供核细胞对体细胞核移植效率的影响

利用体细胞核移植技术克隆动物、生产转基因家畜具有极大的应用潜力。然而,核移植效率低下、克隆后代形态异常等问题仍然制约着体细胞核移植技术的产业化进展。影响体细胞核移植效率的因素很多,该文着重从供核细胞的类型、细胞体外培养、细胞凋亡及转基因操作等方面阐述其对体细胞核移植效率的影响。     

人同源盒基因NKX3.1对前列腺癌细胞的诱导凋亡作用

构建人同源盒基因NKX3.1 cDNA真核表达载体,研究其在前列腺癌细胞PC-3、LNCaP 中的表达及对细胞的促凋亡作用.以人前列腺癌细胞LNCaP细胞中的总RNA为模板,RT-PCR扩增NKX3.1基因全长编码片段,将NKX3.1 cDNA重组到真核表达载体pcDNA3.1（+）中; 将pcDNA3.1-NKX3.1表达载体瞬时转染前列腺癌细胞PC-3和LNCaP 细胞,用RT-PCR和Western印迹检测NKX3.1 cDNA在转录水平和蛋白水平的表达;绘制细胞生长曲线,观察NKX3.1对前列腺癌细胞增殖的抑制作用;用DNA/ladder和流式细胞术检测NKX3.1对前列腺癌细胞凋亡的影响,进一步用RT PCR检测凋亡相关基因caspase3、caspase8、caspase9、Apaf1、survivin和Bcl2表达的变化.人同源盒基因NKX3.1 cDNA真核表达载体pcDNA3.1-NKX3.1经酶切及测序鉴定正确. pcDNA3.1-NKX3.1转染PC-3和LNCaP细胞后,经RT-PCR和Western印迹证明能有效表达NKX3.1.生长曲线显示,前列腺癌细胞转染NKX3.1 cDNA后细胞增殖受到抑制;前列腺癌细胞转染NKX3.1 cDNA 48 h后,DNA电泳呈现具有凋亡特征的DNA ladder;流式细胞术检测出现明显凋亡峰;RT-PCR检测凋亡相关基因.结果显示,caspase3、caspase8、caspase9基因表达明显增加,Bcl2基因表达明显减少.本研究成功构建了真核表达载体pcDNA3.1 NKX3.1, 转染PC3和LNCaP细胞后能有效表达,并对细胞具有诱导凋亡作用     

Effect of temperature and free ammonia on nitrification and nitrite accumulation in landfill leachate and analysis of its nitrifying bacterial community by FISH

The cause of seasonal failure of a nitrifying municipal landfill leachate treatment plant utilizing a fixed biofilm was investigated by wastewater analyses and batch respirometric tests at every treatment stage. Nitrification of the leachate treatment plant was severely affected by the seasonal temperature variation. High free ammonia (NH3-N) inhibited not only nitrite oxidizing bacteria (NOB) but also ammonia oxidizing bacteria (AOB). In addition, high pH also increased free ammonia concentration to inhibit nitrifying activity especially when the NH4-N level was high. The effects of temperature and free ammonia of landfill leachate on nitrification and nitrite accumulation were investigated with a semi-pilot scale biofilm airlift reactor. Nitrification rate of landfill leachate increased with temperature when free ammonia in the reactor was below the inhibition level for nitrifiers. Leachate was completely nitrified up to a load of 1.5 kg NH4-N m(-3)d(-1) at 28 degrees C. The activity of NOB was inhibited by NH3-N resulting in accumulation of nitrite. NOB activity decreased more than 50% at 0.7 mg NH3-N L(-1). Fluorescence in situ hybridization (FISH) was carried out to analyze the population of AOB and NOB in the nitrite accumulating nitrifying biofilm. NOB were located close to AOB by forming small clusters. A significant fraction of AOB identified by probe Nso1225 specifically also hybridized with the Nitrosomonas specific probe Nsm156. The main NOB were Nitrobacter and Nitrospira which were present in almost equal amounts in the biofilm as identified by simultaneous hybridization with Nitrobacter specific probe Nit3 and Nitrospira specific probe Ntspa662.     

AMP-activated protein kinase (AMPK) positively regulates osteoblast differentiation via induction of Dlx5-dependent Runx2 expression in MC3T3E1 cells

This study examined the role of AMPK activation in osteoblast differentiation and the underlining mechanism. An AMPK activator (AICAR or metformin) stimulated osteoblast differentiation with increases in ALP and OC protein production as well as the induction of AMPK phosphorylation in MC3T3E1 cells. In addition, metformin induced the phosphorylation of Smad1/5/8 and expression of Dlx5 and Runx2, whereas compound C or dominant negative AMPK inhibited these effects. Transient transfection studies also showed that metformin increased the BRE-Luc and Runx2-Luc activities, which were inhibited by DN-AMPK or compound C. Down-regulation of Dlx5 expression by siRNA suppressed metformin-induced Runx2 expression. These results suggest that the activation of AMPK stimulates osteoblast differentiation via the regulation of Smad1/5/8-Dlx5-Runx2 signaling pathway.     

Mechanism of activation of human c-KIT kinase by internal tandem duplications of the juxtamembrane domain and point mutations at aspartic acid 816

The patients suffering from acidosis usually sign psychological deficits. The cerebral dysfunction is reportedly caused by an acid-induced functional impairment of GABAergic neurons; however, the role of pyramidal neurons in this process remains unclear. By using electrophysiological method and changing extracellular pH, we investigated the influence of acidic environment on pyramidal neurons in the cortical slices, such as their ability of firing spikes and response to synaptic inputs. A low pH of artificial cerebral spinal fluid elevates the responses of pyramidal neurons to excitatory synaptic inputs and their ability of encoding digital spikes, as well as reduces the signal transmission at GABAergic synapses. The elevated ability of neuronal spiking is associated with the decreases of refractory periods and threshold potentials. Therefore, acidosis deteriorates brain functions through making the activities between cortical pyramidal neurons and GABAergic neurons imbalanced toward the overexcitation of neural networks, a process similar to neural excitotoxicity.     

干旱胁迫下发菜光合作用相关基因差异表达分析

该研究以不同失水处理的发菜为研究材料,以充分吸水状态的发菜为对照,利用高通量测序技术和qRT PCR技术检测了干旱胁迫下发菜光合作用相关基因差异表达规律,并对光合色素和酶活在干旱胁迫下的变化进行了研究。结果表明：(1)发菜在不同程度干旱胁迫下有113个光合作用相关基因差异表达,其中失水30%、75%和100%的发菜分别有44个、74个和91个光合作用相关基因差异表达。(2)随着干旱胁迫程度的加深藻胆素、叶绿素a和类胡萝卜素含量逐渐降低,Rubisco活性随着干旱胁迫程度的增强先上升后下降,GAPDH活性随着干旱胁迫的增强呈现下降的趋势。研究表明,发菜通过光合作用相关基因的差异表达调控光合作用以适应干旱胁迫。该研究结果对进一步研究发菜干旱胁迫响应机制及其耐旱光合机理奠定了基础。     

Signaling-mediated cross-talk modulates swarming and biofilm formation in a coral pathogen Serratia marcescens

Interactions within microbial communities associated with marine holobionts contribute importantly to the health of these symbiotic organisms formed by invertebrates, dinoflagellates and bacteria. However, mechanisms that control invertebrate-associated microbiota are not yet fully understood. Hydrophobic compounds that were isolated from surfaces of asymptomatic corals inhibited biofilm formation by the white pox pathogen Serratia marcescens PDL100, indicating that signals capable of affecting the associated microbiota are produced in situ. However, neither the origin nor structures of these signals are currently known. A functional survey of bacteria recovered from coral mucus and from cultures of the dinoflagellate Symbiodinium spp. revealed that they could alter swarming and biofilm formation in S. marcescens. As swarming and biofilm formation are inversely regulated, the ability of some native α-proteobacteria to affect both behaviors suggests that the α-proteobacterial signal(s) target a global regulatory switch controlling the behaviors in the pathogen. Isolates of Marinobacter sp. inhibited both biofilm formation and swarming in S. marcescens PDL100, without affecting growth of the coral pathogen, indicative of the production of multiple inhibitors, likely targeting lower level regulatory genes or functions. A multi-species cocktail containing these strains inhibited progression of a disease caused by S. marcescens in a model polyp Aiptasia pallida. An α-proteobacterial isolate 44B9 had a similar effect. Even though ∼4% of native holobiont-associated bacteria produced compounds capable of triggering responses in well-characterized N-acyl homoserine lactone (AHL) biosensors, there was no strong correlation between the production of AHL-like signals and disruption of biofilms or swarming in S. marcescens.     

Biofiltration of paint solvent mixtures in two reactor types: overloading by hydrophobic components

Steady-state performance characteristics of a trickle bed reactor (TBR) and a biofilter (BF) in loading experiments with increasing toluene/xylenes inlet concentrations while maintaining a constant loading rate of hydrophilic components (methyl ethyl and methyl isobutyl ketones, acetone, and n-butyl acetate) of 4 g m⁻³ h⁻¹ were evaluated and compared, along with the systems’ dynamic responses. At the same combined substrate loading of 55 g m⁻³ h⁻¹ for both reactors, the TBR achieved more than 1.5 times higher overall removal efficiency (RE_W) than the BF. Increasing the loading rate of aromatics resulted in a gradual decrease of their REs. The degradation rates of acetone and n-butyl acetate were also inhibited at higher loads of aromatics, thus revealing a competition in cell catabolism. A step-drop in loading of aromatics resulted in an immediate increase of RE_W with variations in the TBR, while the new steady-state value in the BF took 6–7 h to achieve. The TBR consistently showed a greater performance than BF in removing toluene and xylenes. Increasing the loading rate of aromatics resulted in a gradual decrease of their REs. The degradation rates of acetone and n-butyl acetate were also lower at higher OL_AROM, revealing a competition in the cell catabolism. The results obtained are consistent with the proposed hypothesis of greater toxic effects under low water content, i.e., in the biofilter, caused by aromatic hydrocarbons in the presence of polar ketones and esters, which may improve the hydrocarbon partitioning into the aqueous phase.     

七星瓢虫生殖的观察

近年来,在我国各地区一个“以虫治虫”的群众运动正在蓬勃展开。实践证明它可作为“综合防治”总方针中的重要一环。以瓢治蚜已经在广大棉区收到较好的效果,深受广大贫下中农的欢迎。本文介绍七星瓢虫(Coccinella septempunctata L.)一些生殖特点,作为瓢虫饲养繁殖时参考。 观察实验是在北京、安阳两个地区进行的,室内和室外观察相结合,加上必要的镜检帮助了解一些机体内部的状况。实验材料七星瓢虫根据观察的需要,有的是室外采集的,有的是室内繁殖饲养的,或是室外采集室内饲养。所用饲料蚜虫,有菜蚜、桃蚜、麦蚜等。在观察实验过程中得到安阳县生防站同志们的热情帮助。