利用合成的基因回路实现细胞水平上尿酸稳态控制的实验研究

目的:利用合成生物学方法构建尿酸介导的基因回路,在细胞水平上研究回路对尿酸稳态的调控作用。方法:以耐辐射异常球菌R1基因组中转录抑制物基因hucR及其结合位点基因hucO为基础,化学合成具有转录抑制功能的基因mUTs及其结合位点的8串联结构hucO8,构建基因回路;转染HeLa细胞,通过检测分泌型碱性磷酸酶(SEAP)的表达量来验证回路的作用原理和对尿酸的感应作用;在此基础上,用优化的黄曲霉菌尿酸氧化酶(Uox)基因smUox替换SEAP基因,转染HeLa细胞,通过检测转染前后培养基中尿酸浓度的变化,验证回路对尿酸的调节作用。结果:分别构建了优化的转录抑制物表达载体pcDNA3.1/V5-mUTs、报告基因表达载体pSEAP-hucO8、优化的黄曲霉菌Uox表达载体phucO8-smUox、pBudCE4.1-smUox,双向共表达载体pBudCE4.1-SEAP-mUTs、pBudCE4.1-mUTs-smUox;单独转染pBudCE4.1-SEAP-mUTs或共转染pSEAP-hucO8和pcDNA3.1/V5-mUTs,通过检测培养基中SEAP的表达量,证明双载体及单载体回路对尿酸的感应作用;用smUox替换SAEP基因后,通过检测转染48 h后培养基中尿酸含量的变化,证明双载体及单载体基因回路均具有一定的尿酸调节能力。结论:在细胞水平上,构建的双载体基因回路(phucO8-smUox、pcDNA3.1/V5-mUTs)和单载体基因回路(pBudCE4.1-mUTs-smUox)均可实现对尿酸的感应及调控作用,在一定范围内通过增加mUTs与hucO8的摩尔比,可以改变回路对尿酸的调控范围及调节程度。     

A novel anti-inflammatory role for spleen-derived interleukin-10 in obesity-induced hypothalamic inflammation

Obesity can be associated with systemic low-grade inflammation that contributes to obesity-related metabolic disorders. Recent studies raise the possibility that hypothalamic inflammation contributes to the pathogenesis of diet-induced obesity (DIO), while another study reported that obesity decreases the expression of pro-inflammatory cytokines in spleen. The following study examines the hypothesis that obesity suppresses the splenic synthesis of the anti-inflammatory cytokine, interleukin (IL)-10, thereby resulting in chronic hypothalamic inflammation. The results showed that due to oxidative stress or apoptosis, the synthesis of splenic IL-10 was decreased in DIO when compared with non-obesity rats. Splenectomy (SPX) accelerated DIO-induced inflammatory responses in the hypothalamus. Interestingly, SPX suppressed the DIO-induced increases in food intake and body weight and led to a hypothalamic pro-inflammatory state that was similar to that produced by DIO, indicating that hypothalamic inflammation exerts a dual effect on energy metabolism. These SPX-induced changes were inhibited by the systemic administration of IL-10. Moreover, SPX had no effect on hypothalamic inflammatory responses in IL-10-deficient mice. These data suggest that spleen-derived IL-10 plays an important role in the prevention of hypothalamic inflammation and may be a therapeutic target for the treatment of obesity and hypothalamic inflammation.     

Effects of prenatal morphine on hypothalamic metabolism of neurotransmitters and gonadal and adrenal activities,during the early postnatal period in the rat

It is noteworthy that exposure to opiates during fetal development results in permanent changes in adults related to morphological, behavioral and biochemical measures; however little is known concerning the effects of such drugs in early postnatal life. We investigated in newborn rats the effects of prenatal morphine-exposure on both—the hypothalamic metabolism of norepinephrine (NE), serotonin (5 HT) and neuropeptide Y (NPY)—the activity of the hypothalamo-pituitary gonadal and adrenal axes. In a previous study performed in newborns of untreated mothers, we reported some sex-dependent changes in the metabolism of NE, 5 HT and NPY in the hypothalamus and an early activation of the gonadostimulating function and of the corticostimulating one. In control newborns from saline-treated mothers, a slight increase in the hypothalamic metabolism of NE (males) and 5 HT (males and females) was observed and it was comparable in both sexes. On the other hand, the hypothalamic content of NPY was unaffected in early postnatal period in newborn males as well as in females. These changes observed on hypothalamic metabolisms are temporally correlated with the early postnatal activation of the corticostimulating function in neonates of both sexes and that of the gonadostimulating one, mainly in males. Prenatal morphine exposure altered the hypothalamic metabolism of 5 HT which was increased mainly in newborn females but did not affect either the metabolism of NE or the NPY content of the hypothalamus. The more drastic effect of the prenatal morphine treatment is the atrophy and hypoactivity of the adrenals in newborns of both sexes at birth time and during the early postnatal period. In contrast morphine did not impair postnatal surge of the plasma testosterone level in male pups as well as late and slight increase of plasma estradiol in female ones.     

一种昆虫呼吸代谢测定方法的改进

为了探索精准、简易的昆虫呼吸代谢测定办法,本文基于Sable小动物呼吸测量系统,比较了应用Sable呼吸测量系统配置的8通道气路转换器呼吸室和采用鲁尔接头注射器作为替代呼吸室测试昆虫的呼吸代谢。结果表明:应用测量系统自带呼吸室和应用替代呼吸室检测棉铃虫蛹O_2消耗量(前者为0.2425(±0.0143) mL/g·h,后者为0.2389(±0.0146) mL/g·h)和CO_2释放量(前者为0.1562(±0.0098) mL/g·h,后者为0.1639(±0.0092) mL/g·h),两种测试方法无显著差异。与采用系统配置8通道气路转换器和自带呼吸室每测试7个样本耗时2.30 h相比较,应用替代呼吸室测试21个样本仅耗时2.75 h,明显节省测试时间。应用替代呼吸室,从呼出二氧化碳动态亦可以区分黑纹粉蝶不同虫态或不同发育状态的呼吸代谢差异。通过对两种测试方法的分析,推荐应用鲁尔接头注射器作为替代呼吸室的改进方法进行昆虫呼吸代谢生理的研究。     

Systems and synthetic biology approaches in understanding biological oscillators

Background: Self-sustained oscillations are a ubiquitous and vital phenomenon in living systems. From primitive single-cellular bacteria to the most sophisticated organisms, periodicities have been observed in a broad spectrum of biological processes such as neuron firing, heart beats, cell cycles, circadian rhythms, etc. Defects in these oscillators can cause diseases from insomnia to cancer. Elucidating their fundamental mechanisms is of great significance to diseases, and yet challenging, due to the complexity and diversity of these oscillators. Results: Approaches in quantitative systems biology and synthetic biology have been most effective by simplifying the systems to contain only the most essential regulators. Here, we will review major progress that has been made in understanding biological oscillators using these approaches. The quantitative systems biology approach allows for identification of the essential components of an oscillator in an endogenous system. The synthetic biology approach makes use of the knowledge to design the simplest, de novo oscillators in both live cells and cell-free systems. These synthetic oscillators are tractable to further detailed analysis and manipulations. Conclusion: With the recent development of biological and computational tools, both approaches have made significant achievements.     

基于地质灾害敏感性的生态安全格局关键区识别与修复——以济南市为例

区域生态安全格局构建对提升生态系统服务功能提供了重要路径,同时统筹各种生态要素进行生态保护与修复分区也是新时期做好生态修复的重要举措。以济南市为例,基于现状生态系统类型分布,聚焦生态本底和地质灾害敏感性的特征,基于形态学空间格局分析方法和自然保护区结合进行生态源地提取。采用夏季降水、植被覆盖度、坡度3个地质灾害敏感性因子修正基本生态阻力面。并采用最小成本路径方法（Least-Cost Path method,LCP）提取生态廊道,构建了市域的生态安全格局。采用电路理论进行生态关键区域（生态"夹点"和生态障碍点）的识别,进一步划分生态修复改善区,并对此提出针对性的生态保护修复策略和工程措施。研究表明：1）市域生态源地的个数为35个,面积为567.15 km²,主要类型为林地和草地。空间上主要分布南部山区。生态廊道818.42 km,平均廊道长度为12.99 km,廊道分布存在较为明显的空间分布差异性,整体呈现出"一屏、一带、三轴"的生态安全格局。2）识别的生态修复关键区包含生态"夹点"25处,历城区生态"夹点"分布最为密集。全市亟需修复的生态障碍点共34处,面积为6.90 km²,主要分布章丘区。生态改善区共识别2994.84 km²,近期亟需修复的面积为96.1 km²,主要分布在长清区、历城区、莱芜区。3）通过对比生态修复关键区和现状土地利用类型,因地适宜的制定了生态修复策略与工程措施布置指引方向。研究结果可为济南市国土空间生态修复规划提供一定的技术支撑,同时也可为其他地质灾害敏感性区域的生态修复规划提供指引。     

The transient and constitutive inflammatory signaling in tumorigenesis

Comment on: Rokavec M, et al. Mol Cell 2012; 45:777-89.     

Activity-dependent local protection and lateral inhibition control synaptic competition in developing mitral cells in mice

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Modifications of neuronal activity in the rat hypothalamic supraoptic nucleus after long-lasting vibrational stimulation

We analyzed the background impulse activity (BIA) generated by neurons of the rat hypothalamic supraoptic nucleus in the norm and under conditions of long-lasting vibrational stimulation (exposure 5, 10, or 15 days). Distributions of neurons by the level of regularity, dynamics of discharge trains, form of histograms of interspike intervals (ISIs), as well as distributions of neurons by the BIA frequency ranges, were studied. We also calculated the mean frequency of impulsation of the neurons under study and the coefficient of variation of ISIs. After vibrational influences, we found modifications of both the internal structure of the recorded spike trains and the mean frequency of impulsation within the entire studied group and different frequency subgroups. Neirofiziologiya/Neurophysiology, Vol. 38, No. 3, pp. 224–230, May–June, 2006.