• 【二硝基苯胺与apicomplexan和运动质体 α-微管蛋白的结合和相互作用。】 复制标题 收藏 收藏
    DOI:10.1021/jm060472+ 复制DOI
    作者列表:Mitra A,Sept D
    BACKGROUND & AIMS: :Despite years of use as commercial herbicides, it is still unclear how dinitroanilines interact with tubulin, how they cause microtubule disassembly, and why they are selectively active against plant and protozoan tubulin. In this work, through a series of computational studies, a common binding site of oryzalin, trifluralin, and GB-II-5 on apicomplexan and kinetoplastid alpha-tubulin is proposed. Furthermore, to investigate how dinitroanilines affect tubulin dynamics, molecular dynamics simulations of Leishmania alpha-tubulin with and without a bound dinitroaniline are performed. The results obtained provide insight into the molecular mechanism by which these compounds interact with tubulin and function to prevent microtubule assembly. Finally, to aid in the design of effective parasitic microtubule inhibitors, several novel dinitroaniline analogues are evaluated. The location of the binding site and the relative binding affinities of the dinitroanilines all agree well with experimental data.
    背景与目标: : 尽管已作为商业除草剂使用多年,但仍不清楚二硝基苯胺如何与微管蛋白相互作用,它们如何引起微管分解,以及为什么它们对植物和原生动物的微管蛋白具有选择性活性。在这项工作中,通过一系列计算研究,提出了米唑林,三氟拉林和GB-II-5在apicomplexan和动质体 α-微管蛋白上的共同结合位点。此外,为了研究二硝基苯胺如何影响微管蛋白动力学,对有和没有结合的二硝基苯胺的利什曼原虫 α-微管蛋白进行了分子动力学模拟。获得的结果提供了对这些化合物与微管蛋白相互作用并阻止微管组装的分子机制的深入了解。最后,为了帮助设计有效的寄生微管抑制剂,评估了几种新型的二硝基苯胺类似物。结合位点的位置和二硝基苯胺的相对结合亲和力都与实验数据吻合良好。
  • 【干扰素 α2b与达卡巴嗪,卡莫司汀,顺铂和他莫昔芬在转移性黑色素瘤中的II期研究。】 复制标题 收藏 收藏
    DOI:10.1097/00008390-199704000-00009 复制DOI
    作者列表:Schultz MZ,Buzaid AC,Poo WJ
    BACKGROUND & AIMS: A phase II trial was conducted to determine the efficacy and toxicity of the addition of interferon-alpha 2b (IFN-alpha) to the chemotherapy combination of dacarbazine (DTIC), carmustine (BCNU), cisplatin and tamoxifen (DBCT), in patients with stage III or IV melanoma. Treatment consisted of DTIC 220 mg/m2 and cisplatin 25 mg/m2 intravenously on days 1-3, BCNU 100 mg/m2 IV on day 1 only, tamoxifen 20 mg orally twice daily and IFN-alpha 5 x 10(6) units/m2 subcutaneously on days 1-5. Cycles were repeated every 4 weeks. All patients received a loading dose of tamoxifen 100 mg orally twice daily for 5 days before the first course of therapy. Of the 24 patients treated, three (13%) achieved a complete response (CR) and six (25%) a partial response (PR), for an overall response rate of 38% (95% confidence interval, 17-58%). Two patients, one who achieved a clinical CR and one a PR, had pathologically confirmed complete responses. Severe myelosuppression occurred in 47% of cycles and constitutional symptoms were common. Overall, the addition of IFN-alpha to the DBCT regimen did not appear to enhance the response rate and may have increased toxicity.

    背景与目标: 进行了II期试验,以确定在达卡巴嗪 (DTIC),卡莫司汀 (BCNU),顺铂和他莫昔芬 (DBCT) 的化疗组合中添加干扰素 α2b (IFN-α) 的疗效和毒性。III或IV期黑色素瘤患者。治疗包括在第1-3天静脉注射DTIC 220 mg/m2和顺铂25 mg/m2,仅在第1天静脉注射BCNU 100 mg/m2,他莫昔芬20 mg每天口服两次,在第1-5天皮下注射IFN-α5 × 10(6) 单位/m2。每4周重复一次周期。所有患者接受负荷剂量的他莫昔芬100 mg口服,每日两次,治疗前5天。在接受治疗的24例患者中,3例 (13% 例) 达到完全缓解 (CR),6例 (25% 例) 达到部分缓解 (PR),总缓解率为38% (95% 置信区间17-58%)。两名患者,其中一名获得了临床CR,一名获得了PR,经病理证实完全缓解。严重的骨髓抑制发生在47% 的周期和全身症状是常见的。总体而言,在DBCT方案中添加IFN-α 似乎并未增强反应率,并且可能具有增加的毒性。
  • 【阿尔法节律的阴暗面: 完全黑暗期间诱导阿尔法调制的fMRI证据。】 复制标题 收藏 收藏
    DOI:10.1111/ejn.12083 复制DOI
    作者列表:Ben-Simon E,Podlipsky I,Okon-Singer H,Gruberger M,Cvetkovic D,Intrator N,Hendler T
    BACKGROUND & AIMS: :The unique role of the EEG alpha rhythm in different states of cortical activity is still debated. The main theories regarding alpha function posit either sensory processing or attention allocation as the main processes governing its modulation. Closing and opening eyes, a well-known manipulation of the alpha rhythm, could be regarded as attention allocation from inward to outward focus though during light is also accompanied by visual change. To disentangle the effects of attention allocation and sensory visual input on alpha modulation, 14 healthy subjects were asked to open and close their eyes during conditions of light and of complete darkness while simultaneous recordings of EEG and fMRI were acquired. Thus, during complete darkness the eyes-open condition is not related to visual input but only to attention allocation, allowing direct examination of its role in alpha modulation. A data-driven ridge regression classifier was applied to the EEG data in order to ascertain the contribution of the alpha rhythm to eyes-open/eyes-closed inference in both lighting conditions. Classifier results revealed significant alpha contribution during both light and dark conditions, suggesting that alpha rhythm modulation is closely linked to the change in the direction of attention regardless of the presence of visual sensory input. Furthermore, fMRI activation maps derived from an alpha modulation time-course during the complete darkness condition exhibited a right frontal cortical network associated with attention allocation. These findings support the importance of top-down processes such as attention allocation to alpha rhythm modulation, possibly as a prerequisite to its known bottom-up processing of sensory input.
    背景与目标: : EEG α 节律在不同皮质活动状态中的独特作用仍存在争议。有关alpha功能的主要理论将感觉处理或注意力分配视为控制其调制的主要过程。关闭和睁开眼睛是一种众所周知的对阿尔法节奏的操纵,尽管在光照过程中也伴随着视觉变化,但可以将注意力从内向向外向分配。为了弄清注意力分配和感觉视觉输入对 α 调制的影响,要求14名健康受试者在光线和完全黑暗的情况下睁开和闭上眼睛,同时同时记录脑电图和功能磁共振成像。因此,在完全黑暗的情况下,睁眼状态与视觉输入无关,而仅与注意力分配有关,从而可以直接检查其在alpha调制中的作用。将数据驱动的山脊回归分类器应用于EEG数据,以确定在两种光照条件下 α 节律对睁眼/闭眼推理的贡献。分类器结果显示,无论在明暗条件下,alpha都有显着贡献,这表明无论是否存在视觉感觉输入,alpha节律调制都与注意方向的变化密切相关。此外,在完全黑暗的条件下,从alpha调制时间过程中得出的fMRI激活图显示出与注意力分配相关的右额叶皮层网络。这些发现支持自上而下的过程的重要性,例如将注意力分配给alpha节奏调制,这可能是其已知的感觉输入自下而上处理的先决条件。
  • 【通过蛋白质组学方法观察到STZ诱导的糖尿病大鼠肾线粒体中alpha-2u球蛋白的下调。】 复制标题 收藏 收藏
    DOI:10.1016/j.ando.2012.09.007 复制DOI
    作者列表:Sun SH,Liu SQ,Cai CP,Cai R,Chen L,Zhang QB
    BACKGROUND & AIMS: AIM:To identify the changes of mitochondrial protein expression in diabetic renal parenchyma and to characterize their molecular functions and biological processes in diabetes. METHODS:Mitochondrial proteins extracted from renal parenchyma mitochondria of streptozotocin-induced diabetic rats and normal rats were separated by two-dimensional polyacrylamide gel electrophoresis and identified by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry. RESULTS:Eleven proteins from 533 visualized protein spots displayed significant different expressions in mitochondria of diabetic kidneys compared with those in normal ones. Among these altered proteins, two proteins with the most obvious changes in protein expression were identified as alpha-2u globulin (mature protein, named A2) and its proteolytically modified form (named A2-fragment) respectively. These proteins were found in mitochondria of male rat renal parenchyma and were proved to be down-regulated in diabetic rats simultaneously. CONCLUSION:Our results suggest that down-regulation of alpha-2u globulin may be associated with an abnormal β-oxidation of long-chain fatty acids during diabetes. The decreased expression of A2-fragment in renal mitochondria of diabetic nephropathy may reduce fatty acid β-oxidation, which leads to a diminished energy supply from mitochondria to kidney tissue and the deposition of a large number of fatty acids in the kidney, ultimately causing and aggravating kidney damage. In conclusion, these findings may be helpful for understanding the molecular mechanism of diabetic nephropathy.
    背景与目标:
  • 【血清甲胎蛋白的半衰期: 肝细胞癌肝切除术后复发和生存的早期预后指标。】 复制标题 收藏 收藏
    DOI:10.1097/SLA.0b013e318273be70 复制DOI
    作者列表:Shim JH,Han S,Lee YJ,Lee SG,Kim KM,Lim YS,Chung YH,Lee YS,Lee HC
    BACKGROUND & AIMS: OBJECTIVE:To explore the prognostic value of the postsurgical half-life (HL) of serum alpha-fetoprotein (AFP). BACKGROUND:There is still a paucity of early surrogate indicators of clinical endpoints after liver resection of hepatocellular carcinoma (HCC). METHODS:The analysis was based on cohorts of 225 (exploration set) and 117 (validation set) treatment-naïve HCC patients undergoing curative liver resection. We defined 3 categories of AFP HL: early complete resolution of AFP, normal HL, and prolonged HL if the HL exceeded 7 days. Overall, probabilities of recurrence and survival were estimated and compared across the AFP HL categories. RESULTS:In the exploration cohort, 48 patients (21.3%) achieved early AFP complete resolution, 116 (51.6%) had normal HL, and 61 (27.1%) had prolonged HL. Long AFP HL was significantly associated with early postoperative recurrence (P < 0.001), as was microvascular invasion. Early recurrence within 2 years of resection was observed in 59% of the patients with prolonged AFP HL compared with only 29.3% of those with normal AFP HL (P < 0.001). A log-rank test followed by multivariate Cox analysis identified an independent function of prolonged AFP HL in predicting shorter recurrence-free survival and overall survival time after HCC resection (hazard ratios, 2.81 and 3.58; P < 0.001). When AFP HL analysis was applied to the validation cohort, the association between prolonged AFP HL and survival endpoints (hazard ratio, 11.63 and 16.39; P < 0.001) was confirmed.
    背景与目标:
  • 【路易体患者的阿尔茨海默氏病和痴呆症的脑脊液可增强体外 α-突触核蛋白原纤维的形成。】 复制标题 收藏 收藏
    DOI:10.1016/j.expneurol.2006.08.012 复制DOI
    作者列表:Ono K,Noguchi-Shinohara M,Yoshita M,Naiki H,Yamada M
    BACKGROUND & AIMS: :Deposition of alpha-synuclein (alphaS) aggregates inside brain cells is a pathological hallmark of several neurodegenerative diseases, including Parkinson's disease (PD), and dementia with Lewy bodies (DLB). Recently, extracellular alphaS was detected in cerebrospinal fluid (CSF) and plasma of humans. We investigated whether CSF influences alphaS aggregation in vitro using fluorescence spectroscopy with thioflavin S and electron microscopy. We found that CSF obtained from Alzheimer's disease (AD) and DLB patients enhanced the alphaS fibril formation compared with tauopathy and non-central nervous system disease. Thus, CSF of AD and DLB characterized by aggregation of Abeta or alphaS might promote falphaS formation.
    背景与目标: : 脑细胞内 α-突触核蛋白 (alpha) 聚集体的沉积是几种神经退行性疾病的病理标志,包括帕金森氏病 (PD) 和路易体痴呆 (DLB)。最近,在人类的脑脊液 (CSF) 和血浆中检测到细胞外alpha。我们使用硫黄素S的荧光光谱法和电子显微镜研究了CSF是否会影响alpha的体外聚集。我们发现,与tauopathy和非中枢神经系统疾病相比,从阿尔茨海默氏病 (AD) 和DLB患者获得的CSF增强了alpha原纤维的形成。因此,以Abeta或alpha聚集为特征的AD和DLB的CSF可能会促进falpha的形成。
  • 【过量表达异源 α-淀粉酶引起的生产和分泌压力导致枯草芽孢杆菌的孢子形成抑制和活动期延长。】 复制标题 收藏 收藏
    DOI:10.1128/AEM.00472-07 复制DOI
    作者列表:Lulko AT,Veening JW,Buist G,Smits WK,Blom EJ,Beekman AC,Bron S,Kuipers OP
    BACKGROUND & AIMS: :Transcriptome analysis was used to investigate the global stress response of the gram-positive bacterium Bacillus subtilis caused by overproduction of the well-secreted AmyQ alpha-amylase from Bacillus amyloliquefaciens. Analyses of the control and overproducing strains were carried out at the end of exponential growth and in stationary phase, when protein secretion from B. subtilis is optimal. Among the genes that showed increased expression were htrA and htrB, which are part of the CssRS regulon, which responds to high-level protein secretion and heat stress. The analysis of the transcriptome profiles of a cssS mutant compared to the wild type, under identical secretion stress conditions, revealed several genes with altered transcription in a CssRS-dependent manner, for example, citM, ylxF, yloA, ykoJ, and several genes of the flgB operon. However, high-affinity CssR binding was observed only for htrA, htrB, and, possibly, citM. In addition, the DNA macroarray approach revealed that several genes of the sporulation pathway are downregulated by AmyQ overexpression and that a group of motility-specific (sigmaD-dependent) transcripts were clearly upregulated. Subsequent flow-cytometric analyses demonstrate that, upon overproduction of AmyQ as well as of a nonsecretable variant of the alpha-amylase, the process of sporulation is severely inhibited. Similar experiments were performed to investigate the expression levels of the hag promoter, a well-established reporter for sigmaD-dependent gene expression. This approach confirmed the observations based on our DNA macroarray analyses and led us to conclude that expression levels of several genes involved in motility are maintained at high levels under all conditions of alpha-amylase overproduction.
    背景与目标: : 转录组分析用于研究由解淀粉芽孢杆菌分泌充分的AmyQ α-淀粉酶过量产生引起的革兰氏阳性细菌枯草芽孢杆菌的整体应激反应。当枯草芽孢杆菌的蛋白质分泌最佳时,在指数生长结束时和固定相进行对照和过量生产菌株的分析。表达增加的基因包括htrA和htrB,它们是CssRS调节子的一部分,它们对高水平的蛋白质分泌和热应激做出反应。在相同的分泌胁迫条件下,与野生型相比,对cssS突变体的转录组谱进行分析,揭示了几个以CssRS依赖性方式改变转录的基因,例如citM,ylxF,yloA,ykoJ和flgB操纵子的几个基因。然而,仅在htrA,htrB和可能的citM中观察到高亲和力的CssR结合。此外,DNA macroarray方法揭示了孢子形成途径的几个基因被AmyQ过表达下调,并且一组运动性特异性 (sigmaD依赖性) 转录本被明显上调。随后的流式细胞仪分析表明,在过量生产AmyQ以及不可分泌的 α-淀粉酶变体后,孢子形成过程受到严重抑制。进行了类似的实验以研究hag启动子的表达水平,hag启动子是sigmaD依赖性基因表达的公认报告基因。这种方法证实了基于我们的DNA宏观阵列分析的观察结果,并使我们得出结论,在所有 α-淀粉酶过度生产的条件下,与运动有关的几个基因的表达水平都保持在高水平。
  • 【包括HDL在内的氧化脂蛋白及其脂质过氧化产物抑制THP-1人巨噬细胞分泌TNF-α。】 复制标题 收藏 收藏
    DOI:10.1016/s0891-5849(97)00061-0 复制DOI
    作者列表:Girona J,La Ville AE,Heras M,Olivé S,Masana L
    BACKGROUND & AIMS: It has been established that oxidized LDL (ox-LDL) modifies cytokine secretion by macrophages, for example, by reducing tumor necrosis factor alpha (TNF-(alpha) m-RNA. However, little is known about the effects of oxidized high density lipoprotein (ox-HDL). This study reports the effects of ox-HDL subfractions 2 and 3 (ox-HDL2, ox-HDL3) compared with that of ox-LDL and some products of oxidation (hydroperoxides and aldehydes) on the secretion of TNF-alpha from THP-1 human monocytes derived macrophages in vitro. HDL2, HDL3 and LDL were oxidized with 10 microM Cu++ for 12 h and/or 24 h. Native and oxidized HDL and LDL were incubated for 24 h with macrophages with or without LPS (10 ng/ml) after which TNF-alpha secretion was measured in the culture medium. Lipid hydroperoxides and apolar aldehydes were also incubated with the cells for 2 h following which the medium was replaced and TNF-alpha secretion measured after a further 22 h of incubation. An inhibition of TNF-alpha by ox-HDL2 (p < .05), ox-HDL3 (p < .05) and ox-LDL (p < .05) from THP-1 macrophages was observed in the presence and absence of LPS. This inhibition remained the same after incubation with ox-HDL 12 h and 24 h. Hydroperoxides of linoleic acid did not modify TNF-alpha secretion by cells while five out of eight aldehydes analyzed (2,4-heptadienal, hexanal, 2-nonenal, 2-octenal, 2,4-decadienal) inhibited TNF-alpha secretion (p < .05). These findings demonstrate that ox-HDL, and some of its lipid peroxidation products, plays a role in the modulation of the inflammatory response by macrophages as previously observed for ox-LDL.

    背景与目标: 已经确定氧化的LDL (ox-LDL) 修饰巨噬细胞的细胞因子分泌,例如,通过减少肿瘤坏死因子 α (TNF-(α) m-RNA。然而,对氧化型高密度脂蛋白 (ox-HDL) 的影响知之甚少。这项研究报告了ox-HDL亚组分2和3 (ox-HDL2,ox-HDL3) 与ox-LDL和某些氧化产物 (氢过氧化物和醛) 对体外THP-1人单核细胞来源的巨噬细胞分泌TNF-α 的影响相比。HDL2,HDL3和LDL用10 microM Cu ++ 氧化12小时和/或24小时。将天然和氧化的HDL和LDL与有或没有LPS (10 ng/ml) 的巨噬细胞孵育24小时,然后在培养基中测量TNF-α 分泌。脂质氢过氧化物和极性醛还与细胞一起孵育2小时,然后更换培养基,再孵育22小时后测量TNF-α 分泌。ox-HDL2对TNF-α 的抑制作用 (p <.05),在存在和不存在LPS的情况下观察到来自THP-1巨噬细胞的ox-HDL3 (p < .05) 和ox-LDL (p <.05)。与ox-HDL孵育12小时和24小时后,这种抑制作用保持不变。亚油酸的氢过氧化物不会改变细胞分泌TNF-α,而五在分析的八种醛中 (2,4-庚二烯醛,己醛,2-壬烯醛,2-辛烯醛,2,4-癸烯醛) 抑制TNF-α 分泌 (p <.05)。这些发现表明ox-HDL及其某些脂质过氧化产物,如先前对ox-LDL所观察到的,在巨噬细胞调节炎症反应中起作用。
  • 【从杆状病毒感染的草地贪夜蛾 (IPLB-SF-21AE) 细胞的高尔基样膜中纯化和鉴定 α-甘露糖苷酶II。】 复制标题 收藏 收藏
    DOI:10.1042/bj3240951 复制DOI
    作者列表:Ren J,Castellino FJ,Bretthauer RK
    BACKGROUND & AIMS: An alpha-mannosidase II-like activity was identified in baculovirus-infected Spodoptera frugiperda (IPLB-SF21-AE) cells. The enzyme responsible was purified from Golgi-type membranes to apparent homogeneity by using a combination of steps including DEAE-cellulose, hydroxyapatite, concanavalin A-Sepharose and gel filtration chromatography. The molecular mass of this purified protein was approx. 120 kDa by SDS/PAGE under reducing conditions and approx. 240 kDa under non-reducing conditions, indicating that the enzyme is a disulphide-linked dimer. Substrates demonstrated to undergo hydrolysis with this enzyme were GlcNAc-Man5-GlcNAc-GlcNAc (non-reduced and reduced) and p-nitrophenyl alpha-d-mannopyranoside. The oligosaccharide substrate was converted into GlcNAc-Man3-GlcNAc-GlcNAc through an intermediate GlcNAc-Man4-GlcNAc-GlcNAc. Treatment of the isolated intermediate oligosaccharide with endoglycosidase H resulted in its conversion into GlcNAc-Man4-GlcNAc. This indicated that it contained the alpha-1,3-linked mannose residue on the alpha-1,6-linked mannose arm and showed that the alpha-1,6-linked mannose residue on the alpha-1,6-linked mannose arm had been preferentially hydrolysed by the mannosidase. The oligosaccharide lacking the beta-1,2-linked GlcNAc residue on the alpha-1,3-linked mannose arm (Man5-GlcNAc-GlcNAc) was not hydrolysed in the presence of the enzyme. Metal ions were not required for enzymic activity on any of the substrates, but Cu2+ was strongly inhibitory. The activity of the enzyme was inhibited at low concentrations of swainsonine, but much higher concentrations of 1-deoxymannojirimycin were required to achieve inhibition. All of these properties are characteristic of mannosidase II enzymes from other eukaryotic tissues. The presence of mannosidase II in lepidopteran insect cells would allow entry of N-linked glycoproteins into the complex processing reaction pathway or into the terminal Man3-GlcNAc-GlcNAc pathway.

    背景与目标: 在杆状病毒感染的草地贪夜蛾 (IPLB-SF21-AE) 细胞中鉴定出 α-甘露糖苷酶II样活性。通过使用DEAE-纤维素,羟基磷灰石,伴刀豆球蛋白a-琼脂糖和凝胶过滤色谱等步骤的组合,将负责的酶从高尔基体型膜中纯化至表观均一。该纯化蛋白的分子量约为。在还原条件下通过SDS/PAGE 120 kDa。在非还原条件下240 kDa,表明该酶是二硫化物连接的二聚体。GlcNAc-Man5-GlcNAc-GlcNAc (非还原和还原) 和对硝基苯基 α-d-甘露吡喃糖苷,证明用该酶水解的底物。通过中间GlcNAc-Man4-GlcNAc-GlcNAc将寡糖底物转化为GlcNAc-Man3-GlcNAc-GlcNAc。用内切糖苷酶H处理分离的中间寡糖导致其转化为GlcNAc-Man4-GlcNAc。这表明它在 α-1,6-连接的甘露糖臂上包含 α-1,3-连接的甘露糖残基,并表明 α-1,6-连接的甘露糖上的 α-1,6-连接的甘露糖残基已优先被甘露糖苷酶水解。在存在该酶的情况下,没有在 α-1,3-连接的甘露糖臂 (Man5-GlcNAc-GlcNAc) 上缺少 β-1,2-连接的GlcNAc残基的寡糖被水解。在任何底物上的酶活性都不需要金属离子,但是Cu2具有强烈的抑制作用。在低浓度的swainsonine下,酶的活性受到抑制,但是需要更高浓度的1-脱氧甘露霉素才能达到抑制作用。所有这些特性都是来自其他真核组织的甘露糖苷酶II酶的特征。鳞翅目昆虫细胞中甘露糖苷酶II的存在将允许N-连接的糖蛋白进入复杂的加工反应途径或末端Man3-GlcNAc-GlcNAc途径。
  • 【通过竞争性聚合酶链反应监测干扰素 α 治疗慢性粒细胞性白血病 (CML) 患者的效率。】 复制标题 收藏 收藏
    DOI: 复制DOI
    作者列表:Hochhaus A,Lin F,Reiter A,Skladny H,Hehlmann R,Goldman JM,Cross NC
    BACKGROUND & AIMS: Interferon alpha (IFN-alpha) induces cytogenetic responses of variable degree in patients with CML. We sought to establish the relationship between BCR-ABL transcript numbers measured by competitive two-step reverse transcription polymerase chain reaction (RT-PCR) and cytogenetic status in CML patients treated with IFN-alpha. All 398 samples from 163 patients investigated by RT-PCR were positive for BCR-ABL transcripts. In order to standardize results for variability in RNA and cDNA quality, we quantified total ABL transcripts in each sample as internal control. The BCR-ABL/ABL ratios correlated with the cytogenetic results. Quantitative nested PCR allowed the detection of residual BCR-ABL transcripts in all complete cytogenetic responders on IFN-alpha. We conclude that competitive PCR with internal controls is a reliable method for monitoring patients on IFN-alpha and reduces the need for repeated marrow investigations.

    背景与目标: 干扰素 α (IFN-α) 诱导CML患者不同程度的细胞遗传学反应。我们试图建立通过竞争性两步逆转录聚合酶链反应 (rt-pcr) 测量的bcr-abl转录数与接受IFN-α 治疗的CML患者的细胞遗传学状态之间的关系。通过rt-pcr调查的来自163名患者的所有398样品的bcr-abl转录物均为阳性。为了标准化RNA和cDNA质量变异性的结果,我们将每个样品中的总ABL转录本量化为内部对照。Bcr-abl/ABL比值与细胞遗传学结果相关。定量巢式PCR可以检测到IFN-α 上所有完整的细胞遗传学反应中的残留BCR-ABL转录本。我们得出的结论是,带有内部对照的竞争性PCR是一种可靠的方法,用于监测患者的IFN-α,并减少了重复进行骨髓检查的需要。
  • 【在伤口愈合的体外模型中,α6β4整联蛋白的表面重新定位和半桥体的组装。】 复制标题 收藏 收藏
    DOI:10.1083/jcb.115.6.1737 复制DOI
    作者列表:Kurpakus MA,Quaranta V,Jones JC
    BACKGROUND & AIMS: :A transmembrane extracellular matrix receptor of the integrin family, alpha 6 beta 4, is a component of the hemidesmosome, an adhesion complex of importance in epithelial cell-connective tissue attachment (Stepp, M. A., S. Spurr-Michaud, A. Tisdale, J. Elwell, and I. K. Gipson. 1990. Proc. Natl. Acad. Sci. USA. 87:8970-8974; Jones, J. C. R., M. A. Kurpakus, H. M. Cooper, and V. Quaranta. 1991. Cell Regulation. 2:427-438). Cytosolic components of hemidesmosomes include bullous pemphigoid (BP) antigens while extracellular components include a 125-kD component of anchoring filaments (CAF) and collagen type VII-containing anchoring fibrils. We have monitored the incorporation of the alpha 6 beta 4 integrins into forming hemidesmosomes in an in vitro wound-healing explant model. In epithelial cells recently migrated from the edges of unwounded sites over bare connective tissue, alpha 6 beta 4 first appears along the entire cell surface. At this stage, these cells contain little or no cytosolic hemidesmosomal components, at least as detectable by immunofluorescence using BP autoantibodies, whereas they are already positive for laminin and CAF. At a later stage, as cells become positive for cytosolic hemidesmosome components such as BP antigens as well as collagen type VII, alpha 6 beta 4 becomes concentrated along the basal pole of the epithelial cell where it abuts the connective tissue of the explant. Polyclonal antibodies to beta 4 do not interfere with the migration of epithelial cells in the explant. However, they prevent assembly of hemidesmosomal complexes and inhibit expression of collagen type VII in cells that have migrated over wound areas. In addition, they induce disruption of established hemidesmosomes in nonmigrating cells of the unwounded area of the explant. Monoclonal antibodies to alpha 6 have a more dramatic effect, since they completely detach epithelial cells in the unwounded area of the explant. Antibodies to CAF also detach epithelial cells in unwounded areas, apparently by inducing separation between epithelium and connective tissue at the lamina lucida of the basement membrane zone. These results suggest a model whereby polarization of alpha 6 beta 4 to the basal surface of the cells, perhaps induced by a putative anchoring filament-associated ligand, triggers assembly of hemidesmosome plaques.
    背景与目标: : 整联蛋白家族的跨膜细胞外基质受体 α6β4是半染色体的组成部分,半染色体是上皮细胞-结缔组织附着中重要的粘附复合物 (Stepp,m.a.,S. Spurr-Michaud,A. Tisdale,J. Elwell,和I. K. Gipson。1990. Proc. Natl. Acad. Sci.美国。87:8970-8974; Jones,j.c.R.,M.Kurpakus,H. M. Cooper和V. Quaranta。1991.细胞调控。2:427-438)。半桥粒的胞质组分包括大疱性类天疱疮 (BP) 抗原,而细胞外组分包括锚定细丝 (CAF) 的125-kD组分和含VII型胶原的锚定原纤维。我们已经在体外伤口愈合外植体模型中监测了 α6β4整合素在形成半桥体中的掺入。在最近从裸露的结缔组织上未受伤部位的边缘迁移的上皮细胞中,α6β4首先出现在整个细胞表面。在这个阶段,这些细胞含有很少或没有胞质半染色体成分,至少可以通过使用BP自身抗体的免疫荧光检测到,而它们已经对层粘连蛋白和CAF呈阳性。在稍后的阶段,随着细胞对胞质半桥粒成分 (例如BP抗原) 以及VII型胶原呈阳性,α6β4沿着上皮细胞的基极集中,并与外植体的结缔组织邻接。针对 β4的多克隆抗体不会干扰外植体中上皮细胞的迁移。但是,它们阻止了半桥粒复合体的组装,并抑制了在伤口区域迁移的细胞中VII型胶原的表达。此外,它们还会诱导外植体未受伤区域的非迁移细胞中已建立的半桥粒的破坏。针对 α6的单克隆抗体具有更显着的作用,因为它们完全脱离了外植体未受伤区域的上皮细胞。针对CAF的抗体也可以通过诱导基底膜区的上皮和结缔组织之间的分离而分离未受伤区域的上皮细胞。这些结果提出了一个模型,其中 α6β4极化到细胞的基底表面,可能是由假定的锚定细丝相关配体诱导的,触发了半桥粒斑块的组装。
  • 【雌激素通过雌激素受体 α 刺激内皮祖细胞。】 复制标题 收藏 收藏
    DOI:10.1111/j.1365-2265.2007.02918.x 复制DOI
    作者列表:Foresta C,Zuccarello D,Biagioli A,De Toni L,Prana E,Nicoletti V,Ambrosini G,Ferlin A
    BACKGROUND & AIMS: CONTEXT:Oestrogens play an important protective role on the vascular system. The endothelial cell layer is a direct target for these hormones, and expresses at least two oestrogen receptors, ER-alpha and ER-beta. Recent studies have shown that vascular healing is significantly modulated by circulating bone marrow-derived cells. A subset of these stem cells, endothelial progenitor cells (EPCs), have recently been described as a population of pluripotent cells within the peripheral blood capable of differentiating into endothelial cells. OBJECTIVE:In the present study we investigated the expression of ER-alpha and ER-beta on human EPCs and the effect that oestrogens have on the function of EPCs in vitro. METHODS:EPCs were isolated and cultured from healthy donors. RT-PCR, western blotting and immunohistochemistry were used to assess expression of ER-alpha and ER-beta. Proliferation and CFU assays were used to assess the response of EPCs to different doses of 17,beta-oestradiol. MAIN OUTCOME MEASURES:Expression of ER-alpha and ER-beta in EPCs, and the effect of 17,beta-oestradiol on proliferation of EPCs. RESULTS:Human EPCs express ER-alpha mRNA and protein. 17,beta-oestradiol increases proliferation of EPCs and CFU in a dose-dependent manner. CONCLUSIONS:Human EPCs express ER-alpha but not ER-beta, and oestrogens can stimulate the proliferation of these cells in vitro. Oestrogens exert these effects at concentrations that are usually reached during stimulation for in vitro fertilization in women, and therefore further studies are needed to clarify the clinical significance of these effects.
    背景与目标:
  • 【胎儿多囊肾疾病中 α-整合素亚基的分布。】 复制标题 收藏 收藏
    DOI:10.1007/s004670050275 复制DOI
    作者列表:Daïkha-Dahmane F,Narcy F,Dommergues M,Lacoste M,Beziau A,Gubler MC
    BACKGROUND & AIMS: :An alteration in cell/matrix interactions is one of the suggested mechanisms leading to cyst formation in polycystic kidney diseases. Most of these interactions are mediated by beta 1-integrins, a subfamily of integrin receptors, formed by the association of the beta 1-chain with different alpha-subunits. To date, no study on alpha-integrin subunit distribution during the early stages of cyst development has been reported. Using immunofluorescence, we analyzed the distribution of alpha-integrin subunits (alpha 1, alpha 2, alpha 3, alpha 5, and alpha 6) and basement membrane proteins in kidneys of fetuses with autosomal dominant (ADPKD) or autosomal recessive polycystic kidney disease (ARPKD). The distribution was compared with that observed in normal fetal and post-natal kidneys, and in fetal cystic dysplasia and Meckel syndrome. Marked increase in alpha 1-integrin staining was observed in normal and cystic collecting duct cells of both polycystic diseases (PKD), compared with normal and cystic controls. The distribution of integrin subunits alpha 2, alpha 3, and alpha 6 was irregular in cyst epithelial cells of PKD and cystic controls. The increased expression of the alpha 1-subunit specifically observed in PKD collecting duct cells may be an early consequence of the genetic defect in ARPKD. In ADPKD it parallels the reported expression of polycystin, the protein product of PKD1. The irregular expression of alpha 2, alpha 3, and alpha 6 integrin subunits observed in all types of cysts suggests that cell/matrix interactions are altered early and may participate in the development of cysts, perhaps by contributing to the deregulation of cell survival in cystic diseases.
    背景与目标: : 细胞/基质相互作用的改变是导致多囊肾疾病中囊肿形成的建议机制之一。这些相互作用中的大多数是由 β1-整合素 (一种整合素受体的亚家族) 介导的,由 β1-链与不同的 α-亚基结合形成。迄今为止,尚无关于囊肿发育早期阶段的 α-整联蛋白亚基分布的研究。使用免疫荧光,我们分析了常染色体显性遗传 (ADPKD) 或常染色体隐性多囊肾 (ARPKD) 胎儿肾脏中 α-整合素亚基 (α1,α2,α3,α5和 α6) 和基底膜蛋白的分布。将分布与正常胎儿和产后肾脏以及胎儿囊性发育不良和梅克尔综合征的分布进行了比较。与正常和囊性对照相比,在两种多囊性疾病 (PKD) 的正常和囊性集合管细胞中观察到 α1-整合素染色显着增加。在PKD和囊性对照的囊肿上皮细胞中,整合素亚基 α2,α3和 α6的分布不规则。在PKD收集导管细胞中特异性观察到的 α1亚基表达增加可能是ARPKD遗传缺陷的早期结果。在ADPKD中,它与pkd1的蛋白质产物多囊蛋白的报道表达相似。在所有类型的囊肿中观察到的 α2,α3和 α6整联蛋白亚基的不规则表达表明,细胞/基质相互作用在早期发生改变,并可能参与囊肿的发展,这可能是通过改变囊性疾病中细胞存活的失调。
  • 【诱发的alpha在主要感觉处理中的可能作用: 猫颅内记录和人类EEG和MEG的共同特性。】 复制标题 收藏 收藏
    DOI:10.1016/s0167-8760(97)00762-9 复制DOI
    作者列表:Schürmann M,Başar-Eroglu C,Başar E
    BACKGROUND & AIMS: :Regarding the evoked potential (EP) as a superposition of evoked EEG rhythms in several frequency ranges, we investigated the following issue: Are distinct evoked rhythms, in particular the alpha (8-15 Hz) response, related to separable physiological processes? Frequency domain analysis of EPs was used to evaluate results of cross-modality experiments, i.e.: responses to auditory stimuli were simultaneously recorded from the auditory cortex (adequate stimulation) and from the visual cortex (inadequate stimulation). Responses to visual stimuli were recorded from the same sites. The results of these experiments and further measurements (EEG and MEG responses in humans, among them multiple sclerosis patients) are consistent in the following respect: The amplitudes of alpha responses are dependent on whether or not a stimulus applied is adequate. Alpha responses may thus be related mostly to primary sensory processing. In contrast, theta responses (4-7 Hz) were observed for adequate as well as inadequate stimuli. They may be related rather to associative and cognitive processing than to primary sensory processing. Thus frequency responses, in particular the alpha response, are not artificial results of digital filtering, but functionally significant brain responses.
    背景与目标: : 关于诱发电位 (EP) 作为诱发EEG节律在几个频率范围内的叠加,我们研究了以下问题: 不同的诱发节律,特别是 α (8-15Hz) 响应是否与可分离的生理过程有关?EPs的频域分析用于评估交叉模式实验的结果,即: 同时从听觉皮层 (适当的刺激) 和视觉皮层 (不充分的刺激) 记录对听觉刺激的反应。从相同的部位记录对视觉刺激的反应。这些实验和进一步测量的结果 (人类的EEG和MEG反应,其中包括多发性硬化症患者) 在以下方面是一致的: α 反应的幅度取决于所施加的刺激是否足够。因此,Alpha反应可能主要与初级感觉处理有关。相反,对于足够和不充分的刺激,观察到theta反应 (4-7Hz)。它们可能与联想和认知加工有关,而不是与主要的感觉加工有关。因此,频率响应 (尤其是alpha响应) 不是数字滤波的人工结果,而是功能上重要的大脑响应。
  • 【Α-硫辛酸抑制糖尿病大鼠P2X受体活性和内脏对结直肠扩张的超敏反应。】 复制标题 收藏 收藏
    DOI:10.1038/s41598-017-04283-7 复制DOI
    作者列表:Hu J,Qin X,Song ZY,Yang PP,Feng Y,Sun Q,Xu GY,Zhang HH
    BACKGROUND & AIMS: :The present study was designed to investigate the roles of P2X3 receptors in dorsal root ganglion (DRG) neurons in colonic hypersensitivity and the effects of alpha-lipoic acid (ALA) on P2X3 receptor activity and colonic hypersensitivity of diabetic rats. Streptozotocin (STZ) was used to induce diabetic model. Abdominal withdrawal reflex (AWR) responding to colorectal distention (CRD) was recorded as colonic sensitivity. ATP-induced current density of colon-specific DRG (T13-L2 DRGs) neurons was measured with whole-cell patch clamp. The expression of P2X3Rs of T13-L2 DRGs was measured by western blot analysis. The results showed that AWR scores significantly increased after STZ injection. P2X3R expression and ATP current density of T13-L2 DRG neurons were enhanced in diabetic rats. Intraperitoneal injection with ALA once a day for 1 week remarkably reduced P2X3R expression and ATP current density in diabetic rats. Importantly, ALA treatment attenuated colonic hypersensitivity in diabetic rats. Our data suggest that STZ injection increases expression and function of P2X3 receptors of colon-specific DRG neurons, thus contributing to colonic hypersensitivity in diabetic rats. Administration of ALA attenuates diabetic colonic hypersensitivity, which is most likely mediated by suppressing expression and function of P2X3 receptors in DRGs of diabetic rats.
    背景与目标: 本研究旨在研究背根神经节 (DRG) 神经元中P2X3受体在结肠超敏反应中的作用以及 α-硫辛酸 (ALA) 对糖尿病大鼠P2X3受体活性和结肠超敏反应的影响。链脲佐菌素 (STZ) 用于诱导糖尿病模型。对结直肠扩张 (CRD) 反应的腹部撤药反射 (AWR) 被记录为结肠敏感性。用全细胞膜片钳测量ATP诱导的结肠特异性DRG (T13-L2 DRGs) 神经元的电流密度。通过蛋白质印迹分析测量T13-L2 DRGs中P2X3Rs的表达。结果显示,注射STZ后AWR评分明显升高。糖尿病大鼠T13-L2 DRG神经元P2X3R表达和ATP电流密度增强。每天一次腹腔注射ALA,持续1周,可显着降低糖尿病大鼠的P2X3R表达和ATP电流密度。重要的是,ALA治疗可减轻糖尿病大鼠的结肠超敏反应。我们的数据表明,STZ注射可增加结肠特异性DRG神经元的P2X3受体的表达和功能,从而导致糖尿病大鼠的结肠超敏反应。ALA的给药可减轻糖尿病结肠超敏反应,这很可能是通过抑制糖尿病大鼠DRGs中P2X3受体的表达和功能来介导的。

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