The effects of quinoa bran dietary fiber on glucose and lipid metabolism and hepatic transcriptome in obese rats.

BACKGROUND: As a complex chronic metabolic disease, obesity not only affects the quality of human life but also increases the risk of various other diseases. Therefore, it is important to investigate the molecular mechanisms and therapeutic effects of dietary interventions that counteract obesity. RESULTS: In this study, we extracted soluble (SDF) and insoluble dietary fiber (IDF) from quinoa bran using an enzymatic method and further investigated their effects on lipid metabolism and blood lipid levels in obese rats. Quinoa bran dietary fiber showed significantly reduced body weight, blood glucose level, total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol levels compared to those in the model group of obese rats. Aspartate aminotransferase and alanine aminotransferase levels were significantly lower in the IDF group, demonstrating that IDF improved liver injury more significantly than SDF, which was consistent with the analysis of liver tissue sections. IDF supplementation significantly improved the oxidation resistance of obese rats by decreasing malondialdehyde and increasing superoxide dismutase and glutathione peroxidase levels compared to the high-fat diet group levels. Transcriptome analysis showed that IDF caused hepatic changes in genes (Ehhadh, PPARα, FADS, CPT1, CPT2, SCD-1, Acadm, and CYP7A1) related to fatty acid degradation, and this result coincided with that of the gene expression validation result. CONCLUSION: Overall, our research offers crucial data for the logical development of dietary fiber from quinoa bran with nutritional purposes. © 2023 Society of Chemical Industry.

Auxin response factors (ARFs) differentially regulate rice antiviral immune response against rice dwarf virus.

There are 25 auxin response factors (ARFs) in the rice genome, which play critical roles in regulating myriad aspects of plant development, but their role (s) in host antiviral immune defense and the underneath mechanism remain largely unknown. By using the rice-rice dwarf virus (RDV) model system, here we report that auxin signaling enhances rice defense against RDV infection. In turn, RDV infection triggers increased auxin biosynthesis and accumulation in rice, and that treatment with exogenous auxin reduces OsIAA10 protein level, thereby unleashing a group of OsIAA10-interacting OsARFs to mediate downstream antiviral responses. Strikingly, our genetic data showed that loss-of-function mutants of osarf12 or osarf16 exhibit reduced resistance whereas osarf11 mutants display enhanced resistance to RDV. In turn, OsARF12 activates the down-stream OsWRKY13 expression through direct binding to its promoter, loss-of-function mutants of oswrky13 exhibit reduced resistance. These results demonstrated that OsARF 11, 12 and 16 differentially regulate rice antiviral defense. Together with our previous discovery that the viral P2 protein stabilizes OsIAA10 protein via thwarting its interaction with OsTIR1 to enhance viral infection and pathogenesis, our results reveal a novel auxin-IAA10-ARFs-mediated signaling mechanism employed by rice and RDV for defense and counter defense responses.

Rice stripe virus NS3 protein regulates primary miRNA processing through association with the miRNA biogenesis factor OsDRB1 and facilitates virus infection in rice.

MicroRNAs (miRNAs) are small regulatory RNAs processed from primary miRNA transcripts, and plant miRNAs play important roles in plant growth, development, and response to infection by microbes. Microbial infections broadly alter miRNA biogenesis, but the underlying mechanisms remain poorly understood. In this study, we report that the Rice stripe virus (RSV)-encoded nonstructural protein 3 (NS3) interacts with OsDRB1, an indispensable component of the rice (Oryza sativa) miRNA-processing complex. Moreover, the NS3-OsDRB1 interaction occurs at the sites required for OsDRB1 self-interaction, which is essential for miRNA biogenesis. Further analysis revealed that NS3 acts as a scaffold between OsDRB1 and pri-miRNAs to regulate their association and aids in vivo processing of pri-miRNAs. Genetic evidence in Arabidopsis showed that NS3 can partially substitute for the function of double-stranded RNA binding domain (dsRBD) of AtDRB1/AtHYL1 during miRNA biogenesis. As a result, NS3 induces the accumulation of several miRNAs, most of which target pivotal genes associated with development or pathogen resistance. In contrast, a mutant version of NS3 (mNS3), which still associated with OsDRB1 but has defects in pri-miRNA binding, reduces accumulation of these miRNAs. Transgenic rice lines expressing NS3 exhibited significantly higher susceptibility to RSV infection compared with non-transgenic wild-type plants, whereas the transgenic lines expressing mNS3 showed a less-sensitive response. Our findings revealed a previously unknown mechanism in which a viral protein hijacks OsDRB1, a key component of the processing complex, for miRNA biogenesis and enhances viral infection and pathogenesis in rice.

Rice Dwarf Virus P2 Protein Hijacks Auxin Signaling by Directly Targeting the Rice OsIAA10 Protein, Enhancing Viral Infection and Disease Development.

The phytohormone auxin plays critical roles in regulating myriads of plant growth and developmental processes. Microbe infection can disturb auxin signaling resulting in defects in these processes, but the underlying mechanisms are poorly understood. Auxin signaling begins with perception of auxin by a transient co-receptor complex consisting of an F-box transport inhibitor response 1/auxin signaling F-box (TIR1/AFB) protein and an auxin/indole-3-acetic acid (Aux/IAA) protein. Auxin binding to the co-receptor triggers ubiquitination and 26S proteasome degradation of the Aux/IAA proteins, leading to subsequent events, including expression of auxin-responsive genes. Here we report that Rice dwarf virus (RDV), a devastating pathogen of rice, causes disease symptoms including dwarfing, increased tiller number and short crown roots in infected rice as a result of reduced sensitivity to auxin signaling. The RDV capsid protein P2 binds OsIAA10, blocking the interaction between OsIAA10 and OsTIR1 and inhibiting 26S proteasome-mediated OsIAA10 degradation. Transgenic rice plants overexpressing wild-type or a dominant-negative (degradation-resistant) mutant of OsIAA10 phenocopy RDV symptoms are more susceptible to RDV infection; however, knockdown of OsIAA10 enhances the resistance of rice to RDV infection. Our findings reveal a previously unknown mechanism of viral protein reprogramming of a key step in auxin signaling initiation that enhances viral infection and pathogenesis.

Insulin receptor substrate 1/2 (IRS1/2) regulates Wnt/ß-catenin signaling through blocking autophagic degradation of dishevelled2.

Wnt signaling plays a pivotal role in cell proliferation, tissue homeostasis, and tumorigenesis. Dishevelled (Dvl) is a central node of Wnt signaling. Insulin receptor substrates (IRSs), as a critical component of insulin signaling, are involved in cell proliferation, metabolism, and cancer development. In this study, we report that IRS1/2 promotes Wnt/ß-catenin signaling by stabilizing Dvl2. We found that IRS1/2 interacts with Dvl2. Overexpression of IRS1/2 increased the protein level of Dvl2 and promoted canonical Wnt signaling, as evidenced by the increased T cell-specific factor 4 transcriptional activity and the up-regulation of expression of CYCLIN D1 and c-MYC, two Wnt target genes critical for cell growth, whereas depletion of IRS1/2 reduced the level of Dvl2 and attenuated Wnt/ß-catenin signaling. Biochemical analyses revealed that IRS1/2 decreased Lys-63-linked ubiquitination of Dvl2 and stabilized Dvl2 protein via suppressing its autophagy-mediated degradation. We further revealed that IRS1/2 blocks autophagy-induced formation of the Dvl2-p62/SQSTM1 complex, resulting in disabled association of Dvl2 to autophagosomes. We demonstrated that IRS1/2 promoted the induction of epithelial-mesenchymal transition (EMT) and cell proliferation in response to Wnt stimulation, whereas depletion of Dvl2 impaired the IRS1/2-mediated EMT and cell growth. Our findings revealed that IRS1/2 promotes EMT and cell proliferation through stabilizing Dvl2.

Preparation of quinoa bran dietary fiber-based zinc complex and investigation of its antioxidant capacity in vitro.

In order to improve the economic utilization of quinoa bran and develop a safe and highly available zinc ion biological supplement. In this study, a four-factor, three-level response surface optimization of quinoa bran soluble dietary fiber (SDF) complexation of zinc was studied. The effect used four factors on the chelation rate was investigated: (A) mass ratio of SDF to ZnSO4.7H2O, (B) chelation temperature, (C) chelation time, and (D) pH. Based on the results of the single-factor test, the four-factor three-level response surface method was used to optimize the reaction conditions. The optimal reaction conditions were observed as mentioned here: the mass ratio of quinoa bran SDF to ZnSO4.7H2O was 1, the reaction temperature was 65°C, the reaction time was 120 min, and the pH of the reaction system was 8.0. The average chelation rate was 25.18%, and zinc content is 465.2 µg/g under optimal conditions. The hydration method rendered a fluffy quinoa bran SDF structure. The intramolecular functional groups were less stable which made the formation of the lone pairs of electrons feasible to complex with the added divalent zinc ions to form a quinoa bran soluble dietary fiber-zinc complex [SDF-Zn(II)]. The SDF-Zn(II) chelate had higher 2,2-diphenylpicrylhydrazyl (DPPH), ABTS+, hydroxyl radical scavenging ability, and total antioxidant capacity. Therefore, metal ion chelation in dietary fiber is of biological importance.

Preparation, structure, and in-vitro hypoglycemic potential of debranched millet starch-fatty acid composite resistant starch.

Currently, the preparation methods and basic physicochemical properties of starch-FA complexes have been widely studied; however, no in-depth research on the regulatory mechanism of the digestive properties of debranched starch-unsaturated FA complexes has been conducted. Therefore, six fatty acids with different carbon chains and different degrees of unsaturation were complexed with de-branched millet starch in this research, using the microwave method. Microwave millet starch-linoleic acid complex (MPS-LOA) had the highest resistant starch (RS) content, and the structure and physicochemical properties of MPS-LOA were determined using various molecular techniques. The results indicate that MPS-LOA had a resistant starch (RS) content of 40.35% and the most notable fluorescence. The characteristic UV peaks of MPS-LOA were blue-shifted, and new IR peaks appeared. The crystalline structure changed to V-type crystals, the crystallinity increased, and the molecular weight decreased. The enthalpy and coagulability of MPS-LOA increased, and the swelling force decreased. Additionally, MPS-LOA showed enhanced α-glucosidase and α-amylase inhibition, and in-vitro hydrolysis kinetics analysis of MPS-LOA showed a hydrolysis index of 53.8 and an extended glycemic index (eGI)I of 54.6, indicating a low eGI food suitable for consumption by people with type II diabetes. These results provide a theoretical basis for the preparation of amylopectin- and starch-based foods with an anti-enzyme structure and a low glycemic index (GI).

5,7,2,5-tetrahydroxy-8,6-dimethoxyflavone up-regulates miR-145 expression and inhibits proliferation of gastric cancer cells.

Introduction: Gastric cancer is a frequently detected malignancy, and its incidence has increased over the past decades in East Asia. The present study investigated the effect of 5,7,2, 5-tetrahydroxy-8,6-dimethoxyflavone (THDMF) on gastric cancer cells and explored the underlying mechanism. The study analysed cell viability changes, apoptotic features, and metastasis potential of treatment with THDMF. Material and methods: MTT colorimetric assay was used for measurement of MKN28, MKN45, and GES-1 cell proliferation and flow cytometry for the detection of apoptosis. Transwell and wound healing assays were used to observe the invasion and migration abilities of MKN28 cells. The expression of p21, MMP2/-9, PI3K, and c-Myc proteins was detected by western blotting. Results: The THDMF treatment significantly (p < 0.05) reduced MKN28 and MKN45 cell proliferation without changing GES-1 cell viability. A significant increase in apoptotic cell population on treatment with THDMF was observed. Treatment of MKN28 cells with THDMF increased the percentage of cells in the G1 phase. Exposure of MKN28 cells to THDMF caused a marked decrease in invasion and migration potential in comparison to control cells. The expression of miR-145 was markedly increased in MKN28 cells on treatment with THDMF. In MKN28 cells expression of c-Myc, PI3K, p-AKT, MMP-2, and MMP-9 was suppressed markedly on exposure to THDMF. The expression of p21 protein in MKN28 cells was markedly promoted on exposure to THDMF. Conclusions: THDMF exhibits anti-cancer effect on gastric cancer cells in vitro by activation of cell apoptosis and arrest of cell cycle. In addition, THDMF promoted miR-145 expression and down-regulation of PI3K/AKT signalling pathway in MKN28 cells. Therefore, THDMF may be utilised as a potential novel therapeutic agent for the treatment of gastric cancer.

The key micronutrient copper orchestrates broad-spectrum virus resistance in rice.

Copper is a critical regulator of plant growth and development. However, the mechanisms by which copper responds to virus invasion are unclear. We previously showed that SPL9-mediated transcriptional activation of miR528 adds a previously unidentified regulatory layer to the established ARGONAUTE (AGO18)-miR528-L-ascorbate oxidase (AO) antiviral defense. Here, we report that rice promotes copper accumulation in shoots by inducing copper transporter genes, including HMA5 and COPT, to counteract viral infection. Copper suppresses the transcriptional activation of miR528 by inhibiting the protein level of SPL9, thus alleviating miR528-mediated cleavage of AO transcripts to strengthen the antiviral response. Loss-of-function mutations in HMA5, COPT1, and COPT5 caused a significant reduction in copper accumulation and plant viral resistance because of the increased SPL9-mediated miR528 transcription. Gain in viral susceptibility was mitigated when SPL9 was mutated in the hma5 mutant background. Our study elucidates the molecular mechanisms and regulatory networks of copper homeostasis and the SPL9-miR528-AO antiviral pathway.

Effect of Fermentation Time on Molecular Structure and Physicochemical Properties of Corn Ballast Starch.

The effect of fermentation treatment on the surface morphology, crystal structure, molecular weight, chain length distribution, and physicochemical properties of corn starch was investigated using natural fermentation of corn ballast. The amylose content in corn ballast starch reduced at first after natural fermentation, then grew, following the same trend as solubility. There were certain erosion marks on the surfaces of fermented corn ballast starch granules. The crystalline structure of corn ballast starch remained the same, i.e., a typical A-type crystalline structure, at different fermentation times; however, the intensities of diffraction peaks were different. The weight-average molecular weight of starch first increased and then decreased after fermentation. The content of low-molecular-weight starch (peak 3) decreased from 25.59 to 24.7% and then increased to 25.76%, while the content of high-molecular-weight starch (peak 1) increased from 51.45 to 53.26%, and then decreased to 52.52%. The fermentation time showed a negative correlation with the viscosity of starch, and the pasting temperature first increased, and then decreased. Natural fermentation can be used as a technical means to produce corn starch products as a result of the experiments' findings, and future experiments will detect and analyze the bacterial structure of corn fermentation broth in order to better understand the molecular mechanism of natural fermentation affecting the structure and physicochemical properties of corn starch.

Multiple functions of Rice dwarf phytoreovirus Pns10 in suppressing systemic RNA silencing.

RNA silencing is a potent mechanism of antiviral defense response in plants and other organisms. For counterdefense, viruses have evolved a variety of suppressors of RNA silencing (VSRs) that can inhibit distinct steps of a silencing pathway. We previously identified Pns10 encoded by Rice dwarf phytoreovirus (RDV) as a VSR, the first of its kind from double-stranded RNA (dsRNA) viruses. In this study we investigated the mechanisms of Pns10 function in suppressing systemic RNA silencing in the widely used Nicotiana benthamiana model plant. We report that Pns10 suppresses local and systemic RNA silencing triggered by sense mRNA, enhances viral replication and/or viral RNA stability in inoculated leaves, accelerates the systemic spread of viral infection, and enables viral invasion of shoot apices. Mechanistically, Pns10 interferes with the perception of silencing signals in recipient tissues, binds double-stranded small interfering RNA (siRNAs) with two-nucleotide 3' overhangs, and causes the downregulated expression of RDR6. These results significantly deepen our mechanistic understanding of the VSR functions encoded by a dsRNA virus and contribute additional evidence that binding siRNAs and interfering with RDR6 expression are broad mechanisms of VSR functions encoded by diverse groups of viruses.

Converting Poisonous Sulfate Species to an Active Promoter on TiO2 Predecorated MnOx Catalysts for the NH3-SCR Reaction.

MnOx-based catalysts possess excellent low-temperature NH3 selective catalytic reduction (NH3-SCR) activity, but the poor SO2/sulfate poisoning resistance and the narrow active-temperature window limit their application for NOx removal. Herein, TiO2 nanoparticles and sulfate were successively introduced into MnOx-based catalysts to modulate the NH3-SCR activity, and the active-temperature window (NO conversion above 80%, T80) was significantly broadened to 100-350 °C (SO42--TiO2@MnOx) compared to that of the pristine MnOx catalyst (ca. T80: 100-268 °C). Combined with advanced characterizations and control experiments, it was clearly shown that the poisonous effects of sulfate on the MnOx catalyst could be efficiently inhibited in the presence of TiO2 species due to the interaction between sulfate and TiO2 to form a solid superacid (SO42--TiO2) species as NH3 adsorption sites for the low-temperature process. Furthermore, such solid superacid (SO42--TiO2) species could weaken the redox ability to inhibit the excessive oxidation of NH3 and thus enhance the high-temperature activity significantly. This work not only puts forward the TiO2 predecoration strategy that converts sulfate to a promoter to broaden the active temperature window but also experimentally proves that the requirement of redox ability and acidity in the MnOx-based NH3-SCR catalyst was dependent on the reaction temperature range.

Effects of High-Temperature, High-Pressure, and Ultrasonic Treatment on the Physicochemical Properties and Structure of Soluble Dietary Fibers of Millet Bran.

OBJECTIVES: The effects of high-temperature, high-pressure, and ultrasonic treatment on the physicochemical properties and structure of soluble dietary fibers in millet bran were studied to provide a comprehensive reference for the utilization of millet bran. METHODS: Different physical methods were used to treat millet bran dietary fibers, and their microstructures and Fourier-transform infrared spectra before and after modification were compared. The physicochemical properties (water-holding capacity, swelling capacity, oil-holding capacity, fat-binding capacity, cation exchange capacity), total antioxidant capacity, and thermal characteristics were also analyzed. RESULTS: There were no significant changes in the chemical groups of millet bran's soluble dietary fibers after modification, but cracks appeared on the surface of the fibers and the structure became loose and porous. Fiber agglomeration was observed, as well as improved thermal stability. After modification, the water-holding capacity, swelling capacity, oil-holding capacity, fat-binding capacity, and cation exchange capacity of millet bran were improved. When compared to the original soluble dietary fibers, ultrasound-treated fibers showed the most substantial improvement in all four capabilities, with increases of 140, 50, 78.1, 65.7, and 37.8%, respectively, compared with the original soluble dietary fibers (P < 0.05). The total antioxidant capacity of the ultrasound-treated fibers was found to be higher than those of the fibers that underwent the other three treatments (P < 0.05). CONCLUSIONS: The physicochemical qualities and structural characteristics of the soluble dietary fibers in millet bran are affected by all three physical modification methods; however, the physicochemical properties of the ultrasound-treated fibers are most significantly improved.

Arabidopsis putative deacetylase AtSRT2 regulates basal defense by suppressing PAD4, EDS5 and SID2 expression.

The silent information regulator protein (Sir2) and its homologs are NAD(+)-dependent deacetylase enzymes that play important roles in a variety of physiological processes. However, the functions of the Sir2 family in plants are poorly understood. Here, we report that Arabidopsis AtSRT2, a homolog of yeast Sir2, negatively regulates plant basal defense against the pathogen Pseudomonas syringae pv. tomato DC3000 (PstDC3000). In response to PstDC3000 infection, the expression of AtSRT2 was down-regulated in a salicylic acid (SA)-independent manner. In addition, knock-out of AtSRT2 (srt2) enhanced resistance against PstDC3000 and increased expression of pathogenesis-related gene 1 (PR1). Conversely, overexpression of AtSRT2 resulted in hypersusceptibility to PstDC3000 and impaired PR1 induction. Consistent with this phenotype, expression of PAD4, EDS5 and SID2, three essential genes in the SA biosynthesis pathway, were increased in the srt2 mutant and decreased in AtSRT2-overexpressing plants. Taken together, these results demonstrate that AtSRT2 is a negative regulator of basal defense, possibly by suppressing SA biosynthesis.

Crystallization and preliminary X-ray analysis of tubulin-folding cofactor A from Arabidopsis thaliana.

Tubulin-folding cofactor A (TFC A) is a molecular post-chaperonin that is involved in the beta-tubulin-folding pathway. It has been identified in many organisms including yeasts, humans and plants. In this work, Arabidopsis thaliana TFC A was expressed in Escherichia coli and purified to homogeneity. After thrombin cleavage, a well diffracting crystal was obtained by the sitting-drop vapour-diffusion method at 289 K. The crystal diffracted to 1.6 A resolution using synchrotron radiation and belonged to space group I4(1), with unit-cell parameters a=55.0, b=55.0, c=67.4 A.

The P2 capsid protein of the nonenveloped rice dwarf phytoreovirus induces membrane fusion in insect host cells.

Insect transmission is an essential process of infection for numerous plant and animal viruses. How an insect-transmissible plant virus enters an insect cell to initiate the infection cycle is poorly understood, especially for nonenveloped plant and animal viruses. The capsid protein P2 of rice dwarf virus (RDV), which is nonenveloped, is necessary for insect transmission. Here, we present evidence that P2 shares structural features with membrane-fusogenic proteins encoded by enveloped animal viruses. When RDV P2 was ectopically expressed and displayed on the surface of insect Spodoptera frugiperda cells, it induced membrane fusion characterized by syncytium formation at low pH. Mutational analyses identified the N-terminal and a heptad repeat as being critical for the membrane fusion-inducing activity. These results are corroborated with results from RDV-infected cells of the insect vector leafhopper. We propose that the RDV P2-induced membrane fusion plays a critical role in viral entry into insect cells. Our report that a plant viral protein can induce membrane fusion has broad significance in studying the mechanisms of virus entry into insect cells and insect transmission of nonenveloped plant and animal viruses.

The effect of sorghum resistance resistant starch-mediated equol on the histological morphology of the uterus and ovaries of postmenopausal rats.

Equol is a metabolite of daidzein and has a higher biological activity than daidzein. Equol, combined with estrogen receptors, can reduce the incidence of diseases such as cardiovascular disease, osteoporosis, and breast cancer; more effectively alleviate the symptoms of perimenopausal syndrome; and improve age-related decline of the uterus and ovaries. Research has shown that food composition can greatly affect the formation of equol in the intestinal tract. In the intestines, the content of nonstarch polysaccharides that can stimulate fermentation is high, thereby allowing intestinal bacteria to quickly and completely transform the daidzein into equol. This study used Sprague Dawley (SD) rats as a model, where menopause was established through direct intragastric administration of formistan. In the 6-week-long experiment, intragastric administration of RS while feeding bean pulp reduced the body weight of postmenopausal rats, reduced the efficiency of feed utilization of rats, and increased the weight of organs such as the uterus and ovaries. Routine blood indexes showed that no adverse reactions were produced by intragastric administration of RS. 16s rDNA sequencing further verified Lactobacillus and Clostridium XIVa, as the bacteria that converted daidzein into equol.

The resistant starch from sorghum regulates lipid metabolism in menopausal rats via equol.

Equol is a metabolite of daidzein and has a higher biological activity than daidzein. High levels of non-starch polysaccharides can stimulate fermentation in the intestine leading to rapid conversion of daidzein into equol that has great potential to reduce obesity in postmenopausal women. In the present study, female Sprague-Dawley rats were used to establish a menopausal model by oral administration of formestane and to compare the protective effect of resistant starch on lipid metabolism, with or without soybean feed. The resistant starch was found to effectively control body weight and adipose tissue quality, while increasing the high-density lipoprotein cholesterol (HDL-C) concentration and lowering the glycerol, triacylglycerols (TG), total cholesterol (TC), and low density lipoprotein cholesterol (LDL-C) concentrations with soybean feed. Equol inhibited the expression of SREBPC1 gene by inhibiting SHP in the liver via transcription factor FXR, thereby inhibiting the synthesis of triglyceride and fatty acid in the liver. PRACTICAL APPLICATIONS: Intake of a certain amount of resistant starch while eating the soy product can better regulate lipid metabolism in menopausal obese rats compared to consumption of resistant starch alone. Studies have shown that resistant starch converts daidzein to Equol by regulating the structure of the intestinal flora and acts as an estrogen in menopausal rats. This research will further expand the health applications of resistant starch and provide useful information for the food industry.

Identification of the Metabolites in Rat Urine after Oral Administration and Elucidation of the Metabolic Process of Naozhenning Granule Using LC-MS.

Naozhenning (NZN) granule, a Chinese herbal formula, is widely used to treat craniocerebral trauma and promote functional recovery. In our previous study, the chemical components, as well as the serum metabolites in the male Sprague-Dawley rats of the NZN granule after oral administration were characterized. In this study, the urine metabolites in the male Sprague-Dawley rats were further investigated by ultrahigh-performance liquid chromatography-Q Exactive hybrid quadrupole-Orbitrap high-resolution accurate mass spectrometry. In order to identify the urine metabolites comprehensively, three sample preparation methods were used, including solid-phase extraction, protein precipitation method and solvent partition. Based on the accurate molecular weight and the fragmentation information from the MS spectra, a total of 76 urine metabolites were identified, which including 17 prototypes and 59 metabolites. The results showed that the detected urine metabolites were different for the different pretreatment methods, as some metabolites could only be detected in the particular pretreatment method. In addition, the metabolic processes of the components from NZN granule to the serum and urine were also elucidated and discussed. The results will provide useful information for further studying the relationship between the chemical components and pharmacological activity of NZN granule.

A non-structural protein encoded by Rice Dwarf Virus targets to the nucleus and chloroplast and inhibits local RNA silencing.

RNA silencing is a potent antiviral mechanism in plants and animals. As a counter-defense, many viruses studied to date encode one or more viral suppressors of RNA silencing (VSR). In the latter case, how different VSRs encoded by a virus function in silencing remains to be fully understood. We previously showed that the nonstructural protein Pns10 of a Phytoreovirus, Rice dwarf virus (RDV), functions as a VSR. Here we present evidence that another nonstructural protein, Pns11, also functions as a VSR. While Pns10 was localized in the cytoplasm, Pns11 was localized both in the nucleus and chloroplasts. Pns11 has two bipartite nuclear localization signals (NLSs), which were required for nuclear as well as chloroplastic localization. The NLSs were also required for the silencing activities of Pns11. This is the first report that multiple VSRs encoded by a virus are localized in different subcellular compartments, and that a viral protein can be targeted to both the nucleus and chloroplast. These findings may have broad significance in studying the subcellular targeting of VSRs and other viral proteins in viral-host interactions.