Objective The purpose of this study was to investigate the effects of gossypol acetic acid (GAA) on protein and mRNA expressions of hMLH1 gene in human tongue carcinoma cell line Tca8113 in vitro in order to discuss the mechanism of tumor suppression of GAA. Methods (1) Western-blot was used to study the effects of GAA on protein expressions of hMLH1 gene in Tca8113 cell line treated by different concentrations of GAA for 48 h. (2) Real-time fluorescence quantitative PCR (RFQ-PCR) was used to investigate the effects on the mRNA expressions of hMLH1 gene in Tca8113 cell line treated by GAA for 48 h. Results (1) Compared with the control group, the results of Western-blot showed that the protein expression of hMLH1 gene was increased after treatment by GAA for 48 h ( <0.05) . (2) The results of RFQ-PCR indicated that the mRNA expression of hMLH1 gene was increased after GAA treatment for 48 h ( <0.05) . Conclusion GAA could up-regulate protein and mRNA expression of hMLH1 in Tca8113 cell line, which indicated that it may be one of the mechanisms of tumor suppression effect of GAA.

Objective:To study the effects of gossypol acetic acid(GAA)on the methylation level and the mRNA expression of hM-LH1 in human tongue cancer Tca8113 cells.Methods:Tca8113 cells were treated by GAA at various doses for 24 h,48 h and 72 h respectively.MTT assay was used to detect the cell proliferation.Nested methylation specific PCR(nMSP)was used to detect methyl-ation level of hMLH1 .Real-time fluorescence quantitative PCR(RFQ-PCR)was applied to investigate the mRNA expression of hM-LH1 gene.Results:GAA inhibited the proliferation of Tca8113 cells dose-and-time dependently,decreased the DNA methylation level of hMLH1(P<0.05)and increased hMLH1 mRNA expression in Tca8113 cells(P<0.05).Conclusion:GAA can suppress proliferation of Tca8113 cells by demethylation of hMLH1 gene and increase of hMLH1 mRNA expression.

Objective:To investigate the application of tracheal intubation by the endoscopic-assisted mouth floor-submandibular approach(EAMFA) for general anesthesia and the operation of craniofacial multiple fractures.Methods:8 patients with complex craniofacial fractures and associated with the contraindication of nasal trachea cannula underwent EAMFA general anesthesia and operation.Blood pressure(BP),saturation of blood oxygen(SPO2),heart rate(HR) and electrocardiogram(ECG) were monitored timely.Results:The intubation and anesthesia were successfull in all patients;BP,SPO2,HR and ECG were normal in all operation procedure in the patients.No complication was observed.Conclusion:EAMFA is an effective anesthesia approach for the surgical treatment of craniofacial multiple fractures.

Objective This study aims to assess the role of DNA methylation changes in tongue cancer through a comprehensive analysis of global DNA methylation alterations during experimental lingual carcinogenesis.Methods C57BL/6J mice were subjected to 16-week oral administration of 4-nitroquinoline-1-oxide(4NQO,50 mg/L).Lingual mucosa samples,being representative of normal tissue(week 0)and early(week 12)and advanced(week 28)tumorigen-esis,were harvested for microarray and methylated DNA immunoprecipitation sequencing(MeDIP-Seq).The mRNA and promoter methylation of transforming growth factor-beta-signaling protein 1(SMAD1)were evaluated with real-time quantitative reverse transcription polymerase chain reaction and Massarray in human lingual mucosa and tongue cancer cell lines.Results The cytosine guanine island(CGI)methylation level observed at 28 weeks surpassed that of both 12 weeks and 0 weeks.The promoter methylation level at 12 weeks exceeded that at 0 weeks.Notably,208 differentially expressed genes were negatively correlated to differential methylation in promoters among 0,12,and 28 weeks.The mRNA of SMAD1 was upregulated,con-current with a decrease in promoter methylation levels in cell lines compared to normal mucosa.Conclusion DNA methylation changed during lingual carcinogenesis.Overexpression of SMAD1 was correlated to promoter hypomethyl-ation in tongue cancer cell lines.