Objective To understand the effect of the organic extracts of tap water deeply treated with O3-BAC on DNA damage. Methods During June to July 2005, water samples were collected from 6 sites in waterworks A treated with O3-BAC, the raw water, the pre-chlorination water, the filtration water, the post-ozonation water, the BAC water and the tap water respectively and 4 sites in waterworks B treated by general treatment, the raw water, the pre-chlorination water, the filtration water, and the tap water respectively. The test was carried out on extracts of water sample from waterworks A with dosage(7.00, 3.00, 1.50, 1.00, 0.75, 0.38 L/plate)and waterworks B with dosage(7.00, 3.00, 1.00 L/plate)using S.typhimurium strains TA98 and TA100. Cytokinesis-blocked micronucleus (CBMN) test, Comet assay were used on extracts of water sample from waterworks A with dosage(3.00, 1.50, 0.75, 0.38 L/plate). Human embryo lung fibroblast (KMB17 strain) p53 ELISA were used with dosage(3.00, 1.00, 0.3 L/plate). Results Ames test showed that in the waterworks A, at the dose of 7.0 L, the revertants of the raw water, pre-chlorination water, the filtration water on TA98-S9 and 7.0 L, 3.0 L/plate, the revertants of the raw water, pre-ozonation water, filtration water on TA98+S9 were twice more than that of solvent control; in waterworks B, at the dose of 7.0 L/plate, the revertants of the raw water, pre-chlorination water, filtration water on TA98-S9 and 7.0 L, 3.0 L/plate the revertants of the tap water on TA98-S9, and 7.0 L/plate pre-chlorination water on TA98+S9 were twice more than that of solvent control. Cytokinesis-blocked micronucleus (CBMN) test showed that in waterworks A, at the dose of 3.0 L/plate, the micronucleus rates of the raw water, filtration water were significant high than that of solvent control(P

Objective : To investigate the role of hepatic long-chain non-coding RNA (lncRNA) H19 in key genes associated with glucose metabolism disorder induced by p,p′-dichlorodiphenyldichloroethylene (p,p′-DDE). Methods: Human embryonic liver CCC-HEL-1 cells were divided into the DMSO group, 0.1 μmol/L p,p′-DDE group, 1 μmol/L p,p′-DDE group, 10 μmol/L p,p′-DDE group, small interference RNA (siRNA)+DMSO group and siRNA+10 μmol/L p,p′-DDE group. The promoter region methylation, mRNA expression and protein expression of hepatocyte nuclear factor 4α (HNF4α), forkhead box transcription factor O1 (FoxO1) and insulin-like growth factor (IGF2) were detected in CCC-HEL-1 cells using the bisulfite method, real-time fluorescence quantitative PCR (qPCR) assay and BCA assay, respectively. The changes in H19 mRNA expression, the methylation of associated genes in the promoter region and transcriptional expression were compared in CCC-HEL-1 among groups. Results: Exposure to p,p′-DDE alone at different doses resulted in an increase in H19 expression, and the H19 mRNA expression was higher in the 10 μmol/L p,p′-DDE group than in the DMSO group [(1.31±0.25) vs. (1.02±0.22); P<0.05]. Lower methylation of the HNF4α gene in the pro moter region [(38.59±32.77)% vs. (61.43±24.64)%; P<0.05] and higher HNF4α mRNA expression [(1.33±0.26) vs. (1.03±0.28); P<0.05] were detected in the 10 μmol/L p,p′-DDE group than in the DMSO group, while no significant differences were detected between the two groups in terms of the methylation of FoxO1 and IGF2 genes in the promoter region, FoxO1 and IGF2 mRNA and protein expression (P>0.05). Following siRNA-induced H19 knockdown, higher methylation of the HNF4α gene in the promoter region [(71.33±22.23)% vs. (38.59±32.78)%; P<0.05], lower HNF4α mRNA expression [(0.71±0.17) vs. (1.33±0.26); P<0.05], higher methylation of FoxO1 gene in the promoter region [(47.73±34.24)% vs. (25.09±25.35)%; P<0.05] and higher IGF2 mRNA [(1.39±0.25) vs. (0.80±0.20); P<0.05] were found in the siRNA+DMSO group than in the 10 μmol/L p,p′-DDE group, and higher IGF2 protein expression was detected in the siRNA+DMSO group than in the DMSO group [(1.03±0.11) vs. (0.74±0.12); P<0.05]. Conclusion Hepatic H19 may alter HNF4α, FoxO1 and IGF2 transcription and expression through mediating the methylation of genes in the promoter region, thereby playing a role in p,p′-DDE-induced glucose metabolism disorders.

Objective: To establish a fluorescence method based on turncated aptamer for the determination of bisphenol A in water. Methods: The bisphenol A truncated aptamer containing 38 bases was selected as a recognition module, and was modified with the fluorophore 6-FAM at the 5'end. The 3'end of the complementary sequence cDNA was modified with the quencher DABCYL. The standard solutions of bisphenol A and interfering compounds were configured. The detection system was established after optimizing the number of bases in cDNA, the concentration ratio of truncated aptamer to cDNA, the incubation temperature and time, and the pH of the buffer. The specificity and recovery experiments were carried out. Results: When the complementary sequence cDNA included 9 bases, the concentration ratio of the truncated aptamer to cDNA was 1:1.5, the pH value of the buffer solution was 7.5, the cDNA was incubated at 55 ℃ for 60 minutes, in the concentration range of 10-75 pmol/L, the linear regression equation was y=2 230.7x+110 825, the correlation coefficient was 0.926. The limits of detection was 3.3 pmol/L. The difference values of fluorescence intensity between tetrabromobisphenol A, estradiol, estriol, bisphenol S and bisphenol A were obviously different, so there was no significant interference to the test result. The recovery rates were 97.8%, 98.8% and 102.3% with the spiked concentrations of 20.0, 40.0 and 60.0 pmol/L. The relative standard deviations were 4.4%, 2.1% and 2.6% (n=5), respectively. Conclusion The fluorescence method based on turncated aptamer has the advantages of easy operation, high sensitivity and specificity, which can be used for the determination of bisphenol A in water.

Objective : To investigate the effects of the exposure of sludge from sewage treatment plants and microplastic extracted from sludge on the oxidative stress levels in zebrafish, so as to put insights into the research into the impact of sludge and microplastics on human health. Methods : Adult wild AB zebrafish were exposed to five groups of sludge (0, 12.5, 25, 50 and 75 g/L) and four groups of microplastics extract from sludge (0, 240, 480, 960/L), with 24 zebrafish in each group. The color, activity and death of zebrafish were observed every day. The contents of superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and malondialdehyde (MDA) were detected 0 h, 24 h, 48 h, 72 h, 96 h and 7 d post-exposure. A two-factor ANOVA was used to analyze the effects of different concentrations and time of exposure on the indicators above. Results : Under 75 g/L sludge exposure, zebrafish began to show mortality at 72 h and all died after 7 d. The zebrafish in the other sludge groups and all microplastic groups had normal color and activity, and no mortality was observed. Sludge concentration interacted with exposure time to affect SOD, CAT, GSH and MDA (P<0.05). With increasing sludge concentration and exposure time, SOD decreased, MDA increased, CAT increased first and then decreased, GSH decreased first and then increased, and GSH continued to decrease since 24 h in the 75 g/L group. The microplastic concentration interacted with exposure time to affect SOD and GSH (P<0.05), but not CAT or MDA (P>0.05). With increasing microplastic concentration and exposure time, SOD and MDA increased, CAT increased first and then decreased, the GSH was slightly increased at 24 h and decreased after 72 h. Conclusion Both sludge and microplastics extracted from sludge can induce oxidative stress damage in zebrafish, and exposure time and concentration can interact to affect oxidative stress levels. The microplastics extracted from sludge have less effect on oxidative stress levels in zebrafish than sludge.

Objective : To investigate the combined effects of triclosan（TCS）and PCB153 on the activity of superoxide dismutase（SOD）and the concentration of malondialdehyde（MDA）in zebrafish liver. Methods : Adult zebrafish were exposed to a series of concentrations of TCS,and the mortality in each group was observed and recorded during the acute toxicity test process. The concentrations in subsequent combined exposure experiments were arranged on the basis of the 96 h-LC50. The factorial design was used to determine the concentrations of combined exposure groups between TCS（0,0.125,0.5 μmol/L）and PCB153（0,0.05,0.2 μmol/L）. After 5,10 and 14 days of exposure,the zebrafish livers were dissected and frozen in each group. The potential interactions of these two compounds were analyzed according to the results of the SOD and MDA. Results : The 96 h-LC50 of TCS exposed to adult zebrafish was 2.64 μmol/L（95%CI：2.37-2.89 μmol/L）. After 5 days of exposure,combined exposure to 0.5 μmol/L TCS+0.2 μmol/L PCB153 caused lower liver SOD activities than single exposure groups and the control group（P<0.05）. After 10 days of exposure,combined exposure to 0.125 μmol/L TCS+0.05 μmol/L PCB153,0.5 μmol/L TCS+0.05 μmol/L PCB153 caused lower liver SOD activities than single exposure groups and the control group（P<0.05）. After 14 days of exposure,combined exposure to 0.5 μmol/L TCS+0.05 μmol/L PCB153,0.5 μmol/L TCS+0.2 μmol/L PCB153 caused higher liver SOD activities than single exposure groups and the control group（P<0.05）. There was an interactive effect between TCS and PCB153 on the liver SOD activity in zebrafish（P<0.05）. There was no significant effect of MDA content in each group. Conclusion Combined exposure to TCS and PCB153 could enhance （inhibit first） the liver SOD activities in zebrafish,and the interaction was synergistic.

OBJECTIVETo evaluate the therapeutic effects, larynx preservation and adverse events of non-surgical combined treatments for laryngeal organ preservation in locally advanced laryngeal squamous cell carcinomas(SCCs).

METHODSForty-six patients with locally advanced laryngeal carcinoma (T2-4, N0-N3) were treated individually with non-surgical combined treatments for larynx preservation (LP). These treatments included concurrent chemoradiotherapy (CCRT)(±epidermal growth factor receptor (EGFR) inhibitor), induction chemotherapy (ICT) followed by CCRT(± EGFR inhibitor), or concurrent radiotherapy and EGFR inhibitor. Radiation therapy was given to a total dose of 60-70 Gy. The Kaplan-Meier method was used to determine the overall survival. Side-effects were evaluated with the established Common Terminology Criteria for Adverse Events (CTCAE) version 3.0 criteria.

RESULTSThe average follow-up time was 31.8 months (range 6-95 months). All patients completed the planned radiotherapy without treatment breaks, and 45(97.8%) of 46 patients completed the planned chemotherapy.The 3-year and 5-year overall survival rates were 87.3%and 67.2%, respectively.The 5-year larynx preservation rate was 100.0%. The 3-year and 5-year progression free survival rates were 95.1% and 87.7%, respectively. The most common acute side effect in grade 3 was oropharyngeal mucositis. After treatment, tracheotomy was still required in 2 patients with glottis cancer for laryngeal edema or stenosis. No patient depended on a percutaneous gastrostomy and experienced speech impairment.

CONCLUSIONPatients with locally advanced laryngeal cancer can be offered non-surgical combined treatments for laryngeal preservation and the high quality of life, showing a higher laryngeal preservation survival rate with minimal toxicities.

Antineoplastic Agents ; therapeutic use ; Carcinoma, Squamous Cell ; drug therapy ; radiotherapy ; Chemoradiotherapy ; Combined Modality Therapy ; Disease-Free Survival ; Head and Neck Neoplasms ; drug therapy ; radiotherapy ; Humans ; Laryngeal Neoplasms ; drug therapy ; radiotherapy ; Larynx ; Organ Sparing Treatments ; Quality of Life ; Survival Rate