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Sensitivity to bitter tastes provides animals with an important means of interacting with their environment and thus, influences their dietary preferences. Genetic variants encoding functionally distinct receptor types contribute to variation in bitter taste sensitivity. Our previous study showed that two nonsynonymous sites, A52V and Q296H, in the TAS2R20 gene are directionally selected in giant pandas from the Qinling Mountains, which are speculated to be the causative base‐pair changes of Qinling pandas for the higher preference for bamboo leaves in comparison with other pandas. Here, we used functional expression in engineered cells to identify agonists of pTAS2R20 (i.e., giant panda's TAS2R20) and interrogated the differences in perception in the in vitro responses of pTAS2R20 variants to the agonists. Our results show that pTAS2R20 is specifically activated by quercitrin and that pTAS2R20 variants exhibit differences in the sensitivity of their response to the agonist. Compared with pTAS2R20 in pandas from other areas, the receptor variant with A52V and Q296H, which is most commonly found in Qinling pandas, confers a significantly decreased sensitivity to quercitrin. We subsequently quantified the quercitrin content of the leaves of bamboo distributed in the Qinling Mountains, which was found to be significantly higher than that of the leaves of bamboo from panda habitats in other areas. Our results suggest that the decreased sensitivity to quercitrin in Qinling pandas results in higher‐quercitrin‐containing bamboo leaves to be tasting less bitter to them and thus, influences their dietary preference. This study illustrates the genetic adaptation of Qinling pandas to their environments and provides a fine example of the functional effects of directional selection in the giant panda., Sensitivity to bitter tastes provides animals with an important means of interacting with their environment and thus, influences their dietary preferences. TAS2R20 variants confer dietary adaptation to high‐quercitrin bamboo leaves in Qinling giant pandas. The decreased sensitivity to quercitrin in Qinling pandas results in higher‐quercitrin‐containing bamboo leaves to be tasting less bitter to them and thus, influences their dietary preference.

The synthesis of α-linolenic acid (ALA) requires the activity of ω-3 fatty acid desaturases (ω-3 FADs). The quality of peanut oil would be much improved if the content of ALA could be increased. A scan of the peanut genome revealed that it harbored 36 FAD genes, mapping to 16 of the species’ 20 chromosomes. A phylogenetic analysis concluded that these genes belonged to six sub-families, namely stearoyl-acyl-acyl carrier protein desaturases (SAD), FAD2, FAD3, FAD4/5, FAD6 and FAD7/8. Of these, FAD3 and FAD7/8 encoded ω-3 FADs, while genes belonging to the other four sub-families encoded ω-6 FADs. Based on RNA-Seq data, each of the 36 FAD genes was shown to be transcribed in non-stressed plants, but there was variation between them with respect to which organs they were transcribed in. Four ω-3 AhFAD3 genes were functionally characterized; when expressed in Arabidopsis thaliana protoplasts, each was localized mainly in the endoplasmic reticulum, while within peanut, the genes were more strongly transcribed in the developing seed than in either the root or the leaf. When constitutively expressed in Arabidopsis thaliana, both the total fatty acid content of the seed and the relative contribution of ALA were increased. The transgenic seedlings also exhibited an improved level of survival when challenged by salinity stress.

High-pressure homogenization (HPH) has the potential to improve the exaction yield of bioactive compound from food and food waste. This study investigated the use of HPH or microfluidization as an alternative assisted method for sulforaphane extraction from raw broccoli seeds. The mean particle size, morphological characteristics, and extraction yields of all samples processed at different HPH pressure levels (3000-23,000 psi) and passes (1–5) were examined. After HPH, the particle size of broccoli seeds was reduced 2–10 times and the particle size distribution pattern also changed from bimodal to unimodal with increasing pressures and number of passes. The highest sulforaphane content obtained with HPH at 5000 psi and 5 passes was 2199 μg sulforaphane/g broccoli seeds, which is 3 times more than the control one. Particular interest was given to the relationship between sulforaphane yield and particle size. Generally, the extraction yield increased with decreasing particle size; however, excessive size reduction did not necessarily result in significant increase in sulforaphane content. In addition, the pressure is not the higher the better (

Several studies have highlighted the underlying role of mesenchymal stem cells microvesicles (MSC-MVs) in acute lung injury (ALI). Hepatocyte growth factor (HGF) derived from MSC-MVs is partly involved in their therapeutic effects; however, the detailed mechanism remains unclear. MVs were isolated from human Wharton's Jelly MSCs. The rat model of ALI was established by intratracheal instillation of bleomycin (BLM). A co-culture model of alveolar epithelial cells or pulmonary endothelial cells and MSC-MVs was utilized. Total protein content in bronchoalveolar lavage fluid (BALF) was determined by bicinchoninic acid method. White blood cell (WBC) and neutrophil in BALF were counted. ELISA was used for the determination of cytokines and HGF in BALF. Apoptosis was determined by TUNEL assay and Annexin V-FITC/PI staining as well as caspase-3 activity detection. HE and Masson staining of lung tissues was used for histopathology analysis. The expression of HGF and proteins involved in the PI3K/AKT/mTOR pathway were measured by quantitative Real-Time PCR (qRT-PCR) and western blotting. Treatment with MSC-MVs significantly inhibited BLM-induced apoptosis and fibrosis in lung tissues and PI3K/AKT/mTOR activation, which was reversed by HGF mRNA deficient MVs. Intriguingly, these effects were completely abrogated by PI3K inhibitor. The therapeutic effect of MSC-MVs in ALI was partly mediated through HGF mRNA.

The NAC transcription factor (TF) is one of the most significant TFs in plants and is widely involved in plant growth, development, and responses to biotic and abiotic stresses. To date, there are no systematic studies on the NAC family in peanuts. Herein, 132 AhNACs were identified from the genome of cultivated peanut, and they were classified into eight subgroups (I–VIII) based on phylogenetic relationships with Arabidopsis NAC proteins and their conserved motifs. These genes were unevenly scattered on all 20 chromosomes, among which 116 pairs of fragment duplication events and 1 pair of tandem duplications existed. Transcriptome analysis showed that many AhNAC genes responded to drought and abscisic acid (ABA) stresses, especially most of the members in groups IV, VII, and VIII, which were expressed at larger differential levels under polyethylene glycol (PEG) and/or ABA treatment in roots or leaves. Furthermore, 20 of them selected in response to PEG and ABA treatment were evaluated by quantitative real-time polymerase chain reaction. The results showed that these genes significantly responded to drought and ABA in roots and/or leaves. This study was helpful for guiding the functional characterization and improvement of drought-resistant germplasms in peanuts.

There are currently many problems related to excessive fertilizer application, low fertilizer-use efficiency and lack of an agricultural labor force for cotton production in China. Therefore, the objective of this paper was to explain the optimal application time for once fertilization based on cotton nutrient accumulation of nitrogen, phosphorus and potassium to provide technical support for simplified fertilization management in cotton production. A 2 yr field experiment and a 1 yr pot experiment were conducted with fertilizer (225, 67.5, and 225 kg ha−1 of N, P2O5, and K2O, respectively) applied once at 0 (FT1), 5 (FT2), 10 (FT3), 15 (FT4), or 20 (FT5) d after the appearance of the first flower and a triple application (preplant 30%, first bloom 40%, and peak bloom 30%) as the conventional control (FT6). The results showed that FT1 exhibited the greatest nutrient accumulation speed for both the average (5.81, 1.22, and 5.74 kg ha−1 of N, P2O5, and K2O, respectively) and the maximum (6.31, 1.44, and 6.24 kg ha−1 of N, P2O5, and K2O, respectively) during the fast accumulation period. Moreover, among the different treatments, FT1 exhibited the greatest nutrient recovery and partial productivity. The results suggest that applying the minimum amount of fertilizer at the appearance of the first flower is optimal for maximizing nutrient utilization while minimizing environmental disturbance.

Because the transcription factor activator protein-1 (AP-1) regulates a variety of protein-encoding genes, it is a participant in many cellular functions, including proliferation, transformation, epithelial mesenchymal transition (EMT), and apoptosis. Inhibitors targeting AP-1 have potential use in the treatment of cancer and other inflammatory diseases. Here, we identify veratramine as a potent natural modulator of AP-1, which selectively binds to a specific site (TRE 5′-TGACTCA-3′) of the AP-1 target DNA sequence and regulates AP-1-dependent gene transcription without interfering with cystosolic signaling cascades that might lead to AP-1 activation. Moreover, RNA-seq experiments demonstrate that veratramine does not act on the Hedgehog signaling pathway in contrast to its analogue, cyclopamine, and likely does not harbor the same teratogenicity and toxicity. Additionally, veratramine effectively suppresses EGF-induced AP-1 transactivation and transformation of JB6 P+ cells. Finally, we demonstrate that veratramine inhibits solar-ultraviolet-induced AP-1 activation in mice. The identification of veratramine and new findings in its specific regulation of AP-1 down stream genes pave ways to discovering and designing regulators to regulate transcription factor.

Triacylglycerols (TAGs) are the most important energy storage form in oilseed crops. Diacylglycerol acyltransferase (DGAT) catalyzes the rate-limiting step of the Kennedy pathway of TAG biosynthesis. To date, little is known about the regulation of DGAT activity in peanut (Arachis hypogaea), an agronomically important oilseed crop that is cultivated in many parts of the world. In this study, seven distinct forms of type 1 DGAT (AhDGAT1.1-AhDGAT1.7) were identified, cloned, and characterized. Comparisons of the nucleotide sequences and gene structures revealed many different splicing variants of AhDGAT1, some of which displayed different organ-specific expression patterns. A representative gene (AhDGAT1.1) was transformed into wild-type tobacco and was shown to increase seed fatty acid (FA) content by 14.7%-20.9%. All seven AhDGAT1s were expressed in TAG-deficient Saccharomyces cerevisiae strain H1246; the five longest AhDGAT1 variants generated high levels of acyltransferase activity and complemented the free fatty acid lethality phenotype in this strain. The alternative splicing that gives rise to AhDGAT1.2 and AhDGAT1.4 creates predicted protein C-terminal truncations. The proteins encoded by these two variants were not active and did not complement the fatty acid sensitivity in H1246. These results were verified by visualization of intracellular lipid droplets using Nile Red staining. Collectively, the results presented here represent the first comprehensive analysis of the peanut DGAT1 gene family, which, unlike in other published plant DGAT1 sequences, shows widespread alternative splicing that may affect the expression patterns and enzyme activities of some members of the gene family.

The Plasmodium falciparum cysteine protease falcipain-2 (FP-2) is an attractive antimalarial target. Here, we discovered that the natural compound NP1024 is a nonpeptidic inhibitor of FP-2 with an IC50 value of 0.44 μmol L–1. The most exciting finding is that both in vitro and in vivo , NP1024 directly targets FP-2 in malaria parasite-infected erythrocytes as a natural fluorescent probe, thereby paving the way for an integration of malaria diagnosis and treatment.

Seed dormancy and germination are the two important traits related to plant survival and reproduction, and crop yield. To understand their regulation mechanism, it is crucial to clarify which genes or which pathways participate in the regulation of these processes. However, little information is available during the procedure of seed dormancy and germination in peanut. In this study, the seeds of the variety Luhua No.14 with non-deep dormancy were selected and its transcriptional changes at three developmental stages: the fresh-harvest (FS), the after-ripened (DS) and the just-germinated seeds (GS), were investigated by comparative transcriptomics analysis. The results showed that genes with increased transcription in DS vs FS comparison were overrepresented for oxidative phosphorylation, glycolysis pathway and tricarboxylic acid cycle (TCA), suggesting that after a period of drying storage, the intermediates stored in dry seeds were rapidly mobilized by glycolysis, TCA cycle, glyoxylate cycle, etc.; the electron transport chain accompanying with respiration has been reactivated to provide ATP for mobilization of other reserves and seed germination. In GS vs DS pairwise, dozens of the up-regulated genes were related to plant hormone biosynthesis and signal transduction, including the majority of components in auxin signal pathway, and brassinosteroid biosynthesis and signal transduction, and some GA and ABA signal transduction genes. During seeds germination, the expression of some EXPANSIN and XYLOGLUCAN ENDOTRANSGLYCOSYLASE was also significantly enhanced. To investigate the effect of different hormone during the procedure of seed germination, the contents and the differential distribution of ABA, GA, BR and IAA in cotyledon, hypocotyl and radicle, and plumule of three seed sections at different developmental stages were also detected. Combining with previous data in other species, a model of regulatory network related to peanut seed germination was developed. This model will helpful to gain further understanding of the mechanisms controlling seed dormancy and germination.

Soil organic matter (SOM) property and the desorption effect by the hydrochemical solution during rainfall or irrigation play key roles in soil carbon dynamics. The quantity and molecular characteristics of the dissolved organic matter (DOM) extracted from the soil at a wheat field in the North China Plain were investigated to understand the chemical release characteristic of the SOM and the responsible factors. The amounts of DOM desorbed from the different extraction solutions were of the following order: Na2SO4 (62.1 mg/kg) > NaCl (58.7 mg/kg) > H2O (55.7 mg/kg)≈NaHCO3 (55.6 mg/kg) > CaCl2(46.2 mg/kg). The comparative differences of spectroscopy parameters including the smaller E2/E3, larger SUVA254, higher HIX, and lower BIX and FI of the Na-derived DOM than those of the Ca-obtained DOM indicate that the sodium electrolytes were favorable for the desorption of the organic matter with larger molecular weight, higher aromaticity, and humification degrees representing the more refractory property of the DOM. The good ternary relationship of the soil capacity of exchangeable cation (CEC), HIX of soil DOM, and soil TOC (rCEC-HIX = 0.78 ± 0.07, p

Leaching dynamics of dissolved organic matter (DOM) from agricultural soil are influenced by the chemistry of irrigation water. This study used multiple spectroscopy methods to investigate the dynamics of DOM in column leaching process by utilizing selected solutions. DOC was leached out by Na solutions more effectively than by Ca, despite the additional contributions of the ligand exchange of SO42− and the hydrolysis of HCO3-. The concentration ranking corresponding to solutions is Na2SO4 > NaHCO3 > NaCl > CaCl2. Furthermore, the aromaticity and molecular weights of DOM leached by Na solutions increased with the leaching, whereas contrary trends were reared by Ca. Thus, specific cation substantially affects DOM concentration and composition. Parallel factor analysis of the three-dimensional fluorescence spectra (EEM-PARAFAC) determined the characteristics of DOM components. (1) The long-wavelength, humic-like components (HLC, i.e., C1, C3, and C5) were largely accumulated at the early water saturating stage but attenuated toward dynamic stability in the salt-solution leaching (SSL) stage. (2) Reactive functional group-related HLC (i.e., C2, and C4) remarkably increased in response to the solutions chemistry. (3) The protein-like component (PLC, i.e., C6) was released with insignificant dynamic differences between the solutions. Despite approximately stable DOC concentration at the late stage of each SSL, HLC exhibited accelerating release in sodium salts but declining trend in calcium salt whereas PLC showed a roughly constant dynamic in the both. Ca-bridging reactions with the detachment of OM-Ca-clay complex prompting HLC release and its formation to stabilize the HLC well explain the contrast dynamics of HLC with Na or Ca leaching. The diffusion process that was defined for the slow, lasting and small concentration of leaching of inactive components could contribute to the steady dynamic of PLC, given its insensitivity to either the salt cation. This study identified distinct release kinetics of DOM compositions from those of DOC by EEM-PARAFAC and correspondingly the potential mechanisms, which offers promising advantages in monitoring DOM transport and fractionation in dynamic leaching process.

A novel series of tetrahydroprotoberberine derivatives (THPBs) were designed, synthesized, and evaluated as selective α1A-adrenergic receptors (AR) antagonists for the treatment of benign prostatic hyperplasia. On the basis of the pharmacophore model of the marketed drug silodosin, THPBs were modified by introducing an indole segment into their core scaffolds. In calcium assays, 7 out of 32 compounds displayed excellent antagonistic activities against α1A-ARs, with IC50 less than 250 nM. Among them, compound (S)-27 had the most potent biological activity; its IC50 toward α1A-AR was 12.8 ± 2.2 nM, which is 781 and 20 times more selective than that toward α1B- and α1D-AR, respectively. In the functional assay using isolated rat tissues, compound (S)-27 inhibited norepinephrine-induced urethra smooth muscle contraction potently (IC50 = 0.5 ± 0.3 nM), without inhibiting the aortic contraction (IC50 > 1000 nM), displaying a better tissue selectivity than the marketed drug silodosin. Additional results of preli...

The tribocorrosion behavior of type 316L stainless steel and CrN coated 316L by PVD was investigated in seawater environment. Tests were carried out in a triboelectrochemical cell using a ball-on-plate tribometer integrated with a potentiostat for electrochemical control. Sliding contact against silicon nitride balls had a significant effect on the open circuit potential (OCP) and corrosion potential ( E corr ) for 316L. However, the OCP for the CrN coating remained relatively stable during the sliding. Tribocorrosion tests performed under various applied potentials demonstrated that the friction coefficient decreased and wear-loss increased as the applied potential increased from −1 V to 0 V. Under cathodic potential of −1 V, abrasive wear form of mechanical wear dominated the wear of 316L stainless steel. As the potential increases, the general corrosion was observed and tribo-corrosive contributions to the wear increased. Plastic deformation dominated the wear process of CrN coating under a cathodic potential. With an increase in potential, the mechanical wear contribution decreased and corrosion-accelerated wear contribution increased. At high anodic potential (0.5 V), pitting corrosion occurred in the wear track and on the outer part of wear track on 316L and CrN coated 316L.

Interfaces between materials with different electronic ground states have become powerful platforms for creating and controlling novel quantum states of matter, in which inversion symmetry breaking and other effects at the interface may introduce additional electronic states. Among the emergent phenomena, superconductivity is of particular interest. Here, by depositing metal films on a newly identified topological semimetal tungsten carbide (WC) single crystal, interfacial superconductivity is obtained, evidenced from soft point-contact spectroscopy. This very robust phenomenon is demonstrated for a wide range of metal/WC interfaces, involving both nonmagnetic and ferromagnetic films, and the superconducting transition temperatures are surprisingly insensitive to the magnetism of thin films. This method offers an opportunity to explore the long-sought topological superconductivity and has potential applications in topological-state-based spin devices.

Seed dormancy and germination are the two important traits related to plant survival, reproduction and crop yield. To understand the regulatory mechanisms of these traits, it is crucial to clarify which genes or pathways participate in the regulation of these processes. However, little information is available on seed dormancy and germination in peanut. In this study, seeds of the variety Luhua No.14, which undergoes nondeep dormancy, were selected, and their transcriptional changes at three different developmental stages, the freshly harvested seed (FS), the after-ripening seed (DS) and the newly germinated seed (GS) stages, were investigated by comparative transcriptomic analysis. The results showed that genes with increased transcription in the DS vs FS comparison were overrepresented for oxidative phosphorylation, the glycolysis pathway and the tricarboxylic acid (TCA) cycle, suggesting that after a period of dry storage, the intermediates stored in the dry seeds were rapidly mobilized by glycolysis, the TCA cycle, the glyoxylate cycle, etc.; the electron transport chain accompanied by respiration was reactivated to provide ATP for the mobilization of other reserves and for seed germination. In the GS vs DS pairwise comparison, dozens of the upregulated genes were related to plant hormone biosynthesis and signal transduction, including the majority of components involved in the auxin signal pathway, brassinosteroid biosynthesis and signal transduction as well as some GA and ABA signal transduction genes. During seed germination, the expression of some EXPANSIN and XYLOGLUCAN ENDOTRANSGLYCOSYLASE genes was also significantly enhanced. To investigate the effects of different hormones during seed germination, the contents and differential distribution of ABA, GAs, BRs and IAA in the cotyledons, hypocotyls and radicles, and plumules of three seed sections at different developmental stages were also investigated. Combined with previous data in other species, it was suggested that the coordination of multiple hormone signal transduction nets plays a key role in radicle protrusion and seed germination.

Farnesoid X receptor (FXR) is a member of nuclear receptor family involved in multiple physiological processes through regulating specific target genes. The critical role of FXR as a transcriptional regulator makes it a promising target for diverse diseases, especially those related to metabolic disorders such as diabetes and cholestasis. However, the underlying activation mechanism of FXR is still a blur owing to the absence of proper FXR modulators. To identify potential FXR modulators, an in-house natural product database (NPD) containing over 4,000 compounds was screened by structure-based virtual screening strategy and subsequent hit-based similarity searching method. After the yeast two-hybrid (Y2H) assay, six natural products were identified as FXR antagonists which blocked the CDCA-induced SRC-1 association. The IC50 values of compounds 2a, a diterpene bearing polycyclic skeleton, and 3a, named daphneone with chain scaffold, are as low as 1.29 and 1.79 μM, respectively. Compared to the control compound guggulsterone (IC50 = 6.47 μM), compounds 2a and 3a displayed 5- and 3-fold higher antagonistic activities against FXR, respectively. Remarkably, the two representative compounds shared low topological similarities with other reported FXR antagonists. According to the putative binding poses, the molecular basis of these antagonists against FXR was also elucidated in this report.

Peanut (Arachis hypogaea L.) is one of the major oil crops and is the fifth largest source of plant oils in the world. Numerous genes participate in regulating the biosynthesis and accumulation of the storage lipids in seeds or other reservoir organs, among which several transcription factors, such as LEAFY COTYLEDON1 (AtLEC1), LEC2, and WRINKLED1 (WRI1), involved in embryo development also control the lipid reservoir in seeds. In this study, the AtLEC1 gene was transferred into the peanut genome and expressed in a seed-specific manner driven by the NapinA full-length promoter or its truncated 230-bp promoter. Four homozygous transgenic lines, two lines with the longer promoter and the other two with the truncated one, were selected for further analysis. The AtLEC1 mRNA level and the corresponding protein accumulation in different transgenic overexpression lines were altered, and the transgenic plants grew and developed normally without any detrimental effects on major agronomic traits. In the developing seeds of transgenic peanuts, the mRNA levels of a series of genes were upregulated. These genes are associated with fatty acid (FA) biosynthesis and lipid accumulation. The former set of genes included the homomeric ACCase A (AhACC II), the BC subunit of heteromeric ACCase (AhBC4), ketoacyl-ACP synthetase (AhKAS II), and stearoyl-ACP desaturase (AhSAD), while the latter ones were the diacylglycerol acyltransferases and oleosins (AhDGAT1, AhDGAT2, AhOle1, AhOle2, and AhOle3). The oil content and seed weight increased by 4.42–15.89% and 11.1–22.2%, respectively, and the levels of major FA components including stearic acid, oleic acid, and linoleic acid changed significantly in all different lines.

We report the observation of local superconductivity induced at the point contact formed between a normal metal tip and WC -- a triple point topological semimetal with super hardness. Remarkably, the maximum critical temperature is up to near 12 K but insensitive to the tip's magnetism. The lateral dimensions of the superconducting puddles were evaluated and the temperature dependencies of superconducting gap and upper critical field were also obtained. These results put constraints on the explanation of the induced superconductivity and pave a pathway for exploring topological superconductivity., Comment: 7 pages, 6 figures

Here we report the discovery of the first ternary molybdenum pnictide based superconductor K2Mo3As3. Polycrystalline samples were synthesized by the conventional solid state reaction method. X-ray diffraction analysis reveals a quasi-one-dimensional hexagonal crystal structure with (Mo3As3)2- linear chains separated by K+ ions, similar as previously reported K2Cr3As3, with the space group of P-6m2 (No. 187) and the refined lattice parameters a = 10.145(5) {\AA} and c = 4.453(8) {\AA}. Electrical resistivity, magnetic susceptibility, and heat capacity measurements exhibit bulk superconductivity with the onset Tc at 10.4 K in K2Mo3As3 which is higher than the isostructural Cr-based superconductors. Being the same group VIB transition elements and with similar structural motifs, these Cr and Mo based superconductors may share some common underlying origins for the occurrence of superconductivity and need more investigations to uncover the electron pairing within a quasi-one-dimensional chain structure.

Oxidative stress and neuroinflammation are highly relevant to the pathological processes of various neurodegenerative diseases including Alzheimer's disease (AD). (+)-2-(1-hydroxyl-4-oxocyclohexyl) ethyl caffeate (HOEC), a novel 5-lipoxygenase inhibitor, was isolated from the whole plant of Incarvillea mairei var granditlora (Wehrhahn) Grierson. In this study, we investigated the protective effect of HOEC on hydrogen peroxide (H2O2) and lipopolysaccharide (LPS) -induced cytotoxicity and neuroinflammation in vitro and in vivo. MTT assay, LDH release assay, morphological observation and Hoechst 33342/PI dual staining followed by EIA, immunofluorescence staining and Western Blotting analysis were performed to elucidate the neuroprotective effect of HOEC. Treatment with HOEC at various concentrations prior to H2O2 exposure significantly enhanced cell viability, decreased LDH release, prevented cell morphologic changes and apoptosis. Instead of PGE2 reduction, HOEC markedly inhibited the production of LTB4 and suppressed the macrophage-mediated neurotoxicity. Western blotting and immunofluorescence staining showed that HOEC inhibited H2O2-induced p38 phosphorylation and NF-κB activation. Neuroprotective effect of HOEC was abolished by a p38 inhibitor. Further in vivo studies of LPS-induced neuroinflammation confirmed the anti-inflammatory effects of HOEC. These findings that HOEC protects SH-SY5Y cells from H2O2 and LPS-induced injury via arachidonic acid network modulation followed by p38 MAPK and NF-κB signaling, might make HOEC be considered as a therapeutic candidate for prevention and treatment of neurodegenerative diseases involving oxidative stress or/and inflammation.

Valepotriates, plant secondary metabolites of the family Valerianaceae, contain various acyloxy group linkages to the valepotriate nucleus and exhibit significant biological activities. Identification of valepotriates is important to uncover potential lead compounds for the development of new sedative and antitumor drugs. However, making their structure elucidation by nuclear magnetic resonance (NMR) experiments is too difficult to be realized because of the overlapped carbonyl carbon signals of acyloxy groups substituted at different positions. Thus, the mass spectrometric profiling of these compounds in positive ion mode was developed to unveil the exact linkage of acyloxy group and the core of valepotriate. In this study, electrospray ionization tandem multistage mass spectrometry (ESI-MS/MS(n)) in ion trap and collision-induced dissociation tandem MS were used to investigate the fragmentation pathways of four types of valepotriates in Valeriana jatamansi, including 5-hydroxy-5,6-dihydrovaltrate hydrin (5-hydroxy-5,6-dihydrovaltrate chlorohydrin), 5,6-dihydrovaltrate hydrin (5,6-dihydrovaltrate chlorohydrin), 5-hydroxy-5,6-dihydrovaltrate and valtrate hydrin (valtrate chlorohydrin). The high-resolution mass spectrum (HRMS) data of all the investigated valepotriates from quadrupole time-of-flight MS/MS were used as a supportive of the fragmentation rules we hypothesized from ion-trap stepwise MS(n). As a result, the loss sequence of acyloxy groups and the abundance of key product ions, in combination with the characteristic product ions corresponding to the valepotriate nucleus, could readily differentiate the four different types of valepotriates. The summarized fragmentation rules were also successfully exploited for the structural characterization of three new trace valepotriates from V. jatamansi. The results indicated that the developed analytical method could be employed as a rapid, effective technique for structural characterization of valepotriates, especially for the trace compounds that could not be identified by NMR techniques. This study may also arouse interest for further structural analysis of other valepotriate-containing type herbal medicines.

Systematic phytochemical investigations on Abies fabri resulted in the isolation of 94 compounds, consisting of 68 terpenoids, six lignans, seven flavonoids, and 13 other miscellaneous chemical constituents. Their structures were elucidated on the basis of spectroscopic methods, and the absolute configurations of three of these previously unknown compounds were determined by Cu-Kα X-ray crystallographic analysis. Twelve previously unreported compounds, one artifact, and one potential artifact were identified, including six triterpenoids, four diterpenoids, two sesquiterpenoids, one lignan, and one phenol. 23-Hydroxy-3-oxolanosta-8,24-dien-26,23-olide showed weak cytotoxic activity against A549 and THP-1 cells with the IC50 values of 5.3 and 5.1 μM, respectively.

The biotransformation of cyclopamine ( 1 ) and its congeners ( 2 , 3 and 4 ) by Cunninghamella echinulata (ACCC 30369) was investigated. The chemical structures of two new congeners ( 2 and 4 ) and nine new metabolites were elucidated by 1D NMR ( 1 H, 13 C and DEPT), 2D NMR (COSY, HMBC, HSQC and NOESY) and HRESIMS analyses and further confirmed by X-ray diffraction studies. Among these compounds, 2 and 4 showed moderate cytotoxicity in HepG2 cells ( 2 , IC 50 = 8.03 ± 1.92 μM) and A549 cells ( 4 , IC 50 = 10.19 ± 2.18 μM). Conversely, the cytotoxicity of the nine metabolites was sharply reduced. Similar to 1 , compound 4 induced a cyclopia phenotype in zebrafish embryos at 20 μM. Moreover, compound 4 was more stable than 1 in an acidic environment.

1β-Hydroxyalantolactone (IJ-5) is a sesquiterpene lactone compound isolated from Inula japonica Thunb (Asteraceae). Sesquiterpene lactones have been shown to modulate many processes that influence inflammatory reactions.The present study examines the protective effect of IJ-5 on atopic dermatitis (AD) in a mouse model induced by 2,4-dinitrochlorobenzene (DNCB).AD-like skin lesions were induced in Balb/c mice by sensitizing once with painting 100 μL of 5% DNCB on their shaved back skin and then challenging with 20 μL of 0.2% DNCB five times on their right ears at 3 d interval starting on day 5 post-sensitization. IJ-5 was administrated intraperitoneally at 10 mg/kg 1 h before each DNCB challenge.IJ-5 treatment attenuated DNCB-induced dermatitis severity and right ear swelling. The serum levels of IgE, IL-4, and IL-6 in IJ-5-treated mice were reduced by 54.7, 56.5, and 53.0%, respectively, while the mRNA levels of TNFα, IL-1, IL-4, and IL-6 in back skin lesions of IJ-5-treated mice were reduced by 47.7, 61.5, 57.5, and 58.5%, respectively, compared with untreated controls. Histopathological examination showed that IJ-5 treatment decreased DNCB-induced hypertrophy, hyperkeratosis, and infiltration of inflammatory cells in both ear and back skins. Moreover, IJ-5 suppressed the expression of TNFα, IL-1, and IL-6 with IC50 values of 6.58, 9.48, and 7.01 μM, respectively, and inhibited the activation of NF-κB pathway in TNFα-stimulated HaCat cells.The present study demonstrates the protective effects of IJ-5 against AD-like skin inflammation and highlights IJ-5 as a potential therapeutic agent for AD.

Viscoelasticity-induced particle migration has recently received increasing attention due to its ability to obtain high-quality focusing over a wide range of flow rates. However, its application is limited to low throughput regime since the particles can defocus as flow rate increases. Using an engineered carrier medium with constant and low viscosity and strong elasticity, the sample flow rates are improved to be 1 order of magnitude higher than those in existing studies. Utilizing differential focusing of particles of different sizes, here, we present sheathless particle/cell separation in simple straight microchannels that possess excellent parallelizability for further throughput enhancement. The present method can be implemented over a wide range of particle/cell sizes and flow rates. We successfully separate small particles from larger particles, MCF-7 cells from red blood cells (RBCs), and Escherichia coli (E. coli) bacteria from RBCs in different straight microchannels. The proposed method could broaden the applications of viscoelastic microfluidic devices to particle/cell separation due to the enhanced sample throughput and simple channel design.

Three new decomposition products of valepotriates, valtrals A-C (1-3), and two known products, baldrinal and homobaldrinal, are formed during the isolation procedure of the ethanol extract of the whole plants of Valeriana jatamansi. Their structures were determined by spectroscopic methods including IR, MS, 1D, and 2D NMR experiments. Compounds 1-3 showed selective cytotoxicity against metastatic prostate cancer (PC-3M) and colon cancer (HCT-8) cell lines.

The amphiphilic graft copolymer poly (lactic-co-glycolic acid)- g -dextran (Dex-PLGA) was successfully synthesized to fabricate micelles for the delivery of paclitaxel with low critical micelle concentration (CMC). The sizes of paclitaxel-loaded Dex-PLGA (Dex-PLGA/PTX) micelles were kept below 100nm with a relatively narrow size distribution. This novel PTX nano-formulation was found to exhibit slightly stronger in vitro cytotoxicity against SKOV-3, OVCAR-8 and MCF-7 cells with Taxol®. However, it could overcome the drug resistance of multi-drug resistant human breast carcinoma cells (MCF-7/Adr cells). The maximum tolerated dose (MTD) of Dex-PLGA/PTX after a single dose was more than 200mg PTX/kg, which were 8-fold higher than that of Paclitaxel Injection. The in vivo antitumor activity results indicated that Dex-PLGA/PTX micelles treatments effectively suppressed the tumor growth and highly reduced the toxicity against animals than Taxol® and could eliminate the SKOV-3 tumor by highly increasing the drug dose. From the Clinical Editor Chemotherapy for cancer has always been hampered the toxic side effect of the drugs. Nanotechnology has helped to produce various drug delivery systems to minimize these side effects. In this article, the authors designed dextran-based micelles loaded with paclitaxel. They showed effective anti-tumor activity in both in vitro and in vivo experiments with significant lower systemic toxicity.

Five pairs of new epimeric lanostane-type triterpenoids, abiespirones A-D (1-4) and G (7), two pairs of new epimeric cycloartane-type triterpenoids, abiespirones E and F (5, 6), and a pair of new epimeric 7(8→9)abeo-spirolanostane abiespirones H (8) with spiro-B/C and -E/F ring systems were isolated from Abies faxoniana as inseparable mixtures of C-23 epimers in a specific proportion. The HPLC plots showed that each purified isomer rapidly equilibrated with the C-23 epimer in solution. The structures of compounds 1-8 were elucidated by analysis of the NMR spectra and single-crystal X-ray diffraction. Compound 6 showed cytotoxicity against three hepatoma cell lines, namely, HepG2, Huh7, and SMMC7721, with IC50 values of 9.8, 7.5, and 10.7 μM, respectively, but exerted low cytotoxicity on normal QSG7701 hepatic cells, indicating its selective cytotoxicity for hepatoma cells. Compound 6 arrests the cell cycle at G2/M and induces cell apoptosis in Huh7 cells. In addition, the generation of reactive oxygen species (ROS) was detected in Huh7 cells when treated with compound 6, and a ROS scavenger partly blocked the effects of compound 6-induced Huh7 cell death, suggesting that compound 6-induced apoptosis is associated with elevated levels of ROS in Huh7 cells.

Seven new diterpenoids lanceolatins, A–G (1–7), together with five known diterpenoids (8–12), were isolated from the branches and leaves of Cephalotaxus lanceolata. The structures of the new diterpenoids were elucidated based on spectroscopic analysis, including 1D, 2D NMR, and HR-ESI-MS. Single crystal X-ray diffraction (CuKα radiation) was employed to confirm the structure of lanceolatin G (7), and its absolute configuration was finally established. Compound 12 showed significant inhibition against human tumor cell lines HCT116 and HepG2 with IC50 values of 0.17 and 0.63 μg mL−1, respectively, whereas compounds 3–5 can inhibit nitric oxide (NO) release in LPS-induced RAW264.7 macrophages with IC50 values of 8.72, 10.79, and 12.73 μM, respectively.

Merrilliaquinone (1), a new racemic quinone was isolated from the branches and leaves of Illicium merrillianum. Chiral separation of 1 gave two enantiomers (−)-merrilliaquinone (1a) and (+)-merrilliaquinone (1b). The structure of 1 was established by comprehensive spectroscopic analysis, and the absolute configurations of 1a and 1b were determined by quantum mechanical calculation of ECD spectra. It is very interesting that 1b had a selective cytotoxicity against human hepatoma cell lines SMMC7721 and HuH7 with IC50 values of 0.91 μM and 1.29 μM, while its IC50 values on normal human hepatic cells QSG7701 and LO2 were 47.79 μM and 36.71 μM, respectively. Moreover, 1a and racemate 1 only exhibited very weak cytotoxicity against SMMC7721 and HuH7 cells with IC50 values of 27.01–37.82 μM. These results implied that the absolute configurations of 1a and 1b possess remarkable influences on their cytotoxicities. Further mechanism studies indicated that 1b dose-dependently induced SMMC7721 cell apoptosis and reduction of mitochondrial membrane potential (MMP) at 1–10 μM. Flow cytometry analysis showed that the 1b-induced SMMC7721 cell apoptosis was associated with cell cycle arrest during the G0/G1 phase.

Six new 2,4-linked exo/endo sesquiterpene lactone dimers, faberidilactones A–E (1–5) and endodischkuhriolin (6), were isolated from Carpesium faberi. The characteristic chemical shift of Ha-3′ and diagnostic negative/positive CE can be used to assign the exo/endo stereochemistry of those dimers. Compounds 1–3, 5 and 6 exhibited potent cytotoxicity with IC50 value in the range of 1.11–8.50 μM against the four human tumor cell lines (CCRF-CEM, K562, HL-60 and HCT116 cells). Moreover, 1, 3 and 5 (exo-SLDs) showed potent NF-κB inhibitory effect with inhibition ratio of 65.98, 53.27 and 71.45%, respectively.

Diacylglycerol acyltransferase (DGAT) catalyzes the final step in triacylglycerol (TAG) biosynthesis via the acyl-CoA-dependent acylation of diacylglycerol. This reaction is a major control point in the Kennedy pathway for biosynthesis of TAG, which is the most important form of stored metabolic energy in most oil-producing plants. In this study, Arachis hypogaea type 2 DGAT (AhDGAT2) genes were cloned from the peanut cultivar 'Luhua 14.' Sequence analysis of 11 different peanut cultivars revealed a gene family of 8 peanut DGAT2 genes (designated AhDGAT2a-h). Sequence alignments revealed 21 nucleotide differences between the eight ORFs, but only six differences result in changes to the predicted amino acid (AA) sequences. A representative full-length cDNA clone (AhDGAT2a) was characterized in detail. The biochemical effects of altering the AhDGAT2a sequence to include single variable AA residues were tested by mutagenesis and functional complementation assays in transgenic yeast systems. All six mutant variants retained enzyme activity and produced lipid droplets in vivo. The N6D and A26P mutants also displayed increased enzyme activity and/or total cellular fatty acid (FA) content. N6D mutant mainly increased the content of palmitoleic acid, and A26P mutant mainly increased the content of palmitic acid. The A26P mutant grew well both in the presence of oleic and C18:2, but the other mutants grew better in the presence of C18:2. AhDGAT2 is expressed in all peanut organs analyzed, with high transcript levels in leaves and flowers. These levels are comparable to that found in immature seeds, where DGAT2 expression is most abundant in other plants. Over-expression of AhDGAT2a in tobacco substantially increased the FA content of transformed tobacco seeds. Expression of AhDGAT2a also altered transcription levels of endogenous tobacco lipid metabolic genes in transgenic tobacco, apparently creating a larger carbon 'sink' that supports increased FA levels.

A total synthesis of Daphnodorin A, a member of the Daphnodorins, was accomplished. Key features of the synthetic strategy include construction of 2-substituted-3-functionalized benzofuran via intramolecular Heck reaction and a mild Barton–McCombie deoxygenation process mediated by triethylborane. The total synthesis provided Daphnodorin A in 19.7% or 5.6% overall yield over 7 or 15 steps.

During a survey on chemical constituents of Abies nukiangensis, seven previously unreported compounds, including six triterpenes (1-6) and one phenol (7) were isolated and characterized, together with 37 known miscellaneous chemical constituents. The structures of compounds 1-7 were established mainly by extensive analysis of the 1D and 2D NMR, as well as HRMS data. The absolute configurations of compounds 1 and 8 were confirmed unambiguously by the Cu-Kα X-ray crystallography. Compounds 3 and 8-10 showed significant anti-hepatitis C virus effects with EC50 values of 3.73, 2.67, 1.33 and 2.25μM, respectively.

Investigation of the 80% ethanol extract of the aerial parts of Psammosilene tunicoide afforded two carboline alkaloids (1–2), a γ-pyrone derivative (3), two p-hydroxy cinamic acid derivatives (4–5), two glycosides (6–7) and six known compounds (8–13). Their structures were established on the basis of spectroscopic analysis (1D-, 2D-NMR, ESI-MS, and HR-ESI-MS). All these compounds were evaluated for cytotoxicity against SMML-7721, A549, HCT116, and MCF-7 cell lines, and for inhibitory activity against NO production in LPS-induced RAW264.7 macrophages. However, only compound 2 had weak cytotoxic activity against the HCT116 cell line at 20 μM, while the others are inactive against four human tumor cell lines and only show weak inhibition activity against NO production at 50 μM.

For decades, skin cancer incidence has increased, mainly because of oncogenic signaling pathways activated by solar ultraviolet (UV) irradiation (i.e., sun exposure). Solar UV induces multiple signaling pathways that are critical in the development of skin cancer, and therefore the development of compounds capable of targeting multiple molecules for chemoprevention of skin carcinogenesis is urgently needed. Herein, we examined the chemopreventive effects and the molecular mechanism of (+)-2-(1-hydroxyl-4-oxocyclohexyl) ethyl caffeate (HOEC), isolated from Incarvillea mairei var. grandiflora (Wehrhahn) Grierson. HOEC strongly inhibited neoplastic transformation of JB6 Cl41 cells without toxicity. PI3K, ERK1/2, and p38 kinase activities were suppressed by direct binding with HOEC in vitro. Our in silico docking data showed that HOEC binds at the ATP-binding site of each kinase. The inhibition of solar UV-induced PI3K, ERK1/2, and p38 kinase activities resulted in suppression of their downstream signaling pathways and AP1 and NF-κB transactivation in JB6 cells. Furthermore, topical application of HOEC reduced skin cancer incidence and tumor volume in SKH-1 hairless mice chronically exposed to solar UV. In summary, our results show that HOEC exerts inhibitory effects on multiple kinase targets and their downstream pathways activated by solar UV in vitro and in vivo. These findings suggest that HOEC is a potent chemopreventive compound against skin carcinogenesis caused by solar UV exposure. Cancer Prev Res; 7(8); 856–65. ©2014 AACR.

The degradation of chlorpyrifos is investigated with the treatments of sonolysis, ozonolysis, and sonolytic ozonolysis at various frequencies. Results show that there exists frequency effect in sonolytic ozonolysis. In sonolytic ozonolysis, the maximum degradation rate is obtained at 495 kHz, and the degradation kinetics is fitted to the first-order kinetics model well. However, the most significant synergistic effect between ultrasonic and ozone is at 124 kHz. The kinetics parameters indicate that chlorpyrifos is much more labile to ultrasonic at 495 kHz, while ozone is much more soluble at 124 kHz. The hydrolysis and oxidation are deduced to contribute to the degradation reaction and the degradation pathway for chlorpyrifos degradation is proposed.

Dual-frequency ultrasonic assisted ozonation was proposed to degrade pesticide wastewater. The frequencies of ultrasound were 28 kHz and low-MHz (208 kHz, 495 kHz, 679 kHz and 890 kHz). The influence of ultrasound frequency was investigated in this paper. The electrical conductivity of deionized water was measured under different conditions, and degradation rate of acephate solution was used to verify degradation effect of the treatment. The results implied that the 208 kHz system was suitable for ozone dissolving and 495 kHz system was suitable for acephate degradation.