to analyze the relationship between standardized parathyroid FI and clinical variables. Chi square test was used to analyze the difference of parathyroid detection rate between NIRF and white light. Results:In the 51 patients, the mean standardized parathyroid FI was greater than the standardized thyroid FI (1.72 ± 0.68 vs. 1.25 ± 0.40, t=6.555, P<0.001). The standardized parathyroid FI was not associated with gender, age, operation type, BMI, preoperative serum Ca 2+, parathyroid hormone and calcitonin (all P>0.05), but it was associated with disease type ( F=2.636, P<0.05). The mean standardized parathyroid FI of SHPT was lower than that of PTC, PTC with nodular goiter or NG(0.70±0.28 vs. 1.86±0.70, 1.69±0.49, 1.64±0.44, t value was 3.023, -1.129,-2.019, respectively, all P<0.05). There was no difference in the standardized parathyroid FI between SHPT and PHPT (1.34±0.18, t=1.218, P>0.05). There was no difference in standardized parathyroid FI between PHPT, PTC, NG, and PTC with NG(all P>0.05). Except for 3 cases of SHPT, 117 parathyroid glands were detected by NIRF and 101 parathyroid glands were detected by white light. The detection rate of parathyroid glands detected by NIRF was higher than that detected by white light (98.32% vs. 84.87%, χ 2=13.974, P<0.001). In SHPT, the detection rate of parathyroid gland by NIRF was 25.00%. Conclusions:Except SHPT, parathyroid FI is not affected by other clinical variables. NIRF can improve the detection rate of parathyroid glands during operation.