氦氖激光对绵羊精清中酶活性效应的研究

本文研究结果表明低剂量的氦氖激光可以提高绵羊精清中GOT和LDH酶的活性,并对其机制作了初步的探讨。     

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.     

Intravital lipid droplet labeling and imaging reveals the phenotypes and functions of individual macrophages in vivo

Macrophages play pivotal roles in the maintenance of tissue homeostasis. However, the reactivation of macrophages toward proinflammatory states correlates with a plethora of inflammatory diseases, including atherosclerosis, obesity, neurodegeneration, and bone marrow (BM) failure syndromes. The lack of methods to reveal macrophage phenotype and function in vivo impedes the translational research of these diseases. Here, we found that proinflammatory macrophages accumulate intracellular lipid droplets (LDs) relative to resting or noninflammatory macrophages both in vitro and in vivo, indicating that LD accumulation serves as a structural biomarker for macrophage phenotyping. To realize the staining and imaging of macrophage LDs in vivo, we developed a fluorescent fatty acid analog-loaded poly(lactic-co-glycolic acid) nanoparticle to label macrophages in mice with high efficiency and specificity. Using these novel nanoparticles, we achieved in situ functional identification of single macrophages in BM, liver, lung, and adipose tissues under conditions of acute or chronic inflammation. Moreover, with this intravital imaging platform, we further realized in vivo phenotyping of individual macrophages in the calvarial BM of mice under systemic inflammation. In conclusion, we established an efficient in vivo LD labeling and imaging system for single macrophage phenotyping, which will aid in the development of diagnostics and therapeutic monitoring. Moreover, this method also provides new avenues for the study of lipid trafficking and dynamics in vivo.Supplementary key words: macrophage, inflammation, lipid droplet, nanoparticle delivery, in vivo imaging, fatty acid analog, bone marrow, systemic inflammation, lipid trafficking, biomarker

Macrophages, a type of immune cells, almost reside in all tissues of body, from the skin to the bone marrow (BM) (1). Macrophages have remarkable plasticity, and they can be activated into specific subtypes by modifying their physiology and functions in response to local environmental cues. Activated macrophages are commonly divided into proinflammatory killing subtype and anti-inflammatory repairing subtype. Proinflammatory macrophages responding to bacteria, IFN-γ, and lipopolysaccharide (LPS) are involved in host defense and inflammation, whereas anti-inflammatory macrophages responding to interleukin-4 (IL-4), IL-10, and IL-13 play a pivotal role in tissue homeostasis and remodeling (2). Increasing evidence indicates that the reactivation of macrophages toward proinflammatory states under diverse kinds of stress is correlated with a plethora of inflammatory diseases, such as atherosclerosis, diabetes, obesity, rheumatoid arthritis, neurodegeneration, and BM failure syndromes (3, 4). Thus, characterization of macrophage activation status and the underlying molecular mechanism in situ will help elucidate their functions in these diseases; however, in vivo analysis of the macrophage activation status in their native multicellular microenvironment is challenging.Although lipid droplets (LDs) have been initially described as intracellular fat storage organelles in adipocytes, increasing studies indicate that myeloid cells also form LDs under inflammation and stress (5, 6). Macrophages, as the effector cells of innate immunity, are found to form LDs to support their host defense when exposed to pathogens, such as parasites, bacteria, and viruses (7, 8, 9, 10, 11). However, abnormal LD accumulation in tissue-resident macrophages correlates with the pathogenesis of various inflammatory diseases. For instance, foam cells in atherosclerotic lesions can maintain the local inflammatory response by secreting proinflammatory cytokines (12, 13, 14). Moreover, LD-accumulating microglia contribute to neurodegeneration by producing high levels of reactive oxygen species (ROS) and secreting proinflammatory cytokines (15). These findings indicate that LD accumulation might be a hallmark of macrophages with proinflammatory functions.In this study, based on the typical activation of in vitro BM-derived macrophages, we find that proinflammatory M_{(LPS + IFN-γ)} macrophages are characterized by LD accumulation, whereas resting macrophages and anti-inflammatory M_(IL-4) and M_(IL-10) macrophages do not contain any LDs. These features also hold for Matrigel plug-recruited macrophages and tissue-resident macrophages in mice. These findings demonstrate that LD accumulation could serve as a morphological index to distinguish proinflammatory macrophages from others.It is feasible to distinguish LD-containing cells using imaging techniques, which has translational potential for identification of proinflammatory macrophages in vivo. However, current techniques for LD visualization are traditional in vitro staining method, and in vivo staining and imaging of LD in individual macrophages remains a challenge. Through nanocarrier screening, we selected the poly(lactic-co-glycolic acid) (PLGA) nanoparticles (NPs) as nanocarrier to deliver the lipophilic carbocyanine dye (DiI_C18(5) solid (1,1''-dioctadecyl-3,3,3'',3''-tetramethylindodicarbocyanine, 4-chlorobenzenesulfonate salt) [DiD]) and lipid staining dye (C1-BODIPY 500/510-C12) into macrophages. Using these dual fluorescence-labeled PLGA NPs, we achieved in situ and in vivo functional identification of single macrophages in various tissues under systemic or local inflammatory stress. Collectively, this study establishes an efficient in vivo labeling and imaging system of intracellular LDs for phenotyping the activation status and functions of individual macrophages in their dynamic niche, which is pivotal for disease diagnosis and preclinical research.     

增施氮肥能够缓解麦田土壤线虫群落对O3浓度升高的响应

在辽宁沈阳农田生态系统国家野外科学观测研究站,利用运行2a的开顶式气室,研究了臭氧(O3)浓度升高和不同氮肥施用水平对土壤线虫群落的影响。结果表明:(1)O3浓度升高降低了成熟期小麦根生物量。O3浓度升高和不同氮肥施用水平的交互作用改变了小麦成熟期土壤微生物生物量碳、氮和水溶性有机碳的含量。低氮条件下,O3浓度升高降低了土壤微生物生物量碳、氮和水溶性有机碳的含量;而高氮条件下则表现出相反的趋势。(2)O3浓度升高和不同氮肥施用水平对土壤线虫总数没有产生显著影响,而在灌浆期,食细菌线虫和食真菌线虫中c-p值为4(Ba4 and Fu4)的功能团对O3浓度升高和不同氮肥施用水平的响应敏感;与对照相比,不同氮处理中,O3浓度升高均降低了灌浆期Ba4功能团线虫的数量。灌浆期,O3浓度升高条件下,与对照相比Fu4功能团线虫数量在高氮条件下表现出增加的趋势,而在低氮条件表现出降低的趋势。(3)O3浓度升高和不同氮肥施用水平的交互作用显著影响了小麦灌浆期线虫的成熟度指数(MI)和结构指数(SI)。与对照相比,线虫成熟度指数和结构指数在低氮条件下随O3浓度升高而降低;而在高氮条件下随O3浓度升高而升高。上述结果表明,氮肥的施用能够缓解O3浓度升高对土壤食物网的扰动。     

低温导致中华蜜蜂后翅翅脉的新变异

在封盖发育期,将中华蜜蜂Apis cerana cerana封盖子放入24℃低温环境中分别处理24、48、72 h,再恢复到正常的发育温度35℃至羽化,或将封盖子放在30℃低温中培养至羽化,发现羽化后的蜜蜂后翅翅脉发生明显变异。发现翅脉变异有3种类型,翅脉增加、翅脉突出和翅脉缺失。后翅翅脉增加的变异只发现一种,基室、缘室共有的径分脉与基室、盘室共有的中脉之间,新增径中横脉。新增的径中横脉发生的程度不同,有的在径分脉和中脉相对的位置相向伸出,有的形成完整的新脉,新增的径中横脉与原径中横脉相似,在末端均有"弱化点"。由于原有径中横脉较短,一直忽略其存在,这次径中横脉的确定,可以判定原来公认的"中脉分叉"应分别是径分脉和中脉延伸出来的两条翅脉。后翅突出的变异有2种,中脉向基室突出,径分脉向基室突出。翅脉突出的程度不同,有的略突出,有的明显突出,有的伸出翅脉较长。中脉突出均发生在中脉的转折点上,且此类翅脉变异,仅发生在雄蜂样本中,在工蜂样本中还未见。后翅翅脉缺失的变异有2种,发生在肘臀横脉和中脉。肘臀横脉"弱化点"处缺失。将中华蜜蜂雄蜂封盖子放入低温30℃恒温培养箱中发育,发现2个羽化的雄蜂样本发生肘臀横脉在"弱化点"处缺失。中脉向盘室伸出的部分发生缺失。翅脉缺失的程度不同,有的明显缩短,有的仅存痕迹,有的缺失大部分或完全缺失。中脉向盘室伸出的部分是东方蜜蜂Apis cerana区别于西方蜜蜂Apis mellifera的主要形态特征之一,如果东方蜜蜂这段翅脉全部缺失,此部位的形态与西方蜜蜂完全相同。这意味着这个翅脉作为区分东方蜜蜂和西方蜜蜂的翅脉特征将面临挑战。对蜜蜂种质资源鉴定,蜜蜂的系统发育和进化生物学等领域的研究将具有重要影响。温度影响翅脉发育的深入研究,为研究蜜蜂翅脉发育过程,揭示翅脉发育调控机制,以及蜜蜂发育的温度适应生理机制提供一个新思路。