目的 通过网络药理学与分子对接探究姜黄素对口腔鳞状细胞癌抑制作用的分子机制.方法 使用SwissTargetPrediction数据库获得姜黄素靶点,检索GeneCards数据库和OMIM数据库获得口腔鳞癌的靶点,二者取交集得到潜在作用靶点.交集靶点导入String数据库构建PPI网络,使用Cytoscape软件进行优化并借助CytoHubba插件得到核心靶点.交集靶点导入Metascape数据库进行GO和KEGG富集分析.借助Autodock软件和Pymol软件进行姜黄素和核心靶点蛋白间的分子对接和可视化.结果 姜黄素在口腔鳞癌中的作用靶点有36个.PPI分析得到的核心靶点为AKT1、STAT3、EGFR、EP300、CCNA2、CDK2.GO分析结果揭示了潜在作用靶点主要涉及蛋白的磷酸化、对炎症的反应、有丝分裂细胞周期的调控等生物学功能.KEGG分析结果揭示了潜在作用靶点主要涉及FoxO信号通路、PI3K-Akt信号通路和趋化因子信号通路等.分子对接提示,姜黄素与核心靶点(AKT1、STAT3、EGFR、EP300、CCNA2、CDK2)之间结合稳定.结论 姜黄素对口腔鳞癌的抑制作用具有多靶点、多通路的特点,这为进一步的研究和药物研发提供了依据.

Objective:To investigate the transcriptome characteristics and clinical significance of molecular classes of gliomas based on Chinese cohorts.Methods:The clinical information and transcriptome sequencing data of 932 patients with gliomas were retrospectively analyzed in the Chinese Glioma Genome Atlas (CGGA) daatabase. The patients were divided into isocitrate dehydrogenase (IDH, include IDH1 and IDH2) mutation with chromosomal 1p/19q co-deletion (Mut_Co) type (182 cases), IDH mutation with chromosomal 1p/19q co-deletion (Mut_nCo) type (315 cases) and IDH wild (WT) type (435 cases). The age of diagnosis, gender, histopathological type, TCGA (The Cancer Genome Atlas) subtype, CpG island methylation phenotype (G-CIMP) and tumor purity were compared among patients with three molecular classes of gliomas. We explored the transcriptome characteristics of three molecular types of gliomas, the biological pathways involved and their differences in cancer marker scores. A total of 885 patients with transcriptomic data and complete follow-up data were included for survival analysis and Kaplan-Meier curves were drawn to compare the difference in median survival of patients with three molecular classes of gliomas.Results:Compared with Mut_Co and Mut_nCo, the proportion of WT patients with diagnosis age ≥43 years, WHO grade Ⅳ, glioblastoma, TCGA classical type and G-CIMP type was higher, and the tumor purity was the lowest, the differences were statistically significant (all P<0.001). Compared with Mut_Co and Mut_nCo, the intra- and local- invasion scores of WT glioma were higher, and the proportion of cell cycle in DNA synthesis phase (S phase) was higher, with statistical significance (all P<0.001). In WT type, the transcription factor PAX3 was highly expressed and the p53 signaling pathway was lowly expressed. Pathway functional enrichment analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that Mut_Co and Mut_nCo types were widely involved in the interaction of ligand receptors stimulating neural tissue and long-term depression, which were also widely involved in Hedgehog protein signal, WNTβ CATENIN signal and other cancer markers; WT type was mainly involved in p53 signaling pathway, adhesion plaque, apoptosis and other biological pathways and cancer hallmarkers. The log-rank test showed that the median survival time of Mut_Co patients was the longest (most of them survived), followed by Mut_nCo (1 258 d), and WT with the shortest survival time (415 d), and the difference was statistically significant ( P<0.001). Conclusion:The three molecularly classified gliomas based on the Chinese cohorts are associated with different clinical features, different prognosis of patients, specific transcriptomic features, and involve specific biological pathways and cancer hallmarkers.

目的 观察MMP-9抑制剂对人口腔鳞癌细胞系CAL27增殖和侵袭能力的影响.方法 选取人口腔鳞癌细胞系CAL27进行培养,用不同浓度的MMP-9抑制剂BB-94处理对数期CAL27细胞,通过RT-PCR检测BB-94在基因水平上对MMP-9的抑制效果.细胞免疫化学实验检测BB-94在蛋白水平上对MMP-9的抑制效果.MTT实验检测MMP-9抑制剂BB-94对人口腔鳞癌细胞增殖能力的影响.划痕实验检测MMP-9抑制剂BB-94对人口腔鳞癌细胞迁移能力的影响;Transwell实验检测MMP-9抑制剂BB-94对人口腔鳞癌细胞侵袭能力的影响.结果 MMP-9抑制剂能有效抑制人口腔鳞癌细胞CAL27的增殖、迁移和侵袭能力.结论 抑制MMP-9的表达可以抑制人口腔鳞癌细胞CAL27的增殖、迁移和侵袭等恶性生物学行为.

目的 探讨口腔白斑(OLK)、口腔鳞状细胞癌(OSCC)与正常口腔黏膜组织中抑癌基因RASSF1、RASSF2的甲基化状态变化与病理学及人口学资料的关系.方法 收集就诊于首都医科大学附属北京口腔医院黏膜科OLK 87例,OSCC 39例,30例正常口腔黏膜作为对照,采用内切酶消化法和实时定量PCR的方法对RASSF1和RASSF2甲基化状态进行研究.结果 在正常口腔黏膜中,RASSF1、RASSF2启动子区处于未甲基化状态,口腔白斑中RASSF1甲基化阳性率为44.83％,RASSF2甲基化阳性率为52.87％,与正常对照相比显著增高(P<0.017);口腔鳞癌RASSF1甲基化阳性率为25.64％,RASSF2甲基化阳性率为43.59％,与正常对照相比显著增高(P<0.017).结论 抑癌基因RASSF1、RASSF2启动子甲基化有成为口腔黏膜癌变早期标志物的可能性.

目的 观察姜辣素(Gingerol)对人口腔鳞癌细胞系CAL-27恶性生物学行为的影响.方法 选取人口腔鳞癌细胞CAL-27细胞系进行培养,用不同浓度的Gingerol处理对数期CAL-27细胞,通过MTT实验检测Gingerol对人口腔鳞癌细胞增殖的影响;克隆形成实验检测Gingerol对人口腔鳞癌细胞的克隆形成能力影响;划痕实验检测Gingerol对人口腔鳞癌细胞的迁移能力的影响;Transwell实验检测Gingerol对人口腔鳞癌细胞的侵袭能力的影响.结果 Gingerol能有效抑制人口腔鳞癌细胞CAL-27的增殖、克隆形成、迁移和侵袭能力,并呈一定的时间和剂量依赖性.结论 姜辣素对人口腔鳞癌细胞CAL-27细胞在增殖、克隆和迁移侵袭等恶性生物学行为方面能起到明显的抑制作用,有作为治疗口腔癌药物的应用前景.

Objective To evaluate the effectiveness of a community-based rehabilitation based on conductive education and self-management measures of risk factors in improving awareness, behavior and functional recovery of stroke survivors.Methods Two communities were selected as either the intervention or control community in Dongcheng District. A 3-month community-based rehabilitation treatment plan and 6-month risk factors self-management program, based on modern rehabilitation theory, conductive education and risk factor self-management for stroke survivors, was successively implemented among 109 patients in intervention community. The effects before and after the intervention were evaluated through comparing stroke survivors in intervention community with themselves and 110 stroke survivors in control community.Results After 3-month rehabilitation treatment, median Fugel-Meyer assessment(FMA) score for stroke survivors in intervention community was enhanced from 57.0(30.0-81.5) to 70.5(40.5-92.0)(P <0.001), median Barthel index(BI) score from 90(65-95) to 95(75-100)(P <0.001) and mean score of function activity questionnaire(FAQ) was decreased from(11.6±8.3) to(10.1±8.4)(P <0.001). After 6-month self-management, the stroke risk factors awareness(P <0.001), warning awareness(P <0.001), rehabilitation awareness(P <0.001), blood glucose control(P =0.033) and adherence to rehabilitation(P <0.001) in intervention community were significantly higher than that in control community.Conclusion Community-based rehabilitation treatment and risk factors self-management program may improve awareness, behavior and functional recovery of stroke survivors.

Objective To investigate the regulation of DNA methylation and histone acetylation in oral squamous cell carcinoma( OSCC) cells on KLF4 gene. Methods SCC-6 cells were divided into four groups, negative group ( without any treatment ) , DAC group ( treated with DAC- DNA methylation inhibitor ) , TSA group ( treated with TSA- histone deacetylase inhibitors) and DAC+TSA group( treated with DAC+TSA) . Quantitative PCR was used to examine the RNA expression of KLF4 in SCC-6 before and after the treatment of DAC or/and TSA. Immunecytochemistry was used to detect KLF4 protein expression before and after the treatment of DAC or / and TSA in SCC-6 cells. Results KLF4 RNA expression in DAC group, TSA group, and DAC+TSA group was significantly higher than that in the negative control ( P<0.01). There was no significant difference between TSA group and DAC +TSA group (P >0. 05). The KLF4 RNA expression in DAC group was higher than that in other three groups (P<0. 01). The protein expression of KLF4 in DAC group was significantly higher than that in other three groups. (P<0. 01). KLF4 expression was not significantly different between the TSA group and DAC+TSA group (P>0. 05), but was significantly higher in TSA and DAC+TSA groups than that in the negative control (P<0. 01). Conclusion DNA methylation may be the main reason for KLF4 silence in SCC-6 cells ,and histone acetylation may also participate in the regulation of KLF4 expression, but DNA methylation and histone acetylation has no synergistic effect on improving the expression of KLF4.

患者，女，61岁，2011年10月26日因发现口腔白斑1周来我科就诊。检查：15到17，27腭侧牙龈分别可见大小1.5cm×2cm、1cm×1.5cm的灰白色斑块，表面略粗糙、微突出于黏膜表面、质韧、边界不规则、界清、无触痛；14到26腭侧龈缘可见条带状的浅白色斑块，部分连接成片、界清、无触痛、均质；17腭侧可见0.3cm×0.3cm充血区，无触痛。临床初步诊断：口腔白斑。血常规无明显异常。组织活检：于17腭侧病损处取病理，病理组织沿长轴一分为二，一半用于病理检测，另一半提取DNA，用于甲基化基因芯片分析和肿瘤抑制基因P16、P15、RASSF1A、RASSF2甲基化状态检测。

Objective: To study the relationship of the promoter methylation of tumor suppressor genes P16 and P15 in oral precancerous lesion with the pathologic and demographic data. Methods: Promoter methylation of P16 and P15 was detected in 87 cases of oral mucosa leukoplakia and 39 cases of oral squamous cell carcinoma( OSCC) tissues using methylation sensitive enzyme and methylation dependent enzyme digestion. 20 cases of normal oral mucosa were used as the controls. Results: No promoter methylation of P16 and P15 was found in the controls. In leukoplakia tissues,hypermethylation of P16 and P15 was 41. 38% and 44. 83% of the cases respectively( vs control,P 0. 001). In oral squamous carcinoma,hypermethylation of P16 and P15 was 20. 51% and 23. 08% of the cases respectively( vs control,P 0. 05). No correlation was found between the methylation and the gender,age,smoking, lesion site,pathology and types of the patients. Conclusion: Promoter methylation of P16 and P15 may be biomarkers of early oral carcinogenesis.

Objective To investigate the possible relationship between the single nucleotide polymorphisms(SNP) of TNF-α and the susceptibility of oral lichen planus(OLP).Methods A total of 151 OPL patients(including 81 erosive OLP and 70 non-erosive OLP) who were identified in our department from April to November 2010 and 143 healthy controls who received physical examination in Tiantan Hospital,Beijing from July to October 2010 were enrolled in this study.The polymorphisms of TNF-α-308 were detected in the 151 OLP cases and 143 healthy controls by using the PCR-RFLP technique.The genotype and the allele frequencies were analyzed between the 2 groups.Results There was no significant difference in the genotypepe and allele frequency between the OLP group and the healthy control(P>0.05),but was found between the erosive OLP group and the health control.When compared with healthy control,the allele frequencies of TNF-α-308 *A was much higher in the erosive OLP group(P<0.05),and subjects carrying the TNF1 genotype had significantly increased risk of erosive OLP(OR=2.268,P<0.05).Conclusion The polymorphisms of TNF-α-308 may be not a single dependent factor associated with the susceptibility and development of OLP,but the TNF-α-308 *A allele might attribute to deteriorate OLP.

Objective To examine the effect of arsenic trioxide(AS2O3)on Nrf2 pathway in the oral squamous cell carcinomacell line SCC-15.Methods The expression of Nrf2,Ho-1 and NQO1 in SCC-15 cells stimulated by different concentrations of AS2O3 was analyzed by accounting the mean optical density(MOD)and Western blotting.ResultsAll tested concentrations of AS2O3 activated the expression of Nrf2,Ho-1 and NQO1 in SCC-15 cells.The expression was dose-dependent.Conclusion AS2O3 activtes the Nrf2 pathway in the oral squamous cell carcinomacell line SCC-15 and ptomotes the expression of antioxidant genes.

Objective To observe the effects of treatment of food impaction(FI)by composite resin restoration and occlusal adjustment and to discuss its clinical features.Methods A total of 124 patients(137 sites)with food impaction were selected and classified into two types.Type 1 Static food impaction(sFI)was treated mainly by light-curing composite resin restoration,type 2 kinetic food impaction(kFI)was treated by occlusal adjustment.Proximal caries and periodontal conditions were recorded.One week and 6 months after the treatment,the patients were re-examined.Results There were 103 sites in sFI and 34 in kFI.The mean DFT and periodontal index of the food impaction sites were more severe than that of the control sites.One week after treatment,food impaction disappeared in 115 sites(83.9%),markedly mended in 22 sites(16.1%).Six months after treatment,120 sites(87.6%)showed no food impaction,17 sites(12.4%)improved.Conclusion Most food impaction can be effectively treated with combined use of composite resin restoration and occlusal adjustment.

OBJECTIVE To explore the expression of epidermal growth factor (EGF), epidermal growth factor receptor (EGFR), c-fos and c-jun in oral lichen planus (OLP). METHODS The levels of gene expression of EGF, EGFR, c-fos and c-jun were detected with reverse transcription-polymerase chain reaction (RT-PCR) in 10 cases of erosive OLP, 12 cases of reticular OLP and 9 cases of normal oral mucosa. Protein contents and distribution of EGF, EGFR, C-fos and C-jun were examined with immunohistochemical technique in different tissues. RESULTS The endogenous levels of EGFR mRNA and protein in erosive OLP were (55.9 +/- 23.1)% and (71.1 +/- 10.1)%, respectively, and significantly higher than those in reticular OLP and normal oral mucosa. The mRNA contents of c-fos and c-jun were significantly higher in erosive OLP than in normal oral mucosa. The positive cell rates of c-fos and c-jun protein in erosive OLP were 1.3 and 1.5 times of those in normal oral mucosa, respectively (P < 0.05). CONCLUSIONS The probability of carcinomatous change of erosive OLP might be higher than that of reticular OLP, in which EGF and EGFR might play an important role.

生长因子是一系列具有多种功能的活性多肽类物质,起到信号蛋白的作用.它们与细胞的生长、分化、增殖等过程直接相关,在机体的组织修复、炎症反应和免疫反应过程中起到极其重要的作用.生长因子种类较多,其中表皮生长因子(epidermal growth factor,EGF)是发现较早、研究最多的生长因子.体外细胞培养和动物试验均提示了EGF具有促进细胞增殖的作用,l临床试验也证实EGF可加快创面愈合速度.进一步研究EGF对创面愈合的基本规律及调控机理将为EGF更好地应用于临床提供坚实的理论依据.