Abstract Background: Conotoxins exhibit great potential as neuropharmacology tools and therapeutic candidates due to their high affinity and specificity for ion channels, neurotransmitter receptors or transporters. The traditional methods to discover new conotoxins are peptide purification from the crude venom or gene amplification from the venom duct. Methods: In this study, a novel O1 superfamily conotoxin Tx6.7 was directly cloned from the genomic DNA of Conus textile using primers corresponding to the conserved intronic sequence and 3’ UTR elements. The mature peptide of Tx6.7 (DCHERWDWCPASLLGVIYCCEGLICFIAFCI) was synthesized by solid-phase chemical synthesis and confirmed by mass spectrometry. Results: Patch clamp experiments on rat DRG neurons showed that Tx6.7 inhibited peak calcium currents by 59.29 ± 2.34% and peak potassium currents by 22.33 ± 7.81%. In addition, patch clamp on the ion channel subtypes showed that 10 μM Tx6.7 inhibited 56.61 ± 3.20% of the hCaV1.2 currents, 24.67 ± 0.91% of the hCaV2.2 currents and 7.30 ± 3.38% of the hNaV1.8 currents. Tx6.7 had no significant toxicity to ND7/23 cells and increased the pain threshold from 0.5 to 4 hours in the mouse hot plate assay. Conclusion: Our results suggested that direct cloning of conotoxin sequences from the genomic DNA of cone snails would be an alternative approach to obtaining novel conotoxins. Tx6.7 could be used as a probe tool for ion channel research or a therapeutic candidate for novel drug development. Background channels transporters duct Methods study O Tx67 Tx Tx6 7 Tx6. 3 elements DCHERWDWCPASLLGVIYCCEGLICFIAFCI (DCHERWDWCPASLLGVIYCCEGLICFIAFCI solidphase solid phase spectrometry Results 5929 59 29 59.2 234 2 34 2.34 2233 22 33 22.3 781 81 7.81% addition 1 5661 56 61 56.6 320 20 3.20 hCaV12 hCaV hCaV1 hCaV1. 2467 24 67 24.6 091 0 91 0.91 hCaV22 hCaV2 hCaV2. 730 30 7.3 338 38 3.38 hNaV18 hNaV hNaV1 8 hNaV1. ND723 ND ND7 23 ND7/2 05 5 0. assay Conclusion development 592 59. 2.3 223 22. 78 7.81 566 6 56. 32 3.2 246 24. 09 9 0.9 73 7. 3.3 ND72 ND7/ 2. 7.8 3.

Abstract Mesenchymal stem cells-derived exosomes (MSCs-exosomes) reportedly possess cardioprotective effects. This study investigated the therapeutic potential and mechanisms of MSCs-exosomes on heart failure (HF). H9c2 cells were used to establish a cardiomyocyte hypertrophy model by angiotensin II (Ang II) treatment. Isolated MSCs-exosomes were identified by transmission electron microscope and CD63 detection. Apoptosis rate was measured by terminal deoxynucleotidyl transferase (TdT) dUTP Nick-End Labeling (TUNEL) assay. Levels of inflammatory factors [interleukin (IL)-1β, IL-4, IL-6, and tumor necrosis factor (TNF)-α] and brain natriuretic peptide (BNP) were determined by ELISA. Expression of apoptosis-related proteins [Bax, B-cell lymphoma-2 (Bcl-2), and caspase 3] and Hippo-Yes-associated protein (YAP) pathway-related proteins [YAP, phosphor (p)-YAP, and tafazzin (TAZ)] was detected by western blotting. Cardiomyocyte hypertrophy of H9c2 cells induced by Ang II was ameliorated by MSCs-exosomes treatment. MSCs-exosomes downregulated Bax and caspase 3 levels and upregulated Bcl-2 level in Ang II-induced H9c2 cells. MSCs-exosomes also reduced the levels of BNP, IL-1β, IL-4, IL-6, and TNF-α in Ang II-induced H9c2 cells. Meanwhile, p-YAP was downregulated and TAZ was upregulated after MSCs-exosomes administration. In conclusion, MSCs-exosomes alleviate the apoptosis and inflammatory response of cardiomyocyte via deactivating Hippo-YAP pathway in HF. cellsderived derived MSCsexosomes MSCs (MSCs-exosomes effects HF . (HF) Hc H c H9c treatment CD CD6 detection TdT (TdT NickEnd Nick End TUNEL (TUNEL assay interleukin IL1β, IL1β ILβ IL 1β, 1β β (IL)-1β IL4, IL4 4, 4 IL-4 IL6, IL6 6, 6 IL-6 TNFα TNF α (TNF)-α BNP (BNP ELISA apoptosisrelated related Bax, [Bax Bcell B cell lymphoma2 lymphoma 2 lymphoma- Bcl2, Bcl2 Bcl , (Bcl-2) HippoYesassociated Hippo Yes associated YAP (YAP pathwayrelated YAP, [YAP pYAP, pYAP p (p)-YAP (TAZ) blotting Bcl- IIinduced IL-1β Meanwhile administration conclusion HippoYAP (HF IL- (Bcl-2 (TAZ (Bcl- (Bcl

Abstract The aim of this study was to investigate the influence of microwave drying on the protein quality of japonica sorghum following an intermittent drying test. Using label-free technology and liquid chromatography-tandem mass spectrometry for proteomic analysis, the effects of microwave drying on sorghum differential protein expression, functional classification, and metabolic pathways were analyzed at the molecular level. After sorghum was dried using a microwave, 85 differential proteins were identified. Among them, 51 showed up-regulated expression while 34 had down-regulated expression. The up-regulation and down-regulation of differential protein expressions significantly changed them, and proteins with larger up-regulated and down-regulated expressions were postulated to affect biological and metabolic processes of sorghum during subsequent processing. Differential proteins were significantly (P ˂ 0.01) involved in metabolic pathways, such as carbon metabolism, glycolysis/gluconeogenesis, carbon fixation in photosynthetic organisms, biosynthesis of amino acids, amino sugar and nucleotide sugar metabolism, and the TCA cycle. For the protein interaction network, glyceraldehyde-3-phosphate dehydrogenase of the down-regulated proteins was postulated to be the key factor affecting the entire metabolic system or signal transduction pathway. Up-regulated proteins, including phosphoglycerate mutase and phosphopyruvate hydratase, as well as down-regulated proteins such as glyceraldehyde-3-phosphate dehydrogenase and fructose-bisphosphate aldolase, not only directly or indirectly affected a variety of metabolic processes, but were specifically closely related to glycolysis and glycometabolism. Overall, this study showed that among the related metabolic pathways, differential protein changes in glycometabolism pathways may have the greatest impact on metabolic processes. The research results discussed herein can provide theoretical support for the industrial application of microwave drying and deep processing of sorghum.

Weaning results in intestinal dysfunction, mucosal atrophy, transient anorexia, and intestinal barrier defects. In this study, the effect of prodigiosin (PG) on the intestinal inflammation of weaned rats was investigated by using 1H-NMR spectroscopy and biochemistry indexes to regulate the intestinal metabolism. After administration for 14 days, the body mass of the PG group was increased by 1.29- and 1.26-fold compared with those of the control and alcohol groups, respectively, using a dose of 200 µg PG·kg-1 body weight per day. PG increased organic acid content and decreased moisture, pH values, and free ammonia in feces. In addition, PG alleviated the intestinal inflammation of weaned rats. The analysis of 1H-NMR signal peak attribution and the model validation of metabolic data of feces contents showed that PG significantly affected the metabolism of small molecular compounds in the intestinal tract of weaned rats. This study presents the promising alternative of using PG to alleviate intestinal inflammation effectively in the intestinal tract of weaned rats.

In July 2010, cacti with typical phytoplasma symptoms were observed in Yangling district, Shaanxi Province, China. Based on amplification of 16S rRNA gene, phytoplasmas were detected by polymerase chain reaction (PCR) driven with universal primer pairs P1/ P7, followed by a nested PCR with universal primer pairs R16F2n/R16R2. Fragments of expected sizes (1.8 kb and 1.2 kb) were obtained from symptomatic samples, but not from the asymptomatic samples. Sequencing results and BLASTn analysis of the 1806 bp products (P1/P7) showed that the phytoplasma belonged to group 16SrII. Phylogenetic analysis to the R16F2n/R16R2 region and virtual restriction fragment length polymorphism (RFLP) indicated that this phytoplasma was a member of subgroup 16SrII-C. This is the first reported occurrence of a peanut witches'-broom phytoplasma infecting plants in northern areas of China.