Unfolded Protein Response and Activated Degradative Pathways Regulation in GNE Myopathy

Although intracellular beta amyloid (Aβ) accumulation is known as an early upstream event in the degenerative course of UDP-N-acetylglucosamine 2-epimerase/N-acetylmannosamine kinase (GNE) myopathy, the process by which Aβdeposits initiate various degradative pathways, and their relationship have not been fully clarified. We studied the possible secondary responses after amyloid beta precursor protein (AβPP) deposition including unfolded protein response (UPR), ubiquitin proteasome system (UPS) activation and its correlation with autophagy system. Eight GNE myopathy patients and five individuals with normal muscle morphology were included in this study. We performed immunofluorescence and immunoblotting to investigate the expression of AβPP, phosphorylated tau (p-tau) and endoplasmic reticulum molecular chaperones. Proteasome activities were measured by cleavage of fluorogenic substrates. The expression of proteasome subunits and linkers between proteasomal and autophagy systems were also evaluated by immunoblotting and relative quantitative real-time RT-PCR. Four molecular chaperones, glucose-regulated protein 94 (GRP94), glucose-regulated protein 78 (GRP78), calreticulin and calnexin and valosin containing protein (VCP) were highly expressed in GNE myopathy. 20S proteasome subunits, three main proteasome proteolytic activities, and the factors linking UPS and autophagy system were also increased. Our study suggests that AβPP deposition results in endoplasmic reticulum stress (ERS) and highly expressed VCP deliver unfolded proteins from endoplasmic reticulum to proteosomal system which is activated in endoplasmic reticulum associated degradation (ERAD) in GNE myopathy. Excessive ubiquitinated unfolded proteins are exported by proteins that connect UPS and autophagy to autophagy system, which is activated as an alternative pathway for degradation.     

Effects of salinity and temperature stress on Ecophysiological characteristics of exotic cordgrass, Spartina alterniflora

Salintiy and temperature are two important ecological factors which affect the distribution and abundance of Spartina alerniflora Loisel. To find out how cordgrass adapts to the environmental conditions in the introduced range, we studied the dynamics of a series of important physiological components including superoxide dismutase (SOD), guaiacol peroxidase (POD), catalase (CAT), malondialdehyde (MDA), soluble sugar and free proline under different salinity and temperature stresses. The results showed that low NaCl concentration (lower than 100 mmol·L⁻¹) enhanced the growth of S. alterniflora. On the contrary, high NaCl concentration (higher than 100 mmol·L⁻¹) inhibited the growth of S. alterniflora. To a certain extent, S. alterniflora was able to be acclimated to the osmotic pressure created by external solution concentration by adjusting the activities of POD, SOD and CAT, and the contents of free proline and soluble sugar. S. alterniflora varied in its responses to environment in different parts of the plant under 5°C and 38°C temperature stress. Compared with roots, leaves accumulated more soluble sugar, and CAT activities in leaves were higher, whereas SOD and POD activities in leaves were much lower than those in roots.     

NaCl胁迫对黄花菜生长和生理特性的影响

为探明黄花菜的耐盐性及其生理机制,该试验以大同黄花菜为材料,采用砂培法,以正常营养液为对照（CK）,用不同浓度（50、100、150、200、250 mmol·L^-1）NaCl溶液浇灌大同黄花菜,分别于处理后5、10、15、20、25 d测定生长指标和生理指标,以明确NaCl胁迫对大同黄花菜生长、膜脂过氧化以及有机渗透调节物质含量的影响。结果表明：（1）随NaCl浓度提高,黄花菜根长和根系鲜质量先增大后减小,其他生长指标则逐渐显著降低,同时地上部含水量变化较小,根系含水量明显增加。（2）黄花菜叶片叶绿素a、叶绿素b和类胡萝卜素含量随NaCl浓度提高均明显降低。（3）随NaCl浓度提高,黄花菜叶片丙二醛含量和POD活性逐渐显著增加;抗坏血酸含量在胁迫后20 d明显增加,25 d时呈先增加后降低的变化趋势,并在150 mmol·L^-1 NaCl胁迫下达到最大值;SOD活性在处理后10 d先增加后降低,在200 mmol·L^-1 NaCl处理下达到最大值,15 d后随NaCl浓度提高而显著增加。（4）随NaCl浓度提高,叶片脯氨酸含量逐渐显著增加,可溶性糖含量明显降低,可溶性蛋白含量在短期内逐渐增加,在胁迫15 d后呈先增加后降低的变化趋势,在150 mmol·L^-1 NaCl下达到最大值。研究发现,NaCl胁迫对黄花菜叶片光合色素合成的抑制和过氧化伤害程度均随浓度增加而增大;植株自身抗氧化能力和渗透调节能力在盐胁迫下明显提高,一定程度上缓解了盐胁迫对其植株的伤害,但仍不足以消除胁迫带来的不利影响,使得黄花菜植株生长受到显著抑制;黄花菜对NaCl胁迫的耐性较强,植株在250 mmol·L^-1高盐胁迫下仍能存活。     

Accumulation and tolerance characteristics of lead in <Emphasis Type="Italic">Althaea rosea</Emphasis> Cav. and <Emphasis Type="Italic">Malva crispa</Emphasis> L.

Two ornamental plants of Althaea rosea Cav. and Malva crispa L. were exposed to various concentrations of lead (Pb) (0, 50, 100, 200 and 500 mg·kg^?1) for 70 days to evaluate the accumulating potential and the tolerance characteristics. The results showed that both plant species grown normally under Pb stress, and A. rosea had a higher tolerance than M. crispa, while M. crispa had a higher ability in Pb accumulation than A. rosea. Besides, lower Pb concentration (50 mg·kg^?1) stimulated the shoot biomass in both plant species. Pb accumulation in plants was consistent with the increase of Pb levels, and the main accumulation sites were the roots and the older leaves. In addition, the photosynthetic pigments content and chlorophyll fluorescence parameters were influenced by Pb stress. In such case, both of the plants could improve the activities of antioxidant enzymes of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase (APX), and the contents of the total soluble sugar and soluble protein, which reached the highest value at Pb 100 mg·kg^?1, as well as the accumulation of the total thiols (T-SH) and non-protein thiols (NP-SH) to adapt to Pb stress. Thus, it provides the theoretical basis and possibility for ornamental plants of A. rosea and M. crispa in phytoremediation of Pb contaminated areas.     

Resistance characteristics of Cedrus deodara and Sabina chinensis to heavy metal accumulation under different atmospheric conditions

Cedrus deodara and Sabina chinensis are widely planted in North China. The needles of C. deodara and S. chinensis were sampled in the urban, suburban, and rural districts of Tianjin where the atmospheric conditions are significantly different according to the environmental monitoring results. The Cu, Mn, Zn, and Pb concentrations in the samples were examined via ICP. The resistance indexes, and the malonic aldehyde (MDA), soluble sugar, and free proline levels were also determined. The Pearson coefficients between the resistance indexes and the heavy metal contents were analyzed to compare the two plants abilities to accumulate heavy metal and their resistance characteristics. The results indicated that the heavy metal concentrations had the following significant trend: urban areas > suburban areas > rural areas. In urban areas, the Mn, Zn, and Pb concentrations in C. deodara were as high as 2024.77 mg·kg^? 1, 2397.07 mg·kg^? 1, and 130.07 mg·kg^? 1, significantly higher than in S. chinensis. The Mn, Zn, and Pb concentrations in C. deodara were extremely significantly positively correlated (P < 0.01), but no significant correlations were noted in S. chinensis. The MDA, soluble sugar, and free proline concentrations in C. deodara increased as the heavy metal contents rose along the urban–rural gradient, and were positively correlated with the plant heavy metal contents. They were much higher than the contents in S. chinensis where no differences were noted among the sampling sites. In conclusion, the heavy metal resistance methods used by C. deodara and S. chinensis are different. C. deodara could absorb and accumulate many heavy metals, mainly through increased physiological resistance to stress. In contrast, S. chinensis resistance relied on avoiding contact with the metals and by reducing absorption. These differences are associated with the biological characteristics of C. deodara and S. chinensis, and are closely connected with their coniferous and morphological differences.     

药用植物浸提液抑制蛋白核小球藻生长的化感效应

研究了11种药用植物浸提液对水华藻种蛋白核小球藻的影响,结果显示:黄连、重楼、贯众、防己4种药用植物的浸提液均有抑藻效应。当药用植物浸提液的相对浓度为1g/L处理时,其半抑制效应时间LT50的排序为:防己重楼黄连贯众。进一步研究防己的相对浓度梯度抑藻效应,结果表明:在相对浓度为2g/L时,实验3d后的藻细胞几乎全部死亡;防己的抑藻效应受贮藏时间和贮藏温度的影响不显著。在所研究的药用植物中,防己的抑藻效果最好,在抑藻方面有较大应用前景,其它3种药用植物对轻度藻类爆发的控制也有潜在应用价值。     

小果叶下珠花气味组成及其两性异型性

大戟科(Euphorbiaceae)植物小果叶下珠(Phyllanthus microcarpus)由种特异性细蛾科(Gracillariidae)昆虫头细蛾(Epicephala)专门为其传粉,具体包括:头细蛾在雄花上积极采粉,然后为雌花授粉并在其内产卵等极其不同的传粉行为。花气味在维持小果叶下珠与传粉头细蛾专性传粉互利共生关系中起着至关重要的作用。采用动态顶空吸附法分别收集小果叶下珠雌花和雄花气味,利用气相色谱-质谱(gas chromatography-mass spectrometry,GC-MS)联用技术分析鉴定其成分,并用峰面积归一法与内标法进行定量,最后通过主成分分析法比较两者间的差异性。结果表明:(1)小果叶下珠花气味中共分离出17种化合物,主要以萜类和脂肪族物质为主;顺-β-罗勒烯在雌花和雄花中含量均最高,为主要气味成分;(2)雌花气味释放量显著高于雄花;(3)雌花和雄花之间气味化学成分存在明显的差异,即具有两性异形性。初步推测花气味出现两性异形性是植物为适应传粉头细蛾极其高度特异的传粉行为(雄花采粉,雌花传粉并产卵)而选择进化的结果。     

荸荠营养器官的发育与解剖学研究

荸荠同化茎起源于肉质主茎倒２或倒３叶的叶腋内。同化茎基部着生二鞘状叶,鞘状叶对早期同化茎穿出土面具保护作用。匍匐茎大多起源于同化茎基部鞘状叶的叶腋内。当植株开始抽生花茎时,地下匍匐茎顶端开始膨大。球茎的膨大是匍匐茎顶端５－６节的基本组织经细胞有丝分裂,增加细胞数目,然后由细胞体积的扩大来实现的。球茎具足够的营养物质供来年顶芽萌发的需要,故属水生植物冬芽的性质。     

Range-wide genetic structure in a north-east Asian spruce (Picea jezoensis) determined using nuclear microsatellite markers

Aim We used microsatellite markers to determine the range‐wide genetic structure of Picea jezoensis and to test the hypothesis that the past population history of this widespread cold‐temperate spruce has resulted in a low level of genetic variation and in imprints of inbreeding and bottlenecks in isolated marginal populations. Location The natural range of the three infraspecific taxa of P. jezoensis throughout north‐east Asia, including isolated marginal populations. Methods We analysed a total of 990 individuals across 33 natural populations using four nuclear microsatellite loci. Population genetic structure was assessed by analysing genetic diversity indices for each population, examining clustering (model‐based and distance‐based) among populations, evaluating signals of recent bottlenecks, and testing for isolation by distance (IBD). Results The 33 populations were clustered into five groups. The isolated marginal groups of populations (in Kamchatka, Kii in Japan and South Korea) exhibited low levels of allelic richness and gene diversity and a complete or almost complete loss of rare alleles. A recent bottleneck was detected in the populations in Hokkaido across to mid‐Sakhalin. The IBD analysis revealed that genetic divergence between populations was higher for populations separated by straits. Main conclusions Picea jezoensis showed a higher level of genetic differentiation among populations (F_ST = 0.101) than that observed in the genus Picea in general. This might be attributable to the fact that historically the straits around Japan acted as barriers to the movement of seeds and pollen. The low levels of genetic diversity in the isolated marginal population groups may reflect genetic drift that has occurred after isolation. Evidence of a significant bottleneck between the Hokkaido and mid‐Sakhalin populations implies that the cold, dry climate in the late Pleistocene resulted in the decline and contraction of populations, and that there was a subsequent expansion followed by a founder effect when conditions improved. The high polymorphism observed in P. jezoensis nuclear microsatellites revealed cryptic genetic structure that organellar DNA markers failed to identify in a previous study.     

Adenosine induces apoptosis in human liver cancer cells through ROS production and mitochondrial dysfunction

Mitochondria are the most important sensor for apoptosis. Extracellular adenosine is well reported to induce apoptosis of tumor cells. Here we found that extracellular adenosine suppresses the cell growth by induction of apoptosis in BEL-7404 liver cancer cells, and identified a novel mechanism that extracellular adenosine triggers apoptosis by increasing Reactive Oxygen Species (ROS) production and mitochondrial membrane dysfunction in the cells. We observed that adenosine increases ROS production, activates c-Caspase-8 and -9 and Caspase effectors, c-Caspase-3 and c-PARP, induces accumulation of apoptosis regulator Bak, decreases Bcl-xL and Mcl-1, and causes the mitochondrial membrane dysfunction and the release of DIABLO, Cytochrome C, and AIF from mitochondria to cytoplasm in the cells; ROS inhibitor, NAC significantly reduces adenosine-induced ROS production; it also shows the same degree of blocking adenosine-induced loss of mitochondrial membrane potential (MMP) and apoptosis. Our study first observed that adenosine increases ROS production in tumor cells and identified the positive feedback loop for ROS-mediated mitochondrial membrane dysfunction which amplifies the death signals in the cells. Our findings indicated ROS production and mitochondrial dysfunction play a key role in adenosine-induced apoptosis of 7404 cells.