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Extensive targeted metabolomic and transcriptomic analyses were used to determine the types of flavonoids present in the fruit of two varieties of Actinidia arguta Sieb. Zucc. cultivated in Northern China. Fractionation and preparative liquid chromatography were used to isolate and purify the potentially biologically active flavonoids chrysin (flavone1), rutin (flavone2), and daidzein (flavone3) from the fruit of Actinidia arguta Sieb. Zucc. The structures of these flavonoids were appraised using mass spectroscopy/high-performance liquid chromatography (MS-HPLC). Serum levels of uric acid (UA), urea nitrogen (BUN), creatinine (Cr), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and liver glycogen were measured in male Kunming mice treated with each flavonoid to determine biological effects of the flavonoids on the measured parameters. The three flavonoids were found to be mainly distributed within three metabolic pathways, namely the flavonoids and flavonol biosynthesis pathway(BP), the flavonoid BP, and the secondary metabolite BP. Through a correlation analysis of the transcriptome and metabolome, regulatory genes were screened for activity in the BPs of the three flavonoids. This is the first study to explore the UA-lowering activity of flavonoids in the fruit of Actinidia arguta Sieb., the main variety cultivated in Northern China. These data provide valuable information for understanding the biosynthetic mechanisms and regulatory pathways of flavonoids in Actinidia arguta Sieb. and contribute to its development as a food and drug homologous functional food.

Fungal immunomodulatory proteins (FIPs) have been identified from a series of fungi, especially in Ganoderma species. However, little is known about the FIPs from G. applanatum. In this study, two novel FIP genes, termed as FIP-gap1 and FIP-gap2, were cloned from G. applanatum, characterized and functionally expressed after codon optimization in Pichia pastoris GS115. Results showed that FIP-gap1 and FIP-gap2 comprised 342-bp encoding peptides of 113 amino acids, which shared a high homology with other Ganoderma FIPs. The yield of recombinant FIP-gap1 and FIP-gap2 increased significantly after codon optimization and reached 247.4 and 197.5 mg/L, respectively. Bioactivity assay in vitro revealed that both rFIP-gap1 and rFIP-gap2 could agglutinate mouse, sheep, and human red blood cells. Besides, rFIP-gap1 and rFIP-gap2 obviously stimulated the proliferation of mouse splenocytes and enhanced IL-2 and IFN-γ release. Cytotoxicity detection indicated that IC50 of rFIP-gap1 towards A549 and HeLa cancer cells were 29.89 and 8.34 μg/mL, respectively, whereas IC50 of rFIP-gap2 to the same cancer cells were 60.92 and 41.05 μg/mL, respectively. Taken together, novel FIP gaps were cloned and functionally expressed in P. pastoris, which can serve as feasible and stable resources of rFIP gaps for further studies and potential applications.

The crude polysaccharide (TASP3) was extracted from the fruit pulp of Annona squamosa and then isolated and purified by the combination of grading-alcoholic precipitation and Sephadex G-200. The structure of purified polysaccharide (GASP3-3-I) was determined based on the physicochemical and instrumental analyses. The results indicated that GASP3-3-I was an acidic heteropolysaccharide and its average molecular weight was 2.28×106Da. The monosaccharide composition was analysed by GC-MS and ion chromatography, respectively. It was revealed that GASP3-3-I was consisted of rhamnose, arabinose, xylose, mannose, glucose, galactose, glucuronic acid and galacturonic acid with a molar ratio of 5.06:45.5:5.26:0.63:6.09:31.76:0.49:5.19. Moreover, periodate oxidation reaction, Smith degrading reaction, methylation, FT-IR and NMR were used to conduct the structural characterization of GASP3-3-I. The results indicated that glycosyl residues of GASP3-3-I were mainly composed of (1→) l-arabinose, (1→6), (1→3) and (1,3→6) d-galactose, (1→) d-xylose, (3→) and (3→6) d-glucose, (1→2) l-rhamnose. The α-glucosidase inhibitory activity assay showed that GASP3-3-I had a certain inhibition on α-glucosidase activity.

The Yorkie homologues YAP (Yes-associated protein) and TAZ (transcriptional coactivator with PDZ-binding motif, also known as WWTR1), effectors of the Hippo pathway, have been identified as mediators for mechanical stimuli. However, the role of YAP/TAZ in haemodynamics-induced mechanotransduction and pathogenesis of atherosclerosis remains unclear. Here we show that endothelial YAP/TAZ activity is regulated by different patterns of blood flow, and YAP/TAZ inhibition suppresses inflammation and retards atherogenesis. Atheroprone-disturbed flow increases whereas atheroprotective unidirectional shear stress inhibits YAP/TAZ activity. Unidirectional shear stress activates integrin and promotes integrin-Gα13 interaction, leading to RhoA inhibition and YAP phosphorylation and suppression. YAP/TAZ inhibition suppresses JNK signalling and downregulates pro-inflammatory genes expression, thereby reducing monocyte attachment and infiltration. In vivo endothelial-specific YAP overexpression exacerbates, while CRISPR/Cas9-mediated Yap knockdown in endothelium retards, plaque formation in ApoE-/- mice. We also show several existing anti-atherosclerotic agents such as statins inhibit YAP/TAZ transactivation. On the other hand, simvastatin fails to suppress constitutively active YAP/TAZ-induced pro-inflammatory gene expression in endothelial cells, indicating that YAP/TAZ inhibition could contribute to the anti-inflammatory effect of simvastatin. Furthermore, activation of integrin by oral administration of MnCl2 reduces plaque formation. Taken together, our results indicate that integrin-Gα13-RhoA-YAP pathway holds promise as a novel drug target against atherosclerosis.

Background and objectives There is increasing evidence that microRNAs (miRNAs) play crucial roles in the regulation of neointima formation. However, the translational evidence of the role of miRNAs in dialysis vascular access is limited. Design, setting, participants, & measurements miRNA expression in tissues was assessed by using venous tissues harvested from ten patients on dialysis who received revision or removal surgery, and ten patients who were predialysis and received creation surgery of arteriovenous fistulas served as controls. To extend these findings, 60 patients who received angioplasty of dialysis access were enrolled and the levels of circulating miRNAs were determined before and 2 days after angioplasty. Clinical follow-up was continued monthly for 6 months. The primary outcome of angioplasty cohort was target lesion restenosis within 6 months after angioplasty. Results In the surgery cohort, the expressions of miR-21, miR-130a, and miR-221 were upregulated in stenotic tissues, whereas those of miR-133 and miR-145 were downregulated. In situ hybridization revealed similar expression patterns of these miRNAs, localized predominantly in the neointima region. Twenty eight patients in the angioplasty cohort developed restenosis within 6 months. The levels of circulating miR-21, miR-130a, miR-221, miR-133, and miR-145 significantly increased 2 days after angioplasty. Kaplan–Meier plots showed that patients with an increase of miR-21 expression level >0.35 have a higher risk of patency loss (hazard ratio, 4.45; 95% confidence interval, 1.68 to 11.7). In a multivariable analysis, postangioplasty increase of miR-21 expression was independently associated with restenosis (hazard ratio, 1.20; 95% confidence interval, 1.07 to 1.35 per one unit increase of miR-21 expression level; P=0.001). Conclusions Certain miRNAs are differentially expressed in the stenotic venous segments of dialysis accesses. An increase in blood miR-21 level with angioplasty is associated with a higher risk of restenosis.

The structural properties and Angiotensin-I converting enzyme (ACE) inhibition activities of a polysaccharide (PGE) extracted from

A polysaccharide named PNP was extracted and purified from

A polysaccharide named PNP was extracted and purified from Pholiota nameko. The total sugar content of PNP was 95.29% and the molecular weight was 1.89 × 103 kDa. The structural features of PNP were investigated by the combination of chemical and instrumental analysis such as UV spectrophotometer, specific rotation determination, FT-IR, methylisation analysis and Congo red. The results showed that the optical rotation of PNP was +120° and that it had a triple-helical structure. Besides, PNP was mainly composed of glucose and mannose at the molar ratio of 4.24:1.00. The backbone of PNP was composed of (1→3)-linked-Glc and (1→3)-linked-Man whereas the branches of (1→3,6)-linked- Glc, (1→3,6)-linked-Man and T- Glc. Consistenting with the results of UV–Vis spectra, FT-IR spectroscopy and 1H NMR, indicated that PNP was a complex of polysaccharides and polyphenols. In vitro antioxidant results suggested that PNP was processed with certain scavenging capacity.

The structural properties and Angiotensin-I converting enzyme (ACE) inhibition activities of a polysaccharide (PGE) extracted from Gastrodia elata Blume were investigated. PGE was extracted using hot water and purified by Sephadex G-200 followed by ultra-filtration. The structural characterisation of PGE was analysed by FT–IR, NMR spectroscopy, specific rotation determination, periodate oxidation-smith degradation, methylation analysis, GC–MS and Congo red test. The results revealed that PGE was composed by glucose, with an average molecular weight of 1.54 × 103 kDa. The structure of PGE was 1→3 and 1→4,6-branched-glucopyranose that had a linear backbone of (1 → 4)-linked-d-glucopyranose (Glcp). ACE-inhibitory activity results showed that PGE was efficient to inhibit ACE and the IC50 value was 0.66 mg/mL.

We determined the effect of Chinese mitten crab (CMC) pen culture on the quantified spatial distribution of the macrozoobenthic community and sediment nutrients in Lake Yangcheng. Redundancy analysis indicated that water temperature, macrophyte occurrence, sediment type, and crab culture were the main environmental factors that influence the spatiotemporal macrozoobenthic distribution. Macrozoobenthic assemblages in the lake were characterized by eutrophic indicator species. In the most polluted estuaries, the abundance and diversity indices of the whole community and abundance of chironomids and oligochaetes were significantly depressed, and sediment carbon (C) and phosphorus (P) were significantly enhanced compared with those in the western, middle (MB), and eastern basin (EB). Crab culture in this lake had significant effects on the species composition of the macrozoobenthic community in one of three CMC culture pens (CP), and generally depressed the abundance of most chironomid and oligochaete species. Significantly increased diversity, evenness, sediment carbon and nitrogen content, and sediment C:P ratio in the CP were found compared with those in the three basins. However, no conspicuous difference in sediment P content between the CP and the two basins of MB and EB was detected. Our results showed that the enhanced diversity and evenness of macrozoobenthos might be associated with the joint effect of macrophyte planting and crab predation, and macrophyte planting may modify the effects of CMC culture by leading to disproportional accumulation of C and N in the sediment relative to P in the CP of the lake.

Background and aims MicroRNA (miR)-10a is a shear-regulated miR with the lowest expression in vascular endothelial cells (ECs) in athero-susceptible regions with oscillatory shear stress (OS). The aim of this study is to elucidate the relationship between EC miR-10a and atherosclerosis and develop a hemodynamics-based strategy for atherosclerosis treatment. Methods A combination of in vitro flow system and in vivo experimental animals was used to examine the functional roles of EC miR-10a and its clinical applications in atherosclerosis. Results En face staining showed that EC miR-10a is down-regulated in the inner curvature (OS region) of aortic arch in rats. Co-administration with retinoic acid receptor-α (RARα)- and retinoid X receptor-α (RXRα)-specific agonists rescued EC miR-10a expression in this OS region. These effects of OS and RARα/RXRα-specific agonists on EC miR-10a expression were confirmed by the in vitro flow system, and were modulated by the RARα-histone deacetylases complex, with the consequent modulation in the downstream GATA6/vascular cell adhesion molecule (VCAM)-1 signaling cascade. Animal studies showed that miR-10a levels are decreased in both aortic endothelium of atherosclerotic lesions and blood plasma from apolipoprotein E-deficient (ApoE−/−) mice. In vivo induction of EC miR-10a by administration of RARα/RXRα-specific agonists protects ApoE−/− mice from atherosclerosis through inhibition of GATA6/VCAM-1 signaling and inflammatory cell infiltration. Conclusions Our findings indicate that down-regulation of miR-10a in aortic endothelium and blood serum is associated with atherosclerosis, and miR-10a has potential to be developed as diagnostic molecule for atherosclerosis. Moreover, EC miR-10a induction by RARα/RXRα-specific agonists is a potential hemodynamics-based strategy for atherosclerosis treatment.

This paper investigates the H∞ filter design problem for a class of T-S fuzzy systems based on Hamiltonian model. First, a new method is proposed to change general nonlinear systems to Hamiltonian systems, which are easier to deal with. Then, by using the structural characteristics of Hamiltonian systems, the Lyapunov-Krasovskii functions are found and the H∞ filter can be designed. The unknown parameters of the filter are obtained by solving a special kind of inequalities. Finally, the simulation of an illustrative example shows that the proposed method is effective.

An Astragalus oligosaccharide (AOS) degraded from Astragalus polysaccharide (APS) and purified by membrane dialysis and silicon gel chromatography is studied in this paper. The structural features of AOS were investigated by a combination of chemical and instrumental analysis, such as monosaccharide analysis, periodate oxidation-Smith degradation, methylation analysis, electrospray ionization mass spectrometry (ESI-MS), Fourier transform infrared (FT-IR) spectrometry and nuclear magnetic resonance (NMR). The results indicated that AOS is an octasaccharide that consists of (3→)-linked-Rha, (1→3)-linked-Rha, (1→3,4)-linked-Araf, (1→3)-linked-Gal, terminal-linked-Gal and terminal-linked-Glc. The effects of AOS on cyclophosphamide-induced immunosuppression were determined by various studies, such as the proliferation of nucleated marrow, red blood cell (RBC) and white blood cell (WBC) populations, growth of the spleen and thymus, and increases in hemoglobin (HGB) concentration and granulocyte-macrophage colony stimulating factor (GM-CSF) level. The results indicated that AOS can restore cyclophosphamide-induced immunosuppression by stimulating the secretion of GM-CSF, which promoted the differentiation of progenitor cells after proliferation.

Objectives A prospective, blinded, randomized trial was performed to evaluate the incidence rates of external ventricular drainage (EVD)-related infection (ERI) after tunneled EVD (T-EVD) and standard EVD (S-EVD). Patients and methods From February 2018 to February 2019, all adult patients admitted to the Union Hospital Neurosurgery Center for EVD placement were eligible for inclusion. After the application of strict exclusion criteria, all enrolled patients were randomly divided into two groups. The patients in Group A received S-EVD, and the remaining patients in Group B received T-EVD. A linear incision was made for T-EVD. The distal end of the catheter was inserted approximately 5 cm until cerebrospinal fluid was readily obtained, and then the catheter was tunneled approximately 4–5 cm from the insertion point. Finally, an external CSF drainage system was connected to the catheter. For the S-EVD patients, we secured the catheter at the original incision site after insertion, and an external CSF drainage system was also connected to the catheter. The rates of ERI were compared between the two patient groups. The odds ratios and χ² test were used to analyze the results. Results One hundred twenty patients were randomly divided into two groups and underwent EVD placement. Among them, 60 patients in Group A received S-EVD, and 60 patients in Group B received T-EVD. Finally, 51 patients in Group A and 50 patients in Group B met all of the study inclusion/exclusion criteria and were thus eligible for inclusion in the evaluation of ERI rates. All clinical features of the two groups were similar. A total of 12 patients’ (11.9%) CSF cultures were positive for infection. Ten (19.6%) patients who underwent S-EVD had CSF-positive cultures, while only 2 (4.0%) patients who underwent T-EVD had CSF-positive cultures (P = 0.034). Additionally, 8 patients in Group A and 1 patient in Group B were complicated with CSF leakage (P = 0.039). Conclusions Compared to S-EVD, T-EVD, when performed according to a previously established perioperative management protocol, resulted in lower infection and CSF leakage rates. We recommend that T-EVD should be preferentially performed when surgeons determine whether a catheter can be removed within 10 days, and the catheter used for EVD should be removed as soon as permitted by the clinical circumstances.

Based on the viscoelastic material property of polyethylene pipe, software ANSYS was used to simulate and analyze the mechanical property of polyethylene pipe butt fusion joints with circumferential surface crack defects. The viscoelastic material creep parameters were characterized as Prony series and 1/4 node singular element was selected for meshing along the boundaries of the crack, then the stress intensity factor of polyethylene pipe butt fusion joints with circumferential surface crack was calculated under the uniform internal pressure. Through the finite element simulation, the result showed that polyethylene pipe were most likely to fracture failure when crack initiated. Thus the viscoelasticity of materials can be ignored when analyzing the stress intensity factor of circumferential surface cracks of polyethylene pipe. the main influencing factor of the circumferential crack defects was the ratio of the crack depth to the thickness of polyethylene pipe.

In plants, a promoter is essential to drive the transcription and expression of genes under stress conditions. The cold-regulated promoter is an important molecular switch involved in transcriptional regulation of a dynamic network of genes associated with cold acclimation processes. However, the structure and functions of the cold-regulated promoter are ambiguous. In this review, we first describe the common type and structures of the cold-regulated promoter, such as the core promoter and transcription factor binding sites, and then discuss the synergistic actions of promoter elements and cold-regulated genes. We also describe the transcriptional responses and cross-talk among cold-regulated genes in the ICE-CBF-COR cold-response pathway. Many stress-inducible genes are known to be regulated by endogenous abscisic acid (ABA), which accumulates during osmotic and cold stress. We discuss the regulation of promoters of cold-inducible genes in ABA-dependent and ABA-independent regulatory systems. We also describe the cross-talk among gene networks regulated by different cis-acting regulatory elements. Finally, we propose potential further research on, and practical applications of, the cold-regulated promoter.

Based on the galactinol synthase (AnGolS1) fragment sequence from a cold-induced Suppression Subtractive Hybridization (SSH) library derived from Ammopiptanthus nanus (A. nanus) seedlings, AnGolS1 mRNA (including the 5′ UTR and 3′ UTR) (GenBank accession number: GU942748) was isolated and characterized by rapid amplification of cDNA ends polymerase chain reaction (RACE–PCR). A substrate reaction test revealed that AnGolS1 possessed galactinol synthase activity in vitro and could potentially be an early-responsive gene. Furthermore, quantitative real-time PCR (qRT-PCR) indicated that AnGolS1 was responded to cold, salts and drought stresses, however, significantly up-regulated in all origans by low temperatures, especially in plant stems. In addition, the hybridization signals in the fascicular cambium were strongest in all cells under low temperature. Thus, we propose that AnGolS1 plays critical roles in A. nanus low-temperature stress resistance and that fascicular cambium cells could be involved in AnGolS1 mRNA transcription, galactinol transportation and coordination under low-temperature stress.

Chrysin-β-D-galactopyranoside was efficiently synthesized, evaluated for its inhibitory activities against H22 cell lines compared with chrysin, the scavenging of hydroxyl radical, DPPH radical and superoxide anion, inhibitory effect against bacteria and fungi. The structures of all compounds were fully characterized by spectroscopic data (NMR, MS). The anti-tumor, antioxidant and antimicrobial activities of chrysin-β-D-galactopyranoside were proved to be enhanced significantly compared with chrysin.

A novel homogeneous polysaccharide fraction (APS) was extracted from Astragalus by hot water and purified by Sephadex G-100 and G-75 column. Its molecular weight was 693kDa. APS and APS with ultra-high pressure treatment exhibited significant inhibitory abilities on a-glucosidase, inhibition rate from high to low in order was 400MPa-APS, 300MPa-APS, 500MPa-APS and APS. The inhibition ​percentage of 400MPa-APS (1.5mg/mL) was 49% (max.). This suggested that the inhibitory activity of APS on a-glucosidase was improved by ultra-high pressure treatment. FT-IR, SEM, CD spectra, atomic force microscope and Congo red test analysis of APS and 400MPa-APS showed ultra-high pressure treatment didn't change the preliminary structure but had an effect on its advanced structure.

Ergosterol-3-O-(β-D-galactopyranosyl-(1→4)-β-D-glucopyranoside) was efficiently synthesized and evaluated for its inhibitory activities against S180 cell lines compared with ergosterol. The structures of all synthesized compounds were fully characterized by spectroscopic data (NMR, MS). The antitumor activity of the new derivative was significantly enhanced compared with ergosterol.

The endothelium consists of a single layer of vascular endothelial cells (ECs) and serves as a selective barrier between the blood and arteries. ECs are constantly exposed to blood flow- and pulsatile blood pressure-induced hemodynamic forces. The cells are able to convert these mechanical stimuli into biochemical signals and then transmit the signals into the cell interior to affect cellular functions. These mechanical stimuli are detected by multiple mechanosensors in ECs that activate signaling pathways through their associated adaptor proteins, eventually leading to the maintenance of vascular homeostasis or the development of the pathogenesis of vascular disorders. These mechanosensors are distributed in different parts of the ECs, including the cell membrane, cell-to-cell junctions, the cytoplasm, and the nucleus. This review attempts to bring together recent findings on these mechanosensors and presents a conceptual framework for understanding the regulation of endothelial mechanosensors in response to hemodynamic forces. With verification by in vitro and in vivo evidence, endothelial mechanosensors have been demonstrated to contribute to health and disease by regulating physiological and pathophysiological processes in response to mechanical stimuli.

Arterial smooth muscle cell (SMC) phenotype and proliferation is regulated by their surrounding collagens, which transform from fibrillar to monomeric type in atherogenesis, and platelet-derived growth factor (PDGF)-BB/interleukin (IL)-1β. This study aims at elucidating the mechanisms by which physical (monomeric vs . fibrillar collagens) and chemical (PDGF-BB/IL-1β vs . vehicle controls) stimuli modulate SMC cycle and proliferation. SMCs were cultured on monomeric vs . fibrillar type I collagens. In parallel experiments, SMCs on fibrillar collagen were co-stimulated with PDGF-BB/IL-1β. These physical and chemical factors induced common SMC cycle signaling events, including up-regulations of cyclin-dependent kinase-4/6 and cyclins A/D1, phosphorylation of retinoblastoma (Rb) and its dissociations with E2F2/3. The physical and chemical inductions of SMC cycle signaling and progression were oppositely regulated by phosphatidylinositol 3-kinase (PI3K)-mediated Akt and p38 mitogen-activated protein kinase (MAPK). Fibrillar collagen degraded p66Shc, whose Ser36-phosphorylation plays important roles in the modulation of SMC cycle. Monomeric collagen and PDGF-BB/IL-1β co-stimulation induced p66Shc expression and Ser36-phosphorylation through β 1 integrin and PDGF receptor-β, respectively. In conclusion, our results demonstrate that fibrillar collagen-regulated p66Shc converges the physical and chemical stimuli to modulate SMC cycle and proliferation through PI3K-mediated Akt and p38 MAPK and their opposite regulation in downstream common cell cycle signaling cascades.

The effect of detonation gas upon rock fracture is the cross-frontier research project of fluid dynamics coupling and rock dynamics of crack dynamic growth, having the important scientific significance and explicit application value. In the process of detonation gas driven fracture growth, the varying law of the denotation gas pressure distribution in cracks is inter-influent with the geometric morphology variations in crack growth. This paper analyzes the denotation gas flowing behaviors, establishes the flowing model for denotation gas in rock cracks and the models for the rock crack dynamic response. Also suggests the fluid-solid coupling model for the denotation gas flowing, rock crack response and the numerical model for simulating crack growth driven by the denotation gas. The simulation program is developed on the large-sized rock engineering finite element software (FINAL platform). This numerical analysis platform is used to carry out the systematic numerical experimental study of several main influencing factors of core problems concerning pressure fracture length.

Rotifer community structure was investigated in Yangcheng Lake in 2008. Dominant species and species diversity indices were determined and QB/T was used to assess water quality. The annual average density and biomass of the rotifers in Yangcheng Lake was 2 894 ± 1 006 ind./L and 12.47 ± 10.28 mg/L, respectively. The highest densities were observed in the western portion of the lake in March, but the highest biomass occurred in inflowing creeks in September. Within a year of monitoring in Yangcheng Lake, 93 species were identified and the dominant species were found to be Polyarthra trigla, Brachionus angularis, Keratella cochlearis, Keratella valga, Brachionus calyciflorus, and Filinia major. Of the species recorded, 75 were pollution indicator species. Density and biomass exhibited significant positive correlations with water temperature (R = 0.209, P = 0.003; R = 0.446, P = 0.000), but the peak density showed two lags in response to chl a. According to the Jaccard similarity index (SJ), the greatest similarity among dominant species occurred between creeks and the eastern part of the lake. The annual average Shannon-Wiener diversity index H′, Margalef richness index D and Pielou evenness index J were 1.96 ± 0.34, 1.61 ± 0.50 and 0.77 ± 0.10, respectively. In all four areas of Yangcheng Lake, β-mesosaprobic species comprised the largest share of pollution indicator species. These data suggest that Yangcheng Lake is mesosaprobic.

Using RT-PCR and rapid amplification of cDNA ends, two new full-length cDNAs of SAD (TaSAD1 and TaSAD2) were obtained from a hardiest winter wheat cultivar (Mironovskaya808). Sequence comparison analysis showed that the deduced amino acid sequences of TaSAD1 and TaSAD2 had high similarity to those of other reported SAD proteins. They were also different each other by some substitutions, insertions and/or deletions involving single amino acid residues or motifs. Based on evolution analysis, it was clear that all SAD genes from Poaceae were closer than those from other genus such as Arabidopsis, Glycine, Triadica, Brassica, Sesamum and Bassia. All SAD genes clustered into two major groups in Poaceae. Meanwhile, TaSAD1 and TaSAD2 were clustered into different groups. The tertiary structure prediction indicated that both TaSAD1 and TaSAD2 proteins were a compact globular protein and their model structures almost were the same.

By acid polyacrylamide gel electrophoresis (A-PAGE) analysis, it was indicated that the electrophoresis mobility of gliadins from Crithopsis delileana (Schult) Roshev (2n=2x=14, KK) had obvious difference with those from common wheat in α, γ and ω region. Using homologous primers, two γ-gliadin genes (gli-Kr1 and gli-Kr2) were isolated from C. delileana, which had been deposited in the GenBank under accession numbers EU283818 and EU283821, respectively. Two γ -gliadin genes of C. delileana had the similar primary structures to the corresponding gene sequences from other wheat related species. The differences were mainly resulted from substitutions, insertions and deletions involving single amino acid residues or motifs of γ-gliadins. The repetitive domains of gli-Kr1 and gli-Kr2 from C. delileana are shorter than most of other sequences. By the alignment of γ-gliadin genes from A, B, D, Am, Au, S, Sl, Ssh, Ss and Sb genomes of Triticum and Aegilops, R genome of Secale (γ-secalin), Ee genome of Lophopyrum and K genome of Crithopsis in Triticeae, phylogenetic analysis indicated that two γ-gliadin genes of C. delileana could be clustered together with a γ-gliadin genefrom Ssh genome of Aegilops by an interior paralleled branch. It was the first time that the γ-gliadin genes encoded by K genome of C. delileana were characterized. These could offer precious information for better understanding the qualities associated with gliadins, the response in coeliac disease and studying the evolutionary relationship of gliadins in Triticeae.

In this paper a novel policy is proposed to solve the deadlock problem in a class of flexible manufacturing systems based on the notion that each shared buffer is partitioned into parts to store different types of products, respectively. A subclass of Petri nets called resource-shared net with buffers (RSNB) is defined. An RSNB is constructed by synthesizing some marked graphs, and each marked graph can model the process of manufacturing one type of product. RSNB cannot only model the concurrent execution of manufacturing processes, but also ensure that the modeled system is live. The process of constructing RSNB is described in detail, and a minimal siphon based necessary and sufficient condition is provided to characterize the liveness of RSNB.

Xylo-oligosaccharides, which are usually produced from xylan by enzymatic hydrolysis, have already become newly developed functional oligosaccharides with many beneficial biomedical and health effects. Xylo-oligosaccharides prepared by enzymatic hydrolysis from xylan were analyzed by thin-layer chromatography and high-pressure liquid chromatography in this study. Furthermore, xylose, xylobiose and xylotriose were separated by silica gel column chromatography. Then, their structures were identified by electrospray ionization mass spectrometry and nuclear magnetic resonance. Experiment promoting the discrepancy in bifidobacterium proliferation showed that xylotriose processed the best, followed by xylobiose and xylo-oligosaccharide. So, the main active ingredients of xylo-oligosaccharides were xylobiose and xylotriose, and the best bifidogenic factor was xylotriose.

Rationale: In atherosclerotic lesions, synthetic smooth muscle cells (sSMCs) induce aberrant microRNA (miR) profiles in endothelial cells (ECs) under flow stagnation. Increase in shear stress induces favorable miR modulation to mitigate sSMC-induced inflammation. Objective: To address the role of miRs in sSMC-induced EC inflammation and its inhibition by shear stress. Methods and Results: Coculturing ECs with sSMCs under static condition causes initial increases of 4 anti-inflammatory miRs (146a/708/451/98) in ECs followed by decreases below basal levels at 7 days; the increases for miR-146a/708 peaked at 24 hours and those for miR-451/98 lasted for only 6 to 12 hours. Shear stress (12 dynes/cm 2 ) to cocultured ECs for 24 hours augments these 4 miR expressions. In vivo, these 4 miRs are highly expressed in neointimal ECs in injured arteries under physiological levels of flow, but not expressed under flow stagnation. MiR-146a, miR-708, miR-451, and miR-98 target interleukin-1 receptor–associated kinase, inhibitor of nuclear factor–κB kinase subunit-γ, interleukin-6 receptor, and conserved helix-loop-helix ubiquitous kinase, respectively, to inhibit nuclear factor–κB signaling, which exerts negative feedback control on the biogenesis of these miRs. Nuclear factor-E2–related factor (Nrf)-2 is critical for shear-induction of miR-146a in cocultured ECs. Silencing either Nrf-2 or miR-146a led to increased neointima formation of injured rat carotid artery under physiological levels of flow. Overexpressing miR-146a inhibits neointima formation of rat or mouse carotid artery induced by injury or flow cessation. Conclusions: Nrf-2–mediated miR-146a expression is augmented by atheroprotective shear stress in ECs adjacent to sSMCs to inhibit neointima formation of injured arteries.

Taking mycelial polysaccharides from Cordyceps gunnii (C. gunnii) as the study subject, the effect of ultrasonic power, time and concentration of polysaccharides on antitumor activity of the polysaccharides was investigated. The ultrasonic processing condition of the polysaccharides was optimized by using orthogonal test design, and determined to be 400 W, 15 min and 1g/L. The change of structures of polysaccharides before and after ultrasonic treatment was also studied. Results show that ultrasonic treatment did not change the characteristic attribute of polysaccharides from C. gunnii. The composition of monosaccharide residues and the category of glycosidic bond have not been changed. But the molecular weight and intrinsic viscosity was reduced, and the alpha-helicity was enhanced after ultrasonic treatment. It was possible that ultrasonic treatment is an effective way for enhancing antitumor activity of polysaccharides.

We investigate the role of shear stress in regulating the gene expression in endothelial cells (ECs) in response to tumor necrosis factor-alpha (TNF-alpha). ECs were kept in static condition or pre-exposed to a high level (HSS, 20 dynes/cm2) or a low level of shear stress (LSS, 0.5 dynes/cm2) for 24 h, and TNF-alpha was added under static condition for 4 h. In static ECs, DNA microarray showed that TNF-alpha caused a significant increase in expression of 102 genes and a significant decrease in expression of 12 genes. Pre-shearing of ECs decreased the TNF-alpha-responsiveness of many pro-inflammatory, pro-coagulant, proliferative, and pro-apoptotic genes, whereas it increased the responsiveness of some antioxidant, anti-coagulant, and anti-apoptotic genes. LSS showed less regulatory effects than HSS on EC gene expression in response to TNF-alpha. The microarray data were confirmed by reverse-transcription polymerase chain reaction for 64 selected genes. Pre-shearing of ECs at HSS significantly inhibited the TNF-alpha-induced p65 and p50 mRNA expressions and nuclear factor-kappaB (NF-kappaB)-DNA binding activity. Inhibition of NF-kappaB activity with the p65-antisense or lactacystin under static condition blocked the expression of most of the genes that are TNF-alpha-inducible and shear stress-down-regulated. Our findings suggest that laminar shear stress serves protective functions against atherogenesis.

Objectives— Vascular endothelial cells (ECs) are influenced by shear stress and neighboring smooth muscle cells (SMCs). We investigated the inflammation-relevant gene expression in EC/SMC cocultures under static condition and in response to shear stress. Materials and Methods— Under static condition, DNA microarrays and reverse-transcription polymerase chain reaction identified 23 inflammation-relevant genes in ECs whose expression was significantly affected by coculture with SMCs, with 18 upregulated and 5 downregulated. Application of shear stress (12 dynes/cm 2 ) to the EC side of the coculture for 6 hours inhibited most of the proinflammatory gene expressions in ECs induced by coculture with SMCs. Inhibition of nuclear factor-κB (NF-κB) activation by the p65-antisense, lactacystin, and N-acetyl-cysteine blocked the coculture-induced EC expression of proinflammatory genes, indicating that the NF-κB binding sites in the promoters of these genes play a significant role in their expression as a result of coculture with SMCs. Chromatin immunoprecipitation assays demonstrated the in vivo regulation of NF-κB recruitment to selected target promoters. Shear stress inhibited the SMC coculture-induced NF-κB activation in ECs and monocytic THP-1 cell adhesion to ECs. Conclusions— Our findings suggest that shear stress plays an inhibitory role in the proinflammatory gene expression in ECs located in close proximity to SMCs.

Objective—Vascular endothelial cells (ECs) are subjected to shear stress and cytokine stimulation. We studied the interplay between shear stress and cytokine in modulating the expression of adhesion molecule genes in ECs.Methods and Results—Shear stress (20 dynes/cm2) was applied to ECs prior to and/or following the addition of tumor necrosis factor (TNF)-α. Shear stress increased the TNF-α–induced expression of intercellular adhesion molecule-1 (ICAM-1) at both mRNA and surface protein levels, but decreased the TNF-α–induced expression of vascular adhesion molecule-1 (VCAM-1) and E-selectin. Transfection studies using promoter reporter gene constructs of ICAM-1, VCAM-1, and E-selectin demonstrated that these shear stress modulations of gene expression occur at the transcriptional levels. After 24-hour preshearing followed by 1 hour of static incubation, the effect of preshearing on TNF-α–induced ICAM-1 mRNA expression vanished. The recovery of the TNF-α–induced VCAM-1 and E-selectin mRNA expressions following preshearing, however, required a static incubation time of >6 hours (complete recovery at 24 hours). Pre- and postshearing caused a reduction in the nuclear factor-κB-DNA binding activity induced by TNF-α in the EC nucleus.Conclusions—Our findings suggest that shear stress plays differential roles in modulating the TNF-α–induced expressions of ICAM-1 versus VCAM-1 and E-selectin genes in ECs.