Whole Transcriptome Analysis Revealed a Stress Response to Deep Underground Environment Conditions in Chinese Hamster V79 Lung Fibroblast Cells.

Background: Prior studies have shown that the proliferation of V79 lung fibroblast cells could be inhibited by low background radiation (LBR) in deep underground laboratory (DUGL). In the current study, we revealed further molecular changes by performing whole transcriptome analysis on the expression profiles of long non-coding RNA (lncRNA), messenger RNA (mRNA), circular RNA (circRNA) and microRNA (miRNA) in V79 cells cultured for two days in a DUGL. Methods: Whole transcriptome analysis including lncRNA, mRNAs, circ RNA and miRNA was performed in V79 cells cultured for two days in DUGL and above ground laboratory (AGL), respectively. The differentially expressed (DE) lncRNA, mRNA, circRNA, and miRNA in V79 cells were identified by the comparison between DUGL and AGL groups. Quantitative real-time polymerase chain reaction(qRT-PCR)was conducted to verify the selected RNA sequencings. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway was analyzed for the DE mRNAs which enabled to predict target genes of lncRNA and host genes of circRNA. Results: With |log2(Fold-change)| ≥ 1.0 and p < 0.05, a total of 1257 mRNAs (353 mRNAs up-regulated, 904 mRNAs down-regulated), 866 lncRNAs (145 lncRNAs up-regulated, 721 lncRNAs down-regulated), and 474 circRNAs (247 circRNAs up-regulated, 227 circRNAs down-regulated) were significantly altered between the two groups. There was no significant difference in miRNA between the two groups. The altered RNA profiles were mainly discovered in lncRNAs, mRNAs and circRNAs. DE RNAs were involved in many pathways including ECM-RI, PI3K-Akt signaling, RNA transport and the cell cycle under the LBR stress of the deep underground environment. Conclusion: Taken together, these results suggest that the LBR in the DUGL could induce transcriptional repression, thus reducing metabolic process and reprogramming the overall gene expression profile in V79 cells.

Dibutyl phthalate aggravated asthma-like symptoms through oxidative stress and increasing calcitonin gene-related peptide release.

Dibutyl phthalate (DBP) is one of the most ubiquitous phthalate esters found in everyday products, and is receiving increased attention as an immunologic adjuvant. However, information regarding DBP-aggravated allergic asthma is still limited. This study used a mouse model sensitized with ovalbumin (OVA) to determine any adverse effects of DBP on allergic asthma. Our results reveal that allergic asthmatic mice exposed to DBP for an extended period had a significant increase in inflammatory cell infiltration; a significant increase in levels of serum immunoglobulin and T helper 2 cell (Th2) and T helper 17 cell (Th17) cytokines in lung tissue; and significant changes in lung histology and AHR, all of which are typical asthmatic symptoms. The levels of oxidative stress and levels of the neuropeptide, calcitonin gene related peptide (CGRP), were also elevated after DBP exposure. Interestingly, blocking oxidative stress by administering melatonin (MT) not only reduced oxidative stress and CGRP levels, but also ameliorated the asthmatic symptoms. Collectively, these results show that DBP exacerbates asthma-like pathologies by increasing the expression of CGRP mediated by oxidative stress.

Development of diagnostic model of lung cancer based on multiple tumor markers and data mining.

OBJECTIVE: To develop early intelligent discriminative model of lung cancer and evaluate the efficiency of diagnosis value. METHODS: Based on the genetic polymorphism profile of CYP1A1-rs1048943, GSTM1, mEH-rs1051740, XRCC1-rs1799782 and XRCC1-rs25489 and the methylations of p16 and RASSF1A gene, and the length of telomere in the peripheral blood from 200 lung cancer patients and 200 health persons, the discriminative model was established through decision tree and ANN technique. RESULTS: ACU of the discriminative model based on multiple tumour markers increased by about 10%; The accuracy rate of decision tree model and ANN model for testing set were 93.00% and 89.62% respectively. The ROC analysis showed the decision tree model's AUC is 0.929 (0.894∼0.964), the ANN model's AUC is 0.894 (0.853∼0.935). However, the classify accuracy rate and AUC of Fisher discriminatory analysis model are all about 0.7. CONCLUSION: The early intelligent discriminative model of lung cancer based on multiple tumor markers and data mining techniques has a higher accuracy rate and might be useful for early diagnosis of lung cancer.

Vitamin E antagonizes ozone-induced asthma exacerbation in Balb/c mice through the Nrf2 pathway.

Millions of people are regularly exposed to ozone, a gas known to contribute significantly to worsening the symptoms of patients with asthma. However, the mechanisms underlying these ozone exacerbation effects are not fully understood. In this study, we examined the exacerbation effect of ozone in OVA-induced asthma mice and tried to demonstrate the protective mechanism of vitamin E (VE). An asthma mouse model was established, and used to identify the exacerbating effects of ozone by assessing cytokine and serum immunoglobulin concentrations, airway leukocyte infiltration, histopathological changes in lung tissues, and airway hyper-responsiveness. We then determined the amount of reactive oxygen species (ROS) accumulated, the extent to which VE induced ROS elimination, and examined the antagonistic effects of VE on the ozone-induced exacerbating effects. This study showed that 1-ppm ozone exposure could exacerbate OVA-induced asthma in mice. More importantly we found that ozone induced oxidative stress in asthmatic airways may lead to the inhibition of Nuclear factor-erythroid 2-related factor 2 (Nrf2), and may subsequently induce even more exaggerated oxidative stress associated with asthma exacerbation. Through VE induced Nrf2 activation and the subsequent increase in Nrf2 target protein expression, this study suggests a novel mechanism for alleviating ozone exacerbated asthma symptoms.

Regulation of ozone-induced lung inflammation by the epidermal growth factor receptor in mice.

Human exposure to the highly reactive oxidant gas Ozone (O3 ) is associated with inflammatory responses in the airway epithelium. The mechanisms responsible have not been fully elucidated. Epidermal growth factor receptor (EGFR) has previously been shown to play a critical role in the pathogenesis of lung inflammation. To define the role of EGFR in O3 -induced lung inflammation in mice. 40 BALB/c mice were exposed to filtered air (FA) or (0.25, 0.5, 1.00 ppm) O3 for 3 h per day for 7 consecutive days. Levels of reactive oxygen species (ROS), EGF, and transforming growth factor α (TGF-α) in the bronchoalveolar lavage fluid (BALF) of mice were measured using ELISA. BALB/c mice were intratracheally instilled with the EGFR kinase inhibitor PD153035 2 h prior to O3 exposure and every other day thereafter. Phosphorylation of EGFR (Y1068) in lung sections was determined using immunohistochemical staining and western blot 24 h after exposure. Inhalation of O3 induced pronounced lung inflammation in a dose-dependent manner. Levels of ROS, TGF-α, and total proteins and cells in the BALF of mice exposed to 0.5 ppm or 1.0 ppm of O3 were markedly elevated relative to those in the BALF of the mice exposed to FA. In addition, exposure to O3 induced EGFR(Y1068) phosphorylation in the airway epithelium. Administration of PD153035 resulted in a significantly reduced lung inflammation as well as EGFR phosphorylation induced by O3 exposure. Inhalation of O3 leads to inflammatory responses that are dependent on the activation the EGFR in the airway epithelium. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 2016-2027, 2016.

Acute exposure of ozone induced pulmonary injury and the protective role of vitamin E through the Nrf2 pathway in Balb/c mice.

Ozone (O3) in the lower atmosphere is generally derived from various sources of human activity. It has become a major air pollutant in China and has been shown to adversely affect the health of humans and animals. We undertook a study to ascertain the molecular mechanism of ozone induced lung injury in mice and tried to demonstrate the protective mechanism of vitamin E. In this study, mice were exposed to clean air and three different concentrations of ozone. Oxidative stress (reactive oxygen species and malondialdehyde) and Th cytokines in the lung, serum IgE, as well as histopathological examination and the airway hyper-responsiveness (AHR) test were used to reflect inflammation and damage to the lungs of ozone-exposed mice. We then chose an effective concentration of ozone and combined treatment with vitamin E (VE) to explore the underlying mechanism of ozone-induced lung damage. The results of immunological and inflammatory biomarkers (total-immunoglobulin (Ig) E and Th cytokines) as well as histopathological examination and AHR assessment supported the notion that high doses of ozone (>0.5 ppm) could induce inflammation and lung injury in mice and that this induction was counteracted by concurrent administration of VE. The elimination of oxidative stress, the reduced Th2 responses and Ig production, and the relief of lung damage were proposed to explain the molecular mechanism of ozone induced lung injury. We also showed that VE, an antioxidant that enhanced the expression of Nrf2 and up-regulated the antioxidant genes HO-1 and NQO1, could decrease the levels of oxidative stress and alleviate ozone-induced lung injury.

Comparison of ultrasound-assisted cloud point extraction and ultrasound-assisted dispersive liquid liquid microextraction for copper coupled with spectrophotometric determination.

In this work, ultrasound-assisted cloud point extraction (UA-CPE) and ultrasound-assisted dispersive liquid liquid microextraction (UA-DLLME) were investigated and compared firstly as ultrasound-assisted liquid phase microextraction methods, which were coupled with spectrophotometer for copper preconcentration and detection. Compared to conventional CPE and DLLME, the extraction patterns were changed and improved by the effect of ultrasound. As novel methods, their applications were expanded and the analytical performance of spectrophotometric determination for copper was considerably improved. The influence factors of UA-CPE and UA-DLLME were studied in detail. Under the optimal conditions, the limits of detection (LODs) for copper were 0.7 µg L(-1) of UA-CPE and 0.8 µg L(-1) of UA-DLLME with sensitivity enhancement factors (EFs) of 17 and 16. The developed methods were applied to the determination of trace copper in real water samples with satisfactory analytical results.

[Toxicity evaluation on the effluent water from a sewage treatment plant in Zhengzhou City].

OBJECTIVE: To explore the acute toxicity and genotoxicity of inlet and outlet water of a sewage treatment plant in Zhengzhou to provide scientific basis for the safety of sewage reuse. METHODS: Inlet water, secondary and tertiary processed outlet water were collected from the sewage treatment plant. Acute toxicity and genotoxicity of the inlet and outlet water were detected by luminescent bacteria toxicity test and Vicia faba root tip cell micronucleus test, respectively. RESULTS: The luminosity inhibition rates of luminescent bacteria by inlet water, secondary and tertiary processed outlet water were (33.96 +/- 7.51)%, (14.32 +/- 7.36)% and (7.24 +/- 5.58)%, and the micronucleus rates were (12.67 +/- 2.08) per thousand, (6.33 +/- 1.53) per thousand and (2.67 +/- 0.58) per thousand, respectively. The pollution levels of these three samples were heavy, mild and scarce, respectively. The inhibition rates of luminescent bacteria by the secondary and tertiary processed water were significantly lower than that of inlet water (F = 12.159, P = 0.008). A similar result was observed for micronucleus rate (F = 56.850, P < 0.001). CONCLUSION: The acute toxicity and genotoxicity of processed water were decreased greatly. However, some potential ecological risks of the processed water still existed for environment.

[Influence of fluorine on expression of androgen-binding protein and inhibin B mRNA in rat testis sertoli cells].

OBJECTIVE: To study the influence of fluorine on the transcription level of androgen binding protein (ABP) and inhibin B (INHB) mRNA in testis sertoli cells of Sprague Dawley rats. METHODS: A method was set up the model to culture the Sertoli cells. Use a series of concentrations of NaF solutions of 2.5, 5.0, 10.0 and 20.0 mg/L to poison the cells and then, measure the relative expression amount of ABP and INHB mRNA by RT-PCR method. RESULTS: (1) Compare the relative expression amount of ABP mRNA of each group of different concentration with the control group. 2.5 mg/L group was higher than that in the control group, and the difference has the statistical significance (P < 0.05). The 5.0 mg/L group was also higher than that of the control group, and the difference has no statistical significance (P > 0.05). (2) Compare the relative expression amount of INH B mRNA of each group of different concentration with the control group. Both the 2.5 mg/L group and the 5.0 mg/L group were higher than that in the control group, and the difference has the statistical significance (P < 0.05). The rest 2 groups were lower than that in the control group and the difference has no statistical significance (P > 0.05). CONCLUSION: In the range of concentrations between 2.5 and 20.0 mg/L, no distinct influence of fluorine on the expression of androgen binding protein (ABP) and inhibin B (INHB) mRNA in testis sertoli cells of Sprague Dawley rats.

[Effects of Rap2b gene on foci formation and wound-healing of NIH3T3 cells].

OBJECTIVE: To explore the effects on foci formation and wound-healing of NIH3T3 cells, and to provid the experimental evidence of its function. METHODS: DNA from human lung cells was extracted and amplification of Rap2b gene was done by PCR. Eukaryotic expression vector pcDNA3. 1-Rap2b was constructed and was stable transfected into NIH3T3 cell followed with foci formation assay and wound-healing assay. RESULTS: The number of the foci formation of NIH3T3 cell transfected by eukaryotic expression vector pcDNA3. 1-Rap2b was increased remarkably in foci formation assay (P < 0.01) and NIH3T3 cells transfected by eukaryotic expression vector pcDNA3. 1-Rap2b were quickly heal up in wound-healing assay. CONCLUSION: The extrinsic expression of Rap2b could transform NIH3T3 cell using foci formation assay and wound-healing assay. Rap2b gene might play an oncogenic role in tumorigenesis.

[Removal effect of riverside pumping project on microcystins in the Yellow River].

OBJECTIVE: To explore removal effect of riverside pumping project on microcystins in the Yellow River. METHODS: In the summer and autumn of 2008, water samples of the Yellow River and five selected tube-wells in "9 x 5" beach of riverside pumping project were collected and microcystin contents in water were determined using ELISA. RESULTS: Microcystin contents in water in the Yellow River were less than microcystin-LR reference value in Drinking Water Standards. The average contents of the autumn of microcystins were more higher than those of the summer (P < 0.01). Microcystins contents of five tube-wells were more lower than those of the Yellow River. Removal effect of riverside pumping project on microcystins has nothing to do with the distance from the trunk of the Yellow River. CONCLUSION: Riverside pumping project has a good removal efficiency on microcystins.

[Study on DNA-protein crosslinks of certain organs of mice induced by microcystin-LR].

OBJECTIVE: To explore the effects of DNA-protein crosslinks (DPC) of liver, kidney and spermery cell induced by microcystin-LR (MR-LR). METHODS: Kunming male mice were treated by eritoneal injection with different doses of MC-LR. The quantities of junction DNA and free DNA of liver, kidney, testicle cell were detected, and the DPC coefficient were calculated, then we can judge the degree of DNA and protein crosslinks. The DPC coefficient equal junction DNA/(junction DNA + free DNA). RESULTS: Of the groups treated with MC-LR, DPC formations in liver cell were observed significantly increased, in comparison it on with the control groups (P < 0.05). DPC formations in kidney cell were observed significantly increased at the doses of 3 microg/kg bw and 6 microg/kg bw MC-LR, in comparison it on with the control groups (P < 0.05), but there was no effect at the dose of 12 microg/kg bw MC-LR, in comparison it on with the control groups (P > 0.05). At the dose of 3 microg/kg bw MC-LR, DPC formation in testicle cell were not observed significantly increased, in compare it on with the control groups (P > 0.05). DPC formation in testicle cell were observed significantly increased at the dose of 6 microg/kg bw and 12 microg/kg bw MC-LR, in compare it on with the control groups (P < 0.05). CONCLUSION: Microcystin-LR could induce DPC formation in liver, kidney and testicle cells of male mice.