Objective To investigate the differences and characteristics of virulence genes carried by Salmonella enteritidis from different sources in Shijiazhuang City.Methods One hundred and twenty-four strains of Salmonella enteritidis isolated from morning markets of raw and poultry stalls,slaughterhouses and food poisoning specimens in Shijiazhuang area were collected.Eight virulence genes (invA,sopE,agfA,spvR,hilA,stn,pefA,shdA) were detected by polymerase chain reaction (PCR).Results Salmonella enteritidis might have different virulence gene profiles.The above eight virulence genes were detected in different strains.The carrying rate of virulence genes invA,sopE,stn,hilA,spvR and pefA in the food poisoning strains was higher than 94％.There was no difference in the carrying rate of 8 virulence gene between the morning raw poultry stalls isolates and the patient strains,but was different with the slaughterhouse strains.Conclusion There were more risks of food poisoning caused by Salmonella enteritidis from morning markets,and the hygiene supervision should be strengthened to prevent and control foodborne disease.

OBJECTIVETo investigate the effect of 1, 2-dichloroethane (1, 2-DCE) on blood brain barrier.

METHODSAcute toxic encephalopathy model was copied with the consecutive static inhalation of 1, 2-DCE. The water content of brain tissue was measured, and the blood brain barrier permeability was detected with lanthanum nitrate. The brain microvascular endothelial cells and neuroglial cells were cultured in vitro, which were administrated with 1, 2-DCE. The cell morphologic structures were observed under light microscope and electron microscope.

RESULTS(1) The extracellular edema was most found in the cerebral tissue and the leakage of lanthanum particles through the barrier were found with the lanthanum tracking method. (2) The water content in cerebral cortex in the moderate and high dose groups was significantly higher than that in the control group and became severer with the increases of the intoxicated time. The water content in cerebral medulla was significantly increased only at 6 hours after the intoxication. (3) The normal morphological structure of brain microvascular endothelial cells and neuroglial cells could be injured by 1, 2-DCE, and the injury to neuroglial cells caused by 1, 2-DCE occurred earlier and severer than that to brain microvascular endothelial cells.

CONCLUSION1, 2-DCE can damage blood brain barrier and induce cerebral edema.

Administration, Inhalation ; Animals ; Blood-Brain Barrier ; drug effects ; Brain ; pathology ; Brain Edema ; chemically induced ; pathology ; Cells, Cultured ; Dose-Response Relationship, Drug ; Endothelial Cells ; drug effects ; pathology ; Ethylene Dichlorides ; toxicity ; Female ; Male ; Rats ; Rats, Sprague-Dawley

AIMTo establish a spectrophotometric method for measurement of the sizes of liposomes for evaluating physical stability of liposomes.

METHODSThe sterically stabilized liposomes (SLs) were prepared by ethanol injection method and extrusion method. The mean cumulant diameters (D) of the vesicles were determined by electron microscopy and dynamic light scattering. On the basis of Rayleigh-Gans-Debye theory, the absorbance at 436 nm per unit lipid concentration (A436 nm/Cp) was measured as a function of vesicle diameter.

RESULTSlog(A436 nm/Cp) was closely related to logD (r2 > or = 0.93, n = 5).

CONCLUSIONThe absorbance of liposomes reflect their relative sizes and can be used to evaluate physical stability of liposomes.

Liposomes ; chemistry ; Particle Size ; Spectrophotometry ; methods

AIMTo prepare sterically stabilized liposomes modified with chimeric TNT-3 monoclonal antibody (chTNT-3) and investigate their immunoreactivity and in vitro targeting.

METHODSAn end-group functionalized polyethylene glycol-lipid derivative (pyridylthiopropionoylamino-polyethylene glycol-hydrogenated soy phosphatidylethanolamine, PDP-PEG-HSPE) was synthesized and incorporated to sterically stabilized liposomes. After mild thiolysis of the PDP groups by dithiothreitol, liposomes were covalently linked with maleimide-derivatized chTNT-3 and formed sterically stabilized immunoliposomes. Coupling efficiency, antibody density, size distribution, immunoreactivity of chTNT-3-modified sterically stabilized liposomes (chTNT-3-SLs) and specific binding properties of the chTNT-3-SLs to fixed Raji cells were determined, separately.

RESULTSHigher initial Ab/PDP-PEG-HSPE molar ratios resulted in higher antibody density on the surface of liposomes but lower coupling efficiency. The optimal coupling efficiency of 71% was obtained while antibody density in liposome was 106 microg antibody/micromol phospholipids (as initial antibody/PDP-PEG-HSPE = 1 : 10). The chTNT-3-SLs had a narrow size distribution after extrusion and the mean size of this immunoliposomes was (115 +/- 33) nm. The immunoreactivity of chTNT-3 can be preserved after efficient attachment of maleimide-derivatized chTNT-3 to the surface of liposomes. But calculated per antibody concentration, the immunoreactivity of chTNT-3-SLs would obviously decrease compared to that of chTNT-3 or chTNT-3 derivatives. Significantly higher binding of chTNT-3-SLs to fixed Raji cells directed by chTNT-3 was obtained compared to other preparations in serial dilutions (P < 0.01).

CONCLUSIONchTNT-3-SLs prepared by PDP-PEG-HSPE method remained most immunoreactivity of chTNT-3 and was able to bind nuclear antigens in vitro.

Antibodies, Monoclonal ; immunology ; Binding Sites ; Cell Line, Tumor ; Drug Carriers ; Drug Delivery Systems ; Humans ; Liposomes ; Lymphoma, B-Cell ; immunology ; pathology ; Necrosis ; Phosphatidylethanolamines ; Polyethylene Glycols

OBJECTIVETo study the effect of necrostatin (Nec-1) on apoptosis induced by aluminum (Al), and approach the mechanism.

METHODSNeural cell death model was made by 4 mmol/L Al treated neuroblastoma cells (SH-SY5Y). Cell viabilities were detected at different concentrations of Al and/or Nec-1. Hoechst 33342/PI double staining was used to observe apoptosis and (or) necrosis that were quantified by flow cytometry using Annexin V/PI double staining. Apoptotic pathway was tested by activities of Caspase-3, Caspase-8 and Caspase-9. In addition, the expression of NF-kappa B and Cyt-c was measured by immunocytochemistry.

RESULTSCell viabilities were significantly decreased with the increasing concentrations of Al (P < 0.05), which could be significantly upregulated by 60 micromol/L Nec-1 (P < 0.05) and were correlated with the concentrations of Nec-1 (P < 0.05, P < 0.01). Apoptosis and necrosis were observed under fluorescent microscope and quantified by flow cytometry, which suggested an increasing trend of apoptotic and necrotic rates (P < 0.05, P < 0.01). Whereas, Nec-1 could not only decrease the necrotic rate but also apoptotic rate as well (P < 0.05, P < 0.01). Data of Nec-1 on caspases activities showed that Nec-1 could not affect Caspase-9 activity (P > 0.05) and Cty-c protein expression as well (P > 0.05). However, Nec-1 could reduce Caspase-8 activity significantly (P < 0.05, P < 0.01) and increase NF-kappa B protein expression (P < 0.05, P < 0.01) and finally decrease Caspase-3 activity (P < 0.05).

CONCLUSIONNec-1 could reduce cell apoptosis induced by Al, through Caspase-8 pathway, and up-regulate the expression of NF-kappa B protein.

Aluminum ; toxicity ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Death ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cytochromes c ; metabolism ; Humans ; Imidazoles ; pharmacology ; Indoles ; pharmacology ; NF-kappa B ; metabolism ; Neuroblastoma

Objective To establish a QAMS method for content determination of six compositions (chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin) from Lonicerae Japonicae Caulis; To verify the feasibility and applicability of this method in quality control of Lonicerae Japonicae Caulis. Methods Chlorogenic acid was set as internal reference substance. The HPLC analysis was performed on a Waters Symmetry C18 column (4.6 mm × 250 mm, 5 μm) with a mobile phase consisted of acetonitrile and 0.4% phosphoric acid solution in gradient elution manner at a flow rate of 1 mL/min. The column temperature was maintained at 35 ℃, and the detection wavelength was set at 327 nm for chlorogenic acid, caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and 236 nm for loganin. Results The relative correction factors of caffeic acid, cryptochlorogenin acid, isochlorogenic acid A, isochlorogenic acid C and loganin were established; there was no obvious difference between calculated value of QAMS and measured value of external standard method. Conclusion The quality control mode of QAMS can be used for multi-index synchronization quality evaluation of the six compositions from Lonicerae Japonicae Caulis.

OBJECTIVETo probe into the effects of familial factors on injury-related behaviors in children.

METHODSInjury-related behaviors and familial factors of 6884 children were investigated with Family Questionnaire and Child Behavior Checklist. Multi-nominal logistic regression analysis was performed.

RESULTSThere were 1670 (24.26%) children having serious injury-related behaviors and 3683 (53.50%) children having moderate injury-related behaviors. Factors contributing to children's injury-related behaviors would include punishment or indifference as well as the mode of parents' education; reintegral type of family; the level of parents' cognition on injuries; unfit location of medicine at home and careless attitudes of parents.

CONCLUSIONThere was close relationship between children's injury-related behaviors and familial factors. To avoid injury-related behaviors and to prevent injury occurrence, the importance of familial factors must be stressed.

Child ; Child Behavior ; psychology ; Family ; psychology ; Female ; Humans ; Logistic Models ; Male ; Surveys and Questionnaires ; Wounds and Injuries ; etiology ; prevention & control ; psychology

OBJECTIVETo clone the gene encoding methyl-accepting chemotaxis signal transduction protein (MCSTP) of Helicobacter hepaticus and analyze the gene structures using bioinformatics methods.

METHODSWith the specific primer of Helicobacter hepaticus MCSTP c1977, MCSTP gene was amplified by PCR from the genomic DNA of Helicobacter hepaticus and ligated to the prokaryotic expression vector pET22b(+). After sequencing, the sequence homology and structural feature of MCSTP gene were analyzed by bioinformatics method.

RESULTSA 99% similarity was identified between MCSTP gene cloned and its counterpart in standard Helicobacter hepaticus strain ATCC51449 genome DNA published by GenBank, with only a replacement of A by T at 1160 bp. A low homology was found in the MCSTP genes between Helicobacter hepaticus, Campylobacter jejuni and Helicobacter pylori by bioinformatics analysis, suggesting the specificity of MCSTP gene in Helicobacter hepaticus among the microbes.

CONCLUSIONThe prokaryotic expression plasmid pET22b(+)/MCSTP is constructed successfully, and the bioinformatics analysis provided evidences and clues for further study of the biological functions and pathogenic mechanism of MCSTP.

Bacterial Proteins ; genetics ; metabolism ; Cloning, Molecular ; Computational Biology ; methods ; Genetic Vectors ; genetics ; Helicobacter hepaticus ; genetics ; isolation & purification ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Methyl-Accepting Chemotaxis Proteins ; Recombinant Fusion Proteins ; genetics ; metabolism ; Signal Transduction

OBJECTIVETo study the effects of benzo[a]pyrene (B[a]P) on capability of learning and memory and the content of amino acid neurotransmitters in hippocampus of rats.

METHODSThirty-two healthy, male SD rats were randomly divided into 4 groups according to their weights after intubated into ventricles: the solvent control group, 2.5, 5.0 and 10.0 mmol/L groups. 10 microl of B[a]P olive oil solutions, of different concentrations 2.5, 5.0 and 10.0 mmol/L, were injected into rats' lateral ventricles, respectively. Rats in the solvent control group were injected into the same volume of olive oil as that in B[a]P group. Rats' capability of learning and memory was tested by Morris water maze. The content of amino acid neurotransmitters in rats' hippocampus were determined by high performance liquid chromatogram with a fluorescence detector.

RESULTSCompared with the controls, the performances of learning and memory of rats decreased significantly in B[a]P treated groups (P<0.01). Levels of glutamate (Glu) were lower significantly in treated groups than that in controls (P<0.01). No significant differences were found in contents of aspartic acid (Asp), glycine (Gly) and aminobutyric acid (GABA) among the four groups.

CONCLUSIONB[a]P can damage rats' spatial learning and memory, and which could be related to decreased contents of excitatory amino acids in hippocampus.

Amino Acids ; metabolism ; Animals ; Benzo(a)pyrene ; toxicity ; Hippocampus ; drug effects ; metabolism ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Neurotransmitter Agents ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of Tiaomaiyin injection on the experimental arrhythmia for analyzing its underlying mechanism in the treatment of cardiovascular disease.

METHODExperimental animals anesthetized with 20% urethane (6 mL x kg(-1)) were evenly randomized into control group, positive control group, low-dose and high-dose Tiaomaiyin group. The rate of ventricular fibrillation (VF) chloroform-induced in mice, and the epoch of ventricular extrasystole (VE), ventricular tachycardia (VT),VF and cardiac arrest (CA), actonitine-induced in rats (1.0 microg x mL(-1) x min(-1)), and vabain-induced in guinea pigs (10 microg x mL(-1) x min(-1)), were detected respectively. The result loas converted into cumulative dosage of actonitine or vabain. In ischemia-reperfusion model in rats, the duration of arrhythmia and activity of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected.

RESULTAfter venous injection of Tiaomaiyin, VF in mice was lower significantly (P < 0.01), VE, VT, VF in rats and VF in guinea pigs were lowered considerably (P <0.05). The duration of arrhythmia in ischemia-reperfusion model was reduced considerably (P < 0.05), and the activity of myocardial SOD was raised significantly (P <0.01).

CONCLUSIONTiaomaiyin shows the reduction of experimental arrhythmia and protect effect to ischemia-reperfusion injury of the heart, which indicates that the effect mechanism may have the relationship with inhabition of lipid peroxidation and damnification of the free radical.

Animals ; Anti-Arrhythmia Agents ; administration & dosage ; isolation & purification ; pharmacology ; Arrhythmias, Cardiac ; etiology ; metabolism ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Electrocardiography ; Guinea Pigs ; Injections ; Malondialdehyde ; metabolism ; Mice ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; etiology ; metabolism ; physiopathology ; Myocardium ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; metabolism