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Drought-induced 19 (Di19) family genes play important roles in plant growth, development, and environmental stress responses. However, little is known about this family in maize. The upstream regulatory network of Di19 genes remains poorly understood in plant stress response, especially. In this study, seven ZmDi19 genes were identified, and sequence alignment, gene structure, and phylogenetic analysis was conducted. According to the phylogenetic analysis, the ZmDi19-5 promoter was cloned and multiple putative stress-responsive cis-acting elements (CAEs) were found in the promoter region. The transient transformation assay indicated that firefly luciferase (LUC)-expressed activity driven by the ZmDi19-5 promoter can be significantly induced by drought stress. A 450 bp core region of ZmDi19-5 promoter was identified, and 28 upstream regulatory proteins were screened using yeast one-hybird (Y1H) system. According to the functional annotation, some genes were related to photosynthesis, light response, and water transport, which may suggest the important roles of these genes in drought response. Particularly, five members that may be involved in drought response exhibited strong binding activity to the core region of the ZmDi19-5 promoter. This study laid an important foundation for further revealing the molecular mechanisms and regulatory network of Di19 genes in drought stress response.

Deficits in coordinated motor behavior and mitochondrial complex V activity have been observed in aged males. Testosterone supplementation can improve coordinated motor behavior in aged males. We investigated the effects of testosterone supplementation on mitochondrial complex V function in the substantia nigra (a brain region that regulates motor activity) in aged male rats. These rats exhibited diminished ATP levels, attenuated mitochondrial complex V activity, and reduced expression of 3 of the 17 mitochondrial complex V subunits (ATP6, ATP8 and ATP5C1) in the substantia nigra. Testosterone supplementation increased ATP levels, mitochondrial complex V activity, and ATP6, ATP8 and ATP5C1 expression in the substantia nigra of the rats. Conversely, orchiectomy reduced mitochondrial complex V activity, downregulated ATP6 and ATP8 expression, and upregulated ATP5C1, ATP5I and ATP5L expression in the substantia nigra. Testosterone replacement reversed those effects. Thus, testosterone enhanced mitochondrial complex V function in the substantia nigra of aged male rats by upregulating ATP6 and ATP8. As potential testosterone targets, these two subunits may to some degree maintain nigrostriatal dopaminergic function in aged males.

BackgroundFrequent occurrence of extreme high temperature is a major threat to crop production. Increasing evidence demonstrates that long non-coding RNAs (lncRNAs) have important biological functions in the regulation of the response to heat stress. However, the regulatory mechanism of lncRNAs involved in heat response requires further exploration and the regulatory network remains poorly understood in maize.ResultsIn this research, high-throughput sequencing was adopted to systematically identify lncRNAs in maize inbred line CM1. In total, 53,249 lncRNAs (259 known lncRNAs and 52,990 novel lncRNAs) were detected, of which 993 lncRNAs showed significantly differential expression (DElncRNAs) under heat stress. By predicting the target genes, 953 common targets shared bycis- andtrans-regulation of the DElncRNAs were identified, which exhibited differential expression between the control and the heat stress treatments. Functional annotation indicated that a number of important biological processes and pathways, including photosynthesis, metabolism, translation, stress response, hormone signal transduction, and spliceosome, were enriched for the common targets, suggesting that they play important roles in heat response. A lncRNA-mediated regulatory network was constructed to visualize the molecular response mechanism in response to heat stress, which represented the direct regulatory relationships of DElncRNAs, differentially expressed miRNAs, target genes, and functional annotations.ConclusionsThis study lays a foundation for further elucidation of the regulatory mechanism for the response to heat stress in the maize inbred line CM1. The findings provide important information for identification of heat-responsive genes, which will be beneficial for the molecular breeding in the cultivation of heat-tolerant maize germplasm.

Adsorption desulfurization of catalytic gasoline (S Zorb) is an important desulfurization measure that is performed to meet the environmental protection requirements before the final product oil is sold in the market. The desulfurization reactor is a gas–solid two-phase flow environment composed of high-temperature and high-pressure hydrogen-oil mixed gas and sorbent particles; erosion prominently occurs on the reactor distribution plate. This study selects the typical gas–solid two-phase flow conditions and defines the erosion mechanism of the gas–solid two-phase flow environment for the plastic material of E347. Moreover, an S Zorb desulfurization reactor model is constructed, the CFD-DEM model is adopted to predict the wall erosion characteristics in a gas–solid two-phase flow environment, typical erosion laws are obtained via calculations. The erosion laws under the influence of variable parameters are studied based on the orthogonal test, the orthogonal test results show the best parameter combination, the parameter combination yields the maximum erosion rate and high erosion area that are 29.9% and 17.3%, respectively, lower than the existing values. Moreover, an optimum scheme of the inner structure parameters of the reactor is determined for reducing erosion rate and area.

The analysis of mycotoxins in foodstuffs is affected by the complexity of the matrix and the extremely low concentration levels. The development of sample pretreatment and analytical methods that enable highly selective enrichment as well as highly sensitive detection is of great significance for food safety. This paper reviews the recent progress in biotoxin analysis methods and summarizes the prospects and development of this field.

Our previous work has shown that atorvastatin exerts anti-inflammatory properties in ischemic stroke, and recent studies have revealed that intestinal microbiota plays a vital role in the pathogenesis of stroke. However, it is not clear whether the anti-inflammatory effects of atorvastatin against ischemic stroke is related to gut function and microbiota. We report herein that atorvastatin significantly ameliorated the defects in sensorimotor behaviors and reduced microglia-mediated neuroinflammation by inhibiting proinflammatory polarization of microglia in the peri-infarct cortex of the mice with permanent middle cerebral artery occlusion (pMCAO). Moreover, atorvastatin reversed microbial composition (characterized by increased abundance of Firmicutes and Lactobacillus and decreased Bacteroidetes abundance), increased fecal butyrate level, promoted intestinal barrier function (elevated protein levels of claudin-1, occludin and mucoprotein 2), as well as regulated intestinal immune function (decreased MCP-1, TNF-α and increased IL-10). Atorvastatin also significantly reduced the level of circulating endotoxin (lipopolysaccharide-binding protein), which is a biomarker of leaky gut. Transplantation of fecal microbiota collected from atorvastatin treated mice potently attenuated neuroinflammation in pMCAO mice, and the anti-inflammatory effects of fecal microbiota transplantation were similar to those of oral atorvastatin administration. These results suggested that the atorvastatin-mediated restoration of gut microbiota, improvement of intestinal barrier function and regulation of intestinal immunity were involved in the anti-inflammatory function in stroke mice.

Pentoxifylline (PTX) is a non-specific phosphodiesterase inhibitor with pleiotropic effects that is routinely used to treat peripheral vascular disease. In this study, we tested whether PTX could also counteract the detrimental effects of aging in the brain. To accomplish that, we treated aged rats with PTX and measured resulting behavioral alterations as well as changes in dopaminergic neurochemical levels, oxidative balance markers, mitochondrial function, nuclear factor erythroid 2-related factor 2 (Nrf2), peroxisome proliferator activated receptor-gamma coactivator 1-alpha (PGC-1α) and downstream gene expression, and cyclic adenosine monophosphate (cAMP) content in the brain. The results demonstrated that PTX improved motor and cognitive deficits and restored levels of dopamine and its metabolites in the brains of aged rats. PTX also reduced malondialdehyde levels and increased the GSH/GSSG ratio, mitochondrial ATP, nuclear Nrf2, and cAMP levels, and upregulated PGC-1α, nuclear respiratory factor 1, and mitochondrial transcription factor A expression in the substantia nigra and hippocampus of aged rats. Thus, increased nuclear Nrf2 levels and upregulation of PGC-1α, which enhance antioxidative capability and promote mitochondrial biogenesis, may be responsible for PTX-induced amelioration of behavioral deficits in aged rats.

Maize (Zeamays L.) is an essential food crop worldwide, but it is highly susceptible to salt stress, especially at the seedling stage. In this study, we conducted physiological and comparative transcriptome analyses of seedlings of maize inbred lines An’nong876 paternal (cmh15) and An’nong876 maternal (CM37) under salt stress. The cmh15 seedlings were more salt-tolerant and had higher relative water content, lower electrolyte leakage, and lower malondialdehyde levels in the leaves than CM37. We identified 2559 upregulated and 1770 downregulated genes between salt-treated CM37 and the controls, and 2757 upregulated and 2634 downregulated genes between salt-treated cmh15 and the controls by RNA sequencing analysis. Gene ontology functional enrichment analysis of the differentially expressed genes showed that photosynthesis-related and oxidation-reduction processes were deeply involved in the responses of cmh15 and CM37 to salt stress. We also found differences in the hormone signaling pathway transduction and regulation patterns of transcription factors encoded by the differentially expressed genes in both cmh15 and CM37 under salt stress. Together, our findings provide insights into the molecular networks that mediate salt stress tolerance of maize at the seedling stage.

A novel method for biphasic extraction and separation of different polysaccharides from Radix Sophorae Tonkinensis (RST) was developed by microwave-assisted aqueous two-phase extraction (MAATPE) in a one-step procedure. Using ethanol/Na2HPO4 system as a green extractant, the effects of the composition of the aqueous two-phase system (ATPS), extraction temperature and extraction time were investigated by single-factor experiments coupled to response surface methodology (RSM) through UV–vis analysis. Under the optimum conditions, extraction yields of two polysaccharides from top and bottom phases were 8.52 ± 0.21% (w/w) and 2.18 ± 0.08% (w/w), respectively. Compared with heating solvent extraction (HSE) and ultrasonic assisted extraction (UAE), MAATPE exhibited higher extraction efficiency. By means of Fourier transform infrared spectroscopy (FT-IR) and high-performance gel permeation chromatography (HPGPC), two polysaccharides obtained from both phases had the characteristics of homogenous heteropolysaccharides, but there were some differences in chemical structures and the molecular weight from each other. Both polysaccharides could be extracted respectively from RST to the top ethanol-rich phase and the bottom salt-rich phase due to their polarity difference. Furthermore, the chemical compositions of two polysaccharides were confirmed by high-performance liquid chromatography (HPLC) analysis after hydrolysis and derivatization, the molar ratios of monosaccharides were glucose: arabinose: galactose: rhamnose: mannose: glucuronic acid: aminogalactose = 63.70: 18.21: 7.00: 5.10: 3.13: 1.78: 1.07 for the polysaccharide in the top phase and 75.24: 3.79: 3.71: 3.37: 2.51: 1.43: 9.95 for those in the bottom phase, respectively. By means of extraction process analysis and scanning electron microscopy (SEM), the exportation of MAATPE mechanism revealed that combining microwave-assisted extraction (MAE) with aqueous two-phase extraction (ATPE) not only improved the extraction yields, but also obtained at least two different polysaccharides. Under microwave field, the ATPS could make the herb cell ruptured, and multiphase mass-transfer was accelerated. The proposed method is a rapid, simple and efficient alternative to biphasic extraction of diverse polysaccharides from natural plants.

A rapid method for simultaneous extraction and separation of multiple alkaloids from Radix Sophorae Tonkinensis (RST) was developed by microwave-assisted aqueous two-phase extraction (MAATPE) using the aqueous two-phase extraction system (ATPS) of ethanol/Na2HPO4 as the extraction solvent. The effects of key factors on extraction yield were investigated by utilizing single-factor experiment coupled to response surface methodology (RSM). The regression model by RSM was significant (p

A novel method for simultaneous determination of pyrethroids residues in Litchi fruit has been developed by HPLC-UV detection using microwave-assisted extraction (MAE) coupled with ultrasonic-assisted dispersive liquid-liquid microextraction (UADLLME). Extraction conditions of MAE and UADLLME were respectively investigated by single-factor experiments and response surface methodology. Optimized experimental conditions included 310μL of chlorobenzene as extraction solvent, 1.3mL of ethanol as dispersive solvent and 3min of extraction time for UADLLME. In the case of MAE, extraction temperature of 70°C, extraction time of 4min and solvent-to-materials ratio of 40:1 were adopted. Results demonstrated that the proposed method had good performance with linearity of 0.0050-4.98mg/L, recovery of 83.3-91.5%, RSDs below 5.6% and detection limit (LOD) of 1.15-2.46μg/L for six pyrethroids, offering higher extraction efficiency and larger enrichment factor. MAE-UADLLME provided a sensitive and efficient alternative to determination of trace amounts of pesticides residues in food samples.

Supplemental material, sj-pdf-2-imr-10.1177_0300060520943452 for Efficacy and safety of human urinary kallidinogenase for acute ischemic stroke: a meta-analysis by Yuanxiang Huang, Binglei Wang, Yue Zhang, Peize Wang and Xiangjian Zhang in Journal of International Medical Research

Supplemental material, sj-pdf-1-imr-10.1177_0300060520943452 for Efficacy and safety of human urinary kallidinogenase for acute ischemic stroke: a meta-analysis by Yuanxiang Huang, Binglei Wang, Yue Zhang, Peize Wang and Xiangjian Zhang in Journal of International Medical Research

In this study, we used high-throughput RNA sequencing to identify mRNAs, long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) that are differentially expressed in the Substantia Nigra (SN) of aged and young rats. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses were used to perform functional annotation of mRNAs that were either differentially expressed themselves (DEMs), targeted by differentially expressed lncRNAs (DELs), or the parents of differentially expressed circRNAs (DECs). A total of 112 DEMs, 163 DELs, and 98 DECs were found in the SN of aged rats. The down-regulated lncRNA NONRATT010417.2 targeted the down-regulated mRNA Myh1, while the down-regulated lncRNA NONRATT015586.2 and the up-regulated lncRNAs NONRATT000490.2 and NONRATT007029.2 all targeted the down-regulated mRNAs Casr and Mis18a. Western blots and RT-qPCR revealed that Myh1, Casr, and Mis18a protein and mRNA expression were significantly reduced in aged rats compared to young rats. This study improves our understanding of the transcriptional alterations underlying aging-related changes in the SN and provides a foundation for future studies of associated molecular mechanisms.

A novel, efficient and specific aptamer-functionalized sorbent was successfully synthesized and applied in the analysis of ultra-trace aflatoxin M1 (AFM1) and its analogues in milk. By using a gold nanoparticle-based colorimetric analysis tool, we could determine that the AFM1-aptamer (M1-aptamer) recognized and bound AFM1 to generate an aptamer-target complex. After analysis and verification, the M1-aptamer on the surface of the carboxyl microspheres were completely modified via EDC/NHs amide formation. The effects of extraction pH, extraction buffer, concentration of Mg2+ and elution solvents were investigated by single-factor experiments to determine the most suitable analysis conditions. Using these conditions, the extraction performance of the M1-aptamer-functionalized sorbent could be studied and an analytical method for AFM1 and its analogues in milk could be established. The results of adsorption equilibrium as well as adsorption selectivity experiments indicated that the M1-aptamer offered a specific adsorption site to selectively adsorb aflatoxins exhibiting a dihydrofuran oxaphthalene scaffold. The adsorption capacity of the aptamer-functionalized sorbent was 233.1 μg/g. A studied method for the determination AFM1 in milk with low matrix effect demonstrated a good linear correlation in the range of 0 to 2.0 μg/L (R2 = 0.9982) and high detection sensitivity (detection limits of 7 ng/kg in milk and 21 ng/kg in milk power). Furthermore, excellent spiked recovery and good repeatability were obtained. The method delivered specific, accurate and reproducible results for enriching and detecting ultra-trace AFM1 and its analogues in milk.

Microwave-assisted aqueous two-phase extraction (MAATPE) was applied to simultaneous extraction of diverse polysaccharides from Lentinus edodes with aqueous two-phase extraction system (ATPS) of ethanol/ammonium sulfate as the biphasic extractant. Key factors in MAATPE process were optimized by response surface methodology, and the optimum conditions were concluded as follows: the ATPS composition of ethanol concentration 26.0% (w/w) and (NH4)2SO4 concentration 19.58% (w/w), extraction temperature 78.7 °C, extraction time 19.55 min and solid-to-solvent ratio 1:50. The extraction yields of the polysaccharides from the top phase (PTP) and the bottom phase (PBP) were 2.12 ± 0.21% and 11.16 ± 0.28%, respectively. By means of FT-IR, HPGPC and HPLC analysis, PTP and PBP were different from each other in chemical structure, their average molecular weight (MW) were respectively 119,533 Da and 3906 Da. Also, the mechanism of MAATPE process was explored by composition analysis and scanning electron microscopy. Both polysaccharides exhibited stronger antioxidant activities, EC50 values of scavenging DPPH, hydroxyl and superoxide radicals for PBP achieved 580 ± 3, 83.1 ± 3.6 and 91.1 ± 0.5 μg/mL as against 560 ± 4, 76.3 ± 2.2 and 89.3 ± 2.4 μg/mL of PTP, respectively. Compared with conventional methods using a mono-phase solvent, MAATPE can not only greatly improve extraction efficiency, but also selectively extract diverse polysaccharides.