Research Progress of DNA-Based Technologies for Postmortem Interval Estimation / 法医学杂志

In criminal investigations, postmortem interval (PMI) is important information to be inferred in homicide investigations, as well as the focus and the difficulty in forensic pathology research. Because the DNA content in different tissues is relatively constant and shows changes regularly with the extension of PMI, it has become a research hotspot of PMI estimation. This paper reviews the recent progress of PMI estimation technologies including DNA-based single cell gel electrophoresis, image analysis, flow cytometry, real-time fluorescence quantitative PCR and high-throughput sequencing, hoping to provide references for forensic medicine practice and scientific research.

The application of transjugular liver biopsy in patients with unexplained liver disease / 中华肝胆外科杂志

Objective:To investigate the clinical value of transjugular liver biopsy (TJLB) in patients with unexplained liver disease complicated with massive ascites or coagulopathy.Methods:A retrospective analysis was performed from patients underwent TJLB in the First Affiliated Hospital of Zhengzhou University, Zhoukou Central Hospital, Shangqiu First People's Hospital and Jincheng People's Hospital from March 2015 to January 2022 due to unexplained liver disease complicated with massive ascites or coagulopathy. A total of 37 patients were included, including 21 males and 16 females, aged (53.5±11.9) years. According to different puncture points, the patients were divided into two groups: transhepatic right vein TJBL and transhepatic middle vein TJBL. The obtained liver tissue sampling effect, puncture times, complications were analyzed.Results:The success rate of TJLB was 97.3%(36/37). Thirty-six patients were able to obtain more than three segments of liver tissue and obtain histological diagnosis, and the pathological diagnosis rate was 100.0%(36/36). The number of puncture times, the amount of hepatic tissue and the number of portal areas in the right hepatic vein group (21 cases) were (3.7±0.9), (3.7±0.7) and (6.5±0.9) respectively, and those in the middle hepatic vein group (15 cases) were (3.7±0.7), (3.7±0.7) and (6.3±0.8) respectively. There were no significant differences between the two groups (all P>0.05). Conclusion:TJLB is safe and feasible for patients with unexplained liver disease complicated with massive peritoneal effusion and coagulopathy. Good liver tissue specimens can be obtained by TJLB from both right hepatic vein and middle hepatic vein.

Association of etheno-DNA adduct and DNA methylation level among workers exposed to diesel engine exhaust / 中华预防医学杂志

Objective@#To investigate the association between etheno-DNA adduct and the promoter of DNA methylation levels of cyclin dependent kinase inhibitor 2A (P16), Ras association domain family 1 (RASSF1A) and O-6-methylguanine-DNA methyltransferase (MGMT) in workers with occupational exposure to diesel engine exhaust (DEE).@*Methods@#We recruited 124 diesel engine testing workers as DEE exposure group and 112 water pump operator in the same area as control group in Henan province in 2012 using cluster sampling. The demographic data were obtained by questionnaire survey; urine after work and venous blood samples were collected from each subject. The urinary etheno-DNA adducts were detected using UPLC-MS/MS, including 1,N6-etheno-2'-deoxyadenosine (εdA) and 3,N4-etheno-2'-deoxycytidine(εdC). The DNA methylation levels of P16, RASSF1A, and MGMT were evaluated using bisulfite-pyrosequencing assay. The percentage of methylation was expressed as the 5-methylcytosine (5mC) over the sum of cytosines (%5mC). Spearman correlation and multiple linear regression were applied to analyze the association between etheno-DNA adducts and DNA methylation of P16, RASSF1A, and MGMT.@*Results@#The median (P25-P75) of urinary εdA level was 230.00 (98.04-470.91) pmol/g creatinine in DEE exposure group, and 102.10 (49.95-194.48) creatinine in control group. The level of εdA was higher in DEE exposure group than control group (P<0.001). DNA methylation levels of P16, RASSF1A and MGMT were 2.04±0.41, 2.19 (1.94-2.51), 2.22 (1.94-2.46)%5mC in exposure group, and 2.19±0.40, 2.41 (2.11-2.67), 2.44 (2.15-2.91)%5mC in control group. DNA methylation levels were lower in exposure group (P values were 0.005, 0.002 and 0.001, respectively). Spearman correlation analysis showed that DNA methylation levels of P16, RASSF1A, and MGMT were negative associated with urinary εdA level (r values were -0.155, -0.137, and -0.198, respectively, P<0.05). No significant correlation was observed between the εdC level and any measured DNA methylation levels (P>0.05) . Multiple linear regression confirmed the negative correlation between εdA and DNA methylation levels of P16, RASSF1A, and MGMT in non-smoking group (β (95%CI) was -0.068 (-0.132--0.003), -0.082 (-0.159--0.004) and -0.048 (-0.090--0.007), P values were 0.039, 0.039 and 0.024, respectively). Moreover, εdC was negative associated with DNA methylation level of MGMT in non-smoking group (β (95%CI) was -0.094 (-0.179--0.008), P=0.032).@*Conclusion@#DEE exposure could induce the increased of εdA and decreased of DNA methylation levels of P16, RASSF1A and MGMT.

Pentacam's anterior segment analysis system evaluates the efficacy of Lotemax in controlling postoperative inflammation following cataract surgery / 眼科新进展

Objeetive To observe the efficacy and safety of 5 g · L-1 Lotemax suspended eye drops in inflammation control after phacoemulsification combined with intraocular lens implantation.Methods Together 60 patients with senile cataract (grade III or more severe) underwent cataract surgery were selected and randomly divided into two groups,including Lotemax group,in which patients were given 5 g · L-1 Lotemax suspended eye drops and Tobramycin eye drops,and Tobradex group,in which patient were alone given Tobradex eye drops.Corneal edema,anterior chamber flare,corneal volume,anterior chamber volume and corneal incision thickness were observed at 1 day,7 days and 30 days after operation,while intraocular pressure was observed at 7 days and 30 days before and after operation.Then the efficacy of Lotemax in control of postoperative inflammation was evaluated by Pentacam's anterior segment analysis system.Results There was no significant difference in corneal edema,anterior chamber flare,corneal volume,anterior chamber volume and corneal incision thickness between the two groups at postoperative 1 day and 30 days (all P ＞ 0.05),but Tobradex group had weak anterior chamber flare and lower corneal volume than Lotemax group (P =0.029,0.019),while the difference in corneal edema,anterior chamber volume and corneal incision thickness was not statistically significant between two groups at postoperative 7 days (P =0.119,0.138,0.101).Moreover,there was no significant difference in intraocular pressure at 7 days and 30 days before and after surgery between the two groups (all P ＞ 0.05).Conclusion Lotemax suspended eye drops can effectively control the inflammation followed by cataract phacoemulsification combined with intraocular lens implantation.

Pentacam's anterior segment analysis system evaluates the efficacy of Lotemax in controlling postoperative inflammation following cataract surgery / 眼科新进展

Objeetive To observe the efficacy and safety of 5 g · L-1 Lotemax suspended eye drops in inflammation control after phacoemulsification combined with intraocular lens implantation.Methods Together 60 patients with senile cataract (grade III or more severe) underwent cataract surgery were selected and randomly divided into two groups,including Lotemax group,in which patients were given 5 g · L-1 Lotemax suspended eye drops and Tobramycin eye drops,and Tobradex group,in which patient were alone given Tobradex eye drops.Corneal edema,anterior chamber flare,corneal volume,anterior chamber volume and corneal incision thickness were observed at 1 day,7 days and 30 days after operation,while intraocular pressure was observed at 7 days and 30 days before and after operation.Then the efficacy of Lotemax in control of postoperative inflammation was evaluated by Pentacam's anterior segment analysis system.Results There was no significant difference in corneal edema,anterior chamber flare,corneal volume,anterior chamber volume and corneal incision thickness between the two groups at postoperative 1 day and 30 days (all P ＞ 0.05),but Tobradex group had weak anterior chamber flare and lower corneal volume than Lotemax group (P =0.029,0.019),while the difference in corneal edema,anterior chamber volume and corneal incision thickness was not statistically significant between two groups at postoperative 7 days (P =0.119,0.138,0.101).Moreover,there was no significant difference in intraocular pressure at 7 days and 30 days before and after surgery between the two groups (all P ＞ 0.05).Conclusion Lotemax suspended eye drops can effectively control the inflammation followed by cataract phacoemulsification combined with intraocular lens implantation.

Method validation of phosphorylated histone H2AX level detection using primary cultured hepatocytes in genotoxic agent screening / 中国药理学与毒理学杂志

OBJECTIVE To establish an in vitro test method and to evaluate the genotoxicity of chemicals using primary cultured mouse hepatocytes and the changes in phosphorylated histone H2AX(γH2AX)expression levels to provide a more reliable marker of the identification of genotoxicity. METHODS Hepatocytes were isolated from BALB/c mice by an improved two-step collagenase diges?tion method and then cultured in sandwich configuration. The primary cultured hepatocytes were treat?ed with various concentrations of four known genotoxic agents bleomycin(BLM),benzo(a)pyrene〔B (a)p〕,styrene and styrene-7,8-oxide(SO)within the range of 40 μmol · L-1 and two non-genotoxic agents azathioprine(Aza)and ciclosporin A(CsA)at different time points within 24 h. The cytotoxicity induced by these toxicants was assessed by CCK-8 assay. Then,the changes in γH2AX expression levels in treated cells were determined by flow cytometry. RESULTS The four genotoxic agents could be detected and two non-genotoxic agents could not be detected by this method. The γH2AX expression level was the highest when hepatocytes were exposed to BLM and SO for 3 h,or B(a)p and styrene for 6 h(P<0.01). The production of γH2AX was 25.67,18.36,12.43 and 14.25 for the four types of genotoxic agents,respectively,and was approximately 19,13,9 and 11 times that of the vehicle control group(P<0.01)at the optimum time point and concentration. There was a significant positive corre?lation between the indicated concentrations of genotoxic chemicals and γH2AX expression levels(P<0.01). In addition,the production ofγH2AX indicated no marked increase in two non-genotoxic agents such as Aza and CsA in comparison with the control group. CONCLUSION This test method can effec?tively distinguish genotoxic agents from non-genotoxic agents,and direct genotoxic agents from indirect genotoxic agents in the absence of S9. γH2AX might be a reliable marker for the identification of the potential genotoxicity of chemicals.

Effect of occupational exposure to toluene diisocyanate on workers' health / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the effect of occupational exposure to toluene diisocyanate (TDI) on the workers' health.</p><p><b>METHODS</b>A total of 76 workers exposed to TDI (exposure group) and 64 management staff members (control group) were selected from a factory as the study subjects. Area sampling was performed for the place with exposure to TDI according to the method in GBZ 159-2004 Specifications of air sampling for hazardous substances monitoring in the workplace, and gas chromatography was applied to measure the concentration of TDI in workplace air. The workers' personal information was collected with questionnaire, pulmonary ventilation function was determined with a portable spirometer, hematological parameters were analyzed by automatic blood analyzer and blood chemistry analyzer, and the indicators of oxidative damage and energy metabolism were measured by the reagent kit provided by Nanjing Jiancheng Bioengineering Institute. SPSS 17 software was applied for statistical analysis.</p><p><b>RESULTS</b>The exposure group had significantly lower forced vital capacity (FVC), forced expiratory volume in 1 second(FEV1.0), and FEV1.0/FVC ratio than the control group (P <0.05). Compared with the control group, the exposure group had significantly higher red blood cell count, platelet distribution width, mean platelet volume, lymphocyte count, and neutrophil count(P<0.01), and significantly lower activities of lactate dehydrogenase(LDH), superoxide dismutase, and succinodehydrogenase (SDH)(P <0.01). In the exposure group, the length of exposure was negatively correlated with the activities of SDH and LDH in the serum (r=-0.319, P <0.05; r=-0.239, P <0.05), and the length of exposure was not found to be correlated with the activity of SOD and pulmonary function indices.</p><p><b>CONCLUSION</b>TDI can induce inflammatory response and lung ventilation function impairment in workers exposed to TDI, as well as oxidative stress and imbalance of energy metabolism. Therefore, it can cause damage to workers' health, and protective measures should be enhanced.</p>

Size distribution characteristics of particulate matter in the top areas of coke oven / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To systematically evaluate the environmental exposure information of coke oven workers, we investigated the concentration and size distribution characteristics of the particle matter (PM) in the top working area of coke oven.</p><p><b>METHODS</b>The aerodynamic particle sizer spectrometer was employed to collect the concentration and size distribution information of PM at a top working area. The PM was divided into PM ≤ 1.0 µm, 1.0 µm < PM ≤ 2.5 µm, 2.5 µm < PM ≤ 5.0 µm, 5.0 µm < PM ≤ 10.0 µm and PM>10.0 µm based on their aerodynamic diameters. The number concentration, surface area concentration, and mass concentration were analyzed between different groups. We also conducted the correlation analysis on these parameters among groups.</p><p><b>RESULTS</b>We found the number and surface area concentration of top area particulate was negatively correlated with particle size, but mass concentration curve showed bimodal type with higher point at PM = 1.0 µm and PM = 5.0 µm. The average number concentration of total particulate matter in the top working area was 661.27 number/cm³, surface area concentration was 523.92 µm²/cm³, and mass concentration was 0.12 mg/m³. The most number of particulate matter is not more than 1 µm (PM(1.0)), and its number concentration and surface area concentration accounted for 96.85% and 67.01% of the total particles respectively. In the correlation analysis, different particle size correlated with the total particulate matter differently. And the characteristic parameters of PM2.5 cannot fully reflect the total information of particles.</p><p><b>CONCLUSION</b>The main particulate matter pollutants in the top working area of coke oven is PM1.0, and it with PM(5.0) can account for a large proportion in the mass concentration of PM. It suggest that PM1.0 and PM(5.0) should be considered for occupational health surveillance on the particulate matter in the top area of coke oven.</p>

Changes of Th17 cytokine in human bronchial epithelial cells induced by coke oven emission / 中国药理学与毒理学杂志

OBJECTIVE To find the infla mmation bio markers induced by coke oven e missions (COE),we investigated the changes of T helper 17 (Th17 )cytokines in hu man bronchial epithelial (16HBE)cells.METHODS 16HBE cells were exposed to organic extracts of COE collected fro m co-king plant at the concentrations of 5,10 and 20 mg·L -1 for 24 h or 5 d to establish short-term and long-term cell models,respectively.Cell viability was measured by MTT assay and infla mmatory da mage was assessed by lactate dehydrogenase assay (LDH).The cytokines in culture supernatant sa mples was detected by co mmercial hu man Th17 cytokine panel kit.RESULTS COE Can induce infla mmation in COE 20 mg·L -1 group and no expression on IL-17 F and IL-1 β.The concentration of IL-10 was 1 .25 ± 0.54,1 .39 ±0.13 and (1 .90 ±0.73)pg·mL -1 in COE 5,10 and 20 mg·L -1 group showing good con-centration-effect relationship (r=0.98,P <0.05 ).IL-23 expression was found only higher at 10 and 20 mg·L -1 and the concentrations were 3.38 ±3.90 and (1 .74 ±2.00 )pg·mL -1 ,respectively.In 16HBE cells treated by COE for 5 d,elevated expression of IL-17A was found in COE 5 and 10 mg·L -1 group,and there was statistically sigificant difference between COE 10 mg·L -1 and DMSO group (P<0.05).Elevated concentration of IL-17F of 10.2 ±1 1 .78 and (6.79 ±7.84)pg·mL -1 was found in COE 5 and 10 mg·L -1 group.The concentration of IL-10 was 1 .71 ±0.02,1 .49 ±0.25 and (2.82 ± 0.33)pg·mL -1 in COE 5,10 and 20 mg·L -1 group,respectively.We found increased IL-1 βexpression with concentration of 2.72 ±0.62,2.25 ±0.33 and (0.93 ±0.21 )pg·mL -1 in COE 5,10 and 20 mg·L -1 group with negative dose-response relationship.We also found more elevated TNF-αlevels in the 5 d than in the 24 h model with no COE specific relationship.CONCLUSION COE induces expression changes of Th17 cytokines profile in 16HBE cells,including IL-23 and IL-1 βfor early and long-term infla mmation,respectively.IL-10 may be a candidate marker for population study on COE induced infla mmatory injury.

Myelin protein zero and its antibody in serum as biomarkers of n-hexane-induced peripheral neuropathy and neurotoxicity effects / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Chronic exposure to n-hexane can lead to peripheral neuropathy that no effective treatment regimen could be applied presently. This study investigated whether myelin protein zero (P0) protein and its antibody could be used to distinguish n-hexane intoxication and protect workers from peripheral neuropathy.</p><p><b>METHODS</b>We compared P0 protein and its antibody among three levels of n-hexane-exposed groups, which included 18 patients with n-hexane-induced peripheral neuropathy as case group, 120 n-hexane-exposed workers as n-hexaneexposed control group, and 147 non-hexane-exposed participants used as control group. ELISA method was applied to detect P0 protein and its antibody.</p><p><b>RESULTS</b>P0 protein in serum was significantly higher in the case group and n-hexane-exposed control group in comparison with the control group (P < 0.01). Compared with the n-hexane-exposed control group, the case group also had significant increase of P0 protein (P < 0.01). After 6 months therapy, P0 protein was observed to decrease significantly in the case group (P < 0.01). The P0 antibody in serum was significantly higher in the n-hexane-exposed control group than in the control group (P < 0.01), but not significantly different between cases and controls.</p><p><b>CONCLUSIONS</b>P0 antibodies in serum may be a short-term effect biomarker for n-hexane exposure. P0 protein in serum may be an early effective biomarker for peripheral nerve neuropathy and its biological limit value needs investigation in the future study.</p>

Endonuclease modified comet assay for oxidative DNA damage induced by detection of genetic toxicants / 中华预防医学杂志

<p><b>OBJECTIVE</b>The aim of this study was to investigate the use of the lesion-specific endonucleases-modified comet assay for analysis of DNA oxidation in cell lines.</p><p><b>METHODS</b>DNA breaks and oxidative damage were evaluated by normal alkaline and formamidopyrimidine-DNA-glycosylase (FPG) modified comet assays. Cytotoxicity were assessed by MTT method. The human bronchial epithelial cell (16HBE) were treated with benzo (a) pyrene (B(a)P), methyl methanesulfonate (MMS), colchicine (COL) and vincristine (VCR) respectively, and the dose is 20 µmol/L, 25 mg/ml, 5 mg/L and 0.5 mg/L for 24 h, respectively. Oxidative damage was also detected by levels of reactive oxygen species in treated cells.</p><p><b>RESULTS</b>Four genotoxicants give higher cytotoxicity and no significant changes on parameters of comet assay treated by enzyme buffer. Cell survival rate were (59.69 ± 2.60) %, (54.33 ± 2.81) %, (53.11 ± 4.00) %, (51.43 ± 3.92) % in four groups, respectively. There was the direct DNA damage induced by test genotoxicants presented by tail length, Olive tail moment (TM) and tail DNA (%) in the comet assay. The presence of FPG in the assays increased DNA migration in treated groups when compared to those without it, and the difference was statistically significant which indicated that the clastogen and aneugen could induce oxidative damage in DNA strand. In the three parameters, the Olive TM was changed most obviously after genotoxicants treatment. In the contrast group, the Olive TM of B(a) P,MMS, COL,VCR in the contrast groups were 22.99 ± 17.33, 31.65 ± 18.86, 19.86 ± 9.56 and 17.02 ± 9.39, respectively, after dealing with the FPG, the Olive TM were 34.50 ± 17.29, 43.80 ± 10.06, 33.10 ± 12.38, 28.60 ± 10.53, increased by 58.94%, 38.48%, 66.86% and 68.21%, respectively (t value was 3.91, 3.89, 6.66 and 3.87, respectively, and all P < 0.05), and the correlation between Olive TM and reactive oxygen species was better than other parameters (r = 0.77, P < 0.05).</p><p><b>CONCLUSION</b>This study indicates that FPG-comet assay appears more specific for detecting oxidative DNA damage induced by genotoxicants exposure, and the application of comet assay will be expanded. The endonuclease modified comet assay will be used widely in the toxicology and molecular epidemiology study.</p>

Study on the pulmonary inflammatory effects induced by inhalation exposure to nanoscale carbon black aerosol in mice / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the carbon black induced effects of lung morphology and pro-inflammation in mice, based on the carbon black aerosol dynamic inhalation exposure model.</p><p><b>METHODS</b>The carbon black aerosol generated by dynamic inhalation device was imported exposure chamber to mice. Scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were used to observe the characters of carbon black. Sixty 9-week-old male BALB/c mice were randomly divided into two control groups, 7 d exposure group and 14 d exposure group. The numbers of four groups of animals were 15, respectively. Mice were exposed to carbon black in the inhalation chamber at (29.33 ± 9.10) mg/m(3) for 6 h/d for continuous exposure 7 d and 14 d, respectively. After 7 d and 14 d exposure, the mice were sacrificed after the last exposure for 24 h. Control mice were killed at 7 d and 14 d. The trachea, lungs, liver, kidneys, and spleen tissues were separated and weighted. Hematoxylin and eosin (HE) staining was used to observe pathological changes of lung by light microscopy. Pulmonary interleukin-8 (IL-8) expression was analyzed by immunohistochemistry. Transmission electron microscopy was used to observe the ultra structure of lung tissue.</p><p><b>RESULTS</b>After 14 d exposure carbon black, the lung coefficient was increased in exposure group compared with control (0.61 ± 0.03 vs 0.79 ± 0.06, t = 6.26, P < 0.01). The spleen coefficient were higher than control(0.39 ± 0.04 vs 0.51 ± 0.06, t = 4.23, P < 0.01) . Other organ coefficients were no significant difference between CB group and control group.Histopathology displayed carbon black particles were deposited in the alveoli and lung bronchial wall in 7 d and 14 d groups. The black carbon particles were deposited within the lung tissue of mice in 14 d group. There were cilia damage, serious damage to the alveolar wall, inflammatory cell infiltration and more hyperemia in 14 d group. Immunohistochemistry showed the level of IL-8 in 7 d (0.272 ± 0.011) and 14 d (0.422 ± 0.065) exposure group were higher than control group in 14 d (0.188 ± 0.041) , F = 31.89, P < 0.01. TEM showed that the lung tissue vision was clear and organelle integrity in the control group. The particles appeared in lung tissue macrophage lysosomes in exposure group, the electron density was consistent with the carbon black particles.</p><p><b>CONCLUSION</b>The dynamic carbon black particles exposure can affect the lung and spleen coefficient, damage integrity of lung morphology and induce inflammation in mice.</p>

Association between intraocular pressure and systemic factor / 中华实验眼科杂志

Background Abnormality of intraocular pressure (IOP) leads to ocular shape change and functional damage.Previous study on the influence factors of IOP mostly focus on eye tissue,but the research of systemic affecting factor is lack.Objective This study was to survey the distribution of IOP and its association with the systemic factors in Chinese population.Methods A cross-section study was designed in this study.The subjects were enrolled from the government employees physical check-up cohort in Guangzhou.IOP was measured using non-contact tonometry.The height,weight and blood pressure were measured based on a presupposed standard protocol.Fundus and slit lamp examinations were performed by trained ophthalmologists.Frequency-doubled perimetry was carried out in the subjects with the suspicious optical disc abnormality,and abnormal perimetry result and operated eye were rejected in the survey.The operative history was learned by questionnaire.Stata 11.0 was used for the data analysis.Results Total 7763 subjects finished all the examination,including 4213 male and 3550 female.Four hundred and fifteen subjects were excluded due to previous intraocular surgery and glaucoma,and the clinical data of the right eyes of 7348 subjects who were eligible for the protocol were brought into statistical analysis.The mean IOP was (15.6±2.8) mmHg in the subjects,and that in the male and female subjects was (15.6±2.8) mmHg and (15.7±2.8) mmHg,respectively,showing a statistically significant difference (t =2.415,P =0.016).A good consistence was found in the IOP between the right eyes and the left eyes (r=0.830,P＜0.001).The IOP value in the subjects presented with a near normal distribution.IOP value was gradually decreased with the increase of age.Multivariate regression suggested a negative correlation between IOP and age or gender (P＜0.001) as well as a positive correlation between IOP and body weight index (BMI),systolic blood pressure or diastolic blood pressure (P ＜0.001).Conclusions Similar to other studies in East Asia population,IOP is associated with age,gender,BMI,systolic and diastolic blood pressure in this survey.

Association between cytokines and trichloroethylene-induced hypersensitivity dermatitis / 中华预防医学杂志

<p><b>OBJECTIVE</b>To detect the cytokines levels in serums of patients with trichloroethylene-induced hypersensitivity dermatitis and explore the effect biomarkers associated with this disease.</p><p><b>METHODS</b>Twenty-two patients with TCE-induced hypersensitivity dermatitis, twenty-two healthy TCE-exposed workers from the same workshops with patients and twenty-two comparable unexposed controls were recruited in this study. Eight cytokines in serums from all subjects were detected using Liquid Suspended Biochip; the correlation among the eight cytokines including interleukin (IL)-1β (IL-1β), IL-5, IL-8, IL-10, interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), macrophage chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1β (MIP-1β) and the correlation between IL-5 and eosinophil count were analyzed.</p><p><b>RESULTS</b>The medians of levels of IL-1β, IFN-γ, IL-5, IL-10, MCP-1, MIP-1β, IL-8 among patients were 0.15, 80.13, 2.95, 6.45, 83.83, 1057.90, 440.22 pg/ml, respectively, which were higher than those among the TCE-exposed workers (0.09, 16.93, 0.11, 0.07, 28.75, 241.07, 28.26 pg/ml, respectively, all P values < 0.01) and unexposed controls (0.09, 3.14, 0.11, 0.07, 25.27, 209.64, 207.34 pg/ml, respectively, all P values < 0.01). The median of level of TNF-α among the patients was 13.26 pg/ml, which was significantly higher than that among TCE-exposed workers (4.87 pg/ml, P < 0.01) but not among unexposed controls; the median of level of IL-5 among the TCE-exposed workers was 0.11 pg/ml, which was significantly higher than that among the unexposed controls (0.11 pg/ml, P < 0.01). The median of levels of IL-8 among the unexposed controls was 207.34 pg/ml, which was significantly higher than that among the TCE-exposed workers (28.26 pg/ml, P < 0.01). In case group, except for correlation of TNF-α and IFN-γ, TNF-α and IL-5, the significant positive correlations were found among any two cytokines (r(IL-1β,IFN-γ) = 0.500, r(IL-1β,TNF-α) = 0.348, r(IL-1β,MCP-1) = 0.537, r(IL-1β,MIP-1β) = 0.477, r(IL-1β,IL-8) = 0.466, r(IL-1β,IL-5) = 0.610, r(IL-1β,IL-10) = 0.626, r(IFN-γ,MCP-1) = 0.460, r(IFN-γ,MIP-1β) = 0.491, r(IFN-γ,IL-8) = 0.322, r(IFN-γ,IL-5) = 0.532, r(IFN-γ,IL-10) = 0.511, r(TNF-α,MCP-1) = 0.325, r(TNF-α,MIP-1β) = 0.283, r(TNF-α,IL-8) = 0.430, r(TNF-α,IL-10) = 0.271, r(MCP-1,MIP-1β) = 0.659, r(MCP-1,IL-8) = 0.526, r(MCP-1,IL-5) = 0.504, r(MCP-1,IL-10) = 0.614, r(MIP-1β,IL-8) = 0.601, r(MIP-1β,IL-5) = 0.451, r(MIP-1β,IL-10) = 0.579, r(IL-8,IL-5) = 0.255, r(IL-8,IL-10) = 0.403, r(IL-5,IL-10) = 0.798, all P values < 0.05). The median of level of IL-5 among the patients with high eosinophils counts was 8.92 pg/ml, which was significantly higher than that among the patients with low eosinophils counts (1.04 pg/ml, P < 0.05).</p><p><b>CONCLUSION</b>The abnormal production of IL-1β, IFN-γ, TNF-α, IL-8, MCP-1, MIP-1β, IL-5 and IL-10 was related with the pathogenesis of hypersensitivity dermatitis induced by TCE. These cytokines could be used as referential indexes in the early health surveillance and clinic disease treatment.</p>

Involvement of p38 MAPK pathway in GLP-1-induced inhibition of apoptosis in human umbilical vein endothelial cells / 生理学报

The aim of the present study was to investigate the effect of glucagon-like peptide-1 (GLP-1) on palmitate-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and the underlying mechanism. HUVECs were cultured in vitro, and then treated by palmitate to induce apoptosis. Meanwhile, GLP-1 was added to explore its effect. After 24 h of the treatments, Caspase-3 activity and DNA fragmentation were measured using ELISA kits. Phospho-p38 mitogen-activated protein kinase (p-p38 MAPK) expression was detected by Western blot. The results showed that incubating HUVECs with 0.125 mmol/L GLP-1 increased Caspase-3 activity and DNA fragmentation. GLP-1 significantly inhibited palmitate-induced increases of Caspase-3 activity and DNA fragmentation in a concentration-dependent manner. Moreover, GLP-1 inhibited the up-regulation of p-p38 MAPK expression induced by palmitate in HUVECs. These results suggest GLP-1 protects HUVECs against lipo-apoptosis, and this effect may be mediated through inhibiting p38 MAPK pathway.

Association between polymorphisms of metabolic genes and telomere length in workers exposed to polycyclic aromatic hydrocarbon / 中华劳动卫生职业病杂志

<p><b>OBJECTIVE</b>To investigate the association between the polymorphisms of metabolic genes and telomere length of genomic DNA in peripheral blood of workers exposed to polycyclic aromatic hydrocarbons (PAHs).</p><p><b>METHODS</b>One hundred and forty five coke-oven workers exposed to PAHs and sixty eight non-exposed medical staffs were recruited in this study. Urinary 1-hydroxypyrene (1-OHP) served as the internal exposure dose of PAHs for all subjects. Relative telomere length (RTL) of genomic DNA in peripheral blood was used as telomere length and measured by real-time PCR. Polymorphisms of metabolic genes were detected by PCR-based methods.</p><p><b>RESULTS</b>Compared with control group, the exposure group shown a decreased RTL (1.10 +/- 0.75 vs 1.43 +/- 1.06, P < 0.05). In the coke-oven workers, after adjusting the sex, age, cigarettes per day and urinary 1-OHP, RTL (1.25 +/- 0.93) of workers with CT genotype at the CYP1A1 3801 T > C was significantly longer than that (0.93 +/- 0.51) of workers with TT genotype (P < 0.05). RTL (0.90 +/- 0.58) of individuals with the Tyr/His genotype at mEH Tyr113His was significantly shorter than that (1.24 +/- 0.90) of individuals with the Tyr/Tyr genotype (P < 0.05). RTL (1.02 +/- 0.64) of individuals with the CT genotype at AHR rs10250822 was significantly shorter than that (1.36 +/- 1.14) of individuals with the CC genotype (P < 0.05). RTL (0.93 +/- 0.54) of individuals with the AT genotype at AHR rs10247158 was significantly shorter than that (1.19 +/- 0.84) of individuals with the AA genotype (P < 0.05).</p><p><b>CONCLUSION</b>The results of present study suggested that PAHs exposure could induce the shorted RTL, CYP1A1, mEH, AHR polymorphisms might influence the change of telomere length of genomic DNA in peripheral blood of workers exposed to PAHs.</p>

Analysis of subgroups of lymphocyte in peripheral blood among dermatitis medicamentosa-like of trichloroethylene patients and healthy exposed workers / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the effects of trichloroethylene (TCE) to lymphocyte subsets among exposed workers, and explore the early immunological effect biomarkers for prevention of hypersensitivity dermatitis induced by TCE.</p><p><b>METHODS</b>Twenty-eight patients with TCE-induced hypersensitivity dermatitis, 56 healthy TCE-exposed workers from the same workshops with patients, and 28 comparable unexposed controls were recruited in this study. The total lymphocyte count and the major lymphocyte subsets including T cell, CD4(+) T cell, CD8(+) T cell, B cell, NK cell in peripheral blood were measured by Flow Cytometer analysis and Standard blood count analysis.</p><p><b>RESULTS</b>The total lymphocyte count and T cell, CD4(+) T cell, CD8(+) T cell among patients (median at 2810.00, 1846.17, 831.87, 904.05 cell counts/µl blood) were significantly increased compared with TCE-exposed workers (median at 2101.00, 1218.59, 643.87, 482.81 cell counts/µl blood, Z = -3.19, -4.96, -3.22, -4.99, P < 0.001) and unexposed controls (median at 1900.00, 1223.60, 558.60, 325.80 cell counts/µl blood, Z = -3.30, -4.46, -3.45, -5.03, P < 0.001), the NK cell and CD3(+)CD4(+)/CD3(+)CD8(+) ratio among patients (median at 255.50 cell counts/µl blood and 1.11) were significantly decreased compared with the unexposed controls (median at 642.60 cell counts/µl blood and 1.96, Z = -3.56 and -3.11, P < 0.01). Meanwhile, for the exposed workers, the CD8(+) T cell (median at 482.81 cell counts/µl blood) was significantly increased and the NK cell and CD3(+)CD4(+)/CD3(+)CD8(+) ratio (median at 318.76 cell counts/µl blood and 1.27) were significantly decreased compared with unexposed controls (median at 325.80 and 642.60 cell counts/µl blood and 1.96, Z = -2.63, -3.52, -2.29, P < 0.05).</p><p><b>CONCLUSION</b>Occupational exposure to TCE could affect the lymphocyte subsets, especially T cell and NK cell. The total lymphocyte count, T cell and CD4(+) T cell might be effect biomarkers for subjects with hypersensitivity dermatitis among TCE-exposed workers.</p>

The effect of 2,5-hexanedione on myelin protein zero expression, and its mitigation using Ginkgo biloba extract / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To investigate the role of myelin protein zero (P(0)) in 2,5-hexanedione (2,5-HD)-induced peripheral nerve injury, and the protective effect of Ginkgo biloba extract (Egb761) on 2,5-HD-induced toxic peripheral neuropathy.</p><p><b>METHODS</b>After 4 weeks of treatment with 2,5-HD at different doses (50, 100, 200, 400 mg/kg) in rats, changes in the levels of P(0) in rat sciatic nerves was investigated, and the effect of Egb761 on 2,5-HD-induced toxic peripheral neuropathy was studied.</p><p><b>RESULTS</b>The blood-nerve barrier (BNB) permeability of the sciatic nerve increased, and the expression of P(0) mRNA and P(0) protein decreased in a dose-dependent manner after treatment with 2,5-HD for 4 weeks. Pretreatment with Egb761 protected against BNB interruption, and inhibited P(0) mRNA and protein reduction during 2,5-HD treatment. Pretreatment with Egb761 significantly reduced loss of body weight (P<0.01) and mitigated gait abnormalities (2.85±0.22) induced by 400 mg/kg 2,5-HD (P<0.01). It also reduced the signs of neurotoxicity induced by 2,5-HD.</p><p><b>CONCLUSION</b>2,5-HD inhibited the expression of P(0) in a dose-dependent manner, and this may be an important mechanism by which toxic peripheral neuropathy is induced by 2,5-HD. Egb761 has a protective effect against 2,5-HD-induced peripheral neurotoxicity in rats.</p>

Hypermethylation of O(6)-methylguanine-DNA methyltransferase in human bronchial epithelial cell induced by organic extracts of coke oven emissions / 中华预防医学杂志

<p><b>OBJECTIVE</b>To elucidate the mechanism of carcinogenesis induced by coke oven emissions by investigating the cell genetic damage index and the methylation of O⁶-methylguanine-DNA methyltransferase (MGMT).</p><p><b>METHODS</b>The human bronchial epithelial cell 16HBE was treated by 1 µmol/L B(a)P for 48 h, and then was exposed continuously to either 1‰ dimethyl sulfoxide (DMSO) or organic extracts of coke oven emission (OE-COE) for five days at the concentrations of 0, 2.5, 5.0, 10.0 and 20.0 µg/ml. The methylation-specific PCR (MSP-PCR), RT-PCR and immunoblotting were applied to detect the methylation status, changes of mRNA and protein of MGMT, respectively. Single cell gel electrophoresis was used to detect DNA damage induced by OE-COE.</p><p><b>RESULTS</b>Compared with the control group (DMSO), there was a significant hypermethylation in all study groups, along with the suppression of mRNA and protein in a dose-dependent manner, and the gradation ratio of them was 1.0, 0.96, 0.96, 0.85, 0.32 and 1.0, 1.0, 1.1, 0.41, 0.52, separately. There was a significant DNA damage with a dose-effect relationship in all study groups (F = 41.22, P < 0.05), and the comet Olive tail moment was (2.98 ± 1.43), (4.76 ± 1.79), (10.09 ± 1.75), (11.38 ± 1.77), (11.67 ± 1.88). The further study found that the index of DNA damage was negatively correlated to the expression of MGMT mRNA and its protein.</p><p><b>CONCLUSION</b>The DNA damage induced by COE might be associated with the suppression of MGMT caused by its hypermethylation.</p>

The effect of 2,5-hexanedione on nerve growth factor in sciatic nerve of rats and VSC4.1 cell / 中华预防医学杂志

<p><b>OBJECTIVE</b>To explore the effect of 2,5-hexanedione (2,5-HD) on the levels of nerve growth factor (NGF) in sciatic nerve of rats and motor-neurons.</p><p><b>METHOD</b>A total of 50 Wistar rats were randomly designed into five groups and intoxicated with 400 mgxkg(-1)xd(-1) 2,5-HD for 0, 7, 14, 21, 28 d. Immunohistochemistry and real-time PCR were used to detect the levels of NGF and NGF mRNA. Motor neuron VSC4.1 cells were administrated with 0, 2.5, 5.0, 10.0, 20.0 mmol/L 2,5-HD for 24 h and 10.0 mmol/L 2,5-HD was chosen to intoxicated VSC4.1 cells for 0, 1, 3, 6, 12, 24, 48 h respectively. Immunofluorescence technique was selected to detect the levels of NGF.</p><p><b>RESULTS</b>The NGF level in sciatic nerve of rats administrated with 400 mgxkg(-1)xd(-1) 2,5-HD showed increase tendency at begin and then decrease after exposure. The NGF mRNA level in 14 d (2(-DeltaDeltaCt)= 3.46), 21 d (2(-DeltaDeltaCt)= 5.28) and 28 d (2(-DeltaDeltaCt)= 3.10) were higher than those in 0 d (2(-DeltaDeltaCt)= 1) and 7 d (2(-DeltaDeltaCt)= 0.78). In vitro tests of VSC4.1 cells showed that NGF levels in 5.0 mmol/L (43.24 +/- 7.52), 10.0 mmol/L (43.48 +/- 10.86) and 20.0 mmol/L (63.13 +/- 10.68) were higher than those in 0 mmol/L (16.32 +/- 4.20)(q values were 19.92, 19.72, 32.78, respectively, P < 0.01) and 2.5 mmol/L (19.78 +/- 2.66) (q values were 17.50, 17.42, 30.63, respectively, P < 0.01) in 24 h and the NGF level in 20.0 mmol/L was higher than those in 5.0 mmol/L (q = 13.04, P < 0.01) and 10.0 mmol/L (q = 11.71, P < 0.01). The NGF levels of VSC4.1 cells with 10.0 mmol/L 2,5-HD in 6 h (18.66 +/- 2.89), 12 h (23.14 +/- 6.08), 24 h (27.66 +/- 6.11) and 48 h (17.25 +/- 3.05) were increased compared with that in 0 h (10.18 +/- 1.81) (q values were 9.64, 15.74, 21.76, 8.50, respectively, P < 0.01), 1 h (9.31 +/- 1.28) (q values were 10.28, 16.17, 21.95, 9.20, respectively, P < 0.01) and 3 h (10.44 +/- 2.13) (q values were 9.25, 15.24, 21.17, 8.10, respectively, P < 0.01), and NGF levels in 12 h and 24 h increased compared with those in 6 h (q values were 5.24, 10.77, respectively, P < 0.01) and 48 h (q values were 7.31, 13.26, respectively, P < 0.01).</p><p><b>CONCLUSION</b>2,5-HD could increase NGF levels in sciatic nerve of rats and motor-neurons, and the dose or time dependent effects were observed in this study.</p>