3D Imaging Detection of HER2 Based in the Use of Novel Affibody-Quantum Dots Probes and Ratiometric Analysis

Patients with breast cancer (BC) overexpressing HER2 (HER2+) are selected for Trastuzumab treatment, which blocks HER2 and improves cancer prognosis. However, HER2+ diagnosis, by the gold standard, immunohistochemistry, could lead to errors, associated to: a) variability in sample manipulation (thin 2D sections), b) use of subjective algorithms, and c) heterogeneity of HER2 expression within the tissue. Therefore, we explored HER2 3D detection by multiplexed imaging of Affibody-Quantum Dots conjugates (Aff-QD), ratiometric analysis (RMA_FI) and thresholding, using BC multicellular tumor spheroids (BC-MTS) (~120 μm of diameter) as 3D model of BC. HER2+, HER2– and hybrid HER2+/? BC-MTS (mimicking heterogeneous tissue) were incubated simultaneously with two Aff-QD probes (anti-HER2 and negative control (NC), respectively, (1:1)). Confocal XY sections were recorded along the Z distance, and processed by automatized RMA_FI (anti-HER2 Aff-QD/ NC). Quantifying the NC fluorescence allowed to predict the fraction of non-specific accumulation of the anti-HER2 probe within the thick sample, and resolve the specific HER2 level. HER2 was detected up to 30 μm within intact BC-MTS, however, permeabilization improved detection up to 70 μm. Specific HER2 signal was objectively quantified, and HER2 3D-density of 9.2, 48.3 and 30.8% were obtained in HER2?, HER2+ and hybrid HER2+/? permeabilized BC-MTS, respectively. Therefore, by combining the multiplexing capacity of Aff-QD probes and RMA_FI, we overcame the challenge of non-specific probe accumulation in 3D samples with minimal processing, yielding a fast, specific spatial HER2 detection and objective quantification.     

利用异源细胞质对小偃6号农艺性状改良的研究

通过以山羊草等２３种不同异源胞质的中国春为母本,以小偃６号为父本,经过七代核置换回交,培育成功了２３种不同细胞质的小偃６号核质杂种。和对照小偃６号比较分析,发现不同的异源细胞质对小偃６号的抽穗期,条锈抗性、结实率等９个农艺性状都有着较明显的影响。     

中华猕猴桃雄性株系"磨山4号"的选育研究

通过对近年来选育的“磨山４号”、“磨山５号”、“武植８２－１２”、“武植８０－４”、“武植８２－５”等５个中华猕猴桃（ＡｃｔｉｎｉｄｉａｃｈｉｎｅｎｓｉｓＰｌａｎｃｈ）雄性株系的花期、花量、花粉生活力、花粉量的观察,并分析与“武植３号”授粉对果实的影响,从综合性状来看,“磨山４号”花期比一般雄株长２个星期左右（约２０天）,有效花粉量高,株型紧凑,授粉后的果实单果重及Ｖｃ的含量均高于其它雄株,为中华猕猴桃适配雄株。     

东京根霉(Rhizopus tonkinense No.9212)用于木薯淀粉生料发酵生产单细胞蛋白的研究

我国华南地区盛产木薯,是制造淀粉的主要原料之一。据测定,鲜木薯含水分70.3％、淀粉21.5％、糖分5.1％、粗蛋白1.1％、粗脂肪0.4％、粗纤维1.1％,灰分0.5％、菜豆苷(又称亚麻苦苷)0.01～0.04％。 木薯的淀粉含量虽很高,但蛋白质含量比较低。若能通过微生物发酵使其中一部分淀粉转化为单细胞蛋白,将可在很大程度上缓解我国饲料蛋白不足的矛盾。淀粉生料发酵,具有能耗低,工艺简便等优点。当前,     

转基因水稻“科丰6号”实时荧光PCR定性定量检测方法研究

旨在建立转基因水稻"科丰6号"外源基因和边界序列的实时荧光PCR检测方法,为科丰6号定性定量检测提供技术支持。根据外源基因和边界序列信息,设计实时荧光PCR探针引物,优化体系,对不同转基因产品和不同转基因含量的"科丰6号"水稻进行检测。结果显示,所设计的引物探针具有很好的特异性,与其他转基因水稻品系、转基因玉米、转基因棉花、转基因番茄和非转基因水稻均无非特异性反应,对转基因水稻"科丰6号"的检测灵敏度达到0.01%。建立的科丰6号实时荧光PCR检测方法重复性好、灵敏度高,能够达到目前国际上转基因产品定量检测的标准,为该水稻品系的定性定量检测提供技术支持。     

A new interpretation of ASM2d for modeling of SBR performance for enhanced biological phosphorus removal under different P/HAc ratios

This study evaluated the prediction capability of Activated Sludge Model No. 2d (ASM2d), for the enhanced biological phosphorus removal (EBPR) performance of a sequencing batch reactor (SBR) receiving variable influent phosphate load. For this purpose, a laboratory-scale SBR was operated with a synthetic feed containing acetate as the sole carbon source. The experiments were conducted in four different Runs to ensure a range of different phosphate/acetate ratios in the influent. Model evaluations were carried out using concentration profiles measured throughout a representative cycle at steady state. An iterative calibration methodology was developed based on sensitivity analysis and applied to four different sets of experimental data on relevant model parameters reflecting SBR performance. ASM2d was able to predict the steady state behavior of the SBR system receiving variable influent phosphate loads only with the recalibrated parameter set. The regular changing pattern of the coefficients could be interpreted with the ability of the SBR system to sustain glycogen accumulating microorganisms, GAOs, which can store substrate under anaerobic conditions without polyphosphate energy, but deriving energy from the degradation of glycogen. Thus they are capable of prevailing at lower P/Ac ratios. The results indicate the need to include glycogen and GAOs as model components for processes involving both phosphate accumulating organisms, (PAOs) and GAOs, in order to obtain a better prediction of X(PHA) and oxygen uptake rate (OUR) profiles in the system.     

Purification and characterization of a Bacillus circulans No. 4.1 chitinase expressed in Escherichia coli

Summary A DNA fragment encoding for 598 amino acids of chitinase protein from Bacillus circulans No. 4.1 was subcloned into pQE-30 expression vector and transformed into Escherichia coli M15 (pREP4). The molecular weight of the expressed protein was approximately 66 kDa. Enzymatic activity of the recombinant protein was assayed after purification using affinity chromatography on a nickel chelating resin. The enzyme hydrolyzed N-acetylchitooligosaccharides mainly to N-acetylchitobiose, and was active toward chitin, carboxymethyl-chitin, colloidal chitin, glycol chitin and 4-methylumbelliferyl-β-d-N, N′-diacetylchitobiose. The pH and temperature optima of the chitinase enzyme were 7.0 and 45 °C, respectively. This enzyme was stable in the pH range of 5.0–9.0 and at temperatures up to 50 °C. In addition, when cleaved by a proteolytic enzyme, the 20-kDa product could retain high chitinolytic activity.     

老年慢性阻塞性肺病患者继发革兰氏阴性杆菌感染的菌型及耐药性分析

目的：探讨老年慢性阻塞性肺疾病（COPD）继发院内革兰氏阴性杆菌感染的发病机理、菌型分布及耐药性。方法：从本院老年COPD继发医院革兰氏阴性杆菌（GNB）肺炎患者痰液中分离的213株GNB进行菌型分类,选用12种常用抗菌药物进行体外MIC药敏试验。结果：213株占COPD继发医院肺炎病原菌总数的69．6％（213／306）。老年COPD患者继发革兰氏阴性菌感染菌种分类为：铜绿假单胞菌（35．7％）、肺炎克雷伯菌（21．1％）、大肠埃希菌（17．4％）、阴沟肠杆菌（11．7％）、嗜麦芽假单胞菌（7．9％）、其他病原菌（6．1％）;药敏结果表明,所有GNB对抗菌药物耐药率均呈上升趋势。结论：铜绿假单胞菌是COPD继发GNB感染的主要致病菌,在临床治疗中必须重视菌型鉴定和药敏试验,合理使用抗生素,才能控制院内感染GNB的发生和日益增高的耐药趋势。     

抗病高淀粉甘薯品种徐薯26的选育及产量形成特点

抗病高淀粉甘薯新品种徐薯26是中国农业科学院甘薯研究所以徐781为母本,通过集团杂交放任授粉、多年多点综合鉴定选育而成。该品种2009年通过国家甘薯新品种鉴定,鉴定编号为国品鉴甘薯2009003。对徐薯26的形态特征、品质性状、抗病性、生产力和产量形成的生理特点等进行了研究,结果表明:徐薯26全生育期长势平稳,薯干品质和食味较好,抗根腐病和蔓割病,薯干和淀粉产量与徐薯18相比增产极显著,生长后期薯块干物质积累较快,经济系数较高,适宜早栽和密植。     

冰川前缘土壤微生物原生演替的生态特征——以乌鲁木齐河源1号冰川为例

为了解冰川前缘土壤微生物在原生演替过程中的生态特征及其影响因素,用空间距离代替时间序列,以乌鲁木齐河源1号冰川终碛堤为起点,沿6个不同演替时期(0,4,15,31,43a和对照)的样带采集土样,以冰川附近发育良好的土壤为对照,测定土壤酶活性、微生物氮矿化与脱氨作用以及微生物生物量。结果表明,土壤脲酶、蛋白酶、酸性磷酸酶、芳基硫酸酯酶、蔗糖酶活性、微生物氮矿化及脱氨作用随演替时间而增加,微生物生物量碳和氮变化呈波动状,趋势不明显。相关分析表明,土壤有机质与酶活、微生物生物量存在极显著正相关(P0.01)。1号冰川前缘微生物多样性指数随着演替时间持续增加,但目前仍未达稳定状态。