Quantitation of Rat Dopamine Transporter mRNA: Effects of Cocaine Treatment and Withdrawal

Dopamine transporter mRNA levels in the rat substantia nigra were quantified using a sensitive nuclease protection assay with a highly homologous human dopamine transporter cDNA clone. The same probe was also used to visualize dopamine transporter mRNA in the substantia nigra by in situ hybridization. Repeated cocaine administration (15 mg/kg, twice a day for 6.5 days) resulted in a greater than 40% decrease in nigral dopamine transporter mRNA levels. In contrast, dopamine transporter mRNA levels were unchanged after either acute treatment (4 h before death) or repeated cocaine treatment followed by a 72-h withdrawal period. Thus, blockade of the dopamine transporter by repeated cocaine administration may result in the down-regulation of dopamine transporter gene expression in dopamine neurons.     

The determination of the pKa of histidine residues in proteins by Raman difference spectroscopy

Sensitive Raman difference spectroscopy was used to monitor the protonation and deprotonation of histidine residues in apo-transferrin. We have shown previously that the behavior of small molecules and/or small molecular groups bound to proteins or other large macromolecules can be studied by Raman difference spectroscopy (Yue, K.T. et al. (1989) J. Raman Spectrosc. 20, 541-545). Using this method, we have measured the Raman difference spectra of human transferrin at different pH values with respect to pH 8.9, titrating its various histidine residues. About 12 +/- 2 of the 19 residues were titrated. The pH difference spectrum of transferrin obtained is very similar to that of histidine in solution, but with clear differences in the 1200-1400 cm-1 region. A titration curve with pKa of 6.08 +/- 0.01 fit the data of histidine in solution and a value of 6.56 +/- 0.02 was found for the average value of the 12 histidine residues inside transferrin. The technique has enough sensitivity at present to monitor a single histidine residue in a 130 kDa molecule and to determine the titration curve of one residue in a 40 kDa protein.     

时域—频域结合分析法—一种分析果蝇求爱歌的新方法

我们设计了一种时域-频域结合分析法,并用此方法分析了6个种群12种果蝇的求爱歌,发现如果将时域与频域的研究结合起来,对求爱歌进行频谱分析,可以定量地揭示出求爱歌的频域特性及其在时域上的细微变化。我们还对果蝇求爱歌的时域模式进行了初步的探讨,发现它们是在同一基本成分上进行调制而产生的,亲缘关系较近的种具有相近的调制方式。在对杂交后代的求爱歌的频谱分析中,我们还发现频谱上的某些特点是能够遗传的。这一新的研究方法为果蝇的进化遗传学和神经遗传的研究提供了一种新的手段。     

氨基酸酯水解酶产生菌的筛选及其合成头孢立新的研究

From ten genera and 146 bacterial strains, 22 strains producing alpha-amino acid ester hydrolase were selected. Among them, AS 1.586 and 41-2 were the best. The optimal conditions for synthesis of cephalexin by pseudomonas aeruginosa 1.204 were investigated. The optimal pH and temperature for enzymatic synthesis reaction was pH 6.8 and 25 degrees C, respectively. By using 1% 7-ADCA, 3% PGME and 4% biomass, about 70% of 7-ADCA was converted to cephalexin under the mentioned conditions.     

Glycosylated modification of MUC1 maybe a new target to promote drug sensitivity and efficacy for breast cancer chemotherapy

Breast cancer, the most common cancer in women, usually exhibits intrinsic insensitivity to drugs, even without drug resistance. MUC1 is a highly glycosylated transmembrane protein, overexpressed in breast cancer, contributing to tumorigenesis and worse prognosis. However, the molecular mechanism between MUC1 and drug sensitivity still remains unclear. Here, natural flavonoid apigenin was used as objective due to the antitumor activity and wide availability. MUC1 knockout (KO) markedly sensitized breast cancer cells to apigenin cytotoxicity in vitro and in vivo. Both genetical and pharmacological inhibition significantly enhanced the chemosensitivity to apigenin and clinical drugs whereas MUC1 overexpression conversely aggravated such drug resistance. Constitutively re-expressing wild type MUC1 in KO cells restored the drug resistance; however, the transmembrane domain deletant could not rescue the phenotype. Notably, further investigation discovered that membrane-dependent drug resistance relied on the extracellular glycosylated modification since removing O-glycosylation via inhibitor, enzyme digestion, or GCNT3 (MUC1 related O-glycosyltransferase) knockout markedly reinvigorated the chemosensitivity in WT cells, but had no effect on KO cells. Conversely, inserting O-glycosylated sites to MUC1-N increased the drug tolerance whereas the O-glycosylated deletant (Ser/Thr to Ala) maintained high susceptibility to drugs. Importantly, the intracellular concentration of apigenin measured by UPLC and fluorescence distribution firmly revealed the increased drug permeation in MUC1 KO and BAG-pretreated cells. Multiple clinical chemotherapeutics with small molecular were tested and obtained the similar conclusion. Our findings uncover a critical role of the extracellular O-glycosylation of MUC1-N in weakening drug sensitivity through acting as a barrier, highlighting a new perspective that targeting MUC1 O-glycosylation has great potential to promote drug sensitivity and efficacy.Subject terms: Breast cancer     

基于微生物模型的合作行为进化研究进展

生物间的合作行为如何在自然选择过程中显示出对欺骗者的优势,一直以来都是进化生物学上的经典问题。实验室构建的具有合作行为的微生物种群是研究这一问题的良好素材。本文综述了几种基于微生物模型的合作行为进化理论,如亲缘选择理论、辛普森悖论、竞争抑制理论、进化博弈理论等,对其中涉及到的微生物模型进行介绍和评价,并展望其研究前景。     

利用Ecopath模型评价鲢鳙放养对千岛湖生态系统的影响

“保水渔业”是中国控制“水华”暴发等生态灾变的措施之一,其结果存在较大的不确定性。在浙江省的新安江水库(千岛湖),“保水渔业”的实施带来了水质改善和渔业增产的双重效果。但在生态系统自组织层面,这种人工干预手段引起的生态系统结构化效应的研究,尚未真正展开。基于2008—2010年千岛湖的生态和渔业资源调查数据,应用EwE(V6.6)构建了2010年千岛湖生态系统的Ecopath模型,并将其与2004和2016年的模型进行对照,分析了千岛湖生态系统在鲢、鳙鱼放养下的变化。千岛湖生态系统在3个年份均为4个整合营养级,营养能流分布成典型的金字塔型,且营养流总量中流向碎屑的占比很大,营养级Ⅰ、Ⅱ的能量被利用得不够充分;除鲢、鳙鱼外大部分鱼类的生物量逐渐下降,浮游植物和碎屑的生物量增多;系统的初级生产力和规模得到了一定提升,但总体的能量转换效率有所降低。在一些和系统成熟度、复原力和稳定性密切相关的参数方面,总初级生产量/总呼吸量增加,Finn氏循环指数和Finn氏平均路径长度逐渐降低,3个年份的连接指数(CI)分别为0.223、0.219、0.263,系统杂食指数(SOI)分别为0.087、0.1...     

沼泽湿地恢复演替过程中朝天委陵菜间隔子性状与分枝强度的关系

间隔子和分株影响克隆植物的空间分布和资源获取,二者之间关系的研究有助于理解克隆植物的生态适应机制。按照恢复时间设置I(5a)、II(15a)、III(25a)3个梯度,研究了永昌北海子国家湿地公园沼泽湿地恢复演替过程中朝天委陵菜(Potentilla supina)间隔子长度、直径与分枝强度之间的关系。结果表明:随着沼泽湿地恢复演替的进行,湿地群落的高度、盖度和生物量逐渐增大,土壤含水量、有机质逐渐增大,土壤容重逐渐降低;湿地群落的优势植物种群由朝天委陵菜转为黑麦草;朝天委陵菜间隔子长度和直径增大,分株数减小;间隔子长度、直径与分枝强度均呈显著负相关关系(P<0.05)。沼泽湿地恢复演替过程中,朝天委陵菜由选择垄断区域资源转向逃避或忍耐不利生境,体现了湿地克隆植物在异质性生境中独特的适应性。     

滇中退化山地不同植被恢复下土壤碳氮磷储量与生态化学计量特征

植被恢复对土壤营养元素的存赋及其生态化学计量特征的影响广受关注,为了深入了解不同植被恢复类型下土壤碳、氮、磷储量与生态化学计量特征,选择滇中地区退化山地飒马场流域具有代表性的4种不同修复阶段的典型植被(荒坡灌草丛、云南松林、针阔混交林和次生常绿阔叶林)为研究对象,分析了不同植被类型下不同深度土壤中有机碳(SOC)、全氮(TN)、全磷(TP)储量和化学计量变化特征。结果表明,退化山地的植被恢复显著改变土壤碳氮磷储存能力和化学计量比,这种改变作用整体上随土壤深度增加而降低。其中,在0—60 cm土层上,SOC储量在次生常绿阔叶林最高,达123.41 t/hm²,其次是针阔混交林(115.69 t/hm²)和云南松林(93.08 t/hm²),荒坡灌草丛(89.56 t/hm²)最低;TN储量针阔混交林(4.91 t/hm²)>次生常绿阔叶林(4.58 t/hm²)>云南松林(4.43 t/hm²)>荒坡灌草丛(3.98 ...