Biomedical subjects

Xiaojuan Xu

Publications and source records attributed to Xiaojuan Xu.

16 recordsLinked to original sources

PubMed2006

Generation and immunogenicity of a recombinant pseudorabies virus expressing cap protein of porcine circovirus type 2.

Porcine circovirus type 2 (PCV2) is associated with post-weaning multisystemic wasting syndrome (PMWS), which is an important economical disease affecting the pig industry worldwide. In order to develop an effective vaccine for PMWS, a recombinant pseudorabies virus (PRV) was generated and tested in piglets in this study. The PCV2 open reading frame 2 (ORF2) gene was inserted into pIECMV plasmid and co-transfected with PRV Tk-/gE-/LacZ+ genome into IBRS-2 cells to generate a recombinant Tk-/gE-/ORF2(+) virus. The expression of PCV2 ORF2 gene in the recombinant virus was confirmed by Western blotting and indirect immunofluorescence assay (IFA). Four-week-old piglets were immunized by the recombinant virus, and the immunogenicity of PRV Tk-/gE-/ORF2(+) was tested by PRV-enzyme-linked immunosorbent assay (ELISA), PRV neutralizing assay, ORF2-ELISA and ORF2 specific lymphocyte proliferation response. PRV Tk-/gE-/ORF2(+) elicited significant humoral immune responses to both PRV and PCV2, and the PCV2-specific lymphocyte proliferation response could be detected on day 49 of this experiment. These findings suggest that the recombinant PRV Tk-/gE-/ORF2(+) may be a potential vaccine against both PCV2 and PRV.

Animals↗

PubMed2006

Active avoidance conditioning in zebrafish (Danio rerio).

The zebrafish represents a potentially useful organism for studying genes involved in learning and memory function in vertebrates, because a number of genetic techniques in zebrafish have been developed to produce a wide variety of genetic mutants. While zebrafish mutants are being developed, behavioral studies on learning and memory function in zebrafish are in urgent need. The present study investigated active avoidance conditioning in normal zebrafish. Zebrafish were trained to swim from a lighted (CS) compartment to a dark compartment to avoid an electrical body shock (US) in a shuttle-box that consisted of a water-filled tank separated by an opaque barrier into two equal compartments. By varying the number of trials per training session and the duration of the intertrial interval, Experiments 1 and 2 showed that, with the CS, US, and intertrial interval being 12s, zebrafish learned avoidance responses within a training session consisting of 30 trials and retained the avoidance responses. Experiment 3 showed that zebrafish learned avoidance responses following the association between the CS of light and the US of shock in the avoidance conditioning paradigm. Using the avoidance conditioning paradigm, Experiment 4 investigated the amnestic effects of N-methyl-D-aspartate receptor antagonist MK-801 and nitric oxide synthase inhibitor L-NAME in zebrafish. Experiment 4 showed that post-training injection of L-NAME significantly impaired retention of avoidance responses while MK-801 did not, confirming previous results with other vertebrates. The results of the present study suggest the similar involvements of neurochemicals in learning and memory among vertebrates. Thus, future studies with zebrafish mutants may identify genes involved in learning and memory in vertebrates.

Animals↗

PubMed2006

Vicarious trial-and-error behavior and hippocampal cytochrome oxidase activity during Y-maze discrimination learning in the rat.

The present study investigated whether more vicarious trial-and-error (VTE) behavior, defined by head movement from one stimulus to another at a choice point during simultaneous discriminations, led to better visual discrimination learning in a Y-maze, and whether VTE behavior was a function of the hippocampus by measuring regional brain cytochrome oxidase (C.O.) activity, an index of neuronal metabolic activity. The results showed that the more VTEs a rat made, the better the rat learned the visual discrimination. Furthermore, both learning and VTE behavior during learning were correlated to C.O. activity in the hippocampus, suggesting that the hippocampus plays a role in VTE behavior during discrimination learning.

Analysis of Variance↗

PubMed2006

Antitumor activities of O-sulfonated derivatives of (1-->3)-alpha-D-glucan from different Lentinus edodes.

Four water-insoluble (1-->3)-alpha-D-glucans, coded L-II1, L-II2, L-II3 and L-II4, with different molecular weights were isolated from four kinds of fruiting bodies of Lentinus Edodes. The four alpha-D-glucans were O-sulfonated to obtain derivatives (SL-II) having degrees of substitution (DS) from 0.9 to 2.1 respectively. The structure of the samples was analyzed by infrared spectra, elemental analysis, and 13C NMR. The weight-average molecular weight (Mw), radii of gyration ( z1/2) and intrinsic viscosity ([eta]) of the native alpha-D-glucans and O-sulfonated derivatives were measured by size-exclusion chromatography combined with laser light scattering (SEC-LLS), LLS, and viscometry in 0.2 M aqueous NaCl and in dimethyl sulfoxide (DMSO) containing 0.25 M LiCl at 25 degrees C respectively. The Mw values of the O-sulfonated derivatives were much lower than those of the native alpha-D-glucans. The experimental results indicate that the O-sulfonated derivatives are water-soluble and exist as an expanded flexible chain in aqueous solution owing to intramolecular hydrogen bonding or interaction between charge groups. The in vivo and in vitro antitumor activities of the native alpha-D-glucans and their O-sulfonated derivatives against solid tumor Sarcoma 180 cells were evaluated and compared. Interestingly, all of the O-sulfonated derivatives exhibited higher antitumor activities than those of the native glucans. The results reveal that the effect of O-sulfonation of the alpha-D-glucan on the improvement of their antitumor activities was considerable.

Animals↗

PubMed2006

Development of a specific latex agglutination test based on a recombinant hemagglutinin protein to detect antibodies to H5 avian influenza viruses.

A latex agglutination test (LAT) was developed for rapid detection of antibodies to H5 avian influenza viruses (AIVs). The hemagglutinin protein of H5 AIV was covalently linked to carboxylated latex by ethyl-dimethyl-amino-propyl carbodiimide to prepare the sensitized latex beads. The LAT was evaluated with the hemagglutination inhibition (HI) assay as the reference test. The H5-LAT showed a sensitivity of 87.0% and specificity of 88.9% in detecting 126 serum samples from experimentally infected chickens and a sensitivity of 82.5% and specificity of 86% in detecting 587 field chicken serum samples from mostly vaccinated chickens. The agreement ratio between H5-LAT and HI was found to be 87.3% and 83.1% for the two groups of samples, respectively. Difficulty with background agglutination in stored chicken sera was overcome by serum pretreatment with either dried chicken liver powder or dilution buffer containing detergent Tween-20. The H5-LAT has advantages over a previously reported whole-virus LAT in terms of biosafety in preparation, chemical stability, and higher specificity. It is a rapid and simple test suitable for field monitoring of antibodies to H5-type AIV.

Animals↗

PubMed2005

Preparation and antimicrobial activity of hydroxypropyl chitosan.

Water-soluble hydroxypropyl chitosan (HPCS) derivatives with different degrees of substitution (DS) and weight-average molecular weight (Mw) were synthesized from chitosan and propylene epoxide under basic conditions. Their structure was characterized by IR spectroscopy, NMR spectroscopy, and elemental analysis, which showed that both the OH groups at C-6 and C-3 and the NH2 group of chitosan were alkylated. The DS value of HPCS ranged from 1.5 to 3.1 and the Mw was between 2.1x10(4) and 9.2x10(4). In vitro antimicrobial activities of the HPCS derivatives were evaluated by the Kirby-Bauer disc diffusion method and the macrotube dilution broth method. The HPCS derivatives exhibited no inhibitory effect on two bacterial strains (Escherichia coli and Staphylococcus aureus); however, some inhibitory effect was found against four of the six pathogenic fruit fungi investigated. Some derivatives (HPCS1, HPCS2, HPCS3, HPCS3-1, and HPCS4) were effective against C. diplodiella and F. oxysporum. HPCS3-1 is the most effective one with MIC values of 5.0, 0.31, 0.31, and 0.16mg/mL against A. mali, C. diplodiella, F. oxysporum, and P. piricola, respectively. Antifungal effects were also observed for HPCS2 and HPCS3-1 against A. mali, as well as HPCS3 and HPCS3-1 against P. piricola. The results suggest that relatively lower DS and higher Mw value enhances the antifungal activity of HPCS derivatives.

Animals↗

PubMed2005

Correlation between antitumor activity, molecular weight, and conformation of lentinan.

A (1-->3)-beta-D-glucan having (1-->6) branching (L-FV-IB) from Lentinus edodes in water was degraded into seven fractions of different molecular weights by ultrasonic irradiation, and each was further fractionated into three parts, by precipitation from water into acetone at room temperature. The weight-average molecular weight (M(w)), radius of gyration ( (z)(1/2)), and intrinsic viscosity ([eta]) of lentinan and its fractions in 0.9% NaCl aqueous solution and dimethyl sulfoxide (Me(2)SO) were determined by size-exclusion chromatography combined with multi-angle laser light scattering (SEC-LLS), LLS, and viscometry. Analysis of M(w), [eta], and (z)(1/2) in terms of known theory for worm-like chains yielded 2240 +/- 100 nm(-1), and 100 +/- 10 nm for molar mass per unit contour length (M(L)), and persistence length (q), respectively, corresponding with theoretical data for triple-helical chains. The [alpha](D) of lentinan in water-Me(2)SO mixtures indicated an order-disorder transition. The results indicated that lentinan exists as a triple helix in 0.9% NaCl aqueous solution and as a single flexible chain in Me(2)SO. Assays in vivo and in vitro against the growth of Sarcoma 180 solid tumor as well as the colorimetric 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method for lentinan showed that the triple-helix sample exhibited a relatively high inhibition ratio. Interestingly, the triple-helix lentinan with M(w) of 1.49 x 10(6) exhibited the highest antitumor activity in vivo, having an inhibition ratio (xi) of 49.5%, close to that of 5-fluorouracil (xi = 50.5%), whereas the bioactivity (xi = 12.3%) of its single flexible chains almost disappeared. The triple-helix conformation plays an important role in enhancing the antitumor effects of lentinan.

Animals↗

PubMed2005

Hippocampal cytochrome oxidase activity of rats in easy and difficult visual discrimination learning.

The present study investigated the functional involvement of the rat hippocampal formation in easy and difficult visual discrimination learning by measuring regional brain cytochrome oxidase (C.O.) activity, an index of neuronal metabolic activity. The results showed that learning was related to C.O. activity in the CA field and the dentate gyrus of the hippocampal formation of rats in the difficult discrimination, whereas learning was related to C.O. activity only in the dentate gyrus of rats in the easy discrimination. The results suggest that difficult visual discrimination learning required greater involvement of the hippocampal formation than easy visual discrimination learning.

Animals↗

PubMed2005

Latex agglutination test for monitoring antibodies to avian influenza virus subtype H5N1.

A latex agglutination test (LAT) based on polystyrene beads sensitized with inactivated avian influenza virus H5N1 particles was developed. Compared with the hemagglutination inhibition test, the sensitivity and specificity of the LAT were 88.8 and 97.6%, respectively, in detecting 830 serum samples from vaccinated chickens. The test has application potential in field practice.

Animals↗

PubMed2005

Effect of molecular mass on antitumor activity of heteropolysaccharide from Poria cocos.

A water-soluble heteropolysaccharide ac-PCM0 from Poria cocos was successfully fractionated using a preparative size exclusion chromatography (SEC) column, and their weight-average molecular mass (M(w)) was characterized by analytical SEC combined with laser light scattering (SEC-LLS). The results indicate that the fractions having relatively high M(w) exhibited higher inhibition ratio in vivo antitumor activity than those having M(w) below 3.29 x 10(4). However, the relatively low molecular mass was beneficial to the in vitro antitumor activity. Moreover, ac-PCM0 has a significantly higher enhancement ratio of the body weight than 5-fluorouracil, and its 50% lethal dose is above 1250 mg/kg, indicating a nontoxic nature.

Animals↗

PubMed2004

Morphologies and conformation transition of lentinan in aqueous NaOH solution.

Molecular morphologies and conformation transition of lentinan, a beta-(1-->3)-D-glucan from Lentinus edodes, were studied in aqueous NaOH solution by atomic force microscopy (AFM), viscometry, multiangle laser light scattering, and optical rotation measurements. The results revealed that lentinan exists as triple-helical chains and as single random-coil chains at NaOH concentration lower than 0.05M and higher than 0.08M, respectively. Moreover, the dramatic changes in weight-average molecular weight Mw, radius of gyration [s2](1/2), intrinsic viscosity [eta], as well as specific optical rotation at 589 nm [alpha]589 occurred in a narrow range of NaOH concentration between 0.05 and 0.08M NaOH, indicating that the helix-coil conformation transition of lentinan was carried out more easily than that of native schizophyllan and scleroglucan, and was irreversible. For the first time, we confirmed that the denatured lentinan molecule, which was dissolved in 0.15M NaOH to be disrupted into single coil chains, could be renatured as triple helical chain by dialyzing against abundant water in the regenerated cellulose tube at ambient temperature (15 degrees C). In view of the AFM image, lentinan in aqueous solution exhibited the linear, circular, and branched species of triple helix compared with native linear schizophyllan or scleroglucan.

Lasers↗

PubMed2004

Chemical structure of aeromonas gum--extracellular polysaccharide from Aeromonas nichidenii 5797.

Aeromonas (A) gum, an extracellular heteropolysaccharide produced by the bacterium Aeromonas nichidenii strain 5797, was studied by 1H and 13C NMR spectroscopy including 2D COSY, TOCSY, 1H, 13C HMQC, HMBC and ROESY experiments after O-deacetylation and Smith degradation. These investigations revealed the presence of an O-acetylated pentasaccharide repeating unit composed of mannose, glucose, xylose and glucuronic acid, and it has the following structure: [Image: see text]

Aeromonas↗

PubMed2004

Construction of recombinant pseudorabies virus expressing NS1 protein of Japanese encephalitis (SA14-14-2) virus and its safety and immunogenicity.

The bivalent genetic engineering vaccine of Japanese encephalitis (JE) and Aujeszkj disease (AD) was developed to provide a novel approach to prevent and control these two diseases. NS1 gene of Japanese encephalitis virus (JEV) SA14-14-2 strain was produced by reverse transcriptase-mediated PCR (RT-PCR) and was cloned into vector pUSK to form recombinant plasmid (designed as pUSK-NS1). A co-transfection experiment was performed in porcine kidney (PK-15) cells with pUSK-NS1 and the genome of the vector virus (PRV TK(-)/gG(-)/LacZ(+) mutant). By plaque purification, PCR detection and southern hybridization, recombinant pseudorabies virus (PRV) expressing NS1 protein of JEV was acquired and named TK(-)/gG(-)/NS1(+). Western blot analysis and ELISA demonstrated the NS1 protein expression. To evaluate the recombinant virus's potential application, we characterized the safety and immune responses in Balb/c mice and swine. The safety test indicated that, when receiving the recombinant virus at a concentration of 10(6.0)pfu, no virulence of the recombinant virus to the mice, piglets and pregnant sows was observed. The vaccinated animals could acquire protective immunity against lethal challenge of the virulent PRV Ea strain and develop a good humoral and cellular immune response against JEV. The above results revealed that the recombinant virus could be a suitable candidate vaccine strain for developing a novel genetic vaccine to combat pseudorabies and Japanese encephalitis in the pig industry.

Animals↗

PubMed2003

The role of telencephalic NMDA receptors in avoidance learning in goldfish (Carassius auratus).

Studies with goldfish (Carassius auratus) have suggested that N-methyl-D-aspartate (NMDA) receptors are concentrated most densely in the telencephalon, a simple structure homologous to the limbic structure of higher vertebrates. The present study investigated the amnestic effects of microinjections of the NMDA receptor antagonist D(-)-2-amino-5-phosphonopentanoic acid (D-AP5) to the goldfish telencephalon on avoidance conditioning. Results showed that microinjections of D-AP5 before training impaired avoidance learning at doses that did not impair performance processes. High-performance liquid chromatography measurements showed that D-AP5 was detected only in the telencephalon following microinjections. Thus, D-AP5 impaired avoidance learning through its interaction with telencephalic NMDA receptors in goldfish. Furthermore, microinjections of D-AP5 to the goldfish telencephalon immediately following training did not impair memory consolidation of avoidance conditioning.

Animals↗

PubMed2002

Chain stiffness of heteropolysaccharide from Aeromonas gum in dilute solution by dynamic light scattering.

Dynamic light scattering measurements have been made on 15 fractions of aeromonas (A) gum, an extracellular heteropolysaccharide produced by the strain Aeromonas nichidenii, with dimethylsulfoxide containing 0.2M lithium chloride as the solvent at 25 degrees C. Data for the translational diffusion coefficient D covering a molecular weight range from 4.5 x 10(5) to 2.1 x 10(6) and ratios of the z-average radius of gyration (z) (1/2) to the hydrodynamic radius R(H) (calculated with previous (z) data) suggest that the polymer behaves like a semiflexible chain in this solvent similar to the stiffness of cellulose derivatives. Thus the D data are analyzed on the basis of the Yamakawa-Fujii theory for the translational friction coefficient of a wormlike cylinder by coarse-graining the heteropolysaccharide molecule. Excluded-volume effects are taken into account in the quasi-two-parameter scheme, as was done previously for (z) and [eta] (the intrinsic viscosity) of A gum in the same solvent. The molecular weight dependence of R(H) is found to be explained by the perturbed wormlike chain with a persistence length of 10 nm, a linear mass density of 1350 nm(-1), an excluded-volume strength parameter of 1.3 nm, and a chain diameter of 2.8 nm. These parameters are in substantial agreement with those estimated previously from (z) and [eta] data, demonstrating that the solution properties (D, (z), and [eta]) of the heteropolysaccharide are almost quantitatively described by the current theories for wormlike chains in the molecular weight range studied.

Aeromonas↗

PubMedDate not supplied

Collapse and association of denatured lentinan in water/dimethlysulfoxide solutions.

Triple helical lentinan, beta-(1-->3)-D-glucan from Lentinus edodes, was denatured in dimethlysulfoxide (DMSO) into single random coils. The DMSO solutions of randomly coiled lentinan were diluted with pure water to different wH (the weight fraction of water in the mixed solvent), and their specific optical rotation [alpha]D, reduced viscosity (lnetar)/c, and hydrodynamic radius Rh were investigated as a function of wH and storage time t. With an increase of wH from 0.1 to 0.2, [alpha]D increased sharply, suggesting that transition of conformation of the macromolecules has occurred. When wH was lower than 0.1, (lnetar)/c of lentinan in water-diluted DMSO exhibited the almost same value as that in pure DMSO and changed hardly with increasing t. Interestingly, (lnetar)/c decreased to reach a minimum with a further increase of wH from 0.1 to 0.25 and then increased with a continuous increase of wH from 0.25 to 0.5. Both (lnetar)/c and Rh of the denatured lentinan in water-diluted DMSO with wH of approximately 0.25 both exhibited a minimum, indicating that collapsed coil chains have occurred. All of the experimental findings revealed that the behaviors of lentinan in water-diluted DMSO solution with wH < 0.1 were consistent with that in good solvent, DMSO. When wH = 0.25, the quality of the mixed solvents became worse, and the dominant intramacromolecular hydrogen-bond interaction enhanced, leading to minimum of viscosity and size of the chains as a result of the collapsed coils. When wH > 0.25, the quality of the mixture weakens further, and the intermolecular hydrogen-bond interaction enhanced and was dominant, leading to aggregation of the collapsed chains.

Dimethyl Sulfoxide↗