The viral proteases have proven to be the most selective and useful for removing the fusion tags in fusion protein expression systems. As a key enzyme in the viral life-cycle, the main protease (M(pro)) is most attractive for drug design targeting the SARS coronavirus (SARS-CoV), the etiological agent responsible for the outbreak of severe acute respiratory syndrome (SARS) in 2003. In this study, SARS-CoV M(pro) was used to specifically remove the GST tag in a new fusion protein expression system. We report a new method to produce wild-type (WT) SARS-CoV M(pro) with authentic N and C termini, and compare the activity of WT protease with those of three different types of SARS-CoV M(pro) with additional residues at the N or C terminus. Our results show that additional residues at the N terminus, but not at the C terminus, of M(pro) are detrimental to enzyme activity. To explain this, the crystal structures of WT SARS-CoV M(pro) and its complex with a Michael acceptor inhibitor were determined to 1.6 Angstroms and 1.95 Angstroms resolution respectively. These crystal structures reveal that the first residue of this protease is important for sustaining the substrate-binding pocket and inhibitor binding. This study suggests that SARS-CoV M(pro) could serve as a new tag-cleavage endopeptidase for protein overproduction, and the WT SARS-CoV M(pro) is more appropriate for mechanistic characterization and inhibitor design.

We examined the spatiotemporal variation in diurnal temperature range (DTR) and discussed the reasons for the changes of DTR in China based on data from 479 weather stations from 1962 to 2011. Results showed that DTR decreased rapidly (0.291°C/decade) from 1962 to 1989 due to slightly decreased T max and significantly increased T min , but the decrease in DTR has stopped since 1990 as T max and T min kept pace with each other. During 1990-2011, DTR remained trendless, with slight increase in the 1990s and slight decrease after 2000. During the whole study period from 1962 to 2011, DTR decreased at a rate of 0.157°C/decade nationally. Spatially, decreases in DTR were greatest in Northeast China and lowest in Southwest China with a transect running from northeast to southwest showing the decreasing trends change from high to low. Seasonally, DTR decreases were greatest in winter and lowest in summer, and the magnitudes of decrease reduced from the north to south of China. The changes in DTR were closely correlated with changes in sunshine duration (SD) in China except the Tibetan Plateau, suggesting that SD decrease is an important contributor to the decrease of DTR through its influence on T max . In addition to the contribution of SD decrease, the increasing of precipitation played an important role in DTR decrease in Northwest China, the most arid region of China. It appeared that changes of cloud cover (CC) were not the reasons for DTR changes in the past 50 years as CC has decreased during the study period.

This study performs a meta-analysis of research that estimates the relationship between FDI and Chinese economic growth. Our sample includes 37 studies and a total of 280 estimates. We include both English-and Chinese-language studies. Our initial "raw" finding is that FDI has had a substantial, positive impact on Chinese economic growth. Furthermore, our results suggest that the effect is not inflated by endogeneity, nor impacted by publication bias. However, the positive effect is found to be smaller for more recent and better designed studies. When we adjust for preferred study and sample characteristics, we find that the estimated economic effect of FDI on Chinese economic growth is much smaller than indicated by the overall literature, and statistically insignificant. This suggests that the cause(s) of the Chinese "economic miracle" likely lie elsewhere.

Reduction in vegetation cover caused by human activities has a great impact on soil temperature. It is important to assess how soil temperature responds to reduction of vegetation height and density. In this paper we first report the trends of mean annual soil surface and air temperatures recorded at the meteorological stations near the Ecological Research Station for Grassland Farming (ERSGF) from 1961 to 2007, then we setup an experiment using reed (Phragmites australis) stalks with different heights and densities to simulate effects of different vegetation height and density on soil and air temperatures. The warming rates of the mean annual soil and air temperatures were 0.043 and 0.041°C a 1 , respectively. Changes of soil temperature were characterized by both increased mean annual maximum and minimum soil temperatures. At the experimental site, mean daily temperature, mean daily maximum soil and air temperatures increased significantly. In contrast, mean daily minimum soil temperature increased significantly while mean daily minimum air temperature decreased significantly as the height and density of reed stalks reduced during the experimental period. Mean diurnal soil temperature ranges were smaller than mean diurnal air temperature ranges. These results highlight that the importance of vegetation cover on soil and air temperatures. There is general consensus that the global climate has changed rapidly, and the global mean surface temperature has increased by 0. 74C between 1906 and 2005 [1]. The warming pattern shows that mean annual minimum temperature has increased almost two orders of magnitude of maximum temperature, i.e. an asymmetric diurnal temperature increase [2,3]. Apart from studies on atmospheric temperature continuously published, more focus on variation of soil temperature and its factors has emerged recently [4][5][6][7][8][9]. Soil temperature is a crucial factor involved in determining/affecting the rates of biochemical reactions and has a strong influence on plant and root growth [2,6]. Diurnal soil temperature range is particularly important in plant growth, such as seed germination and early season growth which are highly correlated with daily maximum temperature of the soil rather than with air temperature [7]. Similar to increased air temperature, soil temperature also increases based on the long-term trend. Hu and Feng [8] reported that soil temperature at 10 cm depth increased 0.031C a 1 from 1967 to 2002 in the contiguous United States. A study recorded 27-year soil temperature at 5 depths showed a significant increase at a grassland in the Netherlands, and the warming rate of soil temperature was higher than air tem-

In order to restore a degraded alkaline grassland, the local government implemented a large restoration project using fences in Changling county, Jilin province, China, in 2000. Grazing was excluded from the protected area, whereas the grazed area was continuously grazed at 8.5 dry sheep equivalent (DSE)/hm 2. In the current research, soil and plant samples were taken from grazed and fenced areas to examine changes in vegetation and soil properties in 2005, 2006 and 2008. Results showed that vegetation characteristics and soil properties improved significantly in the fenced area compared with the grazed area. In the protected area the vegetation cover, height and above-and belowground biomass increased significantly. Soil pH, electrical conductivity and bulk density decreased significantly, but soil organic carbon and total nitrogen concentration increased greatly in the protected area. By comparing the vegetation and soil characteristics with pre-degraded grassland, we found that vegetation can recover 6 years after fencing, and soil pH can be restored 8 years after fencing. However, the restoration of soil organic carbon, total nitrogen and total phosphorus concentrations needed 16, 30 and 19 years, respectively. It is recommended that the stocking rate should be reduced to 1/3 of the current carrying capacity, or that a grazing regime of 1-year of grazing followed by a 2-year rest is adopted to sustain the current status of vegetation and soil resources. However, if N fertilizer is applied, the rest period could be shortened, depending on the rate of application.

Oxidative stress has been implicated in the etiology of multiple gastrointestinal disorders, such as irritable bowel syndrome and inflammatory bowel disease. This study was conducted to evaluate effects of natural product quercetin on diquat-induced oxidative stress in porcine enterocytes and underlying mechanisms. Intestinal porcine epithelial cell line 1 (IPEC-1) cells pretreated with or without quercetin (5 μM, 24 h) were incubated with vehicle or diquat (100 μM) for 6 h. The results showed that diquat treatment induced apoptosis in a caspase-3-dependent manner, as accompanied by elevated reactive oxygen species (ROS) production, increased mitochondrial depolarization, and reduced the abundance of tight junction proteins. These adverse effects of diquat were remarkably abrogated by quercetin administration. Further study indicated that the protective effect of quercetin was associated with elevated protein abundance of nuclear factor erythroid 2-related factor 2 (Nrf2) and increased intracellular glutathione (GSH) content. Interestingly, the beneficial effects of quercetin on diquat-induced oxidative damage were abolished by all-trans-retinoic acid (Atra), a specific inhibitor of Nrf2, indicating a Nrf2-dependent regulation manner. The results show that quercetin attenuates diquat-induced cell injury by promoting protein abundance of Nrf2 and regulating GSH-related redox homeostasis in enterocytes. These findings provide new insights into a function role of quercetin in maintaining intestinal homeostasis.

Scope Inflammatory bowel disease (IBD) is an inflammatory gastrointestinal disorder in which endoplasmic reticulum (ER) stress and dysbiosis of the intestinal microbiota are implicated. Glycine supplementation is reported to reduce inflammatory responses in experimental colitis. However, the underlying mechanisms responsible for the beneficial effects remain unclear. Methods and Results Female C57BL/6 mice are orally administered with glycine (3.5 or 5.2 g kg‐1 body weight) for 14 continuous days. On day 8 post‐glycine supplementation, the mice are orally inoculated with 2 × 109 CFU Citrobacter rodentium (C. rodentium). The results show that glycine alleviates C. rodentium‐induced body weight loss, increased disease activity index and spleen weight, colon length shortening, and colonic hyperplasia. Glycine suppresses the activation and infiltration of inflammatory cells, and secretion of pro‐inflammatory cytokines in the colon tissues. The apoptosis of colon epithelial cells is also abrogated by glycine, which is associated with the inactivation of activating transcription factor 6α (ATF6α)‐C/EBP homologous protein (CHOP) signaling. In addition, glycine administration increases α diversity, restores β diversity, and abolishes the reduction in Lactobacillus, Bifidobacterium, Alistipes, Turicibacter, and Alloprevotella in the colon. Conclusions Glycine supplementation is a nutritional strategy that may ameliorate C. rodentium‐induced colitis by regulating ATF6α‐CHOP‐mediated ER stress and enhancing the abundance of Lactobacillus.

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