Development of a microsphere-based fluorescence immunochromatographic assay for monitoring lincomycin in milk, honey, beef, and swine urine.

The residue of lincomycin (LIN) in edible animal foodstuffs caused by the widespread use of veterinary drugs is in need of rapid, simple, and sensitive detection methods. The present work introduces a fluorescent microsphere immunoassay (FMIA) for detecting LIN in different samples based on the competitive immunoreaction on the chromatography test strip. The residues of LIN in different samples compete with bovine serum albumin (BSA) labeled LIN conjugates on the T-line to bind to the anti-LIN monoclonal antibody labeled fluorescent microspheres (FM-mAbs).

PIGD: a database for intronless genes in the Poaceae.

Background: Intronless genes are a feature of prokaryotes; however, they are widespread and unequally distributed among eukaryotes and represent an important resource to study the evolution of gene architecture. Although many databases on exons and introns exist, there is currently no cohesive database that collects intronless genes in plants into a single database.

Description: In this study, we present the Poaceae Intronless Genes Database (PIGD), a user-friendly web interface to explore information on intronless genes from different plants.

Genome-wide identification and characterization of maize expansin genes expressed in endosperm.

By promoting cell wall loosening, expansins contribute to cell enlargement during various developmental processes. Nevertheless, the role of expansins in the expansion and development of endosperm--a major seed component whose cell size is significantly associated with grain yield--is poorly understood. To explore associated biological processes and the evolution of expansins in maize, we performed a systematic analysis of the expansin gene family encompassing gene structure, phylogeny, chromosomal location, gene duplication, and gene ontology.

Application of RNA interference methodology to investigate and develop SCMV resistance in maize.

Specific fragments of the sugarcane mosaic virus (SCMV) coat protein gene (cp) were amplified by reverse transcriptionpolymerase chain reaction and used to construct a marker free small interfering RNA complex expression vector against SCMV. In planta transformation was performed on maize (Zea mays) inbred line 8112 mediated by Agrobacterium tumefaciens. PCR and Southern blot analyses demonstrated successful integration of the cp segment into the 8112 genome.

Genome duplication and gene loss affect the evolution of heat shock transcription factor genes in legumes.

Whole-genome duplication events (polyploidy events) and gene loss events have played important roles in the evolution of legumes. Here we show that the vast majority of Hsf gene duplications resulted from whole genome duplication events rather than tandem duplication, and significant differences in gene retention exist between species. By searching for intraspecies gene colinearity (microsynteny) and dating the age distributions of duplicated genes, we found that genome duplications accounted for 42 of 46 Hsf-containing segments in Glycine max, while paired segments were rarely identified in Lotus japonicas, Medicago truncatula and Cajanus cajan.

Relationship between tyrosine phosphorylation and protein expression of insulin receptor and insulin resistance in gestational diabetes mellitus.

The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay.

Different evolutionary patterns among intronless genes in maize genome.

Intronless genes, as a characteristic feature of prokaryotes, are an important resource for the study of the evolution of gene architecture in eukaryotes. In the study, 14,623 (36.87%) intronless genes in maize were identified and the percentage is greater than that of other monocots and algae.

A novel maize homeodomain-leucine zipper (HD-Zip) I gene, Zmhdz10, positively regulates drought and salt tolerance in both rice and Arabidopsis.

Increasing evidence suggests that homeodomain-leucine zipper I (HD-Zip) I transcription factors play important roles in abiotic stress responses, but no HD-Zip I proteins have been reported in maize. Here, a drought-induced HD-Zip I gene, Zmhdz10, was isolated from maize and characterized for its role in stress responses. Real-time quantitative PCR showed that expression of Zmhdz10 was also induced by salt stress and ABA.

Cloning, expression, purification and characterization of a bispecific single-chain diabody against fluoroquinolones and sulfonamides in Escherichia coli.

A recombinant bispecific single-chain diabody (scDb), recognizing fluoroquinolones (FQs) and sulfonamides (SAs), was successfully constructed with two single-chain variable fragment antibodies (scFvs). The scDb gene was cloned into the expression vector pJB33, and 6×His-tagged scDb was expressed as soluble bodies in Escherichia coli RV308 host, then purified by one step affinity chromatography of immobilized metal ion affinity chromatography (IMAC). SDS-PAGE and Western blotting analysis of the purified scDb indicated that the prepared scDb was successfully expressed as a ∼60 kDa and the final purity of the scDb protein was up to 95% with yields of approximately 6 mg/L of bacterial culture.

Determination of L-hydroxyproline using hydrophilic interaction chromatography coupled to tandem mass spectrometry with lyophilized concentrated extraction in milk and dairy products.

In this study, a screening and confirmation method for the determination of l-hydroxyproline (Hyp) as a target compound in milk and dairy products using high-performance liquid chromatography with tandem mass spectrometry was developed. The samples were lyophilized after acidic hydrolysis, followed by cleanup with graphitized carbon black to remove pigments. Hyp was separated by a hydrophilic interaction chromatographic column and analyzed by high-performance liquid chromatography with tandem mass spectrometry working with multiple reaction monitoring mode using an electrospray ionization interface in a positive-ion mode.

Production of Monoclonal Antibody and Development of a New Immunoassay for Apramycin in Food.

Apramycin (APR) residue in food of animal origin can cause harmful effects on human health. In this study, a monoclonal antibody (mAb) was successfully produced using APR-BSA as immunogen, which was prepared by using the glutaraldehyde method. mAb 2A2 showed low cross-reactivity (<0.

miR-99a promotes proliferation targeting FGFR3 in human epithelial ovarian cancer cells.

MiRNAs have been reported as important regulators in normal physiological processes, human cancer, and even their roles as therapeutic targets have been proposed. In epithelial ovarian cancer (EOC), the expression of miRNAs is reported to remarkably deregulate, showing that miRNAs are involved in the initiation and progression of this disease. In this study, we found that miR-99a was obviously decreased in EOC tissues, serums and cell lines SKOV-3.

Downregulation of caffeoyl-CoA O-methyltransferase (CCoAOMT) by RNA interference leads to reduced lignin production in maize straw.

Lignin is a major cell wall component of vascular plants that provides mechanical strength and hydrophobicity to vascular vessels. However, the presence of lignin limits the effective use of crop straw in many agroindustrial processes. Here, we generated transgenic maize plants in which the expression of a lignin biosynthetic gene encoding CCoAOMT, a key enzyme involved in the lignin biosynthesis pathway was downregulated by RNA interference (RNAi).

[Design and development of trace Cr(VI) sensor].

Hexavalent chromium detection in medicine capsules is generally analyzed in the laboratory, it is difficult to meet the demand for field detection, and to address this problem, a sensor which can be used for on-site detection of trace amounts of hexavalent chromium was designed. It mainly includes chemically sensitive materials, optical sensing module and signal processing module, the chemical sensitive materials is to achieve the conversion of the hexavalent chromium concentration signal, the optical sensing module is to complete a stable output of the laser light source, and the signal processing module is to complete a photoelectric conversion of the weak fluorescence signal, signal amplification, and data processing and displaying. With using the indigenously developed photoelectric acquisition, conversion and signal processing system to complete the rapid detection of trace amounts of hexavalent chromium, so the miniaturization of testing instruments and on-site detection were achieved.

Molecular evolution of the HD-ZIP I gene family in legume genomes.

Homeodomain leucine zipper I (HD-ZIP I) genes were used to increase the plasticity of plants by mediating external signals and regulating growth in response to environmental conditions. The way genomic histories drove the evolution of the HD-ZIP I family in legume species was described; HD-ZIP I genes were searched in Lotus japonicus, Medicago truncatula, Cajanus cajan and Phaseolus vulgaris, and then divided into five clades through phylogenetic analysis. Microsynteny analysis was made based on genomic segments containing the HD-ZIP I genes.

Determination of zeranol and its metabolites in bovine muscle and liver by a chemiluminescence enzyme immunoassay: compared to an ultraperformance liquid chromatography tandem mass spectroscopy method.

A chemiluminescent enzyme immunoassay (CLEIA) was compared to an ultraperformance liquid chromatography tandem mass spectroscopy (UPLC-MS/MS) procedure for the analysis of zeranol and its metabolites in bovine tissue samples. Apparent recoveries from fortified samples by both methods were comparable at 0.5-4.

Ultra-performance liquid chromatography-tandem mass spectrometry determination and depletion profile of flunixin residues in tissues after single oral administration in rabbits.

An ultra-performance liquid chromatography with tandem mass spectrometric detection (UPLC-MS/MS) method was developed for the detection of flunixin residues in rabbit tissues. The samples were extracted with acidic acetonitrile, defatted with n-hexane, and then purified by HLB solid-phase extraction cartridge. Analysis was carried out on UPLC-ESI-MS/MS working with multiple reaction monitoring (MRM) mode.

Development of a simple method for simultaneous determination of nine subclasses of non-steroidal anti-inflammatory drugs in milk and dairy products by ultra-performance liquid chromatography with tandem mass spectrometry.

A multi-residue analysis method for simultaneous determination of nine subclasses of non-steroidal anti-inflammatory drugs (NSAIDs) in milk and dairy products by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) has been established. The sample was initially extracted and deproteinized with ascorbic acid buffer (0.01M, pH 3) and acetonitrile-ethyl acetate mixture, followed by centrifugation and evaporation, then reconstituted with acetonitrile-0.

Proteomic analysis reveals that proteasome subunit beta 6 is involved in hypoxia-induced pulmonary vascular remodeling in rats.

Background: Chronic hypoxia (CH) is known to be one of the major causes of pulmonary hypertension (PH), which is characterized by sustained elevation of pulmonary vascular resistance resulting from vascular remodeling. In this study, we investigated whether the ubiquitin proteasome system (UPS) was involved in the mechanism of hypoxia-induced pulmonary vascular remodeling. We isolated the distal pulmonary artery (PA) from a previously defined chronic hypoxic pulmonary hypertension (CHPH) rat model, performed proteomic analyses in search of differentially expressed proteins belonging to the UPS, and subsequently identified their roles in arterial remodeling.

Chemiluminescence competitive indirect enzyme immunoassay for 20 fluoroquinolone residues in fish and shrimp based on a single-chain variable fragment.

A chemiluminescent competitive indirect enzyme-linked immunosorbent assay, based on a mutant single-chain variable fragment (scFv), was developed to detect a broad range of fluoroquinolones (FQs) in fish and shrimp matrices. In this study, the best scFvC4A9H1_mut2 was adopted, which showed 10-fold improved affinity to sarafloxacin (SAR), difloxacin (DIF), and trovafloxacin (TRO), while the affinity to other FQs was fully inherited from wild-type scFvC4A9H1. In the optimized generic test, scFvC4A9H1_mut2 in combination with norfloxacin-ovalbumin conjugate and horseradish peroxidase-labeled anti-c-myc 9E10 antibody showed 50 % binding inhibition (IC50) at 0.

Genome-wide identification and expression profiling of ankyrin-repeat gene family in maize.

Members of the ankyrin repeats (ANK) gene family encode ANK domain that are common in diverse organisms and play important roles in cell growth and development, such as cell-cell signal transduction and cell cycle regulation. Recently, genome-wide identification and evolutionary analyses of the ANK gene family have been carried out in Arabidopsis and rice. However, little is known regarding the ANK genes in the entire maize genome.

Development of a highly sensitive chemiluminescence enzyme immunoassay using enhanced luminol as substrate.

In this study, a high sensitivity chemiluminescence enzyme immunoassay (CLEIA) based on novel enhancers was developed. Under optimal conditions, we developed an enhanced chemiluminescence reaction (ECR) catalyzed by horseradish peroxidase (HRP-C) in the presence of 3-(10'-phenothiazinyl) propane-1-sulfonate (SPTZ) and 4-morpholinopyridine (MORP) as enhancers. The limit of detection of the newly prepared chemiluminescent cocktail for HRP was 0.

Development of a rapid chemiluminescent ciELISA for simultaneous determination of florfenicol and its metabolite florfenicol amine in animal meat products.

Background: A rapid one-step chemiluminescent competitive indirect enzyme-linked immunosorbent assay (CL-ciELISA) for florfenicol (FF) and its major metabolite florfenicol amine (FFA) residues in animal meat products has been developed.

Results: The 50% binding inhibition (IC₅₀) values of the method were 0.195 µg kg⁻¹ for FFA and 0.

Genome-wide analysis of the CCCH zinc finger gene family in Medicago truncatula.

In this study, we identified 34 CCCH Znf genes in Medicago truncatula and the results of semi-quantitative RT-PCR revealed that the expression patterns of subfamily VI members were diverse. CCCH-type zinc finger (Znf) proteins are specific transcriptional factors with a typical motif consisting of three cysteine residues and one histidine residue. Increasing evidences have revealed that CCCH Znf proteins participated in the regulation of plant growth, developmental processes and environmental responses.