Antimetastatic effect of prodigiosin through inhibition of tumor invasion

Prodigiosin, a bacterial metabolite, was reported to have immunosuppressive and anticancer activities. In this study, we investigated novel functions of prodigiosin about anti-metastasis and anti-invasion. Prodigiosin dose-dependently inhibited 95-D cells' migration and invasion according to wound healing assay and the Transwell assay. The inhibitive effect could reach about 50% when cells were treated with 5 microM prodigiosin for 12 h. In animal experiment, intraperitoneal administration of 5 mg kg(-1) prodigiosin decreased the number of metastatic nodules by 53% and elevated the survival rate of mice about one-fold comparing with control group. Results of cell aggregation and adhesion assay showed that prodigiosin could promote cell aggregation and simultaneously inhibit cell from adhering to extracellular matrix (ECM). In addition, prodigiosin suppressed RhoA gene expression, hence, decreased protein level of RhoA in 95-D cells, according to RT-PCR assay and Western blot assay. Gel zymogram assay revealed that prodigiosin could suppress the activity of matrix metalloproteinase-2 (MMP-2). These results demonstrate that prodigiosin effectively inhibit tumor metastasis in vitro and in vivo. The action mechanisms of prodigiosin are associated with the promotion of cell aggregation and the inhibition of various steps in cell invasive process, which include the inhibition of cell adhesion and mobility in a RhoA-dependent way and the suppression of MMP-2 ability.     

Pharmacological characterization of novel adenosine ligands in recombinant and native human A2B receptors

The present study was designed to evaluate the effects of novel and recognised compounds at human recombinant A(2B) adenosine receptors expressed in Chinese hamster ovary (hA(2B)CHO), in human embryonic kidney 293 (hA(2B)HEK-293) and at endogenous A(2B) receptors in human mast cells (HMC-1). Saturation binding experiments performed using the new high affinity A(2B) adenosine radioligand [(3)H]-N-benzo[1,3]dioxol-5-yl-2-[5-(2,6-dioxo-1,3-dipropyl-2,3,6,7-tetra hydro-1H-purin-8-yl)-1-methyl-1H-pyrazol-3-yloxy]-acetamide ([(3)H]-MRE 2029F20) revealed a single class of binding sites in hA(2B)CHO, hA(2B)HEK-293 and HMC-1 cells with K(D) (nM) of 1.65+/-0.18, 2.83+/-0.34, 2.62+/-0.27 and B(max) (fmol/mg protein) of 36+/-4, 475+/-50 and 128+/-15, respectively. The pharmacological profile of new compounds, determined in inhibition binding experiments in hA(2B)HEK-293 cells using [(3)H]-MRE 2029F20, showed a rank order of potency typical of the A(2B) receptors with K(i) values in the range 3.2-28nM. In functional assays, recognised agonists and antagonists were studied by evaluating their capability to modulate the cAMP production in hA(2B)CHO and in HMC-1 cells. Novel compounds were able to decrease NECA-stimulated cAMP production in hA(2B)CHO and in HMC-1 cells showing a high potency. New compounds were also able to inhibit cAMP levels in the absence of NECA and in the presence of forskolin stimulation in hA(2B)CHO and in HMC-1 cells. In HEK-293 cells MRE 2029F20 reduced cAMP basal levels with an IC(50) value of 2.9+/-0.3nM. These results suggest that novel compounds are antagonists with an inverse agonist activity in recombinant and native human A(2B) receptors.     

一种复方手消毒液杀菌效果及毒性的实验观察

目的了解一种新型手消毒液杀灭微生物效果与安全性能。方法采用载体定量杀菌实验和动物毒性实验该新型手消毒液进行了实验室研究。结果该消毒液以主要有效成分聚-[2-(2-乙氧基)-乙脂]-氯化胍(含量为0.125%)和聚-环己-胍盐-氯化物(含量为0.375%)的水溶液对布片上大肠杆菌、金黄色葡萄球菌作用5 min,对布片上白色念珠菌作用10 min,杀灭率均达99.99%。将该消毒液原液密封置于37℃放置90 d,杀菌效果与储存前无明显差异。菌悬液内含50%小牛血清对该消毒剂杀菌效果无明显影响。经30人次手的现场消毒实验证明,该洗液对手上自然菌消毒效果比较好,对自然菌杀灭率达到消毒合格要求。该消毒剂对小鼠经口LD50>5 000mg/kg,蓄积系数K>5;该消毒液原液对家兔皮肤刺激实验积分为0,眼刺激实验积分为1.0;对小鼠骨髓嗜多染红细胞微核实验为阴性,精子畸形实验结果为阴性。该消毒剂原液浸泡72 h,对不锈钢、铜无腐蚀,对碳钢和铝基本无腐蚀性。结论该复方消毒剂对细菌繁殖体和真菌杀灭效果较好;属实际无毒级,弱蓄积毒性;对皮肤、眼睛为无刺激性;性能稳定,腐蚀性较低。     

Nucleus accumbens facilitates nociception

We have previously demonstrated an opioid link in nucleus accumbens (NAc) that mediates antinociception produced by a novel ascending pain modulation pathway. For example, noxious stimulation induces heterosegmental antinociception that is mediated by both mu- and delta-opioid receptors in NAc. However, spinal intrathecal administration of the mu-receptor agonist [d-Ala², N-Me-Phe⁴,Gly⁵-ol]-enkephalin (DAMGO) also induces heterosegmental antinociception. The aim of the present study in the rat was to identify the intra-NAc opioid receptors that mediate the antinociceptive effects of spinally administered DAMGO and also to determine the effect of NAc efferent activity on nociception. Intra-NAc administration of either the mu-opioid receptor antagonist Cys²,Tyr³, Orn⁵,Pen⁷amide (CTOP) or the delta-opioid receptor antagonist naltrindole blocked the antinociceptive effect of spinally administered DAMGO on the jaw-opening reflex (JOR). Injection of quaternary lidocaine (QX-314) attenuated the JOR, suggesting that the output of NAc is pronociceptive. In support of this, intra-NAc injection of the excitatory amino acid agonist kainate enhanced the JOR. Thus, it is possible to modulate activity in NAc to bidirectionally attenuate or enhance nociception, suggesting a potential role for NAc in setting nociceptive sensitivity.     

上海市3城区1～6岁儿童饮食行为问题交互式干预的随机对照研究

目的对存在不良饮食行为问题儿童行交互式干预,观察饮食行为的改善情况。方法选择上海市3个城区(长宁区、卢湾区和虹口区)1～6岁常住户籍儿童。编制《上海市儿童饮食行为调查和干预随访问卷》基线版和随访版,经专业培训合格的社区儿保医生确定研究起点(T0)存在饮食行为问题的儿童入组,以密封信封的方式将入组儿童随机分为干预组和对照组。两组根据年龄分为1、2、3、4、5和6岁亚组;根据营养状况分为营养正常、中度营养不良和重度营养不良亚组。干预组在T0时首次干预,并于干预后1(T1)、3(T2)、6(T3)和9个月(T4)分别以随访版内容评估和干预。对照组于T0、T4时点分别行基线版和随访版评估,不行交互式干预。比较干预组和对照组的饮食行为状况,并计算得到1例有益结果需干预的人数(NNT)和不良行为保持率。结果 2009年1～10月共纳入有不良饮食行为问题的儿童490例,至T4时点,随访完全不依从2例,部分不依从26例,失访率为5.7%(28/490例)。462例有效数据纳入分析,干预组245例,对照组217例。①干预组T0时点儿童的饮食行为问题综合评分为(19.7±0.0)分,T4时点为(14.2±0.3)分,呈显著下降(P0.01);对照组T0时点为(19.7±0.5)分,T4时点为(19.6±0.1)分,无显著改变(P0.05)。②干预组T0～T4不同时点各年龄亚组饮食行为问题综合评分随交互式干预的介入均呈下降趋势,各年龄亚组儿童饮食行为综合评分T4较T0时点均显著降低(P0.01),其中3岁亚组△(T0～T4)减分最多,6岁亚组△(T0～T4)减分最少,3和6岁亚组间△(T0-T4)差异有统计学意义(P0.05)。462例存在不良饮食行为问题的儿童干预效果NNT为2.5(95%CI:2.1～3.0),总的不良饮食行为保持率干预组与对照组差异有统计学意义(P0.001)。③干预组营养正常与重度营养不良亚组饮食行为问题综合评分T4较T0显著降低(P0.05)。对照组营养正常和重度营养不良亚组饮食行为问题综合评分T4较T0时点差异无统计学意义(P0.05)。④干预组干预前后各单项饮食行为问题评分T4较T0时点均有显著降低(P0.01);但干预后各项饮食行为问题的△(T0-T4)差异无统计学意义(P0.05)。对照组各项饮食行为问题评分T4较T0时点无显著降低(P0.05)。各单项饮食行为NNT为1.4～2.5。各单项不良饮食行为保持率干预组与对照组差异均有统计学意义(P0.05)。结论合理的饮食行为干预可改善儿童的不良饮食行为,交互式干预模式是适合中国国情的有效干预方法。     

Effect of TNF-α on Raji cells at different cellular levels estimated by various methods

Tumor necrosis factor (TNF)-α, a pleiotropic cytokine, has been shown to induce diverse and opposite effects on lymphoid malignancy depending on TNF receptor system expression. Based on this, we investigated its in vitro dose- and time-related effect on the malignant B-cell line Raji, derived from Burkitt lymphoma patients, at different intracellular levels. The membrane alteration was estimated by lactate dehydrogenase (LDH) release and by flow cytometry; intracellular metabolic energy by determination of the total intracellular LDH activity; total cytosole protein mass by sulforhodamine B assay; and cell growth by incorporation of [³H]thymidine into DNA. Significant increase of LDH through cell membrane alteration was accompanied by decrease of intracellular metabolized energy and total protein mass. TNF-α at lower concentrations (125 and 250 pg/ml) significantly induced cell proliferation in comparison with 1,000 pg/ml of TNF-α, which induced more cell death. TNF-α induced maximal apoptosis rate up to 30% after 24 h, showing more effects for a necrotic form of cell death. Here we reported opposite and diverse effects of TNF-α at different intracellular levels in Raji cells, when applied in different assays, showing characteristics for every cellular compartment.     

Excitatory/inhibitory neuronal metabolic balance in mouse hippocampus upon infusion of [U-13C6]glucose

Hippocampus plays a critical role in linking brain energetics and behavior typically associated to stress exposure. In this study, we aimed to simultaneously assess excitatory and inhibitory neuronal metabolism in mouse hippocampus in vivo by applying ¹⁸FDG-PET and indirect ¹³C magnetic resonance spectroscopy (¹H-[¹³C]-MRS) at 14.1 T upon infusion of uniformly ¹³C-labeled glucose ([U-¹³C₆]Glc). Improving the spectral fitting by taking into account variable decoupling efficiencies of [U-¹³C₆]Glc and refining the compartmentalized model by including two γ-aminobutyric acid (GABA) pools permit us to evaluate the relative contributions of glutamatergic and GABAergic metabolism to total hippocampal neuroenergetics. We report that GABAergic activity accounts for ∼13% of total neurotransmission (V_NT) and ∼27% of total neuronal TCA cycle (V_TCA) in mouse hippocampus suggesting a higher V_TCA/V_NT ratio for inhibitory neurons compared to excitatory neurons. Finally, our results provide new strategies and tools for bringing forward the developments and applications of ¹³C-MRS in specific brain regions of small animals.     

Characterization and Properties of Poly[N‐(2‐cyanoethyl)pyrrole]

PNCPy was prepared by anodic polymerization and its properties in both doped and undoped state were characterized. The doping level of the oxidized material has been found to be larger than that of other conducting polymers; the more relevant electrochemical properties of the doped material were retained after undoping. SEM and AFM data are consistent with a lumpy surface and a multidirectional growing of the polymer chains. Finally, PNCPy has been combined with PEDOT to prepare three‐layer systems with enhanced electroactivity and electrostability. Results suggest that PNCPy is a potential candidate for the fabrication of electric circuit components that are able to block the current flow below a given potential.

     

Novel aldose reductase inhibitors derived from 6-[[(diphenylmethylene)amino]oxy]hexanoic acid

Starting from the recently identified aldose reductase inhibitor 6-[[(diphenylmethylene)amino]oxy]hexanoic acid 1, the following systematic structural modifications were performed: (a) formal substitution of the phenyl rings, (b) isosteric replacement of the benzene core by the heteroarenes pyridine and thiophene, (c) formal reduction of the aromatic substructure and subsequent diminution of the cyclohexyl ring, (d) introduction of methylene spacer between C=N and the phenyl rings, and finally (e) formal ring closure in order to get derivatives of the tricycles fluorenone, xanthone, and thioxanthone, respectively. Out of these series, compounds 22-24 bearing disubstituted phenyl rings exhibit the highest inhibitory activity (IC(50 )value approx. 3 muM) which lie almost in the range of the reference sorbinil.