OBJECTIVETo study the associations of CYP1A1 gene polymorphisms with levels of urinary 1-hydroxypyrene among coke oven workers.

METHODS223 male workers from a coke plant (76, 82 and 65 workers in oven top group, oven-side group and oven-bottom group respectively) and 119 controls without occupational polycyclic aromatic hydrocarbons exposure were selected. The MspI gene polymorphism in CYP1A1 3' flanking region and the genotypes at I462V site in exon 7 of CYP1A1 were detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and allele specific amplification (ASA).

RESULTSThe urinary 1-hydroxypyrene of coke oven workers in oven-top, oven-side and oven-bottom (3.77+/-0.64, 3.57+/-0.49, 3.26+/-0.80 micromol/mol Cr) were significantly higher than controls (2.80+/-1.02 micromol/mol Cr) (P<0.01). The urinary 1-hydroxypyrene was not significantly different among MspI genotypes in CYP1A1 3' flanking region (P>0.05). In oven-top group and oven-side group, the subjects with Val/Val genotype in exon 7 of CYP1A1 had significantly higher urinary 1-hydroxypyrene levels than those with Ile/Val or Ile/Ile genotype, and urinary 1-hydroxypyrene of Ile-Val genotype were also significantly higher than Ile/Ile genotype (P<0.01). Multivariate logistic regression analysis showed that the coke oven workers (OR in oven top group, oven-side group and oven-bottom group was 24.926, 4.226 and 6.729 respectively) and subjects with m2/m2 genotype in CYP1A1 3' flanking region (OR=4.031) or with Val/Val or Ile/Val genotype in exon 7 of CYP1A1 (OR were 5.524 and 3.811) had elevated urinary 1-hydroxypyrene (greater than 95 percentile of control group, 3.876 micromol/mol Cr).

CONCLUSIONBAP concentration of work environment contributes to the elevated urinary 1-hydroxypyrene levels, and the exposed BAP levels were regulated by the CYP1A1 MspI and I462V genotypes. Genetic polymorphism of CYP1A1 gene could be a susceptible biomarker in coke oven workers which was involved in the individual susceptibility on metabolism of PAHs.

Adult ; Benzo(a)pyrene ; adverse effects ; Coke ; Cytochrome P-450 CYP1A1 ; genetics ; Humans ; Male ; Middle Aged ; Occupational Exposure ; adverse effects ; Polymorphism, Genetic ; Pyrenes ; pharmacokinetics ; Urine ; chemistry

OBJECTIVETo develop a quantitative real-time polymerase chain reaction (PCR) for detecting Bartonella henselae.

METHODSAccording to the 16S-23S rRNA intervening sequences (IVS) specific for B. henselae, one pair of primers and one TaqMan-MGB probe were designed. A quantitative real-time PCR was developed with the primers, the probe, and the IVS, a standard template, in DNA sequence detection system (ABI 7900HT).

RESULTSThe standard curve was established with the standard template and the relationship between the value of threshold cycle (Ct) and the DNA copy number was linear (r = 0.997). The sensitivity of this quantitative real-time PCR was about 1000 times higher than that of a common PCR used to detect homologous DNA. By this quantitative real-time PCR, the DNA sample of B. henselae was positively detected but not from other rickettsial or bacterial DNA samples. The variation coefficients of intra- and inter-assay reproducibility were 0.2%-1.9%. Using the real-time quantitative PCR to detect samples from mice that were experimentally infected with B. henselae, the small amount of B. henselae DNA was detected in blood samples on days 2, 3, and 5 and large amount of B. henselae DNA was detected in spleen samples on days 1 and 2 after infection.

CONCLUSIONResults from our study suggested that this quantitative real-time PCR was highly specific, sensitive and with good repeatability for detection of B. henselae. It seemed quite useful for rapid detection of tiny DNA of B. henselae in various samples and laboratory diagnosis of bartonellosis caused by B. henselae.

Animals ; Bartonella Infections ; diagnosis ; Bartonella henselae ; genetics ; DNA, Bacterial ; analysis ; Mice ; Polymerase Chain Reaction ; methods ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo develop a real-time quantitative polymerase chain reaction(PCR) assay for detecting Rickettsia rickettsii.

METHODSThe primers and TaqMan-MGB probe were designed according to the ompB gene of R. rickettsii. A DNA fragment of ompB gene amplified from R. rickettsii by PCR was used as a standard template for the development of the method.

RESULTS5 copies of ompB fragments of R. rickettsii were detected. The genomic DNA of R. rickettsii was detected by the developed quantitative PCR assay. However, the genomic DNA from another rickettsial or bacterial agent was not determined. Through this developed method, the positive results were obtained from the animals and cells, artificially infected with R. rickettsii.

CONCLUSIONThe real-time quantitative PCR assay seemed to be highly sensitive and specific which might be used to rapidly detect R. rickettsia DNA in various samples and to early diagnose patients infected by R. rickettsii.

DNA Primers ; Humans ; Polymerase Chain Reaction ; methods ; Rickettsia rickettsii ; genetics ; Rocky Mountain Spotted Fever ; diagnosis ; Sensitivity and Specificity

OBJECTIVETo study the expression levels of cdk5, p35 and p53 genes in arsenic trioxide (As2O3O)-induced neuron apoptosis and to explore the potential mechanism.

METHODSThe cultured primary rats' neurons were divided into 5 groups, which were exposed to 0, 1, 5, 10 micromol/L As2O3 and dimethyl sulfoxide (DMSO) for 8 h, respectively. The cell viability and cell apoptosis were detected by MTT colouration methods and flow cytometry, respectively. The real-time fluorescence quantitative PCR was used to measured the expression levels of cdk5, p35 and p53 genes.

RESULTSThe cell viability inhibition rates were 16.77%, 19.72% and 27.81% in 1, 5, 10 micromol/L As203 groups, respectively. Compared to the untreated group and DMSO group, the cell apoptosis rates were significantly increased in 5 and 10 micromol/L As2O3 groups (P < 0.05). The expression levels of cdk5, p35 and p53 genes increased with the exposure doses of AsO3. However, there were no significant differences in p35 gene expression between different dose subgroups (P > 0.05). There were significant differences in cdk5 and p53 gene expression between different dose subgroups (P < 0.05). The expression levels of cdk5 gene in 5 and 10 micromol/L As2O3 groups were significantly higher than those in untreated group and DMSO group (P < 0.05). The expression levels of p53 gene in 1, 5 and 10 micromol/L As2O3 groups were significantly higher than that in untreated group (P < 0.05). The expression level of p53 gene in 10 mciromol/L As2O3 group was significantly higher than that in DMSO group (P < 0.05).

CONCLUSIONCdk5, p35 and p53 genes may involve in the process of As2O3-induced neural cell apoptosis.

Animals ; Apoptosis ; drug effects ; Arsenicals ; Cells, Cultured ; Cyclin-Dependent Kinase 5 ; genetics ; metabolism ; Neurons ; drug effects ; metabolism ; Oxides ; toxicity ; Phosphotransferases ; genetics ; metabolism ; Rats ; Tumor Suppressor Protein p53 ; genetics ; metabolism

OBJECTIVETo explore the coincidence of lipid peroxidation and neurobehavioral function changes in coke oven workers.

METHODSOne hundred and thirty-four coke oven workers were divided into three groups: 35 in the oven-bottom group, 49 in the oven-side group and 50 in oven-top group. WHO recommended NCTB was performed on coke oven workers and 36 controls from material conservation department; The contents of total superoxide dismutases (T-SOD), glutathione (GSH) and malondialdehyde (MDA) in blood were determined by test kits.

RESULTSCompared with the controls, the coke oven workers showed lower levels of T-SOD and GSH (P < 0.01), significantly higher MDA levels in blood (P < 0.01), higher score on negative mood state, lower scores on positive mood state, and poorer performance in NCTB test (P < 0.05). Further analysis revealed that there was a weak positive correlation between neurobehavioral function changes and the level of lipid peroxidation with a coefficient lower than 0.25.

CONCLUSIONThe level of lipid peroxidation in coke oven workers' blood increased and coincided with neurobehavioral function impairment.

Adult ; Affect ; Anxiety ; Case-Control Studies ; Coke ; Fatigue ; Glutathione ; blood ; Humans ; Lipid Peroxidation ; Male ; Malondialdehyde ; blood ; Middle Aged ; Occupational Exposure ; adverse effects ; Superoxide Dismutase ; blood ; Young Adult

OBJECTIVETo investigate the changes of mitochondria membrane potential and cytoplasma cytochrome C as the mechanism of neuron apoptosis induced by B(a)P.

METHODSPrimary neurons were dissociated from cerebral cortex of 1 - 3 days old SD rats and cultured with DMEM incubator at 37 degrees C. After 5 days' cultivation, the neurons were added S9 and B(a)P, and the concentrations of treated B(a)P were 0, 10, 20 and 40 micromol/L respectively. After administering of B(a)P, the neurons were cultivated for 40 hours. Apoptosis rate was measured by flow cytometry using Annexin V-FITC and propidium iodide (PI) staining, and the changes in mitochondrial potential (DeltaPsim) were tested with Rhodamine fluorescence (R2123) technique. Preparation of cytosolic extracts by centrifugation. Western blotting analysis was used to evaluate the level of cytochrome C of cytoplasm.

RESULTSThe apoptotic rate of neuron increased in both the middle dose group and the high dose group compared with controls, and had a dose-response tendency with the concentration of B(a)P. Moreover mitochondrial potential decreased in a dose dependent manner. There was a negative correlation between DeltaPsim and the apoptotic rate of neurons (r = -0.763, P < 0.05); Western blotting analysis showed cytoplasmic cytochrome C level increased significantly, which was positively related with neuron apoptosis (r = 0.831, P < 0.01).

CONCLUSIONLoss of mitochondria membrane potential and increase of cytoplasma cytochrome C may be the main cause of neuron apoptosis induced by B(a)P.

Animals ; Apoptosis ; drug effects ; Benzo(a)pyrene ; toxicity ; Cells, Cultured ; Cytochromes c ; metabolism ; Membrane Potential, Mitochondrial ; Mitochondria ; drug effects ; metabolism ; Neurons ; cytology ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo discuss effective operative method for posterior edge separation of lumbar vertebral body.

METHODSTwelve cases of posterior edge separation of lumbar vertebral body were categorized as lateral type and central type, and were operated upon with limited-incision using lamina retractor. All patients were male and with an average age of 30 years (range, 23 to 40 years). The average history of disease was 17.9 months. In lateral type cases, separation bone was in the posterolateral lower edge of L4 vertebral body in 1 case, in the posterolateral upper edge of L5 vertebral body in 3 cases and in the posterolateral upper border of S1 vertebral body in 6 cases, all with ipsilateral disc herniation. Lateral type cases were treated with unilateral fenestration and removal of disc nucleus and posterior bony edge. Separation bone was in the central upper edge of S1 vertebral body with central disc herniation in 2 central type cases. Two central type cases were treated with bilateral fenestration and removal of disc nucleus and posterior bony edge.

RESULTSAll patients were followed up for 12 to 36 months, and the average follow-up was 22.5 months. According to Macnab postoperative evaluation criterion, the results were excellent in 10 cases, good in 2 cases.

CONCLUSIONLimited-incision for posterior edge separation of lumbar vertebral body has characteristic of less trauma and convenience. It is an effective method for removal posterior bony edge.

Adult ; Follow-Up Studies ; Humans ; Lumbar Vertebrae ; diagnostic imaging ; pathology ; surgery ; Male ; Postoperative Period ; Tomography, X-Ray Computed ; Treatment Outcome ; Young Adult

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 explore the effect of polycyclic aromatic hydrocarbons on the neurobehavioral function of coke oven workers.

METHODS200 healthy adult male coke oven workers were selected from a coke plant of a state-owned steel enterprise in Taiyuan City. 88 controls occupationally unexposed to polycyclic aromatic hydrocarbons (PAHs) were selected from the same enterprise. All the subjects participated in this investigation voluntarily in their consent. Concentration of B(a)P in the working environment was monitored by High Performance Liquid Chromatography (HPLC). Urine samples were sampled immediately after working shifts. The level of urinary 1-hydroxypyrene was determined by HPLC. General information of workers correlated with the investigation was collected in a questionnaire according to the same criteria by well-trained investigators. Neurobehavioral core test battery (NCTB) recommended WHO was performed on coke oven workers and controls to test the neurobehavioral changes and the mood state.

RESULTSthe concentration of B(a)P at oven bottom,oven side and oven top were 0.0195 microg/m3, 0.186 microg/m3 and 1.624 microg/m3 respectively, that at oven side and oven top being higher than the one stipulated by the occupational hygiene criterion. Urinary 1-hydroxypyrene was significantly different between the exposure group (3.42 +/- 0.98 micromol/mol creatinine) and control group (2.75 +/- 1.09 micromol/mol creatinine). No significant differences were found between exposure group and control group of age, working years, smoking, drinking and unhealthy food consumption; however, compared to the controls, the scores of total digital span, the forward digital span, and right dotting in the coke oven workers were lower, but that of total dotting was higher, with a statistical significance. According to urinary 1-hydroxypyrene concentration, all the subjects were divided into three groups. (<3.10 micromol/mol creatinine, 3.10 micromol/mol creatinine, >3.87 micromol/mol creatinine). Significant differences of the total digital span, the forward digital span, backward digital span, digit symbol and Benton visual retentions existed in different urinary 1-hydroxypyrene concentration groups and showed a dose-response tendency. Results of multiple stepwise regression analysis and correlation analysis showed that the level of urinary 1-hydroxypyrene affected memory and perception of coke oven workers and negative correlations between the level of urinary 1-hydroxypyrene and changes in neurobehavioral function were found.

CONCLUSIONPAHs mainly causes decrease of memory and perception in coke oven workers.

Adult ; Air Pollutants, Occupational ; analysis ; Coke ; Humans ; Male ; Memory ; Neuropsychological Tests ; Occupational Exposure ; Polycyclic Aromatic Hydrocarbons ; urine

OBJECTIVETo study the relationship between polymorphism of aryl hydrocarbon receptor (AhR) gene in G1661A and the level of urinary 1-hydroxypyrene among coke oven workers.

METHODS295 male subjects were studied, including 214 workers working in coke oven plant and 81 controls working in raw material plant who were not generally exposed to polycyclic aromatic hydrocarbons occupationally. General in-formation of subjects were collected in a specific questionnaire including age, smoking and drinking habits, the history of occupation and so on. The AhR genotypes were detected by allele specific amplification (ASA), and the levels of urinary 1-hydroxypyrene were measured by high performance liquid chromatography with a fluorescence detector.

RESULTSThe frequencies of G/G, G/A and A/A genotype were 52.8% (113/214), 27.6% (59/214) and 19.6% (42/214) in exposed group and 67.9% (55/81), 19.8% (16/81) and 12.3% (10/81) in control group, respectively. No significant difference was found in three genotypes between the exposed and control group. Allele frequencies of G and A were 66.6% (285/428) and 33.4% (143/428) in exposed group and 77.8% (126/162) and 22.2% (36/162) in control group, and no statistical differences were found in allele frequency between exposed and control group. After the length of service and external exposure orders in general linear model were adjusted, results of covariance analysis showed that logarithmic transformed urinary 1-hydroxypyrene levels were (3.62 +/- 0.12), (3.43 +/- 0.12) and (3.44 +/- 0.08) micromol/mol Cr in individuals with A/A, G/A and G/G, respectively. The logarithmic transformed urinary 1-hydroxypyrene levels were (3.24 +/- 0.09) and (3.43 +/- 0.10) micromol/mol Cr in individuals with allele of G and A. No statistical differences were found in level of 1-hydroxypyrene among A/A, G/A and G/G genotype individuals, and between allele G and allele A after external exposure orders and length of service were adjusted.

CONCLUSIONThe polymorphism of aryl hydrocarbon receptor G1661A has no significant impact on levels of urinary 1-hydroxypyrene.

Adult ; Coke ; Genotype ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Pyrenes ; pharmacokinetics ; Receptors, Aryl Hydrocarbon ; genetics ; Urine ; chemistry