Objective To investigate whether the pyroptosis-related Toll-like receptor 4(TLR4)signaling pathway influences the malignant transformation of actinic keratosis(AK)into cutaneous squamous cell carcinoma(SCC).Methods A total of 6 lesion tissue samples each from patients with AK and SCC,as well as 5 normal skin tissue samples from healthy subjects as controls,were collected from the First Affiliated Hospital of Kunming Medical University between August 2020 and August 2021.Quantitative PCR(qPCR)and Western blot analysis were performed to determine the mRNA and protein expression levels of TLR4 pathway-related factors,including TLR4,CPB1,NLRP3,IL-1β,and IL-18.TLR4/TUNEL double immunofluorescence staining was used to evaluate TLR4 expression and the level of cellular pyroptosis in tissue samples.In addition,Western blotting was performed to analyze the expression differences of pyroptosis-related core proteins(pro-caspase-1,cleaved caspase-1/p20,GSDMD,and cleaved N-terminal GSDMD)and TLR4 among the normal keratinocyte cell line HaCaT and SCC cell lines A431 and SCL-1.Results Quantitative PCR and Western blot results showed that the mRNA and protein expression levels of TLR4,CPB1,NLRP3,IL-1β,and IL-18 were significantly higher in SCC tissues than in AK and normal skin tissues(P<0.05).TLR4/TUNEL double immunofluorescence results revealed a progressive increasing trend in TLR4 expression and pyroptosis levels from normal skin to AK and further to SCC(P<0.05).Furthermore,in SCL-1 cells,the expression levels of pro-caspase-1,cleaved caspase-1/p20,cleaved N-terminal GSDMD,and TLR4 were significantly upregulated(P<0.05),whereas in A431 cells,only TLR4 expression was increased(P<0.05),and the levels of other pyroptosis-related proteins were downregulated compared to HaCaT cells(P<0.05).Conclusion The expression of the TLR4 signaling pathway gradually increased from AK to SCC,and its activation status varied among different cutaneous squamous cell carcinoma cell lines.

Objective To find the clinical and histopathological predictive factors for lateral lymph node ( LN ) metastasis in thyroid papillary microcarcinoma( PTMC ).Methods From January 2007 to December 2010,141 patients with PTMC underwent central and lateral LN dissections were enrolled in the study.The data from the cases were analyzed retrospectively to determine the predictive factors for lateral LN metastasis.Results Among the aforementioned cases,37 cases ( 26.2％ ) were PTMC with the presence of multifocality,24 ( 16.9％ ) Hashimoto thyroiditis,84 ( 59.6％ ) central LN metastasis,56 ( 39.7％ ) lateral LN metastasis.Multifocality,central LN metastasis,underlying Hashimoto thyroiditis,and upper pole location were significantly related to lateral LN metastasis in univariate analysis( P＜0.05 ).These four factors were also found to be independent predictive factors for lateral LN metastasis in multivariate analysis( P＜0.05 ).9(6.4％ ) of there patients were found to show skip metastasis in which there was lateral LN metastasis but no central LN metastasis.The upper pole location was statistically significantly associated with skip metastasis.Conclusions Patients with multifocality,central LN metastasis,underlying Hashimoto thyroiditis,and upper pole location should be paid more attention to the status of lateral LN.Skip metastasis occurs in a minority of patients with PTMC.Even if there is no central LN metastasis,patients with upper pole lesion should be searched carefully for the lateral cervical LN metastasis.