Pyrazolo[3,4-d]pyrimidine derivatives containing a Schiff base moiety as potential antiviral agents

A series of pyrazolo[3,4-d]pyrimidine derivatives containing a Schiff base moiety were synthesized, characterised, and evaluated for their activity against tobacco mosaic virus (TMV). Biological assays indicated that several of the derivatives exhibited significant activity against TMV. In particularly, compounds 5y and 5aa displayed excellent inactivating activity against TMV, with half maximal effective concentration (EC₅₀) values of 70.3 and 53.65?μg/mL, respectively, which were much better than that of ribavirin (150.45?μg/mL), and 5aa was superior to ningnanmycin (EC₅₀?=?55.35?μg/mL). Interactions of compounds 5y and 5aa with TMV coat protein (TMV-CP) were investigated using microscale thermophoresis and molecular docking. Compounds 5y and 5aa displayed strong binding capability to TMV-CP with dissociation constant (K_d) values of 22.6 and 9.8?μM, respectively. These findings indicate that pyrazolo[3,4-d]pyrimidine derivatives containing a Schiff base may be potential antiviral agents.     

小RNA病毒和小DNA病毒的三维结构模型

本文综述了小RNA病毒和小DNA病毒的主要结构特征,绘制出了它们的三维结构模型,从中找出了两者间的共同点和差异,为进一步研究两种病毒提供了依据。     

人胃粘膜G、D细胞与肠化生关系的研究

用免疫细胞化学和原位杂交技术探讨G、D细胞及胃泌素mRNA与肠化生的关系。标本来自胃镜活检的胃粘膜。结果显示,在与大肠化生区相邻的胃粘膜,G细胞突变消失,假幽门腺化生也缺乏G细胞,而淖肠化生仍保留少数G细胞;D细胞不仅见于小肠化生,而且也出现在假幽门腺化生以及某些大肠化生区。胃泌素mRNA仅限于G细胞分布区,未出现在大肠化生区和假幽门腺化生区,G细胞及胃泌素mRNA在大肠化生区的消失,可能由于局部杯状细胞分泌的硫酸粘蛋白改变了局部的微环境,从而影响了G细胞的分化与发育,至于假幽门腺化生区G细胞及胃泌素mRNA消失的原因还不清楚,应继续研究。     

Interleukin-33/ST2 signaling promotes production of interleukin-6 and interleukin-8 in systemic inflammation in cigarette smoke-induced chronic obstructive pulmonary disease mice

Interleukin-33 is a newly described member of the interleukin-1 family. Recent research suggests that IL-33 is increased in lungs and plays a critical role in chronic airway inflammation in cigarette smoke-induced chronic obstructive pulmonary disease (COPD) mice. To determine the role of IL-33 in systemic inflammation, we induced COPD mice models by passive cigarette smoking and identified the IL-33 expression in bronchial endothelial cells and peripheral blood mononuclear cells (PBMCs) of them. After isolation, PBMCs were cultured and stimulated in vitro. We measured expressions of interleukin-6 and interleukin-8 in PBMCs in different groups. The expression of IL-33 in bronchial endothelial cells and PBMCs of COPD mice were highly expressed. Stimulated by cigarette smoke extract (CSE), the expression of IL-6 and IL-8 were induced and enhanced by IL-33. PBMCs of COPD mice produced more IL-6 and IL-8 stimulated by CSE and IL-33. Expression of IL-6 and IL-8 were decreased when stimulated by IL-33 together with soluble ST2. The mRNA production of ST2 in IL-33 stimulated PBMCs was increased. Being pretreated with several kinds of MAPK inhibitors, the secretions of IL-6 and IL-8 in PBMCs did not decrease except for the p38 MAPK inhibitor. We found that IL-33 could induce and enhance the expression of IL-6 and IL-8 in PBMCs of COPD mice via p38 MAPK pathway, and it is a promoter of the IL-6 and IL-8 production in systemic inflammation in COPD mice.     

ETA基因作为荧光定量PCR靶基因设计TaqMan探针快速检测铜绿假单胞菌的研究

铜绿假单胞菌是临床上常见致病菌, 传统的检测方法有各种弊端。本研究对该细菌的ETA基因用生物信息学方法加以分析, 选取相对保守且高度特异的DNA序列, 设计一对特异性引物和一个TaqMan探针, 建立FQ-PCR (fluorescence quantitative PCR)检测PA的方法。通过对梯度浓度的铜绿假单胞菌基因组DNA样品进行FQ-PCR检测和对多种细菌的DNA进行扩增, 来检测其灵敏度和验证引物和探针的特异性。试验结果表明, 对比现有的检测方法, 以ETA基因为靶基因, 基于TaqMan探针的快速FQ-PCR检测技术有更高的灵敏度和更好的特异性等优点, 具有很好的研究价值和应用前景。     

Expression of the large isoform of ribulose-1,5-bisphosphate carboxylase/oxygenase activase gene driven by rbcS promoter in Oryza sativa enhances the photosynthetic capacity

In order to investigate the effect of large isoform of ribulose-1,5-bisphosphate carboxylase/oxygenase (RuBPCO) activase (RuBPCO-A) on photosynthesis, cDNA of the enzyme (rca) was transferred to rice cultivars (Oryza sativa f. japonica cv. Nipponbare) under the control of RuBPCO small subunit gene promoter (rbcS) via Agrobacterium tumefaciens-mediated transformation. Transgenic rice plants were identified by polymerase chain reaction (PCR) and Southern and Western blot analyses. Net photosynthetic rate (P _N) values of the T₁ transgenic lines 34 (T34) and 40 (T40) were 45.26 and 46.32 % higher than that of the control plants, respectively. At the same time, their carboxylation efficiency and RuBPCO initial activity, quantum yield of electron transport in photosystem 2 (Φ_PS2), and steady state photochemical fluorescence quenching (q_P) increased. In addition, heading time of the transgenic rice was advanced. Thus increasing the amount of large isoform of RuBPCO-A in the transgenic rice might have a stimulatory effect on both photosynthesis and plant growth.     

药物筛选生物技术与抗细胞凋亡PI3K信号传导路径

细胞信号传导PI3K-Akt路径涉及细胞周期、再生、分化、衰老与凋亡的调控,采用高通量生物技术分析基因表达谱可用于抗肿瘤或抗衰老的基因与药物筛选.化学诱变剂甲磺酸乙酯(ethlmethane sulformate, EMS)处理CHO细胞,筛选到抗10 μM、20 μM浓度lovastatin的E10、E20突变细胞系,将E10细胞再经诱变剂EMS处理,筛选到抗70 μM浓度lovastatin的ZE70细胞系.Igf-2刺激基因突变的肌肉萎缩小鼠肌肉细胞系,Western免疫杂交试验结果显示,PI3K-Akt路径的Akt蛋白质磷酸化功能正常.生长因子Igf-2刺激肌肉细胞系C2C12的cDNA基因芯片实验表明,促细胞再生、分化相关的基因表达上调,细胞凋亡相关的基因表达下调.细胞的基因突变,激素、Igf-2等生长因子刺激,往往是肿瘤发生的病因;但是,Igf-2等可用于抗肌肉萎缩疾病,以及抗细胞凋亡与衰老的药物筛选.     

Pyrophosphate Fructose-6-P 1-Phosphotransferase from Tomato Fruit : Evidence for Change during Ripening

Three forms of pyrophosphate fructose-6-phosphate 1-phosphotransferase (PFP) were purified from both green and red tomato (Lycopersicon esculentum) fruit: (a) a classical form (designated Q₂) containing α- (66 kilodalton) and β- (60 kilodalton) subunits; (b) a form (Q₁) containing a β-doublet subunit; and (c) a form (Q₀) that appeared to contain a β-singlet subunit. Several lines of evidence suggested that the different forms occur under physiological conditions. Q₂ was purified to apparent electrophoretic homogeneity; Q₁ and Q₀ were highly purified, but not to homogeneity. The distribution of the PFP forms from red (versus green) tomato was: Q₂, 29% (90%); Q₁, 47% (6%); and Q₀, 24% (4%). The major difference distinguishing the red from the green tomato enzymes was the fructose-2,6-bisphosphate (Fru-2,6-P₂)-induced change in K_m for fructose-6-phosphate (Fru-6-P), the `green forms' showing markedly enhanced affinity on activation (K<sub>m</sub> decrease of 7-9-fold) and the `red forms' showing either little change (Q₀, Q₁) or a relatively small (2.5-fold) affinity increase (Q₂). The results extend our earlier findings with carrot root to another tissue and indicate that forms of PFP showing low or no affinity increase for Fru 6-P on activation by Fru-2,6-P₂ (here Q₁ and Q₀) are associated with sugar storage, whereas the classical form (Q₂), which shows a pronounced affinity increase, is more important for starch storage.