靶向沉默髓鞘转录因子MyT1对脑胶质瘤细胞恶性生物学行为的影响

目的探讨靶向沉默髓鞘转录因子1(MyT1)对人脑胶质瘤细胞迁移、侵袭和黏附的影响和相关分子机制。方法设计特异性靶向沉默MyT1基因的shRNA,包装慢病毒后感染人脑胶质瘤U-118MG和U-87MG细胞,qPCR和Western blot检测两种细胞中MyT1的表达水平,BrdU实验、细胞划痕实验、Transwell实验和细胞黏附实验分别检测两种细胞的迁移、侵袭和黏附能力的变化,qPCR检测细胞黏附和肿瘤转移相关基因的表达水平。结果在HEK293T细胞中包装了特异性靶向MyT1基因的shRNA慢病毒,并成功感染了U-118MG和U-87MG细胞;两种细胞中MyT1 mRNA和蛋白表达水平均显著下调(均P<0.05),细胞的迁移、侵袭和黏附能力均有一定程度的降低(均P<0.05),细胞黏附相关基因表达水平显著下降,肿瘤转移相关基因表达水平显著上升(均P<0.05)。结论靶向沉默MyT1基因对人脑胶质瘤U-118MG和U-87MG细胞的迁移、侵袭和黏附能力有抑制作用,其机制可能与调控细胞黏附和肿瘤转移相关基因的表达有关。MyT1基因的表达,可能是脑胶质瘤诊疗的一个潜在靶标...     

Eye bank versus surgeon prepared Descemet stripping automated endothelial keratoplasty tissues: Influence on adhesion force in a pilot study

Purpose:To evaluate and compare the biomechanical properties of the eye bank-prepared and surgeon prepared Descemet stripping automated endothelial keratoplasty (DSAEK) tissues.Methods:In this laboratory study, corneal tissues for research were randomly allocated in the following groups: a) surgeon-cut DSAEK and b) eye bank-prepared (pre-cut and pre-loaded) DSAEK. Endothelial cell loss (ECL), immunostaining for tight junction protein ZO-1, elastic modulus, and adhesion force were investigated.Results:ECL was not found to be significantly different between surgeon-cut DSAEK (7.8% ±6.5%), pre-cut DSAEK (8.6% ±2.3%), and pre-loaded DSAEK (11.1% ±4.8%) (P = 0.5910). ZO-1 was expressed equally across all groups. Surgeon-cut DSAEK grafts showed a significantly higher elastic modulus compared to pre-cut and pre-loaded DSAEK groups (P = 0.0047 and P < 0.0001, respectively). Adhesion force was significantly greater in the surgeon-cut DSAEK compared to pre-cut (P < 0.0001) or pre-loaded DSAEK groups (P = 0.0101).Conclusion:The laboratory data on the biomechanics of DSAEK grafts suggests that surgeon-cut DSAEK grafts present higher elastic modulus and adhesion force compared to eye bank-prepared DSAEK grafts.     

丹皮酚对高脂血清诱导的大鼠主动脉内皮细胞损伤的保护作用及其黏附功能的影响

目的观察丹皮酚（Paeonal,Pae）对高脂血清诱导的大鼠单核细胞（mononuclear cell,MC）与大鼠主动脉内皮细胞（rat aortic endothelial cells,RAECs）黏附的影响及对RAECs损伤的保护作用。方法高脂乳剂灌胃大鼠并制备高脂血清;组织块预消化贴壁法培养RAECs;淋巴细胞分离液分离MC;孟加拉玫瑰红活细胞染色法测定MC与RAECs的黏附功能;甲基噻唑基四唑比色法以及台盼蓝死细胞染色法检测Pae对高脂血清诱导的RAECs损伤的保护作用。结果30～70ml／L高脂血清呈剂量依赖性地诱导MC与RAECs的黏附,50ml／L时促进MC与RAECs黏附达最大值;高脂血清与RAECs共育6h即可明显促进黏附作用,24h时达最大诱导效应。Pae在浓度为15～240μmol／L时,呈剂量依赖性地抑制高脂血清诱导的黏附作用;240μmol／L孵育24h时抑制作用最明显;Pae在30,60,120μmol／L浓度时,对高脂血清损伤的大鼠RAECs有明显的保护作用。结论Pae对高脂血清诱导的大鼠MC与RAECs黏附功能有抑制效应,对高脂血清损伤的大鼠RAECs有明显的保护作用。     

Purmorphamine increased adhesion,proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate

ObjectivesGrowth factors play a significant role in cell proliferation and differentiation during different stages of the bone repair. However, several limitations have been brought researchers attention to an osteoinductive small molecule including Purmorphamine. In this study, we aimed to evaluate the effect of Purmorphamine on adhesion, proliferation and differentiation of human dental pulp stem cells (hDPSCs) seaded on beta-tricalcium phosphate (β-TCP) granules.MethodshDPSCs were established from extracted wisdom teeth of healthy volenteers. Cells at passage 3 were seeded on β-TCP in the presence or absence of Purmorphamine. Cell adhesion and proliferation were assessed using scanning electeron microscopy (SEM) and DNA counting assay, respectively, after 1, 3 and 5 days. Then, hDPSCs seeded on β-TCP were subjected to osteogenic medium with or without Purmorphamine. After 7 and 14 days osteogenic diffrentiation capability of hDPSCs were determined using real-time RT-PCR and alkaline phosphatase (ALP) activity assay.ResultsThe significant increase in amount of DNA was observed at day 3 and 5 in the presence of Purmorphamine. SEM imaging also was confirmed the DNA counting assay; in all given time points, hDPSC attachment and growth was significantly higher in the presence of Purmorphamine. ALP activity was increased by Purmorphamine at both 7 and 14 days of induction. Purmorphamine showed to effect on osteopontin expression at earlier stage of osteogenic differentiation, whereas for osteocalcin expression, this effect was more evident at later stage of differentiation.ConclusionPurmorphamine had a promotive effect on adhesion, proliferation and osteogenic differentiation of hDPSCs cultured on β-TCP. The outcome of the current study would help in development of in vitro culture conditions for better osteogenic differentiation of hDPSCs prior to transplantation.     

Expression of DNAM-1 (CD226) on inflammatory monocytes

DNAM-1 is an activating receptor expressed on NK cells and T cells and plays an important role in cytotoxicity of these cells against target cells. Although the role of DNAM-1 in the function of T cells and NK cells has been well studied, the expression and function of DNAM-1 on myeloid cells have been incompletely understood. In this study, we investigated expression of DNAM-1 on monocyte subsets in mouse peripheral blood and found that only inflammatory monocytes (iMos), but not patrolling monocytes (pMos), expressed high levels of DNAM-1. In addition, we found that DNAM-1 was highly expressed on iMos, rather than pMos, also in human. Furthermore, we found that DNAM-1 on inflammatory monocytes was involved in cell adhesion to CD155-expressing cells. Therefore, we propose that expression of DNAM-1 on inflammatory monocytes are evolutionally conserved and act as an adhesion molecule on blood inflammatory monocytes.     

Soluble Adhesion Molecules in Coronary Surgery and Cardiopulmonary Bypass with Pump Prime Aprotinin

Objective : The purpose of the present study was to establish whether pump prime aprotinin could influence soluble adhesion molecules in patients undergoing elective coronary artery bypass surgery. Design : Thirty patients admitted for first-time elective coronary artery bypass surgery were randomized into control or aprotinin groups. Patients in the aprotinin group received 280 mg of aprotinin in the pump prime. Plasma levels of soluble adhesion molecules were analyzed perioperatively. Results : There were no significant changes in plasma sE-selectin after the operation in either group. Plasma sP-selectin increased significantly up to 20 h after reperfusion to the myocardium. Plasma sICAM-1 decreased in the early stage after cardiopulmonary bypass (CPB), then recovered at 4 h after reperfusion and a significant increase in sICAM-1 was observed 20 h later. There were no significant differences between the groups in postoperative changes in sP-selectin ( p = 0.21) and sICAM-1 ( p = 0.91). Conclusion : Pump prime aprotinin did not influence plasma levels of E-selectin, P-selectin and ICAM-1 in the present patients. The present results do not support the concept of an anti-inflammatory effect of pump prime aprotinin.     

Biofilm-formation capability depends on environmental oxygen concentrations in Candida species

BackgroundAlthough oxygen concentrations inside of the human body vary depending on organs or tissues, few reports describe the relationships between biofilm formation of Candida species and oxygen concentrations. In this study, we investigated the biofilm-forming capabilities of Candida species under various oxygen conditions.MethodsWe evaluated the adhesion and biofilm formation of Candida albicans and C. tropicalis under aerobic, microaerobic (oxygen concentration 5%), or anaerobic conditions. We also examined how oxygen concentration affects adhesion/maturation by changing adhesion/maturation phase conditions. We used crystal violet assay to estimate the approximate biofilm size, performed microscopic observation of biofilm morphology, and evaluated adhesion-associated gene expression.ResultsThe adhered amount was relatively small except for a clinical strain of C. tropicalis. Our biofilm-formation analysis showed that C. albicans formed a higher-size biofilm under aerobic conditions, while C. tropicalis favored microaerobic conditions to form mature biofilms. Our microscopic observations were consistent with these biofilm-formation analysis results. In particular, C. tropicalis exhibited more hyphal formation under microaerobic conditions. By changing the adhesion/maturation phase conditions, we represented that C. albicans had favorable biofilm-formation capability under aerobic conditions, while C. tropicalis showed enhanced biofilm formation under microaerobic adhesion conditions. In good agreement with these results, the C. tropicalis adhesion-associated gene expression tended to be higher under microaerobic or anaerobic conditions.ConclusionsC. albicans favored aerobic conditions to form biofilms, whereas C. tropicalis showed higher biofilm-formation ability and promoted hyphal growth under microaerobic conditions. These results indicate that favorable oxygen conditions significantly differ for each Candida species.     

电烧伤大鼠血清诱导单核细胞分泌VEGF对单核-内皮细胞黏附作用的影响

目的:观察电烧伤大鼠血清诱导的单核细胞分泌血管内皮生长因子(VEGF)的水平,探讨其对单核细胞与血管内皮细胞黏附作用的影响。方法:制做电烧伤大鼠模型,制备电烧伤大鼠血清,同时制备正常大鼠血清作为对照。采用夹心ELISA法检测正常大鼠血清和电烧伤大鼠血清中VEGF及其可溶性受体s Flt-1含量。按照随机数字表法将人单核细胞株THP-1细胞分为正常血清组和电烧伤血清组,ELISA法检测2组细胞上清液中VEGF和s Flt-1含量。按照随机数字表法将人单核细胞株THP-1细胞分为正常血清组、烧伤血清组、正常血清+阻断剂组和烧伤血清+阻断剂组。取培养3 h、6 h的THP-1细胞,加入单层血管内皮细胞株EA.hy926细胞,行单核-内皮细胞黏附检测。结果:大鼠电烧伤后血清VEGF水平较正常大鼠显著增加,s Flt-1水平较正常大鼠明显减少。电烧伤血清诱导THP-1细胞分泌VEGF,s Flt-1水平随之减少。电烧伤血清可促进单核细胞与内皮细胞的黏附作用,s Flt-1可抑制电烧伤血清诱导的单核细胞与内皮细胞的黏附作用。结论:电烧伤大鼠血清诱导单核细胞分泌VEGF,从而促进单核-内皮细胞黏附。阻断VEGF的生物学效应,可有效抑制单核-内皮细胞的黏附。