利用大肠杆菌工程菌廉价高效生产聚羟基丁酸酯

利用大肠杆菌生产聚羟基脂肪酸酯是近来国际上生物可降解塑料的研究热点,本研究通过对适宜于聚羟基脂肪酸酯生产的大肠杆菌菌株的选择和碳源利用试验,初步确立了大肠杆菌代谢工程改造生产聚羟基脂肪酸酯的基础。并在此基础上,通过对大肠杆菌磷酸烯醇式丙酮酸葡萄糖转移酶系统的改造和工程菌环境诱导系统的应用,解决了大肠杆菌工程菌无法同时利用多种碳源合成聚羟基脂肪酸酯的难题。发酵试验证明,工程化改造的大肠杆菌利用廉价底物在5L发酵罐中分批培养32h后,菌体终浓度能够达到8.24g/L,聚羟基脂肪酸酯占细胞干重的84.6%。     

Sensitive and specific molecular detection of Burkholderia pseudomallei, the causative agent of melioidosis, in the soil of tropical northern Australia

Burkholderia pseudomallei, the cause of the severe disease melioidosis in humans and animals, is a gram-negative saprophyte living in soil and water of areas of endemicity such as tropical northern Australia and Southeast Asia. Infection occurs mainly by contact with wet contaminated soil. The environmental distribution of B. pseudomallei in northern Australia is still unclear. We developed and evaluated a direct soil B. pseudomallei DNA detection method based on the recently published real-time PCR targeting the B. pseudomallei type III secretion system. The method was evaluated by inoculating different soil types with B. pseudomallei dilution series and by comparing B. pseudomallei detection rate with culture-based detection rate for 104 randomly collected soil samples from the Darwin rural area in northern Australia. We found that direct soil B. pseudomallei DNA detection not only was substantially faster than culture but also proved to be more sensitive with no evident false-positive results. This assay provides a new tool to detect B. pseudomallei in soil samples in a fast and highly sensitive and specific manner and is applicable for large-scale B. pseudomallei environmental screening studies or in outbreak situations. Furthermore, analysis of the 104 collected soil samples revealed a significant association between B. pseudomallei-positive sites and the presence of animals at these locations and also with moist, reddish brown-to-reddish gray soils.     

Mice deficient in involucrin, envoplakin, and periplakin have a defective epidermal barrier

The cornified envelope is assembled from transglutaminase cross-linked proteins and lipids in the outermost epidermal layers and is essential for skin barrier function. Involucrin, envoplakin, and periplakin form the protein scaffold on which the envelope assembles. To examine their combined function, we generated mice deficient in all three genes. The triple knockouts have delayed embryonic barrier formation and postnatal hyperkeratosis (abnormal accumulation of cornified cells) resulting from impaired desquamation. Cornified envelopes form but are ultrastructurally abnormal, with reduced lipid content and decreased mechanical integrity. Expression of proteases is reduced and the protease inhibitor, serpina1b, is highly upregulated, resulting in defective filaggrin processing and delayed degradation of desmoglein 1 and corneodesmosin. There is infiltration of CD4⁺ T cells and a reduction in resident γδ⁺ T cells, reminiscent of atopic dermatitis. Thus, combined loss of the cornified envelope proteins not only impairs the epidermal barrier, but also changes the composition of T cell subpopulations in the skin.     

间充质干细胞向成骨细胞分化的信号通路

间充质干细胞具有向成骨细胞分化的潜能,可体外分离、培养和扩增,是骨组织工程中理想的种子细胞。近年的研究表明间充质干细胞的成骨分化受到多种信号通路的调控,现就其中研究较为深入的MAPK和Notch通路的情况作一简要综述。     

桑叶水提浸膏的抑菌作用研究

研究桑叶水提浸膏对五种食品常见污染菌的抑菌作用。通过滤纸片扩散法测定其相对抑菌活性。结果表明,桑叶水提浸膏对金黄色葡萄球菌有较强的抑制作用,对大肠杆菌作用次之,对枯草芽孢杆菌、荧光假单孢杆菌、巨大芽孢杆菌抑菌效果较差。桑叶水提浸膏对金黄色葡萄球菌的最小抑菌浓度是5%,抑菌作用受pH和温度的影响较大,在酸性条件下抑菌效果较好,低于60℃处理样品对抑菌效果影响不大,但高于80℃处理,样品抑菌活性明显降低。     

Heterooligomerization of human dopamine receptor 2 and somatostatin receptor 2 Co-immunoprecipitation and fluorescence resonance energy transfer analysis

Somatostatin and dopamine receptors are well expressed and co-localized in several brain regions, suggesting the possibility of functional interactions. In the present study we used a combination of pharmacological, biochemical and photobleaching fluorescence resonance energy transfer (pbFRET) to determine the functional interactions between human somatostatin receptor 2 (hSSTR2) and human dopamine receptor 2 (hD2R) in both co-transfected CHO-K1 or HEK-293 cells as well as in cultured neuronal cells which express both the receptors endogenously. In monotransfected CHO-K1 or HEK-293 cells, D2R exists as a preformed dimer which is insensitive to agonist or antagonist treatment. In control CHO-K1 cells stably co-transfected with hD2R and hSSTR2, relatively low FRET efficiency and weak expression in co-immunoprecipitate from HEK-293 cells suggest the absence of preformed heterooligomers. However, upon treatment with selective ligands, hD2R and hSSTR2 exhibit heterodimerization. Agonist-induced heterodimerization was accompanied by increased affinity for dopamine and augmented hD2R signalling as well as prolonged hSSTR2 internalization. In contrast, cultured striatal neurons display constitutive heterodimerization between D2R and SSTR2, which were agonist-independent. However, heterodimerization in neurons was completely abolished in the presence of the D2R antagonist eticlopride. These findings suggest that hD2R and hSSTR2 operate as functional heterodimers modulated by ligands in situ, which may prove to be a useful model in designing new therapeutic drugs.     

哀牢山亚热带常绿阔叶林土壤温度时空分布对模拟增温的响应

为把握森林不同深度的土壤温度对区域气候变暖的响应,评估气候变暖对亚热带森林土壤呼吸的影响,利用在哀牢山亚热带常绿阔叶林中设置的土壤增温和土壤呼吸人工控制实验,对2011—2013年的对照样地和增温样地不同深度的土壤温度实测数据进行了分析,结果表明:区域气候变暖导致的温度升高不改变土壤温度的年变化和日变化特征;干季的增温效应大于雨季;同一深度增温效应具有一定的年变化,但日变化不明显;增温效应在土壤表层较大,1—4月明显,2月最高,增温可达3℃;5 cm深度的平均增温效应在2℃左右,且年变化较小;随着深度的增加,增温效应呈现对数降低趋势;如果以0.5℃为增温效应的阈值,推算可得出:干季的影响深度为3.82 m,雨季可达12.04 m,年均为6.58 m。     

Homeostatic defects in interleukin 18-deficient mice contribute to protection against the lethal effects of endotoxin

Toll-like receptor-4-lipopolysaccharide (LPS)-mediated inflammation is used to delineate signals involved in cross-talk between antigen-presenting cells (APCs) and lymphocytes such as natural killer (NK) cells. Following APC stimulation and cytokine release, NK cells produce interferon (IFN)-γ. High levels of LPS induce endotoxicosis, a systemic inflammatory disease in which IFN-γ causes significant morbidity and mortality. Several studies have highlighted the role of interleukin (IL)-18, IL-1β, IL-17A and IFN-γ in the development of endotoxicosis, but whether these cytokines interact with each other is yet to be determined. Our data demonstrate that IL-18 and IL-17A have important roles in NK cell IFN-γ production during endotoxicosis. Importantly, we provide the first evidence that IL-18 also has a role in IL-17A production by T-cell receptor (TCR)-δ cells. Furthermore, we demonstrate that IL-18-deficient mice have a defect in γδ T-cell homeostasis and IL-1β production, both of which can contribute to the development of disease through induction of IL-17A. These results reveal novel requirements for IL-18 in innate immune cell homeostasis and activation, demonstrating that the role of IL-18 in innate immunity occurs at a level other than activation.     

白腐菌胞外提取液与纤维素酶对玉米秸秆的协同酶解研究

通过评价几株白腐菌固体发酵不同时间的胞外提取液与纤维素酶同时加入对玉米秸秆酶解产糖的影响,得到一个较高糖化效果的胞外降解体系,并对其相关影响因素进行研究,进而初探其协同酶解的机制.结果表明,Echinodontium taxodii 2538固体培养5 d的胞外提取液与纤维素酶形成的胞外降解体系的协同酶解效果较好,酸处...