To observe the alteration in the expression of DNA repair enzymes hOGG1 and hMYHalpha and the change in 8-OHdG levels in the HBx gene-transfected cells HepG2/HBx and to explore the mechanisms of the HBV-associated hepatocellular carcinoma, the gene-transfected cells HepG2/HBx which stably expressed HBx was established, and the effect of HBx on the cell cycle and proliferation of HepG2 was examined. By using the beta-actin as the interior control, real-time polymerase chain reaction (Real-time qPCR) was employed to quantitatively detect the expression of DNA repair enzymes hOGG1 and hMYHalpha in the HepG2/HBx, the control cells HepG2 and HepG2 transfected with pcDNA3.1 vector (HepG2/pDNA3.1). The 8-OHdG levels were determined by HPLC/ECD in the established gene-transfected cells HepG2/HBx and the control cells HepG2 and HepG2/pcDNA3.1. Our results showed that the expression of DNA repair enzyme hMYHalpha in the HepG2/HBx (0.021+/-0.007) was significantly lower than that of HepG2 (0.099+/-0.041) (P<0.05) and HepG2/pDNA3.1 (0.121+/-0.005) (P<0.05). However, the no significant differences existed in the expression of DNA repair enzyme hOGG1 among the three cell strains (P>0.05). The 8-OHdG level in the HepG2/HBx was significantly higher than that in HepG2 and HepG2/pcDNA3.1 (P<0.05). It is concluded that HBx gene may inhibit the expression of DNA repair enzyme hMYHalpha mRNA to impair the ability to repair the intracellular DNA oxidative damage, to increase the oxidative DNA-adduct 8-OHdG and to affect the nucleotide excision repair function, thus participate in the occurrence and development of hepatocellular carcinoma.
DNA Glycosylases/genetics ; DNA Glycosylases/*metabolism ; Deoxyguanosine/analogs & derivatives ; Deoxyguanosine/metabolism ; Hep G2 Cells ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Trans-Activators/*genetics

OBJECTIVETo investigate the application of serum glutathione S-transferase (GST) and urinary 8-hydroxy-2-deoxyguanosine (8-OHdG) as the monitoring biomarkers for coke oven workers exposed to polycyclic aromatic hydrocarbons (PAHs).

METHODS47 male coke oven workers and 31 male control workers were investigated. Urinary 8-OHdG and serum GST were analyzed using high performance liquid chromatography (HPLC) with electrochemical detection and test kit. Urinary 1-hydroxypyrene (1-OHP) as internal exposure of PAHs was also determined simultaneously by alkaline hydrolysis and HPLC.

RESULTSThe values of urinary 1-OHP, serum GST and urinary 8-OHdG were reported as median with interquartile range (P(25)-P(75)). Urinary 1-OHP [5.7 (1.4-12.0) micromol/mol Cr], serum GST [22.1 (14.9-31.2) U/ml], and urinary 8-OHdG [1.9 (1.4-15.4) micromol/mol Cr] in coke oven workers were significantly higher than in control workers [3.0 (0.5-6.4) micromol/mol Cr (P < 0.05), 13.1 (9.5-16.7) U/ml (P < 0.01), and 1.3 (1.0-4.0) micromol/mol Cr (P < 0.05) respectively]. Categorizing by smoking status, significant differences in urinary 1-OHP and serum GST were found only in smokers among coke oven workers compared to control workers (P < 0.01), and 8-OHdG levels only in non-smokers (P < 0.01). Additionally, there was significant correlation between urinary 1-OHP and serum GST activity (r(s) = 0.31, P < 0.01, n = 78). The multiple logistic regression analysis showed that coke oven workers were at the higher risk of having GST activities above 16.7 U/ml (OR = 13.2) and 8-OHdG levels above 1.8 micromol/mol creatinine (OR = 4.4). High body mass index was an independent factor to affect urinary 8-OHdG levels.

CONCLUSIONSThe elevated serum GST activities and increased oxidative DNA damage were found in the coke oven workers. Occupational exposure and smoking interact on each other. Serum GST may be used as a biomarker for assessing the exposure of PAHs. Assay of urinary 8-OHdG may be useful for evaluating the risk of lung cancer in coke oven workers.

Adult ; Case-Control Studies ; Coke ; Deoxyguanosine ; analogs & derivatives ; urine ; Glutathione Transferase ; blood ; Humans ; Male ; Occupational Exposure

Benzene ; poisoning ; DNA Damage ; DNA Repair ; genetics ; Deoxyguanosine ; analogs & derivatives ; genetics ; Humans ; Mutagens ; poisoning ; Poisoning ; genetics

8-hydroxy-2'-deoxyguanosine (8-OHdG) is a sensitive and stable biomarker for evaluating DNA oxidative damage. A rapid and sensitive colloidal gold immunochromatographic strip was developed for 8-OHdG detection by a competitive method. The sample pad (glass cellulose film), bonding pad (glass cellulose film), nitrocellulose film and absorbent pad were pasted on the polyvinyl chloride (PVC) base plate to construct the test strip. Colloidal gold (AuNPs) was prepared by the reduction of chloroauric acid with sodium citrate. 8-OHdG antibody (Ab) was coated on the outer layer of AuNPs to form Ab@AuNPs as a probe. Bovine serum albumin (BSA) and 8-OHdG were conjugated with carbodiimide hydrochloride to prepare an artificial antigen, which was used as the coating antigen of detection line. Goat anti mouse polyclonal antibody IgG was used as the coating antibody of control line. The experimental parameters were optimized including the type of nitrocellulose membrane, the formula of loading solution, and the spraying amount of gold labeled antibody. The results showed that the appropriate nitrocellulose membrane was CN 95. The optimal loading solution included BSA (1%), Tween-20 (3%), sucrose (3%) and NaCl (0.9%). The optimal spraying amount of gold labeled antibody was 4 μL. 8-OHdG can be detected by the strip under visible light, and the level of 8-OHdG in urine can be preliminarily determined by comparing the color intensity of T line and C line. The 8-OHdG concentration in urine was further calculated by the gray value of T line and the threshold of detection was 2.55 μg/L. This colloidal gold immunochromatographic strip is simple, rapid and specific for detecting 8-OHdG in human urine to preliminarily evaluate the human status.
8-Hydroxy-2'-Deoxyguanosine ; Animals ; Antibodies, Monoclonal ; Gold ; Gold Colloid/chemistry* ; Metal Nanoparticles ; Mice ; Sensitivity and Specificity

OBJECTIVE: To discuss seminal plasma oxidative stress and sperm DNA oxidative damage in patients with idiopathic asthenozoospermia. METHODS: Infertile couples were selected from the clinic outpatients of the Reproductive Center of Xiangya Hospital, Central-South University from December 2010 to March 2011. Fresh semen of 28 men with idiopathic asthenozoospermia was collected as an experiment group, and 24 fertile men with normal semen and normal reproductive history served as a control group. Level of reactive oxygen species (ROS) in the seminal plasma was assessed with luminer chemiluminescence method. Density of sperm DNA oxidation product 8-hydroxy-2'-deoxyguanosine (8-OHdG) was assessed with enzyme linked immunosorbent assay. RESULTS: 1) ROS level in the experiment group was higher than that in the control group (P<0.01). There was negative correlation between the percentage of progressive motility spermatozoa and the ROS level in the seminal plasma in the 2 groups (r=-0.72, P<0.01). 2) Density of sperm 8-OHdG in the experiment group was higher than that in the control group (P<0.01). There was negative correlation between the percentage of progressive motility spermatozoa and the density of sperm 8-OHdG (r=-0.73, P<0.01). 3) There was positive correlation between the ROS level in the seminal plasma and the density of sperm 8-OHdG (r=0.77, P<0.01). CONCLUSION There is sperm DNA oxidative damage in patients with idiopathic asthenozoospermia, which may be related with the oxidative stress. Excessive generation of reactive oxygen species may be a cause of low sperm motility in patients with idiopathic asthenozoospermia.
8-Hydroxy-2'-Deoxyguanosine ; Adult ; Asthenozoospermia ; etiology ; genetics ; metabolism ; Case-Control Studies ; DNA Damage ; Deoxyguanosine ; analogs & derivatives ; metabolism ; Humans ; Infertility, Male ; genetics ; Male ; Oxidative Stress ; physiology ; Spermatozoa ; metabolism ; Young Adult

OBJECTIVETo investigate the smoking or age impact on occupational workers in electrical and electronic equipment waste (e-waste) dismantling procedure, using 8-Hydroxy-2'-deoxyguanosine (8-OHdG) in the urine as a biomarker for oxidative damage to DNA.

METHODSThe pre-workshift and post-workshift urinary samples of 64 male workers in e-waste dismantling procedure were detected by solid-phase extraction-high performance liquid chromatography-electrochemical detector (SPE-HPLC-ECD). The data were statistically analyzed by two factors, age or smoking status.

RESULTSThe 8-OHdG levels in non-smokers' urines (n = 42) were higher than those in smokers' urines (n = 22). The levels in pre-workshift urines were detected at (8.25 +/- 4.23) micromol/mol creatinine in non-smokers, while the values were (5.44 +/- 1.18) micromol/mol in smokers. And, the levels in post-workshift were detected at (43.12 +/- 16.19) micromol/mol creatinine in non-smokers, while the values were (14.82 +/- 2.51) micromol/mol in smokers. The 8-OHdG levels in pre-workshift urines were not different between non-smokers and smokers (t = -0.81, P = 0.42), however after 1 day exposure, urinary 8-OHdG levels were significantly increased in non-smokers than those in smokers (t = - 2.33, P < 0.05). On the other hand, the subjects were divided into five groups according to their age. The 8-OHdG levels in pre-workshift urines were (1.86 +/- 0.66), (3.57 +/- 0.54), (8.12 +/- 4.10), (11.39 +/- 3.70) micromol/mol creatinine in < 20 years group (n = 6), 20 -years group (n = 22), 30 - years group ( n = 23), 40 - 49 years group (n = 11) respectively. No effect of age was found on the pre-workshift urinary 8-OHdG levels (F = 0.98, t = 0.41). However, it was found that the post-workshift urinary 8-OHdG levels increased along with the e-waste workers' age (F = 4.81, P = 0.03), and they were (4.19 +/- 2.85), (19.89 +/- 5.26), (28.89 +/- 14.61), (34.94 +/- 12.50) micromol/mol creatinine in < 20 years group, 20 - years group, 30 - years group, 40 - 49 years group respectively.

CONCLUSIONThe urinary 8-OHdG levels in the e-waste dismantling workers might be inhibited by smoking status. The post-workshift urinary 8-OHdG levels increased along with the e-waste workers' age.

Adult ; Age Factors ; Deoxyguanosine ; analogs & derivatives ; urine ; Electronics ; Humans ; Male ; Middle Aged ; Occupational Exposure ; Refuse Disposal ; Smoking ; Young Adult

OBJECTIVETo study the potential association of DNA oxidation and DNA methylation, in vitro cultured cells were exposed to different doses of H2O2, 8-oxo-dG formation, cell DNA 5-mC contents were analyzed to explore the time- dose-response relationship of DNA oxidation and DNA methylation.

METHODSA549 cells were exposed to different doses of H2O2, 8-oxo-dG formation and cell genomic DNA 5-mC contents were analyzed by a high-performance liquid chromatography system and high performance capillary electrophoresis (HPCE), respectively.

RESULTSH2O2 induced the formation of 8-oxo-dG and 5-mC in different characteristics, it need at least 10 days for significant changes in the level of DNA methylation, whereas under the same conditions, changes in the level of DNA oxidation cast only 12 hours. H2O2 induced decreased levels of DNA methylation in A549 cells in a dose-dependent manner. In a certain range of time and dose, it showed a negative correlation between DNA oxidation and DNA methylation.

CONCLUSIONThe study suggests that oxidative DNA could lead to reduced levels of DNA methylation, DNA oxidation may affect the regulation of cellular methylation mechanisms, in the course of chemical mutagenesis, DNA oxidation may be an earlier important molecule event than DNA methylation.

Cell Line ; DNA ; chemistry ; DNA Damage ; DNA Methylation ; Deoxyguanosine ; analogs & derivatives ; chemistry ; Humans ; Hydrogen Peroxide ; toxicity ; Oxidative Stress

Objective: To investigate the values of serum 8-hydroxydeoxyguanosine (8-OHdG) in predicting disease progression and prognosis of patients with sepsis. Methods: The prospective observational research methods were used. A total of 124 patients with sepsis who met the inclusion criteria were admitted to the Department of Emergency of the First Affiliated Hospital of Wenzhou Medical University from April 2015 to July 2016, including 79 males and 45 females, aged (62±15) years. The sepsis-related organ failure assessment (SOFA) scores of all patients on admission and on the second day of admission and their difference (ΔSOFA) were calculated. The patients were divided into non-progression group with ΔSOFA score <2 (n=101) and progression group with ΔSOFA score ≥2 (n=23), and according to the survival during hospitalization, the patients were divided into survival group (n=85) and death group (n=39). Data of patients between non-progression group and progression group, survival group and death group were compared, including the gender, age, days in emergency intensive care unit (ICU), smoking, hypertension, diabetes mellitus, serum white blood cell count, serum C-reactive protein, and serum procalcitonin on admission, and serum 8-OHdG within 24 h of admission. The multivariate logistic regression analysis was used to screen the independent risk factors of disease progression and death during hospitalization in 124 patients with sepsis, the receiver's operating characteristic (ROC) curves were drawn according to the independent risk factors, and the area under the curve (AUC), the best threshold, and the sensitivity and specificity under the best threshold were calculated. The patients were divided into high 8-OHdG group (n=35) and low 8-OHdG group (n=89) according to the best threshold in ROC curve of death during hospitalization. The data including the gender, age, SOFA score on admission, SOFA score on the second day of admission, and ΔSOFA score of patients in the two groups were compared. The survival rates of patients within 90 d of admission in the two groups were compared by the Kaplan-Meier method. Data were statistically analyzed with independent sample t test, Mann-Whitney U test, chi-square test, and Log-rank test. Results: The gender, age, days in emergency ICU, smoking, complicated with hypertension, complicated with diabetes mellitus, serum white blood cell count, serum C-reactive protein, and serum procalcitonin on admission of patients in non-progression group and progression group were similar (P>0.05). The serum 8-OHdG within 24 h of admission of patients in progression group was significantly higher than that in non-progression group (Z=-2.31, P<0.05). Multivariate logistic regression analysis showed that the serum 8-OHdG within 24 h of admission was the independent risk factor for disease progression of 124 patients with sepsis (odds ratio=1.06, with 95% confidence interval of 1.01-1.11, P<0.05). The AUC under the ROC curve of serum 8-OHdG within 24 h of admission to predict disease progression of 124 patients with sepsis was 0.65 (with 95% confidence interval of 0.52-0.79, P<0.05), the optimal threshold was 32.88 ng/mL, and the sensitivity and specificity under the optimal threshold was 52.2% and 79.2%, respectively. The gender, age, days in emergency ICU, smoking, complicated with hypertension, complicated with diabetes mellitus, and serum white blood cell count, serum C-reactive protein, and serum procalcitonin on admission of patients in survival group and death group were similar (P>0.05). The serum 8-OHdG within 24 h of admission of patients in death group was significantly higher than that in survival group (Z=-2.37, P<0.05). Multivariate logistic regression analysis showed that the serum 8-OHdG within 24 h of admission was the independent risk factor for death of 124 patients with sepsis (odd ratio=1.04, with 95% confidence interval of 1.00-1.09, P<0.05). The AUC under the ROC curve of serum 8-OHdG within 24 h of admission to predict death of patients during hospitalization was 0.63 (with 95% confidence interval of 0.52-0.75, P<0.05), the optimal threshold was 32.43 ng/mL, the sensitivity and specificity under the optimal threshold was 51.3% and 84.7%, respectively. The gender and age of patients in high 8-OHdG group and low 8-OHdG group were similar (P>0.05). The SOFA score on admission, SOFA score on the second day of admission, and ΔSOFA score of patients in high 8-OHdG group were significantly higher than those in low 8-OHdG group (with Z values of -2.49, -3.01, and -2.64, respectively, P<0.05 or P<0.01). The survival rate within 90 d of admission of patients in low 8-OHdG group was significantly higher than that in high 8-OHdG group (χ²=14.57, P<0.01). Conclusions: Serum 8-OHdG level is an independent risk factor for disease progression and death in sepsis patients with limited ability for predicting disease progression and prognosis of sepsis of patients. The patients with higher serum 8-OHdG level have higher death risk within 90 d of admission.
8-Hydroxy-2'-Deoxyguanosine ; Aged ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; ROC Curve ; Retrospective Studies ; Sepsis

Objectives: To study the association between metals mixture exposure and DNA oxidative damage using mixture analysis methods, and to explore the most significant exposure factors that cause DNA oxidative damage. Methods: Workers from steel enterprises were recruited in Shandong Province. Urinary metals were measured by using the inductively coupled plasma mass spectrometry method. The level of urinary 8-hydroxy-2'-deoxyguanosine (8-OHdG) was determined by using the ultra-high performance liquid chromatography-mass spectrometry method. Bayesian kernel machine regression (BKMR), elastic net regression and quantile g-computation regression were used to analyze the association between urinary metals and urinary 8-OHdG. Results: A total of 768 subjects aged (36.15±7.40) years old were included in the study. BKMR, elastic net regression and quantile g-computation all revealed an overall positive association between the mixture concentration and increased urinary 8-OHdG. The quantile g-computation results showed that with a 25% increase in metal mixtures, the urinary 8-OHdG level increased by 77.60%. The elastic net regression showed that with a 25% increase in exposure risk score, the urinary 8-OHdG level increased by 26%. The BKMR summarized the contribution of individual exposures to the response, and selenium, zinc, and nickel were significant contributors to the urinary 8-OHdG elevation. Conclusion: Exposure to mixed metals causes elevated levels of DNA oxidative damage, and selenium, zinc, and nickel are significant exposure factors.
Humans ; Adult ; Nickel/toxicity* ; Selenium ; Bayes Theorem ; Metals/toxicity* ; 8-Hydroxy-2'-Deoxyguanosine ; Oxidative Stress/physiology* ; Zinc ; DNA Damage

BACKGROUND: In many studies, oxidative stress markers have been employed to serve as a measure of a disease process or to reflect oxidative status. These oxidative stress markers must have some degree of predictive validity, but full substantiation of this relation is still lacking. This paper presents data on levels of three biomarkers, oxidized low-density lipoproteins (LDL), carbonyl, and 8-hydroxy-2'-deoxyguanosine (8-OHdG), and a number of life style factors associated with oxidative stress in healthy adults. METHODS: For 237 healthy adults aged 40-60 years, a number of life style factors, biochemical characteristics and oxidative status were evaluated. Markers of oxidative stress were measured by an ELISA method. RESULTS: Waist-hip ratio and use of vitamin supplement were associated with serum oxidized LDL (P<0.05). Body mass index and stress had a relationship (P<0.05) with protein carbonyl. Creactive protein was related to serum oxidized LDL (P<0.01). There was no correlation among three oxidative stress markers, oxidized LDL, carbonyl, and 8-OHdG. CONCLUSIONS: The oxidative stress markers used in this study could not be regarded as a general estimate of the healthy individual oxidative status. Further studies focusing on the development of biomarkers to reflect changes in the oxidative status under normal, non-pathological conditions in humans will be required.
Adult ; Biomarkers ; Body Mass Index ; Deoxyguanosine ; Enzyme-Linked Immunosorbent Assay ; Humans ; Life Style* ; Lipoproteins, LDL ; Oxidative Stress* ; Protein Carbonylation ; Vitamins ; Waist-Hip Ratio