Mediating effect of oxidative stress on association between nicotine and unexplained recurrent spontaneous abortion / 环境与职业医学

Background Previous studies have confirmed that nicotine exposure is an independent risk factor for miscarriage, but it is not clear whether nicotine causes unexplained recurrent spontaneous abortion (URSA) through oxidative stress. Objective To explore potential mediating effect of oxidative stress on the relationship between nicotine exposure and URSA. Methods Using a 1∶1 matched case-control study, 88 patients with URSA visiting Beijing Obstetrics and Gynecology Hospital affiliated to Capital Medical University from April to October in 2018 were selected as the case group, and 88 pregnant women without adverse pregnancy outcomes and seeking induced abortion in the outpatient clinic of the same hospital were selected as the control group. The levels of 8-hydroxy-2'-deoxyguanosine (8-OHdG) and 8-iso-prostaglandin F2α (8-iso-PGF2α) in urine were determined by enzyme-linked immunosorbent assay, and the level of urinary nicotine was determined by gas chromatography-mass spectrometry. Conditional logistic regression was used to analyze the associations of nicotine, 8-OHdG, and 8-iso-PGF2α with the risk of URSA. Multiple linear regression was used to analyze the association of nicotine with 8-OHdG and 8-iso-PGF2α. The potential mediating effect of oxidative stress on URSA after nicotine exposure was explored by dichotomous mediating model. Results The median concentrations (creatinine corrected) of nicotine, 8-OHdG, and 8-iso-PGF2α in urine of the case group were 7.78, 4.84, and 44.10 μg·g−1, respectively, while those of the control group were 6.48, 3.34, and 29.39 μg·g−1, respectively. The concentrations of nicotine, 8-OHdG, and 8-iso-PGF2α in urine of the case group were all higher than those of the control group (P < 0.05). The results of conditional logistic regression model showed that after adjusting selected confounding factors, compared with the Q1 groups of nicotine and 8-iso-PGF2α, the OR (95%CI) values of URSA in the Q4 groups were 4.20 (1.33-13.29) and 6.25 (1.66-23.59), respectively. Compared with the Q1 group of 8-OHdG, the OR (95%CI) values of URSA in the Q1, Q2, and Q3 groups were 5.47 (1.43-20.93), 4.24 (1.28-14.07), and 6.36 (1.82-22.28), respectively. The results of multiple linear regression showed that after adjusting confounding factors, there was a positive correlation between urinary nicotine and 8-OHdG in both the case group and the control group, and the b (95%CI) values were 0.76 (0.67-0.86) and 0.81 (0.67-0.95) respectively; there was a positive correlation between urinary nicotine and 8-iso-PGF2α in both the case group and the control group, and the b (95%CI) values were 0.65 (0.55-0.75) and 0.76 (0.64-0.87), respectively. The results of dichotomous mediating analysis showed that the mediating effect of 8-iso-PGF2α and its 95%CI on the relationship between nicotine exposure and URSA was 1.518 (0.749-2.311). Conclusion Internal nicotine exposure is a risk factor for URSA and is positively correlated with oxidative stress, and it may lead to URSA through lipid peroxidation damage.

Can serum 8-hydroxy-2'-deoxyguanosine levels reflect the severity of pulmonary arterial hypertension?

SUMMARY OBJECTIVE: Oxidative stress plays a pivotal role in the pathogenesis of pulmonary arterial hypertension. 8-Hydroxy-2'-deoxyguanosine is a sensitive biomarker that reflects the degree of oxidative damage to DNA. We investigated whether serum 8-Hydroxy-2'-deoxyguanosine is a clinically useful biomarker for the severity of pulmonary arterial hypertension. METHODS: We measured serum 8-Hydroxy-2'-deoxyguanosine levels in 25 patients (age 37±13 years, 68% women) diagnosed with idiopathic pulmonary arterial hypertension, familial pulmonary arterial hypertension, or pulmonary arterial hypertension associated with congenital heart disease. The severity of pulmonary arterial hypertension was evaluated by six-min walking distance, World Health Organization functional class, and serum brain natriuretic peptide levels. Age and gender-matched 22 healthy subjects served as the control group. RESULTS: The comparison of 8-Hydroxy-2'-deoxyguanosine levels between patients and controls was not statistically different [(19.86±9.79) versus (18.80±3.94) ng/mL, p=0.622)]. However, there was a significant negative correlation between 8-Hydroxy-2'-deoxyguanosine levels and six-min walking distance (r= −0.614, p=0.001). Additionally, serum 8-Hydroxy-2'-deoxyguanosine levels in patients with functional class III-IV were significantly higher than those with functional class I-II (functional class III-IV 32.31±10.63 ng/mL versus functional class I-II 16.74±6.81 ng/mL, respectively, p=0.003). CONCLUSION: The 8-Hydroxy-2'-deoxyguanosine levels were significantly correlated with exercise capacity (six-min walking distance) and symptomatic status (functional class), both of which show the severity of pulmonary arterial hypertension in patients.

Study on modern drying processing method for Trichosanthis Pericarpium based on multi-bioactive constituents / 中草药

Objective: To compare the effects of different drying methods on the quality of Trichosanthis Pericarpium, and screen the suitable drying methods for its modern drying processing. Methods: The fresh Trichosanthis Pericarpium collected from Anhui were processed by traditional and modern drying processing methods [hot air drying (40, 50, 60, 70 ℃), microwave vacuum drying (40, 50, 60, 70 ℃; vacuum -0.08 MPa), short-wave infrared drying (50, 60, 70 ℃), vacuum -80 ℃ lyophilization, traditional solar drying, shadow drying]. Combined with the appearance of the samples after drying, the composition and content of the 40 resource chemical compositions [five kinds of flavonoids (rutin, luteoloside, apigenin-7-O-glucuronide, apigenin, tangeretin), three kinds of triterpenoids (cucurbitacin D, cucurbitacin B, cucurbitacin E)] and nutritional nourishing ingredients [two kinds of saccharides (glucose, fructose), eighteen kinds of amino acids (phenylalanine, L-leucine, iso-leucine, L-tryptophan, γ-aminobutyric acid, L-methionine, L-valine, L-proline, L-tyrosine, trans-4-hydroxy-L-proline, L-threonine, L-glutamic acid, L-glutamine, L-serine, L-asparagine, L-citrulline, L-arginine, L-lysine), and twelve kinds of nucleosides (thymidine, 2’-deoxyuridine, adenine, uridine, adenosine, 2’-deoxyinosine, inosine, cytosine, guanine, 2’-deoxyguanosine, cytidine, guanosine)] were evaluated for comprehensively evaluating the quality of the different samples. The best modern drying method for Trichosanthis Pericarpium was preferred by principal component analysis. Results: Among different dry samples, the content of medicinal ingredients and nutrient nourishing ingredients varied greatly, among which fructose and glucose content ranged from 9.78% to 21.32% and 4.46% to 15.63%, respectively. Samples of 70 ℃ microwave vacuum drying had the highest total amount of flavonoids and tetracyclic triterpenoids, while those of 40 ℃ hot air drying treatment were the lowest. Through comprehensive evaluation of 14 kinds of Trichosanthis Pericarpium samples obtained by different drying methods, it was found that samples of 70 ℃ hot air drying, 70 ℃ short-wave infrared drying, vacuum -80 ℃ lyophilization, 50 ℃ microwave vacuum drying and 60 ℃ hot air drying were better than the traditional solar drying. Conclusion: Combined with the appearance of the medicinal properties, color, texture, drying time and functional ingredients, it was recommended that 70 ℃ hot air drying method was the preferred conditions for production based on the current state of the equipment of company. Based on the development of new equipment in the future, short-wave infrared 70 ℃ drying can be used as the development direction of Trichosanthis Pericarpium. The study provided reference for the standardization and quality characteristics of production of Trichosanthis Pericarpium.

The effectiveness of eugenol against cisplatin-induced ototoxicity / A eficácia do eugenol contra a ototoxicidade induzida pela cisplatina

Abstract Introduction: Ototoxicity refers to cellular damage or function impairment developing in the inner ear in association with any therapeutic agent or chemical substance, and still represents the principal side-effect restricting the use of cisplatin. Objective: The aim of this study was to perform a biochemical, functional and histopathological investigation of the potential protective effect of eugenol against cisplatin-induced ototoxicity. Methods: The study was performed with 24 female Sprague Dawley rats. Distortion product otoacoustic emissions tests were performed on all animals, which were randomized into four equal groups. A single intraperitoneal dose of 15 mg/kg cisplatin was administered to cisplatin group, while the eugenol group received 100 mg/kg eugenol intraperitoneal for five consecutive days. 100 mg/kg eugenol was administered to cisplatin + eugenol group for 5 days. On the third day, these rats were received a single dose of 15 mg/kg cisplatin. The control group was given 8 mL/kg/day intraperitoneal saline solution for five days. The distortion product otoacoustic emissions test was repeated 24 h after the final drug administration. All animals were sacrificed, and the cochleas were subsequently used for biochemical and histopathological examinations. Results: Cisplatin caused oxidative stress in the cochlea, impaired the cochlear structure and significantly reduced signal noise ratio levels. Administration of eugenol together with cisplatin reversed these effects and provided functional, biochemical and histopathological protection. Conclusion: The study findings represent the first indication in the literature that eugenol may protect against ototoxicity by raising levels of antioxidant enzymes and lowering those of oxidant parameters.

Resumo Introdução: A ototoxicidade refere-se ao dano celular ou comprometimento da função da orelha interna associado a qualquer agente terapêutico ou substância química e ainda representa o principal efeito colateral que restringe o uso da cisplatina. Objetivo: O objetivo deste estudo foi realizar uma investigação bioquímica, funcional e histopatológica do potencial efeito protetor do eugenol contra a ototoxicidade induzida pela cisplatina. Método: O estudo foi realizado com 24 ratos fêmeas Sprague Dawley. Testes de emissões otoacústicas por produto de distorção foram realizados em todos os animais, os quais foram randomizados em quatro grupos iguais. Uma única dose intraperitoneal de 15 mg/kg de cisplatina foi administrada ao grupo cisplatina, enquanto o grupo eugenol recebeu 100 mg/kg de eugenol intraperitoneal por cinco dias consecutivos. Foram administrados 100 mg/kg de eugenol ao grupo cisplatina + eugenol durante 5 dias. No terceiro dia, estes ratos receberam uma dose única de 15 mg/kg de cisplatina. O grupo controle recebeu 8 mL/kg/dia de solução salina intraperitoneal por cinco dias. O teste de emissões otoacústicas por produto de distorção foi repetido 24 horas após a administração final do medicamento. Todos os animais foram sacrificados e as cócleas foram posteriormente utilizadas para exames bioquímicos e histopatológicos. Resultados: A cisplatina causou estresse oxidativo na cóclea, prejudicou a estrutura coclear e reduziu significativamente os níveis da relação sinal/ruído. A administração de eugenol juntamente com a cisplatina reverteu esses efeitos e forneceu proteção funcional, bioquímica e histopatológica. Conclusão: Os achados do estudo representam a primeira indicação na literatura de que o eugenol pode proteger contra a ototoxicidade, eleva os níveis de enzimas antioxidantes e diminui os níveis dos parâmetros oxidantes.

Effect of diesel exhaust exposure on 8-hydroxy-2' deoxyguanosine level in biological samples of automobile manufacturing workers / 中国职业医学

OBJECTIVE: To investigate the feasibility of using 8-hydroxy-2'deoxyguanosine(8-OHdG) in blood and urine samples as biomarkers for the evaluation of human DNA oxidative damage caused by diesel exhaust(DE). METHODS: A convenient sampling method was used to select 56 male workers exposed to DE in a car manufacturing factory as exposure group, and 52 male workers without exposure to DE were selected as the control group.Urine samples and blood samples were collected from workers in the 2 groups 8 hours after work, and the levels of 8-OHdG in urine and plasma were measured by ultra-high performance liquid chromatography tandem mass spectrometer. RESULTS: The median level of urinary 8-OHdG in the exposure group was higher than that of control group(2.54 vs 2.03 μg/g Cr, P<0.05). The median levels of plasma 8-OHdG in the exposure group and control group showed no statistical significance(32.20 vs 31.40 ng/L, P>0.05). CONCLUSION: The urinary 8-OHdG can be used as a biomarker for evaluating the oxidative damage induced by DE exposure.

Nerium oleander Distillate Can Reduce Oxidative Deoxyribonucleic Acid Damage in Rats Fed with High Cholesterol Diet

Aim: To evaluate the effect of Nerium oleander distillate on the high cholesterol diet(HCD) induced oxidative deoxyribonucleic acid damage via assessing blood 8-hydroxy-2'-deoxyguanosine(8-OHdG) and superoxide dismutase(SOD) levels. Methodolody: Twenty-one male Sprague-Dawley rats were divided equally into three groups. The first group (control group) was fed a normal diet and administered 0.5 ml distilled water via gavage for 90 days. The second and third groups were fed with HCD. The second group was administered 0.5 ml distilled water and the third group was administered 0.5 ml Nerium oleander distillate(0.375 mg/rat) via gavage for 90 days, after being fed the HCD for two weeks. Blood samples were collected, and 8-OHdG and SOD levels were measured. Results: 8-OHdG levels were statistically significantly different in all groups. Highest 8-OHdG levels were determined in the second group whereas Nerium oleander treatment reduced the level of 8-OHdG. In addition, decreased SOD levels were detected in the rats fed with HCD(Groups 2 and 3) when compared to the control group. Conclusion: It may be stated that HCD may cause oxidative damage in deoxyribonucleic acid and Nerium oleander distillate may reduce this damage. Hence, Nerium oleander distillate may show beneficial effects in the treatments of diabetes mellitus, hypercholesterolemia, and metabolic syndrome. In the future, it should investigate the effect of Nerium oleander distillate on different antioxidant pathways.

Serum 8-hydroxy-2-deoxyguanosine level of radiation workers in hospitals / 中华放射医学与防护杂志

Objective To explore the influence of long-term low-dose ionizing radiation on 8-hydroxy-2-deoxyguanosine(8-OHdG) level in the serum of radiation workers in hospitals.Methods 307 age-and sex-matched hospital radiation workers were recruited by stratified random sampling method.After deleting the subjects without dosage information,230 individuals were divided into four groups according to their job title [including diagnostic radiology (n =75),radiotherapy (n =60),nuclear medicine (n =41) and interventional radiology (n =54)].Serum 8-OHdG level was measured by ELISA assay.Results According to the statistical analysis,there was significant difference in the serum 8-OHdG level among four groups (F =9.071,P ＜ 0.05),and the content of serum 8-OHdG was significantly higher in the interventional radiology group than that in the groups of diagnostic radiology,radiotherapy and nuclear medicine (t =-4.473,-3.011,-2.189,P ＜ 0.05).There were significant differences in serum 8-OHdG level among different dose groups and working period groups(F =7.659,3.058,P ＜ 0.05).The serum 8-OHdG levels significantly increased along with exposure dose and working period (r =0.300,0.142,P ＜ 0.05).Conclusions Serum 8-OHdG may be a potential biomarker of oxidative DNA damage in radiation workers exposed to low-dose ionizing radiation.

Simultaneous determination of nucleosides and nucleobases in Cervi Cornu Pantotrichum of sika deer (Cervus nippon) with different processing methods by UPLC / 中草药

Objective To establish the method of simultaneous determination of the content of 13 nucleosides and nucleobases, including cytosine, uracil, adenine, guanine, 6-hydroxypurine, 2,6-dihydroxypurine, uridine, thymine, inosine, guanosine, adenosine, 2′-deoxyguanosine (2′-dG), beta-thymidine, in Cervi Cornu Pantotrichum (CCP) of sika deer (Cervus nippon) by UPLC, and compare the distribution differences of nucleosides and nucleobases in different zones of the CCP with different processing methods. Methods The nucleosides and nucleobases in CCP were extracted by water with assistance of ultrasound. Acquity UPLC HSS T3 column (100 mm × 2.1 mm, 1.8 μm) was used as chromatographic column to separate the nucleosides and nucleobases. Thirteen target compounds were eluted with acetonitrile 100% (eluent A) and water plus 0.006% formic acid (eluent B) at a flow rate of 0.3 mL/min. The column temperature was 30 ℃, and the injection volume was 3 μL and the detection wavelength was 260 nm. Results A total of 13 nucleosides and nucleobases basically reached the baseline separation with a good linearity within linear range (r > 0.999 6). The nucleosides and nucleobases content in wax slices, powder slices, gauze slices of CCP without and with blood were respectively 4.47, 3.95, 2.68 g/kg and 4.14, 3.44, 2.51 g/kg. And those three parts of CCP with boiling and freeze-drying processing were respectively 4.60, 2.95, 2.74 g/kg and 5.06, 4.24, 2.31 g/kg. Conclusion As far as the total content of nucleosides and nucleobases were concerned, the wax slices, powder slices and gauze slices of CCP without blood were all higher than those of CCP with blood. The wax slices, powder slices of CCP with freeze-drying processing were more than those of CCP with boiling processing, while the gauze slices of which were less than those of CCP with boiling processing.

Relationship of PAHs metabolites,DNA oxidative damage and RING2 expression in coke oven workers / 中国职业医学

OBJECTIVE: To investigate the relationship of polycyclic aromatic hydrocarbons( PAH) metabolites,DNA oxidative damage and ring finger protein 2( RING2) expression in coke oven workers. METHODS: A judgment sampling method was used to select 497 coke oven workers in a steel plant as exposure group and 175 water treatment workers in the same plant as control group. The levels of urinary 1-hydroxypyrene, 2-hydroxynathalene, 2-hydroxyfluorene,9-hydroxyphenanthrene and 8-hydroxy deoxyguanosine(8-OHd G) were detected by high performance liquid chromatography.The RING2 expression in whole blood was measured by reverse transcription-polymerase chain reaction. RESULTS: The relative expression of urinary 1-hydroxypyrene,2-hydroxynathalene,2-hydroxyfluorene,9-hydroxyphenanthrene and RING2 in exposure group were higher than that in control group( P < 0. 01). The logistic regression analysis indicated that the higher the level of 1-hydroxypyrene,the higher the risk of high-RING2 expression( P < 0. 05) after adjusting for factors such as sex,age,smoking status,alcohol drinking,2-hydroxynathalene,2-hydroxyfluorene and 9-hydroxyphenanthrene.In 1-hydroxypyrene middle and high level groups,the 8-OHd G concentration of high-RING2 expression workers was significantly higher than those of low-RING2 expression workers( P < 0. 05). CONCLUSION: With the increase of urinary1-hydroxypyrene,the risk of high-RING2 expression was elevated,the degree of DNA oxidative damage was gradually increased.

DNA damage in homocystinuria: 8-oxo-, 8-dihydro-2'-deoxyguanosine levels in cystathionine-ß-synthase deficient patients and the in vitro protective effect of N-acetyl­L­cysteine

Introduction: Homocysteine (Hcy) tissue accumulation occurs in a metabolic disease characterized biochemically by cystathionine ß-synthase (CBS) deficiency and clinically by mental retardation, vascular problems, and skeletal abnormalities. Previous studies indicate the occurrence of DNA damage secondary to hyperhomocysteinemia and it was observed that DNA damage occurs in leukocytes from CBS-deficient patients. This study aimed to investigate whether an oxidative mechanism could be involved in DNA damage previously found and investigated the in vitro effect of N-acety-L-cysteine (NAC) on DNA damage caused by high Hcy levels. Methods: We evaluated a biomarker of oxidative DNA damage in the urine of CBS­deficient patients, as well as the in vitro effect of NAC on DNA damage caused by high levels of Hcy. Moreover, a biomarker of lipid oxidative damage was also measured in urine of CBS deficient patients. Results: There was an increase in parameters of DNA (8-oxo-7,8-dihydro-2'- deoxyguanosine) and lipid (15-F2t-isoprostanes levels) oxidative damage in CBS-deficient patients when compared to controls. In addition, a significant positive correlation was found between 15-F2t-isoprostanes levels and total Hcy concentrations. Besides, an in vitro protective effect of NAC at concentrations of 1 and 5 mM was observed on DNA damage caused by Hcy 50 µM and 200 µM. Additionally, we showed a decrease in sulfhydryl content in plasma from CBS-deficient patients when compared to controls. Discussion: These results demonstrated that DNA damage occurs by an oxidative mechanism in CBS deficiency together with lipid oxidative damage, highlighting the NAC beneficial action upon DNA oxidative process, contributing with a new treatment perspective of the patients affected by classic homocystinuria.

Changes and Significance of Serum 8-hydroxy-2'-deoxyguanosine in Kawasaki Disease Children with Coronary Artery Lesions / 医学研究杂志

Objective To investigate the changes of serum 8-hydroxy-2'-deoxyguanosine(8-OHDG) in Kawasaki disease (KD) children and to explore the importance of 8-OHDG in predicting the severity of coronary artery lesions in Kawasaki Disease.Methods The serum 8-OHDG was measured in KD patients group (n =60),fever patients group (n =12) and health control group (n =12) by ELISA method.Among the KD patients,30 KD patients were in acute stage (10 cases had coronary artery lesions,20 coronary were normal) and 30 patients were in recovery stage.The serum 8-OHDG and brain natriuretic peptide (BNP) were also compared between patients with coronary artery lesions (CALs) and patients with no coronary artery lesions NCALs).ROC curve analysis was used to test the 8-OHDG and BNP predictive values in KD with coronary artery lesions.Results The serum level of 8-OHDG was higher in acute KD patients than recovery KD patients,fever patients and healthy children (P ＜ 0.05) The serum 8-OHDG was higer in CALs patients than the NCALs patients (P ＜ 0.05).The serum BNP was higer in CALs patients than the NCALs patients (P ＜ 0.01).Analysis of the ROC curve showed that serum 8-OHDG had a positive predictive value of 64.3％ and a negative predictive value of 93.8％for distinguishing KD with CALs from KD NCALs when cutoff value was 57.02pg/ml.The area under the curve was 0.820.The serum BNP had a a positive predictive value of 47.6％ and a negative predictive value of 100％ for distinguishing KD with CALs from KD NCALs when cutoff value was 815pg/ml.The area under the curve was 0.745.Conclusion The serum 8-OHDG may have better predictive value than BNP in diagnose KD children with coronary artery lesions.

The significance of 8-hydroxy-deoxyguanosine acid in the diagnosis of nonalcoholic steatohepatitis / 中华内科杂志

Objective To evaluate the significance of serum 8-hydroxy-deoxyguanosine acid ( 8-OHdG) in the diagnosis of nonalcoholic steatohepatitis ( NASH).Methods Patients or healthy subjects were enrolled at the Second Hospital of Tianjin Medical University and the Second People ′s Hospital of Tianjin from May 2013 to December 2015.A total of 41 patients with nonalcoholic fatty liver disease were enrolled in the study , including 20 nonalcoholic simple fatty liver ( NAFL) patients and 21 NASH patients whose diagnosis were proven by liver biopsy.The other 32 healthy subjects were studied as controls.Serum 8-OHdG, ALT, AST and GGT were tested.Nonalcoholic fatty liver disease activity score ( NAS ) and expression of 8-OHdG in liver was investigated between NAFL patients and NASH patients.The correlations between serum 8-OHdG and serum ALT , AST, GGT, and 8-OHdG in liver tissue in NASH group were investigated.In addition , the receiver operating characteristic ( ROC) curve analyses for ALT and 8-OHdG levels were performed in NAFL patients and NASH patients , and the cut-off value was determined.Results Serum 8-OHdG values in healthy controls , NAFL and NASH patients were (0.19 ±0.16) μg/L, (0.22 ±0.16) μg/L, (0.42 ±0.21) μg/L respectively.The serum 8-OHdG and serum ALT, GGT and 8-OHdG in liver tissue were all positively correlated in NASH group with respective correlation coefficient r values as 0.454 7, 0.382 9, and 0.497 6.AUC of 8-OHdG was 0.901 with cut-off value 0.39 μg/L.Its sensitivity was 88.3%and specificity was 81.5%, which were higher than those of ALT.Conclusion The value of serum 8-OHdG would be used as a marker for the diagnosis of NASH.

Study on the expression level of 8-Hydroxy-2'-deoxyguanine in rhesus monkeys of different ages / 中华老年医学杂志

Objectives To investigate the expression of 8-Hydroxy-2'-deoxyguanine(8-oxodG)in white blood cell,plasma and urine of rhesus monkey of different age group.Methods 30 female rhesus macaques at different age(1y,5y,10y,15y,20y,25y)were selected and grouped(n=5,each).10 mL of morning urine and 5 mL of fasting venous blood were collected.The level of 8-oxodG expression in plasma,leukocyte and urine was measured by high-performance liquid chromatography-mass spectrometry(HPLC-MS) method.Results The level of 8-oxodG in leukocytes,plasma and urine was increased along with aging.The level of 8-oxodG was 1.8,1.6 and 1.4 times higher in 25 year group than in 1 year group in plasma,white blood cell and urine,respectively(P＜0.05).The 8-oxodG level was more than 40 times higher in urine than in plasma.Conclusions The expression level of 8-oxodG is increased along with aging.It may be one of the experimental evidence of the aging markers.

Expression and significance of 8-OHdG in primary pterygium / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To detect oxidative DNA damage marker 8-hydroxy-2-deoxyguanosine(8-OHdG)in primary pterygium and normal conjunctival tissues, explore the role of oxidative DNA damage in the pathogenesis of pterygium. <p>METHODS: Totally 35 primary pterygium specimens were collected during surgery and 5 normal conjunctival specimens which above the normal temporal bulbar conjunctiva were collected. The expressions of 8-OHdG in pterygium tissues were detected by immunohistochemical method and compared with the normal conjunctival tissues. The difference of 8-OHdG expression between the two groups was compared. <p>RESULTS: There were 24(69%)pterygium specimens positive for 8-OHdG staining, limited to the nuclei of the epithelial layer. No substantial staining was visible in the subepithelial fibrovascular layers. All normal controls were negative for 8-OHdG staining. The difference of 8-OHdG expression between the two groups was statistically significant(<i>P</i>=0.007). <p>CONCLUSION: The increased levels of 8-OHdG in the pterygium tissues indicate that oxidative DNA damage maybe play an important role in the pathogenesis of pterygium.

Expression of 8-oxodG in Oral Leukoplakia / 沈阳医学院学报

Objective:To study the expression of 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG) in oral leukoplakia for clarifying the role of oxidative DNA damage in the development of oral leukoplakia. Methods:Immunofluorescence labeling method was used to examine the expression of 8-oxodG,a marker of oxidative DNA damage,and the expression of tumor suppressor gene, P53 protein in oral epithelium of normal oral mucosa and biopsy specimens of leukoplakia. Results:In specimens of oral leukoplakia, HE staining showed infiltration of inflammatory cells, hyperkeratosis and epithelial dysplasia. Immunofluorescence labeling study demonstrated that the accumulation of 8-oxodG apparently increased in the oral epithelium of patients with leukoplakia,whereas little or no immunoreactivity was observed in normal oral mucosa. Expression of P53 protein was also observed in oral epithelium of patients with oral leukoplakia. The immunoreactivity of 8-oxodG and P53 was stronger in patients with oral leukoplakia than that in normal oral mucosa (P<0.01) . Moreover,the immunoreactivity increased with the development of disease (r=0.773, P<0.01) . Conclusions:The oxidative DNA damage contributes to the development of oral leukoplakia. 8-oxodG may be a risk predictive marker for oral leukoplakia.

Effect of DNA single strand breaks and 8-OHd G production on cadmium-induced toxicity on human embryonic kidney epithelial cells / 中国职业医学

OBJECTIVE: To explore the effects of cadmium chloride( CdCl_2) on DNA single strand breaks and the production of 8-hydroxy-2'-deoxyguanosine( 8-OHdG) in human embryonic kidney epithelial cells( HEK cells). METHODS: HEK cells in logarithm growth phase were divided into 5 groups and incubated with the different concentrations of CdCl_2( 0. 0,2. 5,5. 0,10. 0 and 20. 0 μmol/L) for 24,48 and 72 hours in vitro. After harvesting the cells,DNA single strand breaks was tested by single cell gel electrophoresis,and the level of 8-OHdG was measured using the enzyme linked immunosorbent assay. RESULTS: The Olive tail moment was statistically significant in the main effect of CdCl_2 exposed HEK cells( P < 0. 01). Among them,when HEK cells were exposed to 5. 0 μmol / L of CdCl_2,the Olive tail moment began to have a statistical significant increasing trend compared with the 0. 0 μmol / L group( P < 0. 05); when CdCl_2 concentration was 2. 5-10. 0 μmol / L,the Olive tail moment lengthened with the increasing dose of cadmium exposure,showing a doseeffect relationship( P < 0. 05). The tail DNA% was statistically significant in the interaction between exposure treatment and exposure time in HEK cells( P < 0. 01). Among them,when CdCl_2 concentration was at 2. 5-10. 0 μmol / L at 24 hours time point and 5. 0-20. 0 μmol / L at 48 hours time point,the tail DNA% raised with the increasing dose of cadmium exposure,showing a dose-effect relationship( P < 0. 05). The tail DNA% at 3 time points of 24,48 and 72 hours after exposure to 20. 0 μmol / L of CdCl_2 in HEK cells increased with the increasing time of cadmium exposure,showing a timeeffect relationship( P < 0. 05). The level of 8-OHdG had statistical significance in the main effect of CdCl_2 exposure treatment in HEK cells( P < 0. 05). Among them,the level of 8-OHdG was first significantly increased only after exposure to 10. 0 μmol / L CdCl_2 compared with the 0. 0 μmol / L group( P < 0. 05). After treatment with Ca Cl2,there was no doseeffect relationship and time-effect relationship found between the cadmium chloride exposure and tail length as well as the tail / head length ratio and 8-OHdG level. CONCLUSION: To a certain extent,CdCl_2 exposure may cause both DNA single strand breaks and 8-OHdG production in HEK cells. Compared with 8-OHdG,the DNA single strand breaks show more significant change with a lower dose of cadmium treatment,which may be related to its higher sensitivity to cadmium toxicity than 8-OHdG.

Urinary 8‑hydroxydeoxyguanosine as a marker of oxidative stress induced genetic toxicity in oral cancer patients.

Context: Recently, non‑communicable diseases have snatched the lead from infectious diseases in causing mortality. Of these, oral cancer accounts for a significant proportion of deaths. Every year in India significant percentage of newly diagnosed malignancy is oral cancer attributed to various reasons. Aims: The aim of this study was to assess the extent of oxidative stress and its effect on modification of DNA by urinary nucleoside 8-hydroxy-2’-deoxyguanosine (8-OHdG) levels in oral cancer subjects. To see the relationship between the nucleoside 8‑OHdG and antioxidant capacity ferric reducing ability plasma (FRAP) in oral cancer subjects. Settings and Design: Case–control study included three groups with 60 volunteers, who were divided into 30 controls, and equal number of clinically diagnosed oral cancer male patients: (Subdivided into newly diagnosed [n = 15] and 1‑year treatment follow‑up oral cancer subjects [n = 15]). Materials and Methods: A random urine sample was used for analysis of 8‑OHdG concentration. Serum triglycerides, lipid peroxidation, protein thiols, and FRAP assay were performed by spectrophotometric technique. Statistical Analysis Used: Student’s t‑test and one‑way analysis of variance were performed for group comparison and Pearson’s correlation analysis were used. A P < 0.05 was considered the optimum level of significance. Results: The urinary 8‑OHdG and serum malondialdehyde levels were significantly elevated in newly diagnosed oral cancer subjects in their 1‑year treatment compared to the control group (P < 0.05). A significant correlation was observed between urinary 8‑OHdG and FRAP in both groups of oral cancer subjects. Conclusions: Urinary 8‑OHdG can be a useful diagnostic marker of oxidative DNA damage in oral cancer subjects.

Relationship Between Mitochondrial DNA Related Factors and Traditional Chinese Medical Syndrome Types in Primary Osteoporosis Patients / 广州中医药大学学报

Objective To investigate the biological characteristics of primary osteoporosis syndrome types from the perspective of mitochondrial DNA ( mtDNA) , thus to reveal the nature of osteoporosis and its traditional Chinese medical syndrome types. Methods A total of 210 osteoporosis women patients meeting the diagnostic criteria, inclusion criteria and exclusion criteria were collected from July of 2011 to October of 2013. The osteoporosis patients were differentiated into the syndrome types of yin deficiency of liver and kidney ( N=67) , yang deficiency of spleen and kidney ( N=70) and qi stagnation and blood stasis ( N=73) . And a total of 69 age-matched post-menopause non-osteoporosis patients were chosen as the control group, which were classified into the syndrome of harmony of Qi and blood. The peripheral blood was sampled for detecting mtDNA copy number with fluorescent quantitatitation PCR and for examining 8-hydroxy-2’-deoxyguanosine ( 8-OHdG) content by enzyme-linked immunosorbent assay (ELISA) . Statistical methods was used to analyze the correlation of bone mineral density (BMD) with mtDNA copy number and 8-OHdG content in different groups. Results The difference of mtDNA copy number was significant between the osteoporosis patients and non-osteoporosis patients (P<0.05), and was also significant among the three syndrome types of osteoporosis patients (P<0.05) . And 8-OHdG content showed the same features between the osteoporosis patients and non-osteoporosis patients (P<0.05) and among the three syndrome types of osteoporosis patients (P<0.05) . The correlation analysis results showed that mtDNA copy number was positively correlated with BMD, while 8-OHdG was negatively correlated with BMD in each group. Conclusion The mtDNA copy number and 8-OHdG content are correlated with the syndrome types of primary osteoporosis patients, and close correlation is shown between spleen-kidney yang deficiency and 8-OHdG, and between liver-kidney yin deficiency and mtDNA copy number.

Iron Indices and Urinary 8-Oxo-7, 8-Dihydro-2’- Deoxyguanosine (8-Oxodg) in Patients with Cervical Intraepithelial Neoplasia.

Aim: To investigate the role of iron status in cervical carcinogenesis through its involvement in the Haber-Weis and Fenton reactions serving as a pathway to carcinogenesis and using 8-oxo-7, 8- dihydro-2’-deoxyguanosine (8-oxodG) as a marker of DNA oxidation in a population where iron deficiency is prevalent. Study Design: It is a cross sectional study. Place of Study: The patients were recruited from the colposcopy clinic of the University College Hospital (UCH), Ibadan and Obafemi Awolowo University Teaching Hospitals Complex, Ile-Ife, Nigeria. The laboratory investigations were carried out at the Haematology and Chemical Pathology laboratories of UCH, Ibadan and Oxidative Stress Group, Department of Cancer Studies and Molecular Medicine, University of Leicester, Leicester, UK. Methodology: Forty-five subjects with CIN and 41 with normal Pap smear result (non-CIN) were recruited. A structured questionnaire was administered to collect information on demographic characteristics, dietary, social and medical history. Fasting blood sample were collected to assess for serum iron, total iron binding capacity and transferrin saturation. Urine was also collected to analyze for creatinine and 8-oxodG. Results: The CIN subjects had more babies; > 5 than non-CIN subjects (P=.003). The individuals with > 5 children were 4 times more likely to have CIN [OR 3.79 (95% CI 1.3-10.33), P=.01]. CIN subjects had higher serum iron and transferrin saturation than non-CIN subjects. Though the mean urinary 8-oxodG level similar between the two groups, there was a trend towards higher levels in individuals with high grade CIN. Conclusion: High serum iron level was linked to frequent ingestion of iron supplement and may contribute to progression of CIN with a potential role for urinary 8-oxodG as a useful bio indicator of altered iron homeostasis and associated DNA damage.

Oxidative damage biomarker levels according to Mn-SOD and GST gene polymorphisms in preschool children / 한국영양학회지

PURPOSE: Genetic polymorphisms in antioxidant defense and detoxification genes may modulate the levels of oxidative stress biomarkers. METHODS: A total of 301 healthy preschool-aged children in the Seoul and Kyung-gi areas were recruited. DNA was extracted from blood for genotyping of manganese superoxide dismutase (Mn-SOD) Val16Ala, glutathione S-transferase (GST) P1 Ile105Val, GSTT1 present/null, and GSTM1 present/null polymorphisms by PCR-restriction fragment length polymorphism or multiplex PCR analyses. In addition to a questionnaire survey, the levels of urinary 8- hydroxyl-2-deoxiguanosine (8-OHdG) and plasma malondialdehyde (MDA) were measured by ELISA. RESULTS: Significantly higher urinary 8-OHdG concentrations were observed in GSTP1 Ile/Val + Val/Val genotype (p = 0.030), and tended to be higher in Mn-SOD Val/Val genotype (p = 0.065). On the other hand, exposure to environmental tobacco smoking (ETS) and interaction between ETS and gene polymorphisms did not significantly influence either urinary 8-OHdG concentrations or serum MDA. CONCLUSION: Based on our findings, GSTP1 Ile/Val gene polymorphisms might modulate the levels of oxidative stress biomarkers in healthy preschool children.