Common genetic risk factors for venous thrombosis in the Chinese population.

Venous thrombosis is a major medical disorder caused by both genetic and environmental factors. Little is known about the genetic background of venous thrombosis in the Chinese population. A total of 1,304 individuals diagnosed with a first venous thrombosis and 1,334 age- and sex-matched healthy participants were enrolled in this study. Resequencing of THBD (encoding thrombomodulin) in 60 individuals with venous thrombosis and 60 controls and a functional assay showed that a common variant, c.-151G>T (rs16984852), in the 5' UTR significantly reduced the gene expression and could cause a predisposition to venous thrombosis. Therefore, this variant was genotyped in a case-control study, and results indicated that heterozygotes had a 2.80-fold (95% confidence interval = 1.88-4.29) increased risk of venous thrombosis. The THBD c.-151G>T variant was further investigated in a family analysis involving 176 first-degree relatives from 38 index families. First-degree relatives with this variant had a 3.42-fold increased risk of venous thrombosis, and their probability of remaining thrombosis-free was significantly lower than that of relatives without the variant. In addition, five rare mutations that might be deleterious were also identified in thrombophilic individuals by sequencing. This study is the largest genetic investigation of venous thrombosis in the Chinese population. Further study on genetics of thrombosis should focus on resequencing of THBD and other hemostasis genes in different populations.

Regulatory T cells protect fine particulate matter-induced inflammatory responses in human umbilical vein endothelial cells.

OBJECTIVE: To investigate the role of CD4(+)CD25(+) T cells (Tregs) in protecting fine particulate matter (PM-) induced inflammatory responses, and its potential mechanisms. METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with graded concentrations (2, 5, 10, 20, and 40 µg/cm(2)) of suspension of fine particles for 24h. For coculture experiment, HUVECs were incubated alone, with CD4(+)CD25(-) T cells (Teff), or with Tregs in the presence of anti-CD3 monoclonal antibodies for 48 hours, and then were stimulated with or without suspension of fine particles for 24 hours. The expression of adhesion molecules and inflammatory cytokines was examined. RESULTS: Adhesion molecules, including vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), and inflammatory cytokines, such as interleukin (IL-) 6 and IL-8, were increased in a concentration-dependent manner. Moreover, the adhesion of human acute monocytic leukemia cells (THP-1) to endothelial cells was increased and NF- κ B activity was upregulated in HUVECs after treatment with fine particles. However, after Tregs treatment, fine particles-induced inflammatory responses and NF- κ B activation were significantly alleviated. Transwell experiments showed that Treg-mediated suppression of HUVECs inflammatory responses impaired by fine particles required cell contact and soluble factors. CONCLUSIONS: Tregs could attenuate fine particles-induced inflammatory responses and NF- κ B activation in HUVECs.

[Association between genetic polymorphisms in pre-miR-146a and pre-miR-196a2 and genetic damage levels among coke oven workers].

OBJECTIVE: To investigate the association of rs2910164 G > C polymorphism and rs11614913 T > C polymorphism in pre-miR-146a and pre-miR-196a2 with genetic damage levels in coke oven workers. METHODS: A total of 575 nonsmoking workers who have worked for more than one year in a coke-oven plant at Wuhan, Hubei Province were enrolled in this study in September to October, 2010. The general characteristics as well as blood and urine samples were collected. The genetic damage levels were detected by cytokinesis-block micronucleus cytom assay and represented as micronucleus (MN) frequencies of binucleate cells in peripheral blood lymphocytes. The rs2910164 G > C polymorphisms in pre-miR-146a and rs11614913 T > C polymorphisms in pre-miR-196a2 were genotyped by using TaqMan assay. The plasma concentrations of benzo[a]pyrene-diolepoxide (BPDE)-albumin adducts were determined by using ELISA. All data were analyzed, the frequency ratio (FR) and 95%CI were calculated. RESULTS: Totally, 575 workers were taken into consideration. The rs2910164 C allele was associated with increased MN frequencies in the coke oven workers (P trend = 0.025), and the MN frequencies were higher in rs2910164 CC genotype carriers (4.38 ± 3.46) than in wild-type rs2910164 GG genotype carriers (4.02 ± 3.09) (FR = 1.18, 95%CI:1.04-1.34). The further stratified analyses by working years, gender, alcohol consumption, and the levels of BPDE-albumin adducts showed that the effects of rs2910164 C allele in increasing MN frequencies were robust in subjects who were males (FR = 1.11, 95%CI:1.02-1.20), nondrinkers (FR = 1.07, 95%CI:1.00-1.14), working years less than 20 (FR = 1.12, 95%CI:1.03-1.22), and workers with lower BPDE-albumin adducts levels (FR = 1.11, 95%CI:1.02-1.21) (P trend = 0.011, 0.044, 0.006 and 0.020, respectively). In addition, the MN frequencies were higher in workers with rs11614913 TC genotype (4.27 ± 2.91) than workers with rs11614913 TT genotype (3.90 ± 3.32) (FR = 1.12, 95%CI:1.02-1.23).Workers carried both rs2910164 GG and rs11614913 TT genotypes were set as a control, and the MN frequencies of workers with both rs2910164 CC and rs11614913 CC genotypes (5.32 ± 4.94) were 1.51 (1.21-1.89) times higher than the control (3.75 ± 3.01). CONCLUSION: The rs2910164 C allele in pre-miR-146a and rs11614913 C allele in pre-miR-196a2 were associated with increased genetic damage levels in coke oven workers.

[The dose response decrease of lung function associated with the urinary polycyclic aromatic hydrocarbons metabolites in coke oven workers].

OBJECTIVE: To analyze the relationship between metabolites of polycyclic aromatic hydrocarbons (PAHs) and lung function in coke oven workers, and to provide scientific basis for further exploring the potential mechanism and developing the preventing strategies of the workers' early lung damage. METHODS: We measured carbon monoxide, sulfur dioxide, benzene soluble matter, particulate matters, and PAHs at different workplaces of a coke oven plant. Detailed information on demography and occupational health condition of 912 workers were collected. We divided these workers into control group and coke oven group according to their workplaces and the different concentrations of COEs in the environment. We detected 10 urinary PAH metabolites and lung function using gas chromatography-mass spectrometry and spirometric tests, respectively. RESULTS: FEV(1.0) (91.12 ± 13.31) and FEV(1.0)/FVC (108.61 ± 20.37) of the coke oven group is significantly lower than the control group (94.16 ± 15.57, 113.45 ± 19.70). In the coke oven group, the hydroxyphenanthrene and 1-hydroxypyrene are negatively correlated with FEV(1.0)/FVC (ß = -0.136, ß = -0.100), Ptrend < 0.05 for all. CONCLUSION: The dose response decrease of lung function is associated with the urinary PAH metabolites in coke oven workers. Indicated that the long exposure to PAHs may cause the early lung damage in coke oven workers, phenanthrene and pyrene may be the main factors.

[The increase of micronuclei frequencies of peripheral blood lymphocyte in plywood workers exposed accumulatively to formaldehyde].

OBJECTIVE: To investigate the effects of occupational exposure to formaldehyde on the micronuclei frequencies in peripheral blood lymphocytes of workers. METHODS: Two hundred thirty six plywood workers were divided into 3 exposure groups (low, middle and high) according to internal exposure biomarker (formaldehyde human serum albumin conjugate, FA-HSA), which was detected by ELISA. The concentrations of formaldehyde (FA) in air of two workshops were measure using the high performance liquid chromatography. Cytokinesis-block micronucleus (CBMN) test was used to detect the micronuclei frequencies of peripheral blood lymphocyte in 236 workers. RESULTS: The average concentrations of FA in the low and high exposure workshops were 0.58 +/- 0.20 and 1.48 +/- 0.61 mg/m3, respectively, there was significant difference (P < 0.01). The average concentrations of serum FA-HAS of workers in two workshops were 69.22 +/- 15.37 and 136.29 +/- 89.49 pg/ml, respectively, there was significant difference (P < 0.01). The results of CBMN test showed that the micronucleus frequencies in low, middle and high exposure groups were 1.94 +/- 1.72, 2.10 +/- 1.92 and 2.10 +/- 1.70 per thousand, respectively, there were no significant differences between groups. However, the micronucleus frequencies in accumulative low, middle and high exposure groups were 1.36 +/- 1.36, 2.31 +/- 1.81 and 2.49 +/- 1.92 per thousand, respectively, there were significant differences between different accumulative exposure groups (P < 0.01). The results of correlation analysis indicated that there was a positive correlation between accumulative exposure levels and micronucleus frequencies (r(s) = 0.321, P < 0.01). The accumulative exposure doses may be a risk factor for high micronucleus frequencies in workers exposed to FA (P(trend) = 0.002). CONCLUSION: FA-HSA levels can serve as an internal exposure biomarker for assessing the exposure level of workers exposed to FA. Accumulative formaldehyde exposure resulted in an increase of micronuclei frequencies of peripheral blood lymphocyte in plywood workers.

[Using the stable HSPA1A promoter-driven luciferase reporter HepG2 cells to assess the overall toxicity of coke oven emissions].

OBJECTIVE: Using the stable HSPA1A (HSP70-1) promoter-driven luciferase reporter HepG2 cells (HepG2/HSPA1A cells) to assess the overall toxicity of coke oven emissions. METHODS: The stable HepG2/HSPA1A cells were treated with different concentrations of coke oven emissions (COEs) collected from the top, side, and bottom of a coke oven battery for 24 h. After the treatments, luciferase activity, cell viability, malondialdehyde (MDA) concentration, Olive tail moment, and micronuclei frequency were determined, respectively. RESULTS: The bottom COEs induced significant increases (P < 0.01) in relative luciferase activity up to 1.4 times the control level at 0.15 µg/L. The low dose of side COEs (0.02 µg/L) led to a significant increase (P < 0.01) in relative luciferase activity that progressively increased to 2.1 times the control level at 65.4 µg/L. The top COEs produced a strong dose-dependent induction of relative luciferase activity up to over 5 times the control level at the highest concentration tested (202 µg/L). In HepG2/HSPA1A cells treated with the bottom COEs, relative luciferase activity was positively correlated with MDA concentration (r = 0.404, P < 0.05). For the three COEs samples, positive correlations were observed between relative luciferase activity and Olive tail moment and micronuclei frequency. CONCLUSION: The relative luciferase activity in HepG2/HSPA1A cells can sensitively reflect the overall toxicity of COEs. The stable HepG2/HSPA1A cells can be used for rapid screening of the overall toxicity of complex air pollutants in the workplace.

[Characteristics of particulate matter pollution in coke oven plant].

OBJECTIVE: To explore the characteristics of particulate matter pollution in coke oven plant, so as to provide scientific data for establishing occupational exposure limits for coke oven emissions. METHODS: Concentrations of CO, SO2, BSM, BTEX (concentrations of benzene, toluene and xylene were determined in this study), PM10, PM2.5, 16 selected PAHs in PM10 and PM2.5 were determined in the work environment of a coke oven plant in Wuhan. The work environment was divided into the adjunct area, the bottom of, the side of and the top of coke oven. RESULTS: The concentrations of CO, SO2, BSM, BETX, PM10, PM2.5, PAHs in PM10 and PM2.5 were significantly related to working environmental categories, respectively, and were increasing as the adjunct area < bottom < side < top (P (trend) < 0.05). PM10 was statistically significantly correlated with CO, SO2, benzene, BTEX and BSM (0.705, 0.823, 0.664, 0.624 and 0.734, respectively). PM2.5 was statistically significantly correlated with CO, SO2, benzene, BTEX and BSM (0.635, 0.916, 0. 680, 0.553 and 0.726, respectively). BSM was statistically significantly correlated with benzene (0.689). The ratios of PM2.5 to PM10 between different work environments were not significantly different in one-way ANOVA (P > 0.05). The distribution of aromatic rings and the concentrations of total benzo[a] pyrene equivalents in PM10 and PM2.5 were not statistically different between work environments. CONCLUSION: The concentrations of particulate matter was related with other contents of coke oven emissions in coke work environment, and the contents and types of PAHs in PM10 and PM2.5 were similar.

[Effects of smoking on the concentrations of urinary 10 metabolites of polycyclic aromatic hydrocarbons in coke oven workers].

OBJECTIVE: To explore the effects of smoking on urinary 10 metabolites of polycyclic aromatic hydrocarbons (PAHs) in the coke oven workers. METHODS: Occupational health examination was performed on 1401 coke oven workers in one coking plant, their urine were collected respectively. The concentrations of the ten monohydroxy polycyclic aromatic hydrocarbons in urine were detected by gas chromatography/mass spectrometry. The 1401 workers were divided into four groups, namely control, adjunct workplaces, bottom and side, top group according to their workplaces and the different concentrations of PAHs in the environment. The concentrations of the ten monohydroxy polycyclic aromatic hydrocarbons between smokers and nonsmokers in each workplace group were compared using analysis of covariance, respectively. RESULTS: The levels of concentrations of the sixteen polycyclic aromatic hydrocarbons we detected at control were significantly higher than those at other areas (P < 0.05). Comparing the ten monohydroxy polycyclic aromatic hydrocarbons levels between smokers and nonsmokers, the levels of 1-hydroxynaphthalene and 2-hydroxynaphthalene among smokers were higher than nonsmokers with statistically significance in control, adjunct workplaces, bottom and side and top groups (P < 0.05). However, the levels of 1-hydroxypyrene had no statistically significant differences between the four areas. CONCLUSION: Urinary 1-hydroxynaphthalene and 2-hydroxynaphthalene may be used as biomarkers for the impact of smoking on monohydroxy polycyclic aromatic hydrocarbons in the coke oven workers.

A novel use of neural network model to determine the effects of multibiomarker on early health damage among Chinese steel workers.

Coke-oven workers are exposed to many kinds of pollutants that can cause health damage even lead to carcinogenesis. Therefore, it is critical to identify biomarkers that predict early health damage in these exposed individuals in molecular epidemiological studies. We applied an artificial neural network (ANN) model to the identification of such predictors in a study of coke-oven workers. The study included 330 steel-factory workers who were exposed to different levels of polycyclic aromatic hydrocarbons (PAHs) in the workplace and their levels of early health damage were determined by cytokinesis-block micronuclei (CMN), heat shock protein 70 (Hsp70) expression, benzo(a)pyrene diolepoxide-albumin adduct (BPDE-AA), and olive tail moment (OTM). The ANN model was built to predict the early health damage index, and the receiver operating characteristic (ROC) curve was used to illustrate the judged criteria and the ANN model. Trend Chi-square modeling was also performed. We found that there were 55 subjects with early health damage among 330 workers based on the multibiomarker criteria using the 95 percentile of the control group as the cut-off value, while there were 22-35 positive subjects if screening by any single biomarker. The Cochran-Armitage trend test for these findings were statistically significant (Z = 3.21, P = 0.0013). Six variables were selected to simulate the ANN model. The area under ROC (AUROC) was 0.726 ± 0.037 (P < 0.001), and the predictors included workplace, cholesterol, waistline, and others. Therefore, collective using CMN frequency, Hsp70 level, BPDE-AA level, and OTM with equal weights to make an initial screening test for early health damage in coke-oven workers is feasible and superior to any single biomarker. The determinants of the effects of multibiomarker on early health damage screening can be identified by the ANN model and ROC curve method.

[Relevance between single nucleotide polymorphisms in nicotinic acetylcholine receptor genes and DNA damage levels in coke oven workers].

OBJECTIVE: To investigate the relevance between single nucleotide polymorphisms (SNPs) in CHRNA3-CHRNB4-CHRNA5 gene cluster and DNA damage levels in the coke oven workers. METHODS: A total of 309 male subjects from a steel plant in Northern China were divided into high exposure group (154 workers) and low exposure group (155 workers) according to the concentrations of plasma benzoapyrene-r-7, t-8, t-9, c-10-tetrahydrotetrol-albumin (BPDE-Alb) adducts. DNA damage levels of peripheral blood lymphocytes were examined by comet assay. Allelic discrimination of SNPs in CHRNA3-CHRNB4-CHRNA5 gene cluster were detected using Matrix-assisted laser desorption/ionization mass spectrometry. RESULTS: The Olive tail moment (OTM) (1.23 ± 1.05) of peripheral blood lymphocytes in the high exposure group was significantly higher than that (0.80 ± 1.07) in the low exposure group (P < 0.01). In the high exposure group, OTM (1.64 ± 0.17) of subjects with rs667282CC genotype was significantly higher than those (0.95 ± 0.13 or 1.09 ± 0.11) of subjects with rs667282CT genotype or rs667282TT + CT genotype (P < 0.01); OTM (1.60 ± 0.17) of subjects with GG genotype at rs12910984 site was significantly higher than those (0.92 ± 0.13 or 1.07 ± 0.10) of subjects with AG genotype or AG + AA genotype (P < 0.01); OTM (1.35 ± 0.17 or 1.64 ± 0.17) of the TA/TA or CG/CG diplotype carriers was significantly higher than that (0.89 ± 0.13) of TA/CG diplotype carriers (P < 0.05). However, the relevance between genotypes of these SNPs and the DNA damage levels was not found in the low exposure group. CONCLUSION: The rs667282CC genotype and rs12910984GG genotype on CHRNA3-CHRNB4-CHRNA5 gene cluster are associated with increased DNA damage levels in the high PAHs exposure group.

The role of RANTES as a crucial downstream cytokine in calcineurin-dependent VSMC apoptosis stimulated by INFgamma and CD40L.

Many studies have suggested that VSMC (vascular smooth muscle cell) apoptosis plays a key role in destabilization and rupture of atherosclerotic plaques. Therefore protection for VSMCs from apoptosis is a promising approach to stabilize 'vulnerable' lesions. However, the mechanisms as to why VSMCs in the fibrous cap often appear as profilerated in early stages, but turn apoptotic in advanced stages, are still unknown. In the present study, using RNAi (RNA interference) technology and a CaN (calcineurin) antagonist, the correlation between CaN and RANTES (regulated upon activation, normal T-cell expressed and secreted) in cultured rat apoptotic VSMCs stimulated by IFNgamma (interferon gamma; 20 ng/ml) and CD40L (CD40 ligand; 100 ng/ml) was investigated. RANTES released from VSMCs in each group was measured by ELISA and its mRNA in VSMCs was determined by RT (reverse transcription)-PCR. The total activity and expression of CaN in VSMCs were detected by the zymochemistry method and Western blot analysis respectively. From the results of the present study it can be hypothesized that an elevated CaN concentration in endochylema, by the CD40-CD40L signal pathway, induces VSMC apoptosis accomplished by the overexpression of RANTES. Therefore RANTES is a potential target for treating vulnerable atherosclerotic plaques owing to its crucial downstream regulating role in CaN-dependent VSMC apoptosis.

[Association of cooking oil fumes exposure and oxidative DNA damage among occupational exposed populations].

BACKGROUND: Previous investigations indicate that cooks are exposed to polycyclic aromatic hydrocarbons (PAH) from cooking oil fumes (COF). However, Emission of PAH and their carcinogenic potencies from cooking oil fumes sources have not been investigated among cooks. AIMS: To investigate the urinary excretion of a marker for oxidative DNA damage, 8-hydroxydeoxyguanosine (8-OHdG), in different groups of cooks and different exposure groups, and to study the association between 8-OHdG and 1-hydroxypyrene (1-OHP), a biological marker for PAH exposure. METHODS: Urine samples were collected from different groups of cooks (n = 86) and from unexposed controls (n = 36), all are male with similar age and smoking habits. The health status, occupational history, smoking, and alcohol consumption 24 hours prior to sampling was estimated from questionnaires. The urinary samples were frozen for later analyses of 8-OHdG and 1-OHP by high performance liquid chromatography. RESULTS: Excretion in urine of 8-OHdG were similar for controls (mean 1.2 µmol/mol creatinine, n = 36), and for those who had been in the kitchen room with exhaust hood operation (mean 1.5 µmol/mol creatinine, n = 45). COF exposed cooks without exhaust hood operation had increased excretion of 8-OHdG (mean 2.3 µmol/mol creatinine, n = 18). The difference between this group and the unexposed controls was significant. The urinary levels of ln 1-OHP and ln 8-OHdG were still significantly correlated in a multiple regression analysis. CONCLUSION: Results indicate that exposure to PAH or possibly other compounds in COF may cause oxidative DNA damage.

[Study on the association of -689C/T polymorphism in the PPARgamma2 promoter with myocardial infarction].

OBJECTIVE: To investigate the association of -689C/T polymorphism in the peroxisome proliferator-activated receptor-gamma2 (PPARgamma2) promoter with myocardial infarction (MI). METHODS: This is a case-control study, which included 194 subjects with MI and 693 subjects without MI in nondiabetic Han population in Wuhan. Polymerase chain reaction-restriction fragment length polymorphism was used to determine the -689C-->T substitution. RESULTS: The CC,CT, and TT genotype frequencies of -689C/T polymorphism were 88.1%,11.9%,and 0.0 in MI patients and 93.1%,6.6%,and 0.3% in controls, respectively (CC vs. CT+TT, P=0.025). The -689T allele was an independent risk factor for MI (OR=2.125, 95%CI: 1.206-3.744, P=0.009) after adjusting for age,sex,waist circumference,body mass index, smoking, alcohol drinking, physical activities, systolic blood pressure, diastolic blood pressure, fasting blood glucose, total cholesterol, triglyceride, level. The -689T allele carriers had significantly higher TC levels than noncarriers [(5.05+/-1.16) mmol/L vs. (4.78+/-1.05) mmol/L, P=0.004] in the total population. CONCLUSION: The PPARgamma2 promoter -689C/T polymorphism is associated with an increased risk of MI.

Associations of polymorphism of P22(phox) C242T, plasma levels of vitamin E, and smoking with coronary heart disease in China.

BACKGROUND: The C242T polymorphisms of P22(phox) and plasma vitamin E have been associated with the risk of coronary heart disease (CHD) in several studies, but the results have been inconsistent. In this study, we sought to examine potential interactions between P22(phox) genotypes, plasma vitamin E concentrations, and smoking in relation to CHD risk. METHODS: We determined C242T genotype frequency in the P22(phox) gene and plasma levels of vitamin E in 565 Chinese patients with CHD and 609 control subjects. Logistic regression was used to control for potential confounders in multivariate analyses. RESULTS: Compared with subjects with a CC genotype, subjects with a CT or TT genotype had a lower risk of CHD (multivariate-adjusted odds ratio [OR] 0.45, 95% CI 0.28-0.74, P = .001). Plasma concentrations of vitamin E were lower in case patients than in control subjects (multivariate-adjusted OR 0.65, 95% CI 0.50-0.86, P = .025). Compared with nonsmokers with a CC genotype, nonsmokers with a CT or TT genotype had a decreased risk of CHD (OR 0.25, 95% CI 0.12-0.53), but smokers with a CT or TT genotype had an increased risk (OR 2.04, 95% CI 0.74-5.61, P for interaction = .039). Smokers with a lower vitamin E concentration had a >3.0-fold greater risk of CHD than did nonsmokers with a higher vitamin E concentration (OR 3.52, 95% CI 2.36-5.24, P for interaction = .041). CONCLUSIONS: These data provide evidence that P22(phox) genotypes are significantly associated with CHD risk in a Chinese population and suggest potential interactions among smoking, P22(phox) genotypes, and vitamin E in relation to CHD risk.

Significant positive correlation of plasma BPDE-albumin adducts to urinary 1-hydroxypyrene in coke oven workers.

OBJECTIVE: To investigate the application of BPDE-albumin adducts as monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs) and to explore possible relationship between BPDE-albumin adducts and urinary 1-hydroxypyrene (1-OHP) levels in them. METHODS: Thirty-seven coke oven workers from a coke plant and 47 controls without the occupational exposure to PAHs were recruited in this study. The levels of plasma BPDE-albumin adducts and urinary 1-OHP were analyzed using high performance liquid chromatography. RESULTS: The median levels of BPDE-albumin adducts (42.10 fmol/mg albumin) and urinary 1-OHP (5.46 micromol/mol creatinine) were significantly higher in coke oven workers than in controls (14.16 fmol/mg albumin, 2.96 micromol/mol creatinine, respectively; P<0.01). Multiple logistic regression analysis showed that coke oven workers were at higher risk of having BPDE-albumin adduct levels above 25.30 micromol/mg albumin (OR=1.79, P<0.01) and urinary 1-OHP levels above 4.13 micromol/mol creatinine (OR=2.45, P<0.05). There was a positive correlation between the levels of BPDE-albumin adducts and urinary 1-OHP in all subjects (rs=0.349, P<0.01). CONCLUSION: BPDE-albumin adduct is a useful biomarker for monitoring long-term exposure to PAHs, and plasma BPDE-albumin adducts level is significantly correlated to urinary 1-OHP levels in coke oven workers.

[Relationship between heat shock protein 72 and DNA genetic damage in peripheral blood lymphocytes of coke oven workers].

OBJECTIVE: To investigate the relationship between heat shock protein 72 (Hsp72) and DNA genetic damage in peripheral blood lymphocytes of coke oven workers and the role of Hsp72 in protection of cells from genetic damage induced by coke oven emissions. METHODS: Two hundred and sixty-seven coke oven workers and thirty controls without occupational PAHs exposure were investigated. Benzo[a]pyrene concentrations in the ambient air individually collected were assayed by high performance liquid chromatography (HPLC). Western Blot was used to measure Hsp72 levels and Comet assay was used to evaluate DNA damage degree. Personal information was collected by questionnaire. RESULTS: The Hsp72 level (G+/-S(G)) and olive comet tail moment (G+/-S(G) of peripheral blood lymphocytes in high-exposure workers (1.24 +/- 0.42 and 4.49 +/- 1.24) were significantly higher than those in low-exposure workers (1.01 +/- 0.35 and 2.99 +/- 1.10, P < 0.05) and control (0.85 +/- 0.34 and 2.40 +/- 1.00, P < 0.05) respectively. The Hsp72 median level of all subjects was used as the limit to divide subjects into high Hsp72 level group and low Hsp72 level group. The rate with high Hsp72 level was 36.7%, 43.1% and 58.3% in control, low exposure and high exposure workers respectively and had a rising tendency following exposure level (P = 0.003). In high Hsp72 level group Hsp72 level in high exposure workers was significantly higher than that in control (P < 0.05), and there was a rising tendency along with the increase of exposed levels. But the olive comet tail moment had no significant difference among three exposed groups (P > 0.05). In low Hsp72 level group there no difference among three exposed groups about Hsp72 levels. The olive comet tail moment in high exposure workers was significantly higher than that in low exposure workers and control (P < 0.01) and high exposure workers in Hsp72 positive group and there was a rising tendency along with the increase of exposed levels. Hsp72 levels had strong negative correlation with the olive comet tail moment (r = -0.503, P < 0.01) in high exposure workers. CONCLUSION: The coke oven emissions can induce hsp72 expression. Hsp72 play a role of protecting cells from DNA damage induced by coke oven emissions.

[Influences of excision repair cross complementation group 4 genetic variations on DNA damage in lymphocytes among coke oven workers].

OBJECTIVE: To investigate the relationship between excision repair cross complementation group 4 ERCC4 gene polymorphisms and DNA damage in lymphocytes of coke oven workers and controls. METHODS: Two hundred and forty-six coke oven workers and one hundred and twenty-seven controls were recruited in the study, and peripheral vein blood was drawn after over night fasting. Comet assay was used to evaluate DNA damage, and TaqMan-MGB probes were used to analyze ERCC4 genetic variations including the three Tagged-single nucleotide polymorphisms (Tag SNPs), referred to rs744154, rs3136079 and rs31870 which were picked out from Hapmap database. Then haplotypes were reconstructed by PHASE2.0.2 software. RESULTS: The lymphocytes Olive TM value of coke oven workers was significantly higher than that of controls (1.26+/-1.12 vs 0.52+/-0.97, P<0.01). Among coke oven workers, no significant difference was found between the Olive TM of those with different genotypes or haplotype pairs at ERCC4 gene (P>0.05). However, in the control group, the TG genotype carriers had higher Olive TM than the TT and GG genotype carriers (0.26+/-0.96 vs 0.66+/-0.98 and 0.66+/-0.51, P<0.05), and the CTG/CTG haplotype pairs carriers had the highest Olive TM (0.69+/-1.01), and no CTG haplotype carriers had the lowest Olive TM (0.25+/-0.80), and the difference was borderline (P=0.08). CONCLUSION: The gene polymorphism at ERCC4 gene has no effects on the DNA damage of lymphocytes in coke oven workers, but the TG genotype carriers has lower DNA damage in the control. DNA damage is influenced by the interaction of genetic and environmental factors.

Elevated levels of urinary 8-hydroxy-2 -deoxyguanosine, lymphocytic micronuclei, and serum glutathione S-transferase in workers exposed to coke oven emissions.

To investigate associations among occupational exposure to coke oven emissions (COEs), oxidative stress, cytogenotoxic effects, change in the metabolizing enzyme glutathione S-transferase (GST), and internal levels of polycyclic aromatic hydrocarbons (PAHs) in coke oven workers, we recruited 47 male coke oven workers and 31 male control subjects from a coke oven plant in northern China. We measured the levels of 1-hydroxypyrene (1-OHP) and 8-hydroxy-2 -deoxyguanosine (8-OHdG) in urine, micronucleated binucleated cells (BNMNs) in peripheral blood lymphocyte, and GST in serum. Our results showed that the group exposed to COEs had significantly increased levels of 1-OHP [median 5.7; interquartile range (IQR), 1.4-12.0 micromol/mol creatinine] compared with the control group (3; 0.5-6.4 micromol/mol creatinine). In addition, the median levels (IQR) of 8-OHdG, BNMNs, and GST were markedly increased in the exposed [1.9 (1.4-15.4) micromol/mol creatinine; 6 (2-8) per thousand ; 22.1 (14.9-31.2) U/L, respectively] compared with controls [1.3 (1.0-4.0) micromol/mol creatinine, 2 (0-4) per thousand; and 13.1 (9.5-16.7) U/L, respectively]. These results appeared to be modified by smoking. However, multivariate logistic regression analysis revealed that exposure to COEs had the highest odds ratio among variables analyzed and that smoking was not a significant confounder of the levels of studied biomarkers. Overall, the present findings suggest that COE exposure led to increased internal PAH burden, genetic damage, oxidative stress, and GST activity. The consequences of the changes in these biomarkers, such as risk of cancer, warrant further investigations.

Brain-derived neurotrophic factor promotes growth and migration of multiple myeloma cells.

The evolution of multiple myeloma (MM) depends on complex signals from the bone marrow microenvironment, which support proliferation and survival of malignant plasma cells. Previous study defined a brain-derived neurotrophic factor-tyrosine kinase receptor B (BDNF/TrkB) axis in myeloma and autocrine growth stimulation by BDNF in various tumor cells. We examined the biological effects of BDNF on MM cells. Using a reverse transcriptase-polymerase chain reaction, Western blotting, and immunohistochemistry, we observed that both BDNF and its high-affinity receptor TrkB are expressed by MM cell lines (RPMI 8226, U266, and KM3) and primary MM cells. Functional studies revealed that BDNF was a potent growth factor for MM. BDNF (5-500 ng/mL) had strong proliferative effects on both MM cell lines and primary MM cells, shown by [(3)H]thymidine incorporation assay. BDNF (12.5-200 ng/mL) also induced migration of MM cells, as indicated by the Transwell migration assay. Together, our data indicate that BDNF is a potent myeloma growth and chemotactic factor and suggest that the BDNF/TrkB pathway is a potential therapeutic target in MM.