雷帕霉素诱导自噬抑制舌鳞癌细胞增殖和迁移

目的探讨自噬诱导对舌鳞癌细胞增殖和迁移能力的影响。方法应用雷帕霉素(Rap)诱导上调舌鳞癌Tca8113细胞自噬活性,Western blot检测诱导后Tca8113细胞自噬标记蛋白Beclin1和LC3的表达变化;MTT法检测自噬诱导对细胞增殖的影响;Wound-healing检测诱导后细胞迁移能力改变;SPSS 19.0统计软件进行数据分析。结果 Rap诱导6h即可上调舌鳞癌Tca8113细胞自噬标记蛋白LC3-Ⅱ表达,24h时LC3-Ⅱ表达最强,自噬活性最高(P<0.05)。与对照组相比,Rap诱导后细胞生长明显抑制(P<0.05),迁移速率减慢。结论上调舌鳞癌细胞自噬活性可以抑制其增殖和迁移。     

哺乳动物雷帕霉素靶蛋白复合物1介导的自噬对骨代谢的调控

哺乳动物雷帕霉素靶蛋白复合物(mTORC)1是哺乳动物雷帕霉素靶蛋白形成的一种复合物,在细胞合成代谢过程中起重要作用,其参与调控的自噬作用近年来受到广泛关注。自噬是细胞降解损坏的蛋白质或细胞器并将其循环利用的过程。随着对mTORC1/自噬效应的研究逐渐深入,其在骨代谢方面的调控作用愈发凸显。本文就mTORC1介导的自噬通路在成骨细胞、破骨细胞等骨相关细胞方面的作用及其机制进行综述,为骨代谢的生物学机制和骨组织疾病的研究提供新思路。     

哺乳动物雷帕霉素靶蛋白在牙周炎大鼠相关肾损伤中的作用

目的 本研究旨在初步探讨哺乳动物雷帕霉素靶蛋白(mammalian target of rapamycin,mTOR)在牙周炎大鼠相关肾损伤中的作用。方法 20只雄性Wistar大鼠被随机分为对照组和牙周炎组,每组各10只。牙周炎组采用0.2 mm正畸丝结扎法构建牙周炎动物模型,对照组不作结扎处理。8周后,分别记录两组大鼠牙龈出血指数、牙周袋探诊深度及松动度,并通过Micro-CT分析牙槽骨吸收情况;采用HE及PAS染色进行组织学分析。qRT-PCR和Western Blot分别检测大鼠肾组织内的mTOR、phosphor-mTOR(p-mTOR)和BECN1(Beclin-1)的mRNA及蛋白表达情况。结果 与对照组对比,牙周炎组大鼠的牙龈组织出现明显肿胀、颜色及质地的改变,牙龈出血指数、牙周袋探诊深度及松动度均明显增加,牙槽骨吸收明显;肾小球鲍曼氏囊腔明显扩张,基底膜增厚,肾小管间质充血明显,刷状缘破坏;肾脏功能未见明显差异;牙周炎组的mTOR的mRNA及蛋白表达(p-mTOR/mTOR)均明显高于对照组,Beclin-1的mRNA及蛋白表达均明显低于对照组。结论 mTOR可能通过抑制自噬过程在大鼠牙周炎相关肾损伤中发挥作用。     

雷帕霉素促进上颌窦黏膜细胞成骨分化

目的:探讨雷帕霉素对上颌窦黏膜细胞成骨分化的影响。方法:上颌窦黏膜细胞经不同浓度的自噬激动剂雷帕霉素处理后诱导成骨分化,然后进行碱性磷酸酶活性检测、茜素红染色以及自噬基因和成骨基因转录水平的检测,评估上颌窦黏膜细胞成骨分化和自噬水平的改变。结果:碱性磷酸酶活性、钙结节形成量以及成骨基因的表达随成骨诱导逐渐增加,并且100 nmol/L的雷帕霉素处理组表现出较强的成骨能力和自噬水平。结论:合适浓度的雷帕霉素可促进上颌窦黏膜细胞的成骨分化,这一过程可能与自噬途径相关。     

抑制脉管畸形的相关靶向药物研究进展

脉管畸形是一种先天性疾患,主要发生在头颈部,其无法自行消退,且随着患者的生长而逐渐加重。脉管畸形的传统治疗方式包括：激光治疗、硬化治疗、介入栓塞、手术切除等。但是对于一些范围较大的病变,传统治疗方式效果不佳。随着分子遗传学的发展,基因突变目前被认为是脉管畸形发生的根本原因,基因突变引起的相关通路的活化进一步促进了脉管畸形病变的进展。低流速脉管畸形主要涉及磷脂酰肌醇3-激酶（phosphatidylinositol 3-kinase,PI3K）/蛋白激酶B(protein kinase B,AKT)/哺乳动物雷帕霉素靶蛋白（mammalian target of rapamycin,mTOR）通路的激活;而高流速的脉管畸形主要涉及大鼠肉瘤（rat sarcoma,RAS）/快速加速纤维肉瘤（rapidly accelerated fibrosarcoma,RAF）/促分裂原活化蛋白激酶激酶（mitogen-activated protein kinase kinase,MAPKK）/细胞外信号调节激酶（extracellular-signalregulated protein kinas...     

NOD2上调自噬抑制舌鳞癌细胞增殖和迁移

目的:探讨NOD2在雷帕霉素(Rap)诱导下上调舌鳞癌SCC-15细胞自噬对增殖和迁移能力的影响。方法:NOD2表达载体转染SCC-15细胞上调NOD2表达,NOD2-shRNA载体转染SCC-15细胞,抑制NOD2表达;应用Rap诱导SCC-15细胞正常组、NOD2上调组和抑制组的细胞自噬活性,采用Western blot检测自噬标记蛋白LC3和Beclin-1表达变化;MTT法检测NOD2对自噬诱导后细胞增殖的影响;划痕实验检测NOD2对自噬诱导后细胞迁移能力的变化。结果:Rap诱导NOD2上调组细胞中LC3-II和Beclin-1表达显著增强(P<0.05)、诱导NOD2抑制组细胞中LC3-II和Beclin-1表达显著降低(P<0.05)。与对照组相比,Rap诱导NOD2上调组细胞生长明显抑制(P<0.05),迁移速率减慢;诱导NOD2抑制组细胞增殖加快(P<0.05),迁移速率增快。结论:NOD2可上调舌鳞癌SCC-15细胞自噬抑制其增殖和迁移。     

哺乳动物的雷帕霉素靶蛋白及其底物在口腔鳞状细胞癌中的表达

目的　探讨哺乳动物的雷帕霉素靶蛋白(mTOR)及其底物对口腔鳞状细胞癌生长的影响。方法　提取 RNA,通过RT-PCR和Western印迹分析观察mTOR及其底物p70 S6激酶(p70 S6k)的α1、α2、β1、β2不同亚型及起始因子4E结合蛋白1(4EBP1)在口腔鳞状细胞癌中的表达。结果　RT-PCR与Western印迹的结果一致。随着肿瘤恶性度的提高,mTOR、p70 S6K的表达明显增加,而4EBP1的表达则下降。结论　mTOR及其底物表达水平的强弱与口腔鳞状细胞癌的分化程度密切相关,它可能成为未来癌症治疗的重要靶向蛋白。     

沉默Beclin1基因抑制自噬促进舌鳞状细胞癌细胞增殖及侵袭转移

目的 探讨抑制自噬基因Beclin1表达对舌鳞状细胞癌(以下简称舌鳞癌)细胞增殖及侵袭转移的影响,了解舌鳞癌侵袭转移的调控机制.方法 利用RNAi技术,针对人cDNA序列设计Beclin1干扰序列,用脂质体Lip02000包裹后转染至舌鳞癌UM2细胞.实验设空白对照组、脂质体2000(Lip02000)组、阴性siRNA组和Beclin1 siRNA组,通过蛋白印迹法检测Beclin1、LC3的表达变化;CCK-8法检测细胞增殖能力变化;Transwell小室检测细胞迁移和侵袭能力改变;SPSS13.0统计软件进行数据分析.结果 转染Beclin1 siRNA对UM2细胞Beclin1有显著敲减作用(P＜0.05),自噬标记蛋白LC3-Ⅱ的表达明显降低(P＜0.05),细胞增殖较对照组增快(P＜0.05),细胞迁移和侵袭转移能力明显增强.结论 沉默Beclin1表达可下调舌鳞癌细胞自噬水平,并促进舌鳞癌细胞增殖和侵袭能力.     

苯妥英钠对大鼠骨髓间充质干细胞向血管内皮细胞分化的影响

目的：研究苯妥英钠（PHT）对大鼠骨髓间充质干细胞（rBMSCs）向血管内皮细胞（rVECs）分化的影响。方法：建立rBMSCs 与 rVECs 共培养组及 rBMSCs 单独培养组,各组分别添加不同浓度的 PHT（0、20、40μg／ml）,培养14 d 后 real-time PCR 检测各组细胞中 ICAM-1、VCAM-1、KDR 和 RUNX2基因的 mRNA 表达水平。结果：添加 PHT 后,各组细胞中 ICAM-1、KDR 和 RUNX2基因的 mRNA 表达均上调。添加相同浓度 PHT 时,共培养组较单独培养组中 rBMSCs 的 ICAM-1、KDR 和RUNX2基因的 mRNA 表达量增高,而 VCAM-1基因的 mRNA 表达量降低。结论：PHT 可能促进大鼠骨髓间充质干细胞向血管内皮细胞分化。     

血管瘤与脉管畸形的鉴别诊断研究进展

血管瘤和脉管畸形是婴幼儿比较常见的血管疾病,过去由于对两者的分类和诊断比较混乱,给临床治疗带来诸多不便,也给患者增加了不必要的痛苦。1982年,Mulliken和Glowacki提出了脉管性疾病的生物学新分类,将血管瘤和脉管畸形归类为2种性质完全不同的病变。血管瘤是多发于婴幼儿的良性肿瘤,大多数可以自行消退;而脉管畸形是血管结构的发育异常,不会自行消退,随患者的生长发育持续增长。因此,对于确诊的脉管畸形,应尽早采取干预措施。这就要求临床医师能及时、准确、有效地鉴别诊断血管瘤和脉管畸形。本文就目前关于血管瘤和脉管畸形的鉴别诊断进展作一综述。     

牙龈卟啉单胞菌感染对大鼠血管平滑肌细胞细胞间黏附分子-1表达的影响

目的 观察牙龈卟啉单胞菌ATCC 33277感染对大鼠血管平滑肌细胞（VSMC）细胞间黏附分子-1（ICAM-1）表达的影响。方法 建立体外牙龈卟啉单胞菌感染大鼠VSMC模型,逆转录-聚合酶链反应（RT-PCR）检测ICAM-1基因的表达。结果 牙龈卟啉单胞菌感染VSMC 8、16、24 h后,ICAM-1表达明显增多,与空白对照组比较,差异有统计学意义（P<0.05）。感染16 h达高峰,感染8 h与感染16、24 h比较,差异有统计学意义（P<0.05）。结论 牙龈卟啉单胞菌感染可引起VSMC ICAM-1高表达,这提示牙周致病菌可能参与血管壁的炎症反应,在动脉粥样硬化的发生、发展中有重要的意义。     

Platelet-poor plasma stimulates the proliferation but inhibits the differentiation of rat osteoblastic cells in vitro

Introduction: Recent studies have shown that the use of platelet preparations in bone and implant surgery might stimulate bone formation. However, the biological mechanisms are not well understood. Moreover, few studies have attempted to evaluate the effect of platelet-poor plasma (PPP), which is a product of the platelet-rich plasma preparation process.
Objective: Thus, this study investigated the behavior of osteoblasts isolated from fetal rat calvaria cultivated in the presence of homologous PPP.
Material and Methods: PPP was obtained by centrifugation of the rat mother's blood and used in replacement of fetal calf serum, which is classically used in primary culture procedures. Proliferation was measured by an MTT assay at 24, 48, and 72 h. Real-time PCR was performed to study the expression of Runx2, Dlx5, and osteocalcin (OC) on days 0 (4 h), 1, 3, 7, and 12.
Results: Alkaline phosphatase (ALP) biochemical activity was evaluated on days 0 (4 h), 1, 3, 7, and 12. Observations by phase-contrast microscopy showed that osteoblasts were able to differentiate until the mineralization of the matrix in the presence of PPP. PPP enhanced the proliferation significantly compared with the control group ( P ≤0.001). PCR results showed that Runx2, Dlx5, and OC were expressed by cells in the experimental group at lower levels compared with the control group. Biochemical assay of ALP showed a lower activity in the experimental group compared with the control group ( P <0.001).
Conclusion: These results suggest that, in the presence of homologous PPP, rat osteoblastic cells are able to maintain their phenotype, with a higher rate of proliferation. However, PPP seems to inhibit osteoblastic differentiation.     

淫羊藿素对 SD 大鼠骨髓基质细胞增殖与成骨分化的影响

目的：研究淫羊藿素（ICT）对体外培养 SD 大鼠骨髓基质细胞（rBMSCs）增殖与成骨分化的影响。方法：体外分离培养 rBMSCs,传代至第4代作多向分化鉴定。分别以10－9、10－8、10－7、10－6、10－5 mol／L ICT 刺激 rBMSCs 后3、6、9 d,分别用 cck-8及碱性磷酸酶 ALP 试剂盒检测 rBMSCs 的增殖及 ALP 活性;10－9 mol／L ICT 处理 rBMSCs 后21 d 作茜素红（AR）染色以判断钙结节的形成。结果：原代培养的 rBMSCs 贴壁生长、呈梭形,能多向分化;ICT 明显抑制了 rBMSCs 的增殖;但增高其 ALP 活性、钙结节形成。结论：ICT 以剂量依赖方式抑制 rBMSCs 的增殖但促进其分化和矿化。     

Role of autophagy in head and neck cancer and therapeutic resistance

Head and neck squamous cell carcinomas (HNSCC) are one of the most common cancers worldwide, accounting for almost 50% of all malignancies in developing nations. Autophagy is a catabolic process involving turnover of long‐lived proteins and organelles and is an important mechanism for cell survival under stress conditions. Autophagy has been shown to play a pivotal role in etio‐pathogenesis of several cancers. Autophagy and apoptosis may be triggered by common upstream signals, and sometimes this results in combined autophagy and apoptosis, or defective apoptosis rendering immortalized epithelial cells highly tumorigenic. Autophagy has been found to buffer metabolic stress and may help in cell survival; however, inhibiting autophagy under conditions of nutrient limitation can restore cell death to apoptosis‐refractory tumors. Therefore, autophagy acts as a double‐edged sword in cancer therapeutics. Role of autophagy in pathophysiology and as a potential cancer therapeutics is a subject of intensive research. This review will focus on the role of autophagy and how it contributes to the pathogenesis and overcoming therapeutic resistance in HNSCC.     

Up-regulation of interleukin-6 and vascular endothelial growth factor-A in the synovial fluid of temporomandibular joints affected by synovial chondromatosis

Our aim was to explore important inflammatory mediators for synovial chondromatosis in the temporomandibular joints (TMJs) by analysing synovial fluid. Samples were collected from 10 patients with unilateral synovial chondromatosis of the TMJ. Control samples were obtained from 11 subjects with no symptoms in the TMJ. Concentrations of aggrecan, interleukin (IL)-2, IL-4, IL-5, IL-6, IL-8 (CXCL8), IL-10, interferon (IFN)-γ, tumour necrosis factor (TNF)-α and vascular endothelial growth factor (VEGF)-A were measured in the samples of synovial fluid, and the results in the two groups compared. The tissues from the affected TMJ were examined histologically and immunohistochemically. Of the proteins evaluated, the concentrations of aggrecan, IL-6, and VEGF-A were significantly higher in the group with synovial chondromatosis. The immunohistochemical analysis showed that the synovial cells around the osteocartilaginous nodules were vigorously expressing VEGF-A.