人牙周膜成纤维细胞体外培养条件的优化

目的：优化体外人牙周膜成纤维细胞(HPLF)培养条件，以提高原代HPLF培养成功率。方法：采用玻璃及塑料培养瓶、DMEM、RPMI1640培养液以及胎牛血清、小牛血清，分别比较原代牙周膜成纤维细胞游出率。结果：塑料培养瓶细胞游出率以及细胞游出最短时间均优于常用玻璃培养瓶；两种不同血清比较，胎牛血清的组织块贴壁以及细胞游出率均明显优于小牛血清；RPMI1640比DMEM培养液更适合于HPLF培养。结论：商品塑料培养瓶、胎牛血清、RPMIl640是提高体外HPLF培养成功率的关键。     

Circulating histocompatibility antigen (HLA) gene products may help differentiate benign from malignant indeterminate pulmonary lesions

Background

This study explores the potential diagnostic utility of soluble Human Leukocyte Antigen (sHLA) molecules differentially released by lung adenocarcinoma and benign lung lesions.

抑制缺氧诱导因子-1α表达对多发性骨髓瘤RPMI8226细胞裸鼠体内成瘤性的影响

目的 利用RNA干扰(RNAi)技术抑制多发性骨髓瘤(MM)细胞系RPMI8226细胞中缺氧诱导因子-1α(hypoxia-inducible factor,HIF-1α)表达,探讨对RPMI8226细胞在裸鼠体内成瘤性的影响.方法 利用短发夹RNA(shRNA)表达载体pSilencer 2.1-U6 hygro构建RNAi载体,在RPMI8226细胞中抑制HIF-1α的表达.应用RT-PCR技术和Western blot方法 ,鉴定RNAi后HIF-1α的表达.ELISA法测定细胞上清中血管内皮生长因子(VEGF)表达的改变.流式细胞术检测RNAi前后细胞周期的改变.研究缺氧(PO2为1%)条件下,抑制HIF-1α的表达对其靶基因VEGF和葡萄糖转运蛋白(Glut-1)表达的影响.利用异种移植瘤模型观察HIF-1α干扰后对肿瘤生长的影响.结果 RNAi后,HIF-1α的表达在mRNA和蛋白水平均明显降低.常氧条件下,RNAi组(RPMI8226-i1和RPMI8226-i2)和未干扰组(RPMI8226-c和RPMI8226)细胞上清中VEGF浓度无明显差异;缺氧条件下四组细胞上清中VEGF浓度分别为(506.0±53.2)、(494.7±63.1)、(984.4±61.9)和(938.2±62.2)pg/ml.RPMI8226-i1和RPMI8226-i2组较RPMI8226-c和RPMI8226组明显降低(P＜0.05).G0/G1期累积受到抑制,S期细胞比例增加.缺氧条件下VEGF和Glut-1等靶基因的表达明显下调(P＜0.05).裸鼠皮下成瘤性实验表明,抑制HIF-1α的表达,能够明显抑制肿瘤的生长.结论 RNAi抑制RPMI8226细胞HIF-1α表达,可能通过抑制肿瘤的血管新生和糖代谢过程来发挥肿瘤抑制作用.     

The effect of pharmacological PI3Kγ inhibitor on eotaxin-induced human eosinophil functions

BackgroundAsthma is characterized by chronic inflammation caused by activation of immune cells including Th2 lymphocytes and eosinophils. Phosphoinositide 3-kinase (PI3K) γ deficient asthmatic mice did not develop lung eosinophilia, although the detailed mechanisms are not well known. A CC chemokine eotaxin (CCL11) plays a prominent role in developing eosinophilic inflammation through CCR3. In this study, we tested the roles of PI3Kγ in eotaxin-induced eosinophil functions using a pharmacological inhibitor.MethodHuman peripheral blood eosinophils were isolated by CD16-negative selection method. The effect of AS605240, synthetic PI3Kγ inhibitor on eotaxin-induced adhesion, chemotaxis, and degranulation were studied using intracellular adhesion molecule-1 (ICAM-1)-coated plates, Boyden chamber system, ELISA for eosinophil-derived neurotoxin (EDN) levels in the culture supernatant, respectively. CCR3 expression levels and extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation were assessed by flowcytometry. Involvement of PI3Kγ in spontaneous apoptosis was studied using flowcytometry.ResultsAlthough AS605240 did not affect the eosinophil spontaneous apoptosis, eotaxin-induced chemotaxis, adhesion to ICAM-1 coated plate, and EDN release were inhibited by AS605240. AS605240 also inhibited the eotaxin-induced ERK1/2 phosphorylation without down-regulation of surface CCR3 expression.ConclusionThese results indicate that PI3Kγ inhibitor attenuates eotaxin-induced eosinophil functions by suppressing the downstream signaling of CCR3 without significant cytotoxicity. PI3Kγ plays an important role in the development of eosinophilic inflammation and blockade of PI3Kγ might be a therapeutic strategy for treatment of eosinophil-related diseases including asthma.     

衣霉素对RPMI-8226细胞分化的影响

目的:探讨衣霉素对RPMI-8226细胞分化的机制.方法:用衣霉素处理RPMI-8226细胞.APAAP法测定细胞表面CD49e的表达率;ELISA法测定培养上清液中免疫球蛋白轻链的变化,Western blot法检测XBP-1u蛋白表达水平.结果:衣霉素处理后RPMI-8226细胞表面CD49e表达率增高,细胞培养上...     

米托蒽醌对人骨髓瘤细胞RPMI8226增殖与凋亡的影响

本研究探讨米托蒽醌对人骨髓瘤细胞RPMI8226的增殖抑制作用及其分子机制。将米托蒽醌作用于骨髓瘤细胞,用MTT法检测细胞增殖抑制率,光学显微镜和电子显微镜观察细胞形态及超微结构变化,流式细胞术检测细胞凋亡率和细胞周期分布,半定量RT-PCR检测BCL-2、BAX及caspase-3 mRNA表达量的变化,Western blot检测BCL-2、BAX和caspase-3 mRNA蛋白表达量变化。结果表明,米托蒽醌可抑制RPMI8226细胞增殖,抑制率呈时间（r=0.924）和浓度（r=0.947）依赖性增强。米托蒽醌作用24 h后在光学显微镜下可见米托蒽醌组细胞数量减少,排列紊乱,细胞形态变得不规则,可见凋亡细胞;在48 h后电子显微镜下可见细胞典型凋亡改变,凋亡小体形成。流式细胞术检测结果显示,在米托蒽醌组RPMI8226细胞凋亡率明显增高（P〈0.05）;米托蒽醌作用48 h后,在低浓度1.0μg/ml组RPMI8226细胞阻滞于G2/M期（P〈0.05）,在高浓度2.0μg/ml组RPM I8226细胞阻滞于S期（P〈0.05）;米托蒽醌作用48 h,BCL-2 mRNA与BCL-2蛋白表达量减少（P〈0.05）,BAX、caspase-3mRNA及蛋白表达量增加（P〈0.05）。结论：米托蒽醌可诱导RPM I8226细胞凋亡,此细胞凋亡可能与细胞周期阻滞,激活细胞内、外源性凋亡的通路有关。     

人芽囊原虫在不同培养基中生长状况的观察

目的筛选培养人芽囊原虫的最适培养基。方法将同一株人芽囊原虫阳性粪便标本以2×105细胞/管接种至RPMI1640、199和LES培养基中,加入20%小牛血清及青、链霉素,pH值为7.5,放置厌氧罐中于37℃恒温培养,每24h计数,每6d转种1次。观察人芽囊原虫在3种培养基中的存活时间、虫体密度和虫体形态。结果人芽囊原虫在RPMI1640培养基中存活时间最长、虫体密度最高,虫体以空泡型多见;在LES培养基中存活时间最短、虫体密度最低,但虫体形态清晰、规则;在199培养基中存活时间和虫体密度均介于前两者之间。结论RPMI1640培养基适宜人芽囊原虫的生长繁殖,为人芽囊原虫体外培养的首选培养基;LES培养基中虫体形态清晰、规则,可用于人芽囊原虫的形态学研究;199培养基也可用于人芽囊原虫的体外培养,但不作为首选。     

The carpus in the conflict between stability and mobility

Summary The stability of the carpus is determined by a precise interaction of the osseous and ligamentous elements. The main load of the carpus responsible for the adaptation of concerned tissues is a longitudinal compression caused by the force of the muscles of the forearm with their insertions to the metacarpus and fingers. As a consequence, the proximal row is distracted due to the wedge-shaped form of the hamatum together with the capitatum. It is the function of the mainly oblique orientated ligaments to take these transverse tensional forces, supported by the circular arrangement of the retinacula. Here, the ligaments are described in respect of their precise course into three groups. The distribution of material which is needed for bearing the enormous static and dynamic forces is minimised by the integration in a system of oblique fibre bundles which guaranties that all elements, osseous as well as ligamentous, are involved in the force distribution. This is the prerequisition for the minimalisation of osseous material. On the other hand, damage of only one element can cause severe consequences to the stability of the whole system.

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Der Carpus im Konflikt zwischen Stabilit?t und Mobilit?t

Zusammenfassung Die Stabilit?t des Carpus wird vom exakten Zusammenspiel seiner kn?chernen und ligament?ren Elemente bestimmt. Als haupts?chliche und damit für die Gewebeanpassung verantwortliche Beanspruchung ist eine longitudinale Kompression anzusehen, wie sie durch den Zug der Unterarmmuskeln bei nahezu jeder st?rkeren Aktivit?t entsteht. Durch die keilf?rmige Anordnung der Knochen der distalen Reihe wird die proximale Reihe einer Querdehnung ausgesetzt, die durch schr?ge radio- und ulnokarpale B?nder aufgenommen wird. Auch die Retinacula sind als zirkul?re Sicherung aufzufassen. Der Bandapparat wird hier aufgrund des Verlaufes der einzelnen Faserbündel in 3 Gruppen eingeteilt, wobei besonders auf die inteross?ren B?nder Bezug genommen wird. Der zur Aufnahme der betr?chtlichen statischen und dynamischen Kr?fte notwendige Materialaufwand wird dadurch minimiert, da?über die schr?g zu den Bewegungsebenen eingestellten Faserbündel des komplexen Bandapparats der gesamte Carpus jeweils in den Kraftflu? einbezogen wird. Dies ist die Voraussetzung der weitgehenden Beweglichkeit der Handwurzel. Die strenge funktionelle Interaktion des differenzierten Bandapparats mit den kn?chernen Elementen des Carpus bedingt allerdings, da? schon die Sch?digung eines einzigen Elements schwerwiegende Folgen für die Stabilit?t des Gesamtkomplexes hat.

     

蛇床子素诱导人多发性骨髓瘤 RPMI －8226细胞凋亡及其机制研究

目的：研究蛇床子素（Osthole）的抗骨髓瘤作用,并探讨其抗骨髓瘤的作用机制。方法：处于对数增殖期人多发性骨髓瘤细胞 RPMI －8226随机分为空白对照组及10、20、40、80、120、160和200μmol／L Osthole组,用 MTT 法检测细胞增殖率,采用 Western blot 法检测凋亡相关蛋白表达量。结果：细胞培养24、48和72h后,不同浓度 Osthole 组细胞增殖数显著低于空白对照组,且作用呈剂量依赖性。不同浓度 Osthole 组细胞凋亡相关蛋白 PARP 被切割明显高于对照组,p53蛋白的表达量明显高于对照组,抗凋亡蛋白 Mcl －1表达明显下降。结论：Osthole 能明显抑制人多发性骨髓瘤细胞 RPMI －8226细胞增殖,分子机制可能是诱导其细胞凋亡,其诱导凋亡的机制有待于进一步深入研究。     

Trypanosoma brucei brucei: Patterns of glycolysis at 37°C in vitro

In the absence of mammalian cells, freshly isolated monomorphic bloodstream forms of Trypanosoma brucei brucei maintain a constant and high level of aerobic glycolysis in vitro for at least 4 h at 37°C when suspended in RPMI medium 1640 containing 20% heat-inactivated and dialyzed fetal calf serum and 25 mM Hepes at an initial pH of 8. In the absence of nutrients other than glucose, salts and protein, some cell death and a decrease in the rate of glycolysis are observed. In the absence of protein, extensive cell death and a decrease in the rate of glycolysis are seen. These observations may be useful in the design of short-term in vitro metabolic studies with T. b. brucei.