The apple 14-3-3 protein MdGRF11 interacts with the BTB protein MdBT2 to regulate nitrate deficiency-induced anthocyanin accumulation

Nitrogen is an important factor that affects plant anthocyanin accumulation.In apple,the nitrate-responsive BTB/TAZ protein MdBT2 negatively regulates anthocyanin biosynthesis.In this study,we found that MdBT2 undergoes posttranslational modifications in response to nitrate deficiency.Yeast two-hybrid,protein pull-down,and bimolecular fluorescence complementation(BiFC)assays showed that MdBT2 interacts with MdGRF11,a 14-3-3 protein;14-3-3 proteins compose a family of highly conserved phosphopeptide-binding proteins involved in multiple physiological and biological processes.The interaction of MdGRF11 negatively regulated the stability of the MdBT2 protein via a 26S proteasome-dependent pathway,which increased the abundance of MdMYB1 proteins to activate the expression of anthocyanin biosynthesis-related genes.Taken together,the results demonstrate the critical role of 14-3-3 proteins in the regulation of nitrate deficiency-induced anthocyanin accumulation.Our results provide a novel avenue to elucidate the mechanism underlying the induction of anthocyanin biosynthesis in response to nitrate deficiency.

Metabolomics and its application in the evaluation of toxicity of traditional Chinese medicine

Metabolomics is a newly developed discipline following genomics,transcriptomics,and proteomics.It is based on the theory of systems biology,supported by modern spectroscopy analysis technology and is analyzed by the external stimulation of organisms.By analyzing the overall changes of metabolites with low molecular weight in organism after being stimulated by the outside world,we can understand the physiological and pathological state of organism,and finally clarify the essence of biological changes of organism.In recent years,metabolomics has developed rapidly.It has been widely used in early toxicity screening of lead compounds,preclinical toxicity of drugs and toxicity evaluation of clinical drugs.The in-depth interpretation of metabolomics results is also developing.This article reviewed the progress of metabolomics technology and its application in the study of toxicity evaluation of traditional Chinese medicine.

Expression of Endogenous Retrovirus ev/J gp85 Gene and Analysis of Its Immunoreactivity in Comparison with Exogenous Viral Protein

The envelope gene gp85 of ev/J, a new family of endogenous avian retroviral sequences identified recently, has the most extensive nucleotide sequence identity ever described with ALV-J avian leukosis virus. This report described expression of ev/J envelope gene gp85 derived from commercial meat-type chicken using the Invitrogen Bac-to-Bac baculovirus expression system. The antigenicity and immunoreactivity of the recombinant endogenous gp85 gene product (SU) were analyzed by indirect immunofluorescence, Western blot, indirect and blocking Enzyme-Linked ImmunoSorbent Assay (ELISA) using JE9 monoclonal antibody (MAb) against the envelope protein of ALV-J (ADOL-4817), positive mouse antiserum against the ev/J gp85 SU and sera from chicken naturally infected with ALV-J. The results showed that the ev/J gp85 SU can bind specifically to JE9 MAb and antiserum from chicken naturally infected with ALV-J, and the binding reactivity between exogenous ALV-J gp85 SU and natural positive chicken serum against exogenous ALV-J can be blocked by positive mouse serum against the ev/J gp85 SU. It is concluded that recombinant endogenous gp85 gene product (SU) has close immunological relatedness to the envelope protein of exogenous ALV-J (ADOL-4817 and IMC10200 strain).

Prescription optimization and characterization of Galangin-PLGA nanoparticles

Objective:To make Galangin-PLGA nanoparticles(GL-PLGA NPs)were prepared using the biodegradable material polylactic-coglycolic acid(PLGA)as the carrier material,and the formulation was optimized.Methods:Nanoparticles were prepared by modified emulsification-solvent evaporation method and the content of galangin was determined by HPLC.With particle size and entrapment efficiency as the indexes,single factor and orthogonal test were used to optimize the formulation,and the quality of the optimized GL-PLGA NPs was evaluated from the surface configuration,particle size distribution,Zeta potential and entrapment efficiency.Results:The optimal prescription conditions of GL-PLGA NPs were:the volume ratio of organic phase to aqueous phase was 1:8,the concentration of PVA was 1.5%,the concentration of PLGA was 1.0%,and the drug concentration was 0.2%.Under these conditions,the average particle size of GL-PLGA NPs was 249±1.32nm,polydispersity index(PDI)was 0.059,Zeta potential was-4.86mV,and the entrapment efficiency was 75.3%.The results of electron microscopy showed that the nanoparticles were spherical,uniform particle size and good dispersion.Conclusion:The preparation method is simple and stable,and GL-PLGA NPs with suitable particle size and high entrapment efficiency can be obtained.

Adiponectin Receptor 1 Single Nucleotide Polymorphism Is Highly Associated with Hypertriglyceridemia in Asian Male—A Novel Genetic Screening to Reduce Risk of Cerebrovascular Disease

Background: Adiponectin is involved in regulating both glucose and fatty acid. Associations of the known adiponectin receptors 1 (ADIPOR1) single nucleotide polymorphism (SNP) with diabetes have been demonstrated while hypertriglyceridemia is frequently associated with cerebrovascular disease (CVD) among diabetes. Triglyceride metabolism was also reported to be different between genders and estrogen was observed to interfere with adiponectin effects via ADIPOR1. It seems important to investigate whether the ADIPOR1 SNP variants may be significant determinants in triglyceride metabolism and hence be a risk of CVD in specific gender. Methods: A survey was performed on random self-reported healthy subjects aged 35 and above with their biochemical data collected. Genotyping for ADIPOR1 SNP (rs1342387) was carried out using?TaqMan Genotyping Assays. Interviews were also conducted regarding stress, adverse diet behavior and exercise. Multivariable logistic regression analyses were performed to identify the strongest contributing variables. Findings: The ADIPOR1 minor allele carrier (T/T and T/C) had significantly (p?= 0.02) higher TG mean compared to homozygous (C/C) major alleles. TG difference was significantly higher in male (p?= 0.02) with a larger difference in mean, whereas the difference disappeared among female (p?= 0.32). Multivariate logistic regression analyses were performed by defining abnormal TG based on NCEP criteria of metabolic syndrome, and when all life style variables were entered in the model with ADIPOR1, only the ADIPOR1 inmale showed significant (p?= 0.03) and very high association with abnormal TG (Exp(β) 16.31). Discussion: The findings provide sample evidence of a relation between ADIPOR1 SNP minor allele carrier and high TG concentrations in male. The association of abnormal TG and ADIPOR1 is much stronger than that compared to life style. The implications of this survey may be further extend to identifying the genetic risk of abnormal TG at young age and reduce the CVD incidence by early intervention.

一步酸化法制备抗菌肽胶束及其抗菌性能研究

提出了简化抗菌肽纳米粒子制备流程的“一步酸化法”.利用[1-(4-氨基苯基)-1,2,2-三苯基]乙烯(TPE-NH2)引发Nε-三氟乙酰基-L-赖氨酸环内酸酐[Lys(Tfa)NCA]和L-谷氨酸-γ-苄酯环内酸酐(BLG NCA)单体的开环共聚,然后选择性脱除三氟乙酰保护基,制备了无规共聚多肽TPE-P(Lys28-stat-BLG16).随后,采取一步酸化法直接制备了共聚多肽胶束溶液.抗菌实验结果表明,该抗菌肽胶束对大肠杆菌及金黄色葡萄球菌的最低抑菌浓度分别为256和64μg/mL,最低杀菌浓度分别为400和100μg/mL.此外,由于其四苯乙烯(TPE)端基具有聚集诱导发光效应,该抗菌肽胶束还有望用于可视化探究抗菌机理.

Biodegradable Polypeptide-based Vesicles with Intrinsic Blue Fluorescence for Antibacterial Visualization

Fluorescence imaging has been an indispensable tool to provide dynamic information about the localization and quantity of organisms.Meanwhile,due to the intrinsic hollow structure and modularized biofunctionalities,polymer vesicles have been widely applied in biomedical field.However,most polymer vesicles are embedded with organic fluorophores for fluorescence imaging,which have certain drawbacks such as leakage and possible cytotoxicity.Here,we present a biodegradable polypeptide-based vesicle with intrinsic blue fluorescence without introducing any fluorophore for real-time visualization of antibacterial process.Through modular design to integrate multiple functional fragments,poly(ε-caprolactone)-block-poly(tryptophan)-block-poly(lysine-stat-phenylalanine)[PCL25-b-PTrp2-b-P(Lys13-statPhe4)]was synthesized,where PCL chains form the hydrophobic membrane,P(Lys-stat-Phe)and PTrp provide intrinsic fluorescence and broadspectrum antibacterial activity.It is noteworthy that the fluorescence emission was shifted from invisible ultraviolet range of amino acids to visible range(emission maximum at 436 nm),which makes it possible to visualize the antibacterial process.In addition,through utilizing the intrinsic fluorescence of vesicles,confocal fluorescent imaging of vesicles with bacteria validated the specific adhesion of vesicle towards bacteria,and the bacterial death through membrane disruption.Overall,we provided a novel approach to developing biodegradable fluorescent polypeptide-based vesicles for real-time visualization of antibacterial process.