OBJECTIVE: To investigate the effects of gender and age on the changes in expression of radiation-responsive genes in irradiated human peripheral blood samples. METHODS: Human peripheral blood samples from 30 adult donors was acutely exposed to ⁶⁰Co γ-rays at doses of 0,0.5,1,2,3,4,6 and 8 Gy with the dose rate of 1 Gy/min. Irradiated peripheral blood samples were subsequently incubated at 37-C for 12 h. The effects of gender and age on changes in radiation-responsive gene expressions were analyzed by quantitative real-time polymerase chain reaction (qPCR). RESULTS: The relative mRNA expression levels of 18 radiation-responsive genes were significantly up-regulated in a dose-dependent manner at 12 h after the irradiation (R²=0.63-0.97,P < 0.05). However,the relative mRNA expression levels of MDM2,XPC,FDXR,DDB2,ASTN2 and TNFSF4 showed approximately fivefold variabilities among different individual samples which were irradiated with the same does. Radiation-induced transcriptional changes of BAX,TNFRSF10B,ASTN2,TNFSF4,POLH and GADD45A in female samples were significantly higher than that in male samples (P < 0.05 or P < 0.01). Expression levels of MDM2,FDXR,DDB2 and RPS27L showed differences between four age groups after exposure to 0.5-8 Gy of γ-rays (P < 0.05 or P < 0.01). CONCLUSION: The changes in expression of radiation-responsive genes in blood samples demonstrate significant inter-individual variations which were mainly related to the gender and age of the individual donors.

OBJECTIVE: Comparison of Cistanche phenylethanoid glycosides (CPhGs) liposomes and CPhGs on-BSA-induced-liver-fibrosis-in rats. METHODS: CPhGs liposomes were prepared by the vacuum freeze-drying method. Rats with liver fibrosis were randomly divided into different groups:the blank control,model,blank liposome,CPhGs and CPhGs liposome groups.Serum liver functions (AST,ALT) and liver fiber fibrosis (LN,HA,PC III and IV-C) of rats in each group were compared among the groups.Liver fibrosis was detected using the Masson staining procedure. Real-time fluorescent quantitative PCR (qPCR) was used to detect mRNA expression of α-sma、and collagen I. Western blot method was used to detect the protein expression of α-sma、Collagen I in the liver tissue of rats. RESULTS: The indexes of AST,ALT,LN,HA,PC III,IV-C in serum of rats in the CPhGs liposome group were lower than those in the CPhGs and the blank liposome groups. Compared with the CPhGs group,the CPhGs liposome group showed reduced mRNA expression of collagen I and collagen Ⅲ in the liver tissues (P < 0.01),and reduced protein expression of α-SMA and collagen I (P < 0.05). CONCLUSION: CPhGs liposomes showed better protection on the BSA-induced liver fibrosis in rats than the bulk drug and the blank liposomes. The mechanism may be related to the reduction of α-SMA and collagen I synthesis.

OBJECTIVE: To screen methylated differential genes in normal human periphery blood lymphocytes that were exposed to ⁶⁰Co γ-rays. METHODS: Methylation level of genes were detected by Illumina 450K chip in human periphery blood lymphocytes exposed to 0,0.5 and 2 Gy. The molecular function and bio-pathway of methylated differential genes were analyzed by GO enrichment analysis. RESULTS: There were 1 311 hypermethylation genes and 286 hypomethylation genes both of in the 0.5 Gy and the 2.0 Gy γ irradiation groups. According to the degree of enrichment,our data indicate that the biological process,cell component and molecular function of methylated differential genes were significantly enriched on cell process (enrichment degree=5.86)、cell (enrichment degree=7.48) and binding (enrichment degree=5.27). After further analysis,the results show that methylated differential genes were enriched on terms related to transcription and transcript factor activities which have been identified in other studies. In addition,methylated differential genes also enriched on nucleoside binding (GO:0001882). Some genes related to DNA repair and maintaining chromosome structure were also found such as RAD50,RAD54L,INIP,HIST1H4K and SMC1B. CONCLUSION: Ionizing radiation can change the methylation level of genes in normal human periphery blood lymphocytes. In future studies,genes related to DNA repair and maintainence of chromosome structure will be investigated to determine whether they can be novel radiation damage bio-marker or not.

OBJECTIVE: B7-H1 is a target of tumor immunotherapy. A polymorphism rs4143815C/G in the 3'-untranslated region of B7-H1 can affect its binding to miR-570,which in turn influences the expression of B7-H1. The aim of this study was to investigate the association between the rs4143815 polymorphism and risk of colorectal cancer (CRC). METHODS: Peripheral venous blood was collected from 215 patients with CRC and 236 age- and gender-matched healthy controls. Tumor tissues and adjacent normal tissues were collected from 65 patients with CRC. The rs4143815 polymorphism was genotyped by direct sequencing and B7-H1 mRNA levels were examined by quantitative PCR. The association between the rs4143815 polymorphism in the B7-H1 gene and the risk of CRC was analyzed using chi-square test. RESULTS: Three genotypes of the rs4143815 polymorphism in the B7-H1 gene were detected in both cases and controls,that is CC,CG and GG genotype. The B7-H1 mRNA expression was observed in CRC tissues and adjacent normal tissues. The rs4143815 GG and CG/GG genotypes were associated with a significantly increased risk of CRC compared with the CC genotype[GG vs. CC:adjusted OR=1.99,95%CI (1.19,3.34),P=0.008;CG/GG vs. CC:adjust OR=1.58,95%CI (1.06,2.36),P=0.02]. Similarly,the G allele was associated with a significantly increased risk of CRC compared with the C allele[adjusted OR=1.48,95%CI(1.14,1.93),P=0.004]. The expression of B7-H1 mRNA in CRC tissues was significantly higher than that in adjacent tissues (P=0.02). Moreover,the B7-H1 mRNA level in the rs4143815 GG genotype carriers was significantly higher than that in the rs4143815 CC genotype carriers (P=0.03). CONCLUSION: The rs4143815 GG genotype in the B7-H1 gene may increase the expression of B7-H1 mRNA and result in an increased risk of CRC.

OBJECTIVE: To compare biological pathways involved in the differential expression of proteins and differential proteins in heart tissues from rats after their exposure to normal and herbal cigarette smoke. METHODS: Twenty-four Sprague-Dawley rats were divided into two groups:1) normal cigarette smoke exposure and 2) herbal-containing cigarette smoke exposure (Jinsheng 4). The tandem mass tags (TMT) quantitative proteomics assay was used. After 3 months of smoke exposure,the differentially expressed proteins in the heart tissues from the two groups were screened. The gene ontology (GO) and the Kyoto Gene and Genomic Encyclopedia (KEGG) database were used to identify the differential proteins and bioinformatics analyses were conducted. RESULTS: A total of 43 proteins with different expressions of more than 1.2-fold were identified. The GO analyses of differentially expressed proteins indicate that these proteins were involved with molecular functions,cellular components and biological processes. KEGG enrichment analyses of differentially expressed proteins in the Jinsheng 4 group show that differentially expressed proteins were involved in six biological pathways. CONCLUSION: This study identifies differential expression of proteins, differential proteins and their biological pathways in heart tissues from rats after their exposure to herbal and common cigarette smoke. Our study provides clues for harm reduction among cigarette smokers.

OBJECTIVE: To investigate the expression and biological significance of LINC00472 in the glycidyl methacrylate (GMA)-induced malignant transformation of the human bronchial epithelial cells (16HBE). METHODS: DMSO was used as the solvent control group,and the experimental group was treated with 8 μg/mL GMA for 72 hours,repeated for 3 times and then subcultured. The 10th,20th and 30th generation cells,corresponding to the pre-transformation,mid-term,and late period,were collected from the treated and the corresponding control groups. The Arraystar HumanLncRNA microarray was used to analyze the changes in expression of LINC00472 in the different cell generations. The prediction of target genes and their functions were conducted using bioinformatics databases. Real-time quantification PCR (qPCR) was used to detect the relative expression levels of LINC00472 and the predicted target genes. RESULTS: The microarray results show that LINC00472 in the 3 generations from the GMA-treated groups were:down-regulated by 2.30-fold,down-regulated by 3.57-fold,and up-regulated by 2.32-fold,respectively. The qPCR results confirmed that from the microarray analysis. In particular,the expression of LIN1000472 at the 10-generation was not statistically different from the control. Consistently,the relative expression of the target gene miR-24-3p was statistically significant (P < 0.05),but the multiples were less than 2-fold. CONCLUSION: Expression of LINC00472 in the GMA-induced malignant transformation of 16HBE cells increased from the early to the late cell generations. However,whether LINC00472 and miR-24-3p interact in this transformation process and their mechanisms of action need further study.

OBJECTIVE: To study the induction of DNA oxidation and methylation in nano-graphene oxide (NGO)-exposed 16HBE cells,the quantities of 8-OHdG and DNA 5-mC were determined. METHODS: 16HBE cells were exposed to 1.25,2.5,5 or 10 μg/mL NGO solution for 24 h,8-OHdG and genomic DNA 5-mC were analyzed using high-performance liquid chromatography and high performance capillary electrophoresis,respectively. RESULTS: NGO induced the formation of 8-OHdG and 5-mC in different characteristics. With 2.5,5,10 μg/mL NGO,8-OHdG quantities in the treated groups were significantly higher than that in the control group (P < 0.05 or P < 0.01),and in a dose-dependent manner. However,significant reduction of DNA methylation was observed using the same doses,by 37.1%,49.7% and 46.3%,respectively,compared to that in the control group (P < 0.05 or P < 0.01). The two endpoints were significantly and negatively correlated (r=-0.69,P < 0.01). CONCLUSION: Our data indicate that NGO exposure in cells caused increased DNA oxidation but decreased DNA methylation. It is possible that cellular DNA oxidation may affect DNA methylation.

OBJECTIVE: To investigate the effect of sodium selenite in combination with resveratrol on anti-oxidant and cyclin in lung cancer A549 cells,and on cell survival and apoptosis. METHODS: A human lung adenocarcinoma cell line,A549,was established and divided into several groups:control,sodium selenite,resveratrol,and combined sodium selenite and resveratrol groups. Methyl thiazolyl tetrazolium (MTT) method was performed to assess proliferation of A549 cells. Cell apoptosis rate and ROS were detected by flow cytometry. Glutathione (GSH) and malondialdehyde (MDA) were detected by Microplate Reader. The protein expression of P-Rb was detected using Western blotting. RESULTS: Compared with the control group,treatment with the combination of sodium selenite and resveratrol significantly reduced cell survival and increased apoptosis (P < 0.01). Within these cells,the content of GSH was significantly reduced (P < 0.05), and MDA and ROS were significantly increased (P < 0.05). The protein expression of P-Rb was down-regulated (P < 0.05). CONCLUSION: Treatment with the combination of sodium selenite and resveratrol inhibited proliferation,induced apoptosis which were associated with increased oxidative damage and down-regulating protein expression of P-Rb.

OBJECTIVE: To study the effect of cerium oxide nanoparticles (CeO₂) on radiosensitivity of A549/DDP. METHODS: The half maximal inhibitory exposure dose and concentration of CeO₂ were determined by inhibitory rate of proliferation of A549/DDP. The cell colony-forming experiment was performed to observe changes of cellular surviving fractions between the radiation group after treated with different concentration of CeO₂ groups and radiation only group. A549/DDP cells were grouped as negative control(control),radiation control (rad),and the low,medium and high concentration of CeO₂ groups (final concentrations were 0.000 5,0.05 and 5 μmol/L). Apoptosis,cell cycle and intracellular pH were determined using by flow cytometry. RESULTS: The half maximal inhibitory exposure dose of A549/DDP induced by γ-rays was 25 Gy,which was determined by inhibitory rate of A549/DDP proliferation. At the survival fraction of 2 Gy (SF₂),the ratios of a/b and the sensitive enhancement ratios (SER) were obtained by the cell colony-forming experiment. Comparing with the rad group,the value SF₂ of low concentration CeO₂ group was decreased. The ratio of a/b and the value of SER of the low concentration CeO₂ group were both increased. From the flow cytometry analyses,the percent of apoptosis in the low concentration CeO₂ group was increased (t=5.42,P < 0.05);S-phase arrest of the low,medium and high concentration CeO₂ groups were obvious (t=3.14,4.08,6.81,P < 0.05). The value of pH in cells of the low and high concentration CeO₂ groups were raised (t=2.86,4.93,P < 0.05). CONCLUSION: All the results indicate that low concentrations of CeO₂ enhanced radiosensitivity of A549/DDP.

OBJECTIVE: To investigate the effect of PM_2.5 on DNA damage in human bronchial epithelial cells (HBE). METHODS: HBE cells were exposed to PM_2.5 (0,8,20,50 μg/mL) for 24 h and DNA damage was detected using the single cell gel electrophoresis (SCGE) assay. For another investigation,cells were treated with 10 and 50 μg/mL,untreated cells were used as negative controland cells treated with 10 μmol/L Cr⁶⁺ were used as positive control. mRNA expressions of DNA damage repair genes,including hOGG1 and hMTH1,were detected using real-time quantitative PCR (qPCR) and protein expressions using Western blot. RESULTS: SCGE data show that the tail DNA content,tail length,tail distance significantly increased in the treated groups compared with the control group (P < 0.05 or P < 0.01). The qPCR data show that,compared with the control group,expression of hOGG1 mRNA increased by 75.0%,132.0%,214.0% after exposure to PM_2.5 at 10,50 μg/mL and positive control Cr⁶⁺,respectively. Expression of hMTH1 mRNA increased by 61.0%,144.0% and 75.0%,respectively. The Western blot data show that,compared with the control group,expression of hOGG1 protein increased by 47.6%,64.0%,47.0% after exposure to PM_2.5 at 10,50 μg/mL and positive control Cr⁶⁺,respectively. Expression of hMTH1 protein increased by 20.5%,49.8%,20.9%,respectively. CONCLUSION: Our investigation shows that PM_2.5 induced DNA strand breaks which caused dose-dependent increase of expression in DNA damage repair genes.

OBJECTIVE: To study the effects of cigarette smoke exposure and chronic stress on learning and memory in rats. METHODS: 40 SPF SD rats,10 in each group,50% males,were randomly divided into non-exposed control,cigarette smoke,chronic stress and the combined groups. Rats in the cigarette smoke group were exposed to the virus by respiratory statics,once a day,10 cigarettes each time for 1 h,for 12 weeks. In the chronic stress group,one chronic stress was randomly given every day from five chronic stress programs,and each chronic stress mode was applied discontinuously for 12 weeks. The combined group was treated with chronic stress and smoked 10 cigarettes a day,for 12 weeks. In the 2,4,8,10,12 weeks,the Morris water maze experiment in rats was conducted,constituting the escape latency of rats,test of 4 group rats' learning and memory ability,infected after 12 weeks,abdominal aortic blood,wring the brain,rat body mass,brain,brain viscera coefficient,brain tissue pathology changes,and determination of Cort concentrations,ACH,CHAT content and ACHE activity in serum. RESULTS: Compared with the control group,Cort concentrations in the chronic stress group was significantly increased (P < 0.05),indicating that the rats in the chronic stress group were under stress. Compared with the chronic stress group,the latency of escape at week 2 in the combined group was decreased and showed an antagonistic effect (P < 0.05). Compared with the control group,the latencies of escape at week 10 in the three treated groups were increased (P < 0.05). Pathology results from the cigarette smoke group show rat neurons mild nuclear pyknosis,meningeal vascular moderate hyperemia and mild bleeding,glial cell hyperplasia,chronic stress group rats neurons,visible light nuclear pyknosis and congested,joint intervention group change is most obvious,joint intervention group rat neurons moderate nucleus pyknosis,meningeal congestion and edema of the mild,neurons around the cavity formation. Compared with the control group,the ACH content of rats in the three treated groups were decreased (P < 0.05),while the CHAT content of rats in the stress group decreased and ACHE activity increased (P < 0.05). CONCLUSION: The effects of combined cigarette smoke and chronic stress were interactive on the learning and memory ability of rats. In addition,learning and memory in rats were reduced in the three treated groups. Furthermore,the synthesis of ACH was impaired which was associated with reduced learning and memory ability of rats. The combined exposure did not interact with ACH,CHAT content and ACHE activity. The specific mechanism of its joint action needs to be further studied.

OBJECTIVE: To evaluate genotoxicity of glycidyl methacrylate (GMA) in Chinese hamster lung cells (V79) using the micronucleus assay. METHODS: V79 cells were treated with different doses of GMA (2.25,4.5,9.0,18.0,36.0 μg/mL) for 3 h with or without an in vitro activation system (S9). Controls include a blank and a DMSO group. Another treatment group was treated for 24 h without S9. Cell replication indices and binuclear micronucleus rates were determined. RESULTS: Without S9,the replication index of the treated V79 cells did not change significantly but the incidence of micronuclei in binuclear cells increased significantly in a dose-dependent manner. With the addition of S9,the rate of micronucleated cells in the high-dose group was not statistically significant. CONCLUSION: In the concentration range of 2.25-36.0 μg/mL,GMA induced micronuclei in V79 cells,indicating that GMA is genotoxic.

OBJECTIVE: The Ames tests were used to investigate the mutagenicity of PM_2.5 samples which were collected from Shenzhen and Taiyuan. METHODS: The histidine-deficient Salmonella typhimurium strains TA97,TA98,TA100,TA102 and TA1535 were used. From the collected samples,four doses of PM_2.5 (40,100,200,and 400 μg per dish),a negative control and a positive control were tested. Each test group was treated with S9 or without S9. RESULTS: After treatment with PM_2.5 samples from Shenzhen,there were significant differences in mutagenicity (P < 0.01) between +S9 and -S9 groups in TA98,TA100 and TA102.Similar results were observed with PM_2.5 samples from Taiyuan in TA97,TA98 and TA100. These results indicated that the PM_2.5 samples from Shenzhen and Taiyuan contained mutagenic chemicals which did not cause mutation in TA1535. CONCLUSION: Overall,our results indicate that PM_2.5 samples from Shenzhen and Taiyuan contained directly and indirectly acting mutagens.