The aquaporin MePIP2;7 improves MeMGT9-mediated Mg2 + acquisition in cassava.

Aquaporins are important transmembrane water transport proteins which transport water and several neutral molecules. However, how aquaporins are involved in the synergistic transport of Mg2+ and water remains poorly understood. Here, we found that the cassava aquaporin MePIP2;7 was involved in Mg2+ transport through interaction with MeMGT9, a lower affinity magnesium transporter protein. Knockdown of MePIP2;7 in cassava led to magnesium deficiency in basal mature leaves with chlorosis and necrotic spots on their edges and starch over-accumulation. Mg2+ content was significantly decreased in leaves and roots of MePIP2;7-RNA interference (PIP-Ri) plants grown in both field and Mg2+ -free hydroponic solution. Xenopus oocyte injection analysis verified that MePIP2;7 possessed the ability to transport water only and MeMGT9 was responsible for Mg2+ efflux. More importantly, MePIP2;7 improved the transportability of Mg2+ via MeMGT9 as verified using the CM66 mutant complementation assay and Xenopus oocytes expressing system. Yeast two-hybrid, bimolecular fluorescence complementation, co-localization, and co-immunoprecipitation assays demonstrated the direct protein-protein interaction between MePIP2;7 and MeMGT9 in vivo. Mg2+ flux was significantly elevated in MePIP2;7-overexpressing lines in hydroponic solution through non-invasive micro-test technique analysis. Under Mg2+ -free condition, the retarded growth of PIP-Ri transgenic plants could be recovered with Mg2+ supplementation. Taken together, our results demonstrated the synergistic effect of the MePIP2;7 and MeMGT9 interaction in regulating water and Mg2+ absorption and transport in cassava.

The Structure and Function of Microbial Community in Rhizospheric Soil of American Ginseng (Panax quinquefolius L.) Changed with Planting Years.

The changes of microbial communities of rhizospheric soil in different ages are speculated to cause soil-borne diseases and replanting problem in American ginseng (Panax quinquefolius L.) cultivation. This study analyzed the physicochemical properties and microbial communities of rhizospheric soil during the planting of American ginseng in the Wendeng area of Weihai, China. The water content and organic matter content of American ginseng rhizospheric soil decreased year by year. A decline in the diversity of bacteria and fungi was observed in the rhizospheric soils planting American ginseng compared with the traditional crop wheat in the control group. During the later planting stage, the abundances of Proteobacteria, Actinobacteria, and Basidiomycota were lower, whereas that of Acidobacteria, Firmicutes, and Mucoromycota were higher. Through the correlation analysis between environmental factors and microbial community, it was found that the content of soil phosphorus was significantly positively correlated with the root rot pathogen Fusarium. The results of functional prediction showed that the decrease of secondary metabolite synthesis of rhizospheric soil bacteria and the increase of plant pathogenic fungi may be the important reasons for the increase of diseases in the later stage of American ginseng planting. This study revealed the evolution of rhizosphere microbial community and function in the process of American ginseng planting, which is valuable for planting management.

Poplar bark lipids enhance mouse immunity by inducing T cell proliferation and differentiation.

Research on the composition and application of immune enhancers in livestock and poultry breeding has been gaining interest in recent years. Poplar bark lipids (PBLs), which are extracted from poplar tree bark, are natural substances known to efficiently enhance the immune response. To understand the chemical makeup of PBLs and their underlying mechanism for enhancing the immune system, we extracted PBLs from poplar bark using petroleum ether and subjected these extracts to chemical analysis. To evaluate PBLs effect on the immune system mice were treated with different doses of PBL via gavage and sacrificed 4 weeks later. PBLs were shown to be rich in vitamin E, unsaturated fatty acids, and other immune-potentiating compounds. Treatment with PBLs increased the spleen index and stimulated spleen and thymus development. In addition, PBLs increased the number of CD3+CD4+ cells in the peripheral blood and the ratio of CD4+/CD8+ cells while decreasing the number of CD3+CD8+ cells. Moreover, PBLs significantly increased IL-4 and IFN-γ levels in mouse serum and TLR4 mRNA and protein expression in the spleen. Taken together these results demonstrate that PBLs exert their immune-potentiating effects by promoting spleen and thymus development, T lymphocyte proliferation and differentiation, and immune factor expression. These immune-potentiating effects may be related to the activation of TLR4. This study provides a theoretical basis for the development of PBLs as an immune adjuvant or feed additive in the future.

New sesquiterpenoids with COX-2 inhibitory activity from the medical plant Physalis. alkekengi L. var. franchetii.

Phytochemical investigations on Physalis. alkekengi L. var. franchetii, a widespread traditional Chinese medicine, led to the isolation and identification of three new sesquiterpenoids physalisitins A-C (1-3). Their structures were elucidated by NMR and HRESIMS analysis, and their absolute configurations were determined by quantum chemical NMR and ECD calculations, as well as by comparing their optical rotation values with those known analogues. All of the isolated compounds were evaluated for their cyclooxygenase-2 (COX-2) inhibitory activity. Compounds 1-3 dose-dependently inhibited the COX-2 enzyme with IC50 values of 3.22 ± 0.25, 6.35 ± 0.84, and 11.13 ± 1.47 µM, respectively.

New Cyclic Diarylheptanoids from Platycarya strobilacea.

Five new cyclic diarylheptanoids (platycary A-E, compounds 1-5) and three previously identified analogues (i.e., phttyearynol (compound 6), myricatomentogenin (compound 7), and juglanin D (compound 8)) were isolated from the stem bark of Platycarya strobilacea. The structures of these compounds were determined using NMR, HRESIMS, and electronic circular dichroism (ECD) data. The cytotoxicity of compounds 1-5 and their ability to inhibit nitric oxide (NO) production, as well as protect against the corticosterone-induced apoptosis of Pheochromocytoma (PC12) cells, were evaluated in vitro using the appropriate bioassays. Compounds 1 and 2 significantly inhibited the corticosterone-induced apoptosis of PC12 cells at a concentration of 20 µΜ.

A new epicatechin glucopyranoside derivative from Styrax suberifolius.

A new derivative of epicatechin glucopyranoside, (2R,3R)-3,7,4'-trihydroxy-5,3'-dimethoxyflavan 7-O-ß-d-glucopyranoside (1), together with three mononuclear phenolic acid esters, methyl orsellinate (2), ethyl orsellinate (3) and methyl ß-orcinolcarboxylate (4) were isolated from the bark of Styrax suberifolius. The structures of 1-4 were determined on the basis of extensive analysis of NMR and MS spectra combined with chemical hydrolysis. The antifungal activities of the isolated compounds against three plant pathogenic fungi, Alternaria solani, Fusarium oxysporum and Phomopsis cytospore were evaluated using radial growth inhibition assay. Compounds 2, 3 and 4 exerted selective inhibitory activities against the tested fungi. Among of them, methyl ß-orcinolcarboxylate (4) exhibited obvious inhibitory effect against P. cytospore, with an inhibition rate of 86.72% at 100 µg/ml.

Ambrosia artemisiifolia as a potential resource for management of golden apple snails, Pomacea canaliculata (Lamarck).

BACKGROUND: Ambrosia artemisiifolia, an invasive weed in Europe and Asia, is highly toxic to the golden apple snail (GAS; Pomacea canaliculata) in laboratory tests. However, little is known about the chemical components of A. artemisiifolia associated with the molluscicidal activity or about its potential application for GAS control in rice fields. This study evaluated the molluscicidal activities of powders, methanol extracts, and individual compounds from A. artemisiifolia against GAS in rice fields and under laboratory conditions. RESULTS: Ambrosia artemisiifolia powders did not negatively affect the growth and development of rice but they reduced damage to rice caused by GAS. Extracts had moderate acute toxicity but potent chronic toxicity. The 24-h 50% lethal concentration (LC50 ) of the extracts against GAS was 194.0 mg L-1 , while the weights, lengths and widths of GAS were significantly affected by exposure to a sublethal concentration (100 mg/mL). Psilostachyin, psilostachyin B, and axillaxin were identified as the most active molluscicide components in the aerial parts of A. artemisiifolia, and the 24-h LC50 values of these purified compounds were 15.9, 27.0, and 97.0 mg/L, respectively. CONCLUSION: The results indicate that chemical compounds produced by A. artemisiifolia may be useful for population management of GAS in rice fields. © 2017 Society of Chemical Industry.

New 30-Noroleanane Triterpenoid Saponins from Holboellia coriacea Diels.

Three new 30-noroleanane triterpenoid saponins, akebonoic acid 28-O-ß-d-glucopyranosyl-(1''→6')-ß-d-glucopyranosyl ester (1), akebonoic acid 28-O-(6''-O-caffeoyl)-ß-d-glucopyranosyl-(1''→6')-ß-d-glucopyranosyl ester (Holboelliside A, 2) and 3ß,20α,24-trihydroxy-29-norolean-12-en-28-oic acid 3-O-(6'-O-caffeoyl)-ß-d-glucopyranoside (Holboelliside B, 3) were isolated from the stems of Holboellia coriacea Diels, together with five known compounds, eupteleasaponin VIII (4), 3α-akebonoic acid (5), quinatic acid (6), 3ß-hydroxy-30-norhederagenin (7) and quinatoside A (8). The structures of these compounds were determined on the basis of spectral and chemical evidence. Compounds 1-5 were evaluated for their inhibitory activity against three human tumors HepG2, HCT116 and SGC-7901 cell lines in vitro.

A rapid and comprehensive quality assessing method of Yin-Qiao-Jie-Du tablets using UHPLC-QTOF-MS in combination with multivariate statistical analysis.

Yin-Qiao-Jie-Du (YQJD) tablet is a well-known non-prescription Chinese patent drug widely used for the prevention and treatment of diseases in China. However, documented studies for assessing and controlling the quality of YQJD tablet are limited. In this article, a rapid and comprehensive method for the quality assessment and control of YQJD tablets was developed, via qualitative and quantitative analysis of the major active ingredients of YQJD tablets by ultra-high performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UHPLC-QTOF-MS) in combination with multivariate statistical analysis. The samples of YQJD tablets were triturated and then extracted with methanol, and the extracts were analyzed by UHPLC-QTOF-MS. A good separation was achieved within 30 min by using an Agilent RRHD SB-C18 (150 mm × 2.1mm I.D., 1.8 µm) column with a gradient elution of water (0.5% formic acid) and acetonitrile. Forty-one major chromatographic peaks were obtained, and 31 of them were identified according to the obtained data from the measurements of accurate mass, isotopic composition, and collision-induced dissociation. Among them, 21 were confirmed by the comparison of retention times, MS, and MS/MS information with those of their references. Quantitative analysis was performed based on the peak areas of extracted ion chromatograms of the exact pseudo-molecular ions (with a 0.01 Da window), and the result of method validation demonstrated that the proposed method possessed desirable specificity, linearity, precision, and accuracy. Subsequently, the method was utilized to analyze the major constituents in YQJD tablets of 18 batches produced by different manufacturers. The obtained contents of the 21 confirmed ingredients were applied for orthogonal partial-least squares to latent structures discriminant analysis (OPLS-DA). An assessment of the quality consistency and stability as well as tracing the origins of the investigated YQJD tablets were successfully achieved by OPLS-DA. All of these results demonstrated that the UHPLC-QTOF-MS method developed in this study was a rapid and desirable approach for differentiating the origin, discriminating the authenticity, and assessing and controlling the quality of YQJD tablets.

Physiological characterization of maize tolerance to low dose of aluminum, highlighted by promoted leaf growth.

MAIN CONCLUSION: Effects of a low aluminum (Al) dose were characterized. The Al supplement inhibited root growth but enhanced leaf growth in maize lines with different Al sensitivities. High levels of Al are phytotoxic especially in acidic soils. The beneficial effects of low Al levels have been reported in some plant species, but not in maize. Maize is relatively more sensitive to Al toxicity than other cereals. Seedlings, at the three leaf stage, of four Chinese maize foundation parent inbred lines with different Al tolerances, were exposed to complete Hoagland's nutrient solution at pH 4.5 supplemented with 48 µM Al(3+) under controlled growth conditions, and then the Al stress (AS) was removed. The leaf and root growth, root cell viability, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), ions (K(+), Ca(++) and Mg(++)), photosynthetic rate and chlorophyll, protein and malondialdehyde contents in tissues were assayed. In conclusion, a low Al dose inhibits root growth but enhances leaf growth in maize. The Al-promoted leaf growth is likely a result of increased protein synthesis, a lowered Ca(++) level, and the discharge of the growth-inhibitory factors. The Al-promoted leaf growth may be a 'memory' effect caused by the earlier AS in maize. Al causes cell wall rupture, and a loss of K(+), Ca(++) and Mg(++) from root cells. CAT is an auxiliary antioxidant enzyme that works selectively with either SOD or POD against AS-related peroxidation, depending on the maize tissue. CAT is a major antioxidant enzyme responsible for root growth, but SOD is important for leaf growth during AS and after its removal. Our results contribute to understanding how low levels of Al affect maize and Al-resistant mechanisms in maize.

Resin glycosides from Porana duclouxii.

A new intact resin glycoside (3) and two glycosidic acids (1 and 2), all having a common trisaccharide moiety and (11S)-hydroxytetradecanoic acid or (3S,11S)-dihydroxytetradecanoic acid as the aglycone, were obtained from the roots of Porana duclouxii. Their structures were elucidated by spectroscopic analyses and chemical correlations. These compounds represent the first examples of resin glycosides from the genus Porana.

Functional characterization of the copper-transporting P-type ATPase gene of Penicillium janthinellum strain GXCR.

Copper (Cu)-transporting P-type ATPase (CTPA) genes have been documented to play an important role in resistance to heavy metals. However, our understanding of roles of CTPA genes of the filamentous fungi was based only on sequence similarity prediction before. In a previous study, we isolated a Penicillum janthinellum strain GXCR of higher tolerance to Cu (200 mM). In this study, we cloned the partial cDNA of CTPA gene, named PcpA, from the strain GXCR. Sequence alignment indicated that the cloned cDNA sequence has the highest identity (94.4%) with a predictive CTPA gene of Aspergillus clavatus. The PcpA-encoded protein, termed PcpA, has classical functional domains of CTPAs, and shows differences from reported CTPAs in some specific sequence motifs and transmembrane regions. Expression of the PcpA was induced by extracellular Cu, cadmium (Cd), and silver (Ag). PcpA RNA interference (RNAi) mutants with a reduced level of PcpA mRNA were more sensitive to Cu, iron, Cd, and Ag than the wild-type (WT) strain GXCR. When grown in the presence of Cu, iron, and Cd, intracellular Cu and iron contents in the PcpA RNAi mutant were significantly (P<0.05) lower than those in the WT; However, intracellular Cd content in the mutant was significantly (P<0.05) higher than that in the WT. Taken together, it can be concluded that the PcpA functions in Cu uptake and homeostasis, iron uptake, and Cd export from the cytosol to the extracytosol.

[Clinical evaluation of therapeutic effects of "Cooling the blood and invigorating blood circulation" method on hepatitis hyperbilirubinemia].

BACKGROUND: To explore the effect of treating hepatitis hyperbilirubinemia with Chinese traditional "cooling the blood and invigorating blood circulation" method. METHODS: Seventy-two patients were randomly divided into two groups. The control group (n=36) was treated with hepatocyte growth factor (HGF) 80-120 mg daily on the basis of routine western medicine treatment. The therapy group (n=36) was treated with "cooling the blood and invigorating blood circulation" decoction on the basis of routine western medicine treatment. After using the medicine for 28 days, alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), albumin (ALB) in plasma were measured to compare the effects of the two methods. RESULTS: The effective rate of the therapy group was 80.55%, that of the control group was 50%. There was significant difference in efficacy between the two groups (P<0.05). CONCLUSION: "Cooling the blood and invigorating blood circulation" method may be effective and safe in the treatment of hepatitis hyperbilirubinemia.