酪氨酸羟化酶基因载体－pCMVTH 质粒的构建及在大鼠骨骼肌内的表达

为用转基因方法治疗巴金森氏病大鼠模型,本研究采用分子克隆技术,将合成多巴胺的关键酶－酪氨酸羟化酶（ＴＨ）的基因,克隆进入以巨细胞病毒ＣＭＶ为启动子的载体质粒内,经限制性内切酶定位分析证实该重组的ＤＮＡ质粒的可靠性。携带ＴＨ基因的ＰＣＭＶＴＨ质粒以ＬＩＰＯ－ＦＥＣＴＩＮ介导,在培养的原代骨骼肌细胞中高效表达。本研究为进一步用转基因的细胞植入脑内以治疗巴金森氏病打下一定基础。     

Activation of a mechanosensitive BK channel by membrane stress created with amphipaths

Some BK channels are activated in response to membrane stretch. However, it remains largely unknown which membrane component transmits forces to the channel and which part of the channel senses the force. Recently, we have shown that a BK channel cloned from chick heart (named SAKCa channel) is a stretch activated channel, while deletion of a 59 amino acids splice insert (STREX) located in the cytoplasmic side, abolishes its stretch-sensitivity. This finding raised a question whether stress in the bilayer is crucial for the mechanical activation of the channel. To address this question we examined the effects of membrane perturbing amphipaths on the stretch activation of the SAKCa channel and its STREX-deletion mutant. We found that both anionic amphipath trinitrophenol (TNP) and cationic amphipath chlorpromazine (CPZ) could dose-dependently activate the channel by leftward shifting the voltage activation curve when applied alone. In contrast, TNP and CPZ compensated each other's effect when applied sequentially. These results can be understood in the framework of the bilayer couple hypothesis, suggesting that stress in the plasma membrane can activate the SAKCa channel. Interestingly, the STREX-deletion mutant channel has much less sensitivity to the amphipaths, suggesting that STREX acts as an intermediate structure that can indirectly convey stress in the membrane to the gate of the SAKCa channel via an unidentified membrane associated protein(s) that can detect or transmit stress in the membrane.     

Biodiversity and ecosystem stability across scales in metacommunities

Although diversity–stability relationships have been extensively studied in local ecosystems, the global biodiversity crisis calls for an improved understanding of these relationships in a spatial context. Here, we use a dynamical model of competitive metacommunities to study the relationships between species diversity and ecosystem variability across scales. We derive analytic relationships under a limiting case; these results are extended to more general cases with numerical simulations. Our model shows that, while alpha diversity decreases local ecosystem variability, beta diversity generally contributes to increasing spatial asynchrony among local ecosystems. Consequently, both alpha and beta diversity provide stabilising effects for regional ecosystems, through local and spatial insurance effects respectively. We further show that at the regional scale, the stabilising effect of biodiversity increases as spatial environmental correlation increases. Our findings have important implications for understanding the interactive effects of global environmental changes (e.g. environmental homogenisation) and biodiversity loss on ecosystem sustainability at large scales.     

Elevated sodium chloride concentrations enhance the bystander effects induced by low dose alpha-particle irradiation

Previous studies have shown that high NaCl can be genotoxic, either alone or combined with irradiation. However, little is known about the relationship between environmental NaCl at elevated conditions and radiation-induced bystander effects (RIBE). RIBE, which has been considered as non-targeted bystander responses, has been demonstrated to occur widely in various cell lines. In the present study, RIBE under the elevated NaCl culture condition was assessed in AG 1522 cells by both the induction of gamma-H2AX, a reliable marker of DNA double-strand break (DSB) for the early process (<1h post irradiation), and the generation of micronuclei (MN), a sensitive marker for relative long process of RIBE. Our results showed that in the absence of irradiation, NaCl at elevated concentration such as 8.0, 9.0 and 10.0g/L did not significantly increase the frequency of gamma-H2AX foci-positive cells and the number of foci per positive cell comparing with that NaCl at a normal concentration (6.8g/L). However, with 0.2cGy alpha-particle irradiation, the induced fraction of gamma-H2AX foci-positive cells and the number of induced gamma-H2AX foci per positive cell were significantly increased in both irradiated and adjacent non-irradiated regions. Similarly, the induction of MN by 0.2cGy alpha-particle irradiation also increased with the elevated NaCl concentrations. With N(G)-methyl-l-arginine, an inhibitor of nitric oxide synthase, the induced fraction of foci-positive cells was effectively inhibited both in 0.2cGy alpha-particle irradiated and adjacent non-irradiated regions under either normal or elevated NaCl conditions. These results suggested that the cultures with elevated NaCl medium magnified the damage effects induced by the low dose alpha-particle irradiation and nitric oxide generated by irradiation was also very important in this process.     

Dynamics of Membrane Tethers Reveal Novel Aspects of Cytoskeleton-Membrane Interactions in Axons

Mechanical properties of cell membranes are known to be significantly influenced by the underlying cortical cytoskeleton. The technique of pulling membrane tethers from cells is one of the most effective ways of studying the membrane mechanics and the membrane-cortex interaction. In this article, we show that axon membranes make an interesting system to explore as they exhibit both free membrane-like behavior where the tether-membrane junction is movable on the surface of the axons (unlike many other cell membranes) as well as cell-like behavior where there are transient and spontaneous eruptions in the tether force that vanish when F-actin is depolymerized. We analyze the passive and spontaneous responses of axonal membrane tethers and propose theoretical models to explain the observed behavior.     

Activation of a mechanosensitive BK channel by membrane stress created with amphipaths

Some BK channels are activated in response to membrane stretch. However, it remains largely unknown which membrane component transmits forces to the channel and which part of the channel senses the force. Recently, we have shown that a BK channel cloned from chick heart (named SAKCa channel) is a stretch activated channel, while deletion of a 59 amino acids splice insert (STREX) located in the cytoplasmic side, abolishes its stretch-sensitivity. This finding raised a question whether stress in the bilayer is crucial for the mechanical activation of the channel. To address this question we examined the effects of membrane perturbing amphipaths on the stretch activation of the SAKCa channel and its STREX-deletion mutant. We found that both anionic amphipath trinitrophenol (TNP) and cationic amphipath chlorpromazine (CPZ) could dose-dependently activate the channel by leftward shifting the voltage activation curve when applied alone. In contrast, TNP and CPZ compensated each other's effect when applied sequentially. These results can be understood in the framework of the bilayer couple hypothesis, suggesting that stress in the plasma membrane can activate the SAKCa channel. Interestingly, the STREX-deletion mutant channel has much less sensitivity to the amphipaths, suggesting that STREX acts as an intermediate structure that can indirectly convey stress in the membrane to the gate of the SAKCa channel via an unidentified membrane associated protein(s) that can detect or transmit stress in the membrane.     

Effects of community structure on the species–area relationship in China's forests

The species–area relationship (SAR) is the oldest and most frequently documented law in ecology. In a community, the SAR is regulated by the abiotic environment and biotic interactions and depends on the individual–spatial distribution of species (ISD) and the species–abundance distribution (SAD). In this study, we explored the effects of aggregation of ISDs and unevenness of SADs on SARs in forests of China by comparing the empirical and simulated SARs of 32 nested plots distributed along an extensive latitudinal gradient. Both aggregation and unevenness affected the shape of SARs significantly: ISDs accounted for 12.6 ± 4.0% of the incremental increase in species richness with area, and SADs accounted for 18.7 ± 3.8 and 23.5 ± 3.9% under the broken‐stick model and even abundance model, respectively. Effects of both aggregation and unevenness decreased as temperature increased, suggesting that individuals of a species were spatially more aggregated than random, and the individuals among species were more discrepant from the null distribution (broken‐stick model and even abundance model in this study), in the cold than in the warm areas. Taken together, our results demonstrate that ISDs and SADs within communities can shape SARs, but these effects vary along latitudinal gradients, and are likely mediated by temperature.     

Isolation and Characterization of Two Novel Antibacterial Cyclic Hexapeptides from Streptomyces alboflavus 313

Two novel cyclic hexapeptides, designated NW-G08 (1) and NW-G09 (2), were isolated from the fermentation broth of Streptomyces alboflavus 313. Their structures were elucidated on the basis of extensive spectroscopic analysis, MS experiments, and chemical analysis. Their antibacterial activities against several strains of bacteria were evaluated by micro-broth dilution method. NW-G08 (1) and NW-G09 (2) were highly antibacterial against Gram-positive but not active against Gram-negative bactaria. The minimum inhibitory concentrations (MICs) of 1 against Bacillus cereus, Bacillus subtilis, and Staphylococcus aureus were 6.25, 6.25, and 1.56?μg/ml, respectively. It is worth noting that 1 and 2 exhibited much higher activities against methicillin-resistant Staphylococcus aureus (MRSA) than ampicillin, which implies that they might be potential candidates for the development of novel drugs against resistant pathogenic bacteria.     

Dielectric Properties Based Detection of Heavy Metal Contaminated Soil in the Frequency Range from 10 MHz to 1 GHz

Physical parameters based electromagnetic methods are promising technologies to detect contaminated sites. In these methods, the dielectric property is a key parameter. In this paper, we studied the dielectric characteristics of heavy metal contaminated soil. The chromium contaminated soil was made into samples, and the open-ended coaxial line was adopted as the measurement method. Experiments were conducted in the frequency band between 10 MHz and 1 GHz. The results showed that the complex permittivity, including the real part and the imaginary part, changes as the ionic content changes. Especially, at low frequencies (<50 MHz), the complex permittivity increases significantly with the increase of ionic content. In addition, it also could be seen that the water content of the soil also affects the complex permittivity. We proposed to adopt the drying method or the Time Domain Reflection method to determine the water content. The dielectric parameters are most affected by the ionic content after knowing the water content. Therefore, it is feasible to detect heavy metal contaminated sites based on dielectric properties.