一种快速简便检测TGF—β生物活性的细胞光度比色法

转化生长因子是一类影响细胞生长和表型的生物活性多肽。它在不同靶细胞和微环境下,不仅具有刺激或抑制细胞生长、诱导细胞分化和其它多种生物学效应,还能引起一些正常细胞失去接触抑制,转化成为锚着不依赖生长。通常利用这种特性使长在软琼脂上的成纤维细胞形成细胞集落来检测TGF-β活性。此法操作繁琐,周期长至一周以上。另一种检测TGF-β活性方法是利用TGF-β能抑制貂肺     

GSK-3β as a driving force in ovarian cancer

Ovarian cancer is one of the most lethal malignancies in women. Identification of new therapeutic targets would provide opportunities for developing potentially more effective treatment regimes. In the July issue of Cell Research, Cao et al. reports that glycogen synthase kinase-3β （GSK-3β） plays an important role in positively regulating the proliferation of human ovarian cancer cells, and thus it may represent such a target [ 1 ]. GSK-3β is a serine/threonine kinase that is known to be involved in regulation of β-catenin signaling, where it participates in the formation of a multi-component destruction complex that promotes the phosphorylation and subsequent degradation of β-catenin. Given that overactive β-catenin signaling is involved in many forms of human cancer, this classic mode of GSK-3β action should qualify it as a ＂tumor suppressor＂. Intriguingly, however, two recent studies have implicated that GSK-3β may actually play a pro-tumor role in pancreatic and colorectal cancers [2, 3]. Since ovarian tumors often exhibit increased expression of GSK-3β, these recent findings prompted Cao et al. to examine the potential role of GSK-3β in ovarian cancer cells.     

Cu（Ⅱ） Potentiation of Alzheimer Aβ1-40 Cytotoxicity and Transition on its Secondary Structure

Mounting evidence has shown that dyshomeostasis of the redox-active biometals such as Cuand Fe can lead to oxidative stress,which plays a key role in the neuropathology of Alzheimer's disease(AD).Here we demonstrate that with the formation of Cu(Ⅱ)·Aβ1-40 complexes,copper markedly potentiatesthe neurotoxicity exhibited by β-amyloid peptide (Aβ).A greater amount of hydrogen peroxide was releasedwhen Cu(Ⅱ)·Aβ1-40 complexes was added to the xanthine oxidase/xanthine system detected by potassiumiodide spectrophotometry.Copper bound to Aβ1-40 was observed by electron paramagnetic resonance(EPR) spectroscopy.Circular dichroism (CD) studies indicated that copper chelation could cause a structuraltransition of Aβ.The addition of copper to Aβ introduced an increase on β-sheet as well as α-helix,whichmay be responsible for the aggregation of Aβ.We hypothesized that Aβ aggregation induced by copper maybe responsible for local injury in AD.The interaction between Cu~(2 ) and Aβ also provides a possible mechanismfor the enrichment of metal ions in amyloid plaques in the AD brain.     

抗GPC3单链抗体的制备及其在肝细胞癌靶向检测中的应用

磷脂酰肌醇蛋白聚糖3 (Glypican-3,GPC3) 是Glypican家族的关键成员,它在肝细胞癌 (HCC) 的发展、血管生成和转移中起着重要作用。大多数HCC过表达GPC3,而在正常的成人肝脏和良性肝脏病变中几乎检测不到GPC3,因此其可以作为HCC高度特异性的诊断标志和理想的治疗靶标。文中将前期筛选得到的靶向GPC3的单克隆抗体1B11轻重链基因通过柔性肽 (Linker) 连接,经原核表达纯化,成功获得抗GPC3单链抗体1B11-scFv。通过蛋白印迹法鉴定验证了单链抗体的表达和装配正确;流式细胞术检测其与GPC3阳性细胞株具有与全长抗体相当的结合活性;将单链抗体与近红外荧光探针Cy5.5-NHS偶联,发现其在荷瘤裸鼠体内可精准靶向肿瘤部位,提示其具有实现探针引导下的多方位肝癌精准手术导航的潜力。     

Caspase-1 activation and mature interleukin-1β release are uncoupled events in monocytes

AIM:To investigate whether caspase-1 activation/intracellular processing of pro-interleukin-1β(pro-IL-1β) and extracellular release of mature IL-1β from activated monocytes are separable events.METHODS:All experiments were performed on fresh or overnight cultured human peripheral blood monocytes(PBMCs) that were isolated from healthy donors.PBMCs were activated by lipopolysaccharide(LPS) stimulation before being treated with Adenosine triphosphate(ATP,1 mmol/L),human α-defensin-5(HD-5,50 μg/mL),and/or nigericin(Nig,30 μmol/L).For each experiment,the culture supernatants were collected separately from the cells.Cell lysates and supernatants were both subject to immunoprecipitation with anti-IL1β antibodies followed by western blot analysis with anti-caspase-1 and anti-IL-1β antibodies.RESULTS:We found that pro-IL-1β was processed to mature IL-1β in LPS-activated fresh and overnight cultured human monocytes in response to ATP stimulation.In the presence of HD-5,this release of IL-1β,but not the processing of pro-IL-1β to IL-1β,was completely inhibited.Similarly,in the presence of HD-5,the release of IL-1β,but not the processing of IL-1β,was significantly inhibited from LPS-activated monocytes stimulated with Nig.Finally,we treated LPS-activated monocytes with ATP and Nig and collected the supernatants.We found that both ATP and Nig stimulation could activate and release cleaved caspase-1 from the monocytes.Interestingly,and contrary to IL-1β processing and release,caspase-1 cleavage and release was not blocked by HD-5.All images are representative of three independent experiments.CONCLUSION:These data suggest that caspase-1 activation/processing of pro-IL-1β by caspase-1 and the release of mature IL-1β from human monocytes are distinct and separable events.     

金华猪3个繁殖性状主基因的分布及其效应的研究

采用PCR RFLP技术分析了金华猪 3个品系的雌激素受体基因 (ESR)、促卵泡素 β亚基基因 (FSHβ)、催乳素受体基因 (PRLR)在金华猪 3个品系中的分布情况 ,并采用SPSS程序分析了ESR、FSHβ和PRLR 3个基因对金华猪Ⅰ系母猪繁殖性状的影响。结果表明 :ESR基因的 3个基因型AA、AB、BB在金华猪Ⅰ系、Ⅱ系和Ⅲ系中的频率分别为 0 0 2 37(AA)、0 36 0 9(AB)、0 6 15 4 (BB) ,0 (AA)、0 5 333(AB)、0 4 6 6 7(BB)和 0 2 90 9(AA)、0 6 (AB)、0 10 91(BB) ;PRLR基因的 3个基因型AA、AB、BB在金华猪Ⅰ系、Ⅱ系和Ⅲ系的频率分别为 0 2 5 6 0 (AA)、0 392 9(AB)、0 35 12 (BB) ,0 2 (AA)、0 2 6 6 7(AB)、0 5 333(BB)和 0 12 73(AA)、0 4 90 9(AB)、0 3818(BB) ,而FSHβ基因仅在金华猪Ⅰ系中存在多态性 ,且有利基因B的频率很低 ,仅为 0 0 12 0。ESR基因对金华母猪经产胎次产活仔数、所有胎次的产活仔数和总产仔数影响显著 (P <0 0 5 ) ,对金华母猪经产胎次的总产仔数的影响接近显著 (P =0 0 6 1)。B基因对产活仔数的加性效应为经产胎次 0 70头 胎 ,所有胎次 0 6 6头 胎 ;对总产仔数的加性效应为经产胎次 0 90头 胎 ,所有胎次 0 84头 胎。PRLR基因对金华母猪的各项繁殖性状都没有显著影响 (P     

TGF-β/Smads在肺癌中的表达研究

转化生长因子TGF-β／Smads是细胞增殖的抑制因子。它可以促进细胞外基质的增生和原发性肿瘤的转移。选择20例肺癌组织标本及8种不同转移能力的肺癌细胞系,通过荧光免疫组织化学染色法进行RⅡ和Smsds蛋白定位及表达分析。初步探讨了TGF-β／Smads信号传导通路与肺癌发生和转移的关系。