Inhibition of lung cancer cells and Ras/Raf/MEK/ERK signal transduction by ectonucleoside triphosphate phosphohydrolase-7 (ENTPD7).

BACKGROUND: The aim of this study was to investigate the effects and mechanisms of ectonucleoside triphosphate phosphohydrolase-7 (ENTPD7) on lung cancer cells. METHODS: The expression characteristics of ENTPD7 and its effect on the survival of lung cancer patients were analyzed by referring to The Cancer Genome Atlas (TCGA). Streptavidin-peroxidase (SP) staining was performed to detect the ENTPD7 protein in tumor tissues and adjacent tissues. Plasmid transfection technology was also applied to silence ENTPD7 gene. Crystal violet staining and flow cytometry were performed to determine cell proliferation and apoptosis. Tumor-bearing nude mice model was established to investigate the effect of sh-ENTPD7 on tumors. RESULTS: The results showed that patients with low levels of ENTPD7 had higher survival rates. ENTPD7 was up-regulated in lung cancer tissues and cells. Down-regulation of the expression of ENTPD7 inhibited proliferation but promoted apoptosis of lung cancer cell. Silencing ENTPD7 also inhibited the expression levels of Ras and Raf proteins and the phosphorylation of mitogen-activated protein kinase (MEK) and extracellular signal-regulated kinase (ERK). Tumor-bearing nude mice experiments showed that silencing ENTPD7 had an inhibitory effect on lung cancer cells. CONCLUSIONS: ENTPD7 was overexpressed in lung cancer cells. Down-regulating ENTPD7 could inhibit lung cancer cell proliferation and promote apoptosis via inhibiting the Ras/Raf/MEK/ERK pathway.

[Intervention effects of selenium yeast on fine particulate matters-induced lung injury in rats].

OBJECTIVE: To evaluate if selenium yeast could inhibit the rat lung injury induced by ambient fine particulate matter( PM_(2. 5) ). METHODS: Fifty-six male SpragueDawley rats were randomly allocated in seven groups( n = 8 each). Saline control group, the rats were exposed to 0. 9% saline by instillation. PM_(2. 5) exposure group, rats were exposed to PM_(2. 5) by intra-tracheal instillation every other day for three times with the accumulated dose of 40 mg/kg. Selenium yeast treatment groups, three groups of rat were exposed to PM_(2. 5) . Then the rats were given low, middle and high dose of selenium yeast, and the doses were 8. 75, 17. 5 and 35 mg/kg, respectively. High dose selenium yeast control group, rats were given high dose of selenium yeast only. Solvent control group, therats were given 1% carboxymethyl cellulose. Saline and PM_(2. 5) were given in the first week. In the second week, selenium yeast and solvent were given by gavage. The rats were sacrificed 24 hours after the last gavage. The bronchoalveolar lavage fluid( BALF)was collected to count the neutrophils numbers and analyze the markers related to inflammation, oxidative stress and cell damage. The lung lobe that was not been lavaged was processed for light microscopic examination. RESULTS: The proportions of neutrophils in BALF and the pathologic scores of lung in PM_(2. 5) - exposed groups were significantly higher than control( P < 0. 05). Selenium yeast treatment caused decrease in tumor necrosis factor-α( TNF-α), interleukin-1ß( IL-1ß), lactate dehydrogenase( LDH), total protein( TP), alkaline phosphatase( AKP) and malondialdehyde( MDA) compared with the only PM_(2. 5) exposure group. Meanwhile, the dose-dependent increase in totalsuperoxide dismutase( T-SOD) and glutathione peroxidase( GSH-Px) activities were observed. There were no significant differences among the groups of saline control group, high dose selenium yeast control group and solvent control group. CONCLUSION: Selenium yeast treatment may protect against acute injury induced by PM_(2. 5) in rat lung.

[Effects of vitamin E and ω-3 fatty acids on protecting ambient PM_(2. 5)-induced cardiovascular injury].

OBJECTIVE: To observe whether vitamin E( Ve) and ω-3 polyunsaturated fatty acids( ω-3 FA) could prevent the fine particulate matter( PM_(2. 5))-induced cardiovascular injury and explore the potential mechanism. METHODS: The SD rats were assigned randomly to 8 groups, those were control group, PM_(2. 5)group, Ve treatment groups( 3, 10, 30 mg/( kg·d)) and ω-3 FA treatment groups( 10, 30 and 90 mg/( kg·d)). The rats were pretreated with different concentration of Ve and ω-3 FA separately for 14 days, then were exposed to ambient PM_(2. 5) by intratracheal instillation( 10 mg/kg BW). All the rats were sacrificed after the last PM_(2. 5) exposure, then the arterial blood, lungs and cardiac tissues were collected. The expressions of tumor necrosis factor-α( TNF-α), interleukin-1ß( IL-1ß), interleukin-6( IL-6) in serum, bronchoalveolar lavage fluid and supernatant of cardiac tissue were detected by ELISA kits. The levels of malondialdehyde( MDA), superoxide dismutase( SOD) and glutathione-peroxidase( GSH-Px) in serum and myocardium were also measured. RESULTS: Compared with the severe injury of rats in PM_(2. 5) exposure group, the rats in Ve or ω-3 FA groups had a slighter injury in lung and cardiac tissue with the increase of Ve and ω-3 FA. Similarly, the levels of IL-1ß, IL-6 in bronchoalveolar lavage fluid had a decreasing trend with the increase of Ve and ω-3 FA compared with the PM_(2. 5) exposure groups. Meanwhile, the expressions of TNF-α in Ve and ω-3 FA high dose groups were significantly reduced when compared with the PM_(2. 5) exposure group( P <0. 05). In addition, the MDA levels in serum were markedly decreased and the activities of SOD were significantly increased compared with the PM_(2. 5)exposure group( P < 0. 05 or P < 0. 01) whereas the SOD activities were elevated only in the ω-3 FA high dose groups( P < 0. 05). Meanwhile, the levels of IL-6 and TNF-α in serum had an obvious decrease compared with the PM_(2. 5) exposure group( P < 0. 01). Similarly, compared with the PM_(2. 5)exposure group, the expressions of MDA were markedly decreased and the activities of SOD and GSH-Px in myocardium were significantly increased( P < 0. 05 or P < 0. 01) in the Ve treatment group. In addition, the activities of GSH-Px was found higher only in the ω-3 FA high treatment group compared with the PM_(2. 5)exposure group( P < 0. 05). Meanwhile, the levels of IL-1ß and TNF-α in cardiac tissue had an obvious decrease trend with the increase of Ve and ω-3 FA. CONCLUSION: PM_(2. 5) exposure may increase inflammatory response and oxidative stress, supplementation with Ve and ω-3 FA could prevent the PM_(2. 5)-induced inflammatory reaction and oxidative stress damage by increasing the activities of SOD and GSH-Px.

Investigation of selenium pretreatment in the attenuation of lung injury in rats induced by fine particulate matters.

Selenium (Se) is vital for health because of its antioxidative and anti-inflammation functions. The aim of this study was to determine if dietary selenium could inhibit the rat lung injury induced by ambient fine particulate matter (PM2.5). Sprague-Dawley rats were randomly allocated in seven groups (n = 8). The rats in PM2.5 exposure group were intratracheally instilled with 40 mg/kg of body weight (b.w.) of PM2.5 suspension. The rats in Se prevention groups were pretreated with 17.5, 35, or 70 µg/kg b.w. of Se for 4 weeks, respectively. Then, the rats were exposed to 40 mg/kg b.w. of PM2.5 in the fifth week. The bronchoalveolar lavage fluid (BALF) was collected to count the neutrophil numbers and to analyze the cytokines (tumor necrosis factor-α (TNF-α), interleukin-1ß (IL-1ß), soluble intercellular adhesion molecule-1 (sICAM-1)) related to inflammation, the markers related to oxidative stress (total antioxidant capacity (T-AOC), total superoxide dismutase (T-SOD), glutathione peroxidase (GSH-Px), and malondialdehyde (MDA)), and the indicators related to cell damage (lactate dehydrogenase (LDH), total protein (TP), alkaline phosphatase (AKP)). The lung lobe that has not undergone bronchoalveolar lavage was processed for light microscopic examination. The results showed that the proportions of neutrophils in the BALF and the pathologic scores of the lung in PM2.5-exposed groups were higher than that in the control group (P < 0.05). Se pretreatment caused a dose-dependent decrease in TNF-α, IL-1ß, sICAM-1, LDH, TP, AKP, and MDA when compared with the PM2.5-only exposure group. Meanwhile, the dose-dependent increase in T-AOC, T-SOD, and GSH-Px activities were observed in rats pretreated with Se. In conclusion, Se pretreatment may protect rat lungs against inflammation and oxidative stress induced by PM2.5, which suggests that Se plays an important role as a kind of potential preventative agent to inhibit the PM2.5-induced lung injury.

Effects of Fine Particulate Matter (PM2.5) on Systemic Oxidative Stress and Cardiac Function in ApoE(-/-) Mice.

AIM: In this study, we aimed to explore the toxic mechanisms of cardiovascular injuries induced by ambient fine particulate matter (PM2.5) in atherosclerotic-susceptible ApoE(-/-) mice. An acute toxicological animal experiment was designed with PM2.5 exposure once a day, every other day, for three days. METHODS: ApoE(-/-) and C57BL/6 mice were randomly categorized into four groups, respectively (n = 6): one control group, three groups exposed to PM2.5 alone at low-, mid-, and high-dose (3, 10, or 30 mg/kg b.w.). Heart rate (HR) and electrocardiogram (ECG) were monitored before instillation of PM2.5 and 24 h after the last instillation, respectively. Cardiac function was monitored by echocardiography (Echo) after the last instillation. Biomarkers of systemic oxidative injuries (MDA, SOD), heart oxidative stress (MDA, SOD), and NAD(P)H oxidase subunits (p22phox, p47phox) mRNA and protein expression were analyzed in mice. The results showed that PM2.5 exposure could trigger the significant increase of MDA, and induce the decrease of heart rate variability (HRV), a marker of cardiac autonomic nervous system (ANS) function with a dose-response manner. Meanwhile, abnormal ECG types were monitored in mice after exposure to PM2.5. The expression of cytokines related with oxidative injuries, and mRNA and protein expression of NADPH, increased significantly in ApoE(-/-) mice in the high-dose group when compared with the dose-matched C57BL6 mice, but no significant difference was observed at Echo. In conclusion, PM2.5 exposure could cause oxidative and ANS injuries, and ApoE(-/-) mice displayed more severe oxidative effects induced by PM2.5.

Rat lung response to ozone and fine particulate matter (PM2.5) exposures.

Exposure to different ambient pollutants maybe more toxic to lung than exposure to a single pollutant. In this study, we discussed the inflammation and oxidative stress responses of rat lung caused by ozone and PM2.5 versus that of rats exposed to saline, ozone, or single PM2.5 . Wistar rats inhaled 0.8 ppm ozone or air for 4 h and then placed in air for 3 h following intratracheal instillation with 0, 0.2 (low dose), 0.8 (medium dose), 3.2 (high dose) mg/rat PM2.5 dissolved in sterile saline (0.25 mL/rat), repeated twice per week for 3 weeks, the cumulative doses of PM2.5 in animals were 1.2, 4.8, and 19.2 mg. Rats were sacrificed 24 h after the last (sixth) exposure. The collected bronchoalveolar lavage fluid (BALF) was analyzed for inflammatory cells and cytokines. Lung tissues were processed for light microscopic and transmission electron microscopic (TEM) examinations. Results showed that total cell number in BALF of PM2.5 -exposed groups were higher than control (p < 0.05). PM2.5 instillation caused dose-trend increase in tumor necrosis factor alpha (TNF-α), interleukin-6, lactate dehydrogenase, and total protein of BALF. Exposure to ozone alone only caused TNF-α significant change in above-mentioned indicators of lung injury. On the other hand, ozone could enhance PM2.5-induced inflammatory changes and pathological characters in rat lungs. SOD and GSH-Px activities in lung were reduced in PM2.5-exposed rats with and without prior ozone exposure compared to control. To determine whether the PM2.5 and ozone affect endothelium system, iNOS, eNOS, and ICAM-1 mRNA levels in lung were analyzed by real-time PCR. These data demonstrated that inflammation and oxidative stress were involved in toxicology mechanisms of PM2.5 in rat lung and ozone potentiated these effects induced by PM2.5. These results have implications for understanding the pulmonary effects induced by ozone and PM2.5.

[Effects of airborne fine particulate matters on pulmonary inflammation injury and Th17/Treg balance of sub-chronic exposure mice].

OBJECTIVE: To explore effects of fine particulate matters (PM 2.5) onpulmonary inflammation, and the changes of Th17/Treg balance as well as related cytokines. METHOD: Thirty-two C57BL/6 male mice were randomly divided into 4 groups including 1 saline control group and 3 PM2.5 exposure groups (1.5, 7.5 and 15 mg/kg BW, respectively). Each group received intratracheal instillation twice per week for 3 consecutive months. 24 hours after the last exposure, anaesthetize the mice with chloral hydrate, bronchoalveolar lavage fluid (BALF) was collected for inflammatory cells and cytokines analysis. The Th17- and Treg-related cytokines in BALF was measured by enzyme linked immunosorbent assay (ELISA). The level of the specific transcription factors of Th17 and Treg in lung tissue was determined by real-time PCR. Unlavaged left lung were fixed with 4% paraformaldehyde for histopathological detection. RESULTS: Th17-related cytokine IL-17 increased, but Treg-related cytokine IL-10 decreased significantly in BALF at 7.5 and 15 mg/kg BW PM2.5 exposure groups compared with control group (P < 0.05). Consistently, the relative mRNA expression of ROR-gammat (specific transcription factors of Th17) increased in a dose-response way, the relative mRNA expression of Foxp3 + (specific transcription factors of Treg) decreased in a dose-response way. CONCLUSION: Sub-chronic PM2.5 exposure caused persistent inflammation, immune injury and disordered the Th17/Treg imbalance as well as related cytokines.

[Effects of ozone and fine particulate matter (PM2.5) on rat cardiac autonomic nervous system and systemic inflammation].

OBJECTIVE: To study the effects of ozone and fine particulate matter (PM2.5) on rat cardiac autonomic nervous system and systemic inflammation and the interconnected relationship of these two pathways. METHODS: 48 Wistar rats were randomly assigned to 8 groups (n = 6). Rats were intratracheal instillation of 0.2, 0.8 and 3.2 mg/rat PM2.5 as low-dose, medium-dose and high-dose PM2.5 alone exposure groups. Ozone combined PM2.5 groups were exposed to 0.8 ppm ozone 4 hours and then instilled 0.2, 0.8 and 3.2 mg/rat PM2.5 separately. Rats only inhaled 0.8 ppm ozone for 4 hours as ozone alone exposure group. Control rats were instilled with saline. Each exposure protocol applied twice a week for three continuous weeks. Systolic blood pressure and electrocardiograph (ECG) morphology of rats were recorded at 24 hours post-exposure of the last exposure. Serum TNF-alpha, IL-6 and CRP were analyzed. Right ventricles were used as pathological examination with HE staining. RESULTS: HRV indices in PM2.5 alone and combined exposure groups showed significant difference compared to control. However, heart rate only reduced significantly in ozone combined PM2.5 groups. Blood pressure rose significantly in combined groups and high-dose PM2.5 alone exposure. TNF-alpha and IL-6 increased significantly in PM2.5 alone/and ozone groups. CRP showed obvious dose-dependent relationship in PM2.5 alone and combined groups. Cardiac histopathology results revealed that PM2.5 deposition and myocardial inflammation were also observed in high-dose PM2.5 rats. CONCLUSION: Ozone can enhance the cardiac autonomic nervous system and systemic inflammation induced by PM2.5.

Preexposure to PM2.5 exacerbates acute viral myocarditis associated with Th17 cell.

BACKGROUND: It is increasingly recognized that exposure to ambient fine particles (PM(2.5)) is a risk factor for the development of cardiovascular events. This study was to explore the link between PM(2.5) exposure and viral myocarditis in the functional mechanism of Th17 cells. METHODS: Male BALB/c mice were administered an intratracheal (i.t.) instillation of 10 mg/kg b.w. PM(2.5) particles. Twenty-four hours later, the mice were injected intraperitoneally (i.p.) with 100 µl of coxsackievirus B3 (CVB3) diluted in Eagle's minimal essential medium (EMEM). Seven days after the treatment, pulmonary and cardiac tissues were examined. RESULTS: The results showed that preexposure to PM(2.5) increased the cardiac and pulmonary injuries and viral replication in the heart of CVB3-infected mice along with an increase in CD4(+) IL-17(+) cells in the spleen and heart. The mRNA expressions of interleukin-17A (IL-17A), perforin, transforming growth factor-ß (TGF-ß) and RORγt were up-regulated in PM(2.5)-pretreated mice than that in the virus-treated mice. Additionally, compared to virus-treated mice, the cardiac protein expressions of IL-17A and matrix metalloproteinases-2 (MMP-2) were increased, but interferon-γ (IFN-γ) and metalloproteinases-1 (TIMP-1) were decreased in PM(2.5)-pretreated mice. Interestingly, PM(2.5) caused IFN-γ decreased, whereas CVB3 caused a dramatic increase in IFN-γ. Subsequently, preexposure to PM(2.5) induced a slight increase of IFN-γ in the sera of CVB3-infected mice. CONCLUSIONS: These results demonstrated that PM(2.5) exposure exacerbated virus-induced myocarditis possibly through the increase in Th17-mediated viral replication, perforin response and imbalance of MMP-2/TIMP-1. These findings provided supportive evidence for the epidemiological research that ambient particles could increase the occurrence and development of cardiovascular diseases.

The biological effects of individual-level PM(2.5) exposure on systemic immunity and inflammatory response in traffic policemen.

BACKGROUND: Ambient fine-particle particulate matter (PM2.5) exposure is associated with the decline in pulmonary function, prevalence of coronary heart disease and incidence of myocardial infarction. The study is to observe the effects of ambient PM2.5 on the cardiovascular system and to explore the potential inflammatory and immune mechanisms. METHODS: The subjects included 110 traffic policemen in Shanghai, China, who were aged 25-55 years. Two-times continuous 24 h individual-level PM2.5 measurements were performed in winter and summer, respectively. The inflammatory marker (high-sensitivity C-reactive protein, hs-CRP), immune parameters (IgA, IgG, IgM and IgE) and lymphocyte profiles (CD4 T cells, CD8 T cells, CD4/CD8 T cells) were measured in blood. The associations between individual-level PM2.5 and inflammatory marker and immune parameters were analysed by multiple linear regression. RESULTS: The average concentration of 24 h personal PM2.5 for participants was 116.98 µg/m(3) and 86.48 µg/m(3) in winter and summer, respectively. In the main analysis, PM2.5 exposure is associated with the increases in hs-CRP of 1.1%, IgG of 6.7%, IgM of 11.2% and IgE of 3.3% in participants, and decreases in IgA of 4.7% and CD8 of 0.7%, whereas we found no statistical association in CD4 T cells and CD4/CD8 T cells. In the adjusted model, the results showed that the increase of PM2.5 was associated with the changes of inflammatory markers and immune markers both in winter and summer. CONCLUSIONS: Traffic policeman have been a high-risk group suffering inflammatory response or immune injury because of the high exposure to PM2.5. These findings provided new insight into the mechanisms linking ambient PM2.5 and inflammatory and immune response.

Effects of ozone and fine particulate matter (PM(2.5)) on rat system inflammation and cardiac function.

In order to understand the toxic mechanisms of cardiovascular system injuries induced by ambient PM(2.5) and/or ozone, a subacute toxicological animal experiment was designed with exposure twice a week for 3 continuous weeks. Wistar rats were randomly categorized into 8 groups (n=6): 1 control group, 3 groups exposed to fine particulate matters (PM(2.5)) alone at 3 doses (0.2, 0.8, or 3.2 mg/rat), 1 group to ozone (0.81 ppm) alone and 3 groups to ozone plus PM(2.5) at 3 doses (0.2, 0.8, or 3.2 mg/rat). Heart rate (HR) and electrocardiogram (ECG) was monitored at approximately 24-h both after the 3rd exposure and the last (6th) exposure, and systolic blood pressure (SBP) was monitored at approximately 24-h after the 6th exposure. Biomarkers of systemic inflammation and injuries (CRP, IL-6, LDH, CK), heart oxidative stress (MDA, SOD) and endothelial function (ET-1, VEGF) were analyzed after the 6th exposure. Additionally, myocardial ultrastructural alterations were observed under transmission electron microscopy (TEM) for histopathological analyses. Results showed that PM(2.5) alone exposure could trigger the significant increase of CRP, MDA, CK, ET-1 and SBP and decrease of heart rate variability (HRV), a marker of cardiac autonomic nervous system (ANS) function. Ozone alone exposure in rats did not show significant alterations in any indicators. Ozone plus PM(2.5) exposure, however, induced CRP, IL-6, CK, LDH and MDA increase, SOD and HRV decrease significantly in a dose-response way. Meanwhile, abnormal ECG types were monitored in rats exposed to PM(2.5) with and without ozone and obvious myocardial ultrastructural changes were observed by TEM. In conclusion, PM(2.5) alone exposure could cause inflammation, endothelial function and ANS injuries, and ozone potentiated these effects induced by PM(2.5).

[Investigation on the characteristics and space-time distribution of fine particles in the atmosphere of residential area in Shanghai City].

OBJECTIVE: To observe the ambient fine particle pollution and the trend of its space-time distribution in residential areas in Shanghai, and to explore the effects of vehicle exhaust emission on the ambient fine particle pollution. METHODS: Two residential areas A and B were selected for monitoring the pollution of fine particles. Area A is a normal residential area and area B is closed to a main road with heavy traffic. Four monitoring sites were set in the distance of 0 m, 50 m, 100 m and 200 m to the roadside and on a place 1.5 - 1.8 m above the ground. The concentration of fine particles in the air were measured in April, July, October 2010 and Jan 2011 for 1l0 days in each month in both areas using SIDEPAK AM510 (TSI, USA) fine particle monitors. RESULTS: The pollution of fine particle was varied in different seasons (spring > winter > autumn > summer) and at different time (with two peaks at 8:00 AM and 19:00 PM, corresponding to the rush hours). The pollution of fine particles is higher in residential area B than that in area A. The concentration of fine particles was reduced with the increase of the distance to the roadside. CONCLUSION: The level of fine particles in residential areas is comparatively high in Shanghai, and the vehicle exhaust emissions have significant effects on the concentration of fine particles in the atmosphere of residential area.

Imbalance of Th1 and Th2 cells in cardiac injury induced by ambient fine particles.

The study was to explore the potential immunoregulatory mechanisms linking fine particles and cardiac injury. Wistar kyoto (WKY) rats were exposed by intratracheal instillation to fine particles with the doses of 0.0, 1.6, 8.0 and 40.0mg/kg b.w., respectively. The exposure was conducted once a day, for three consecutive days. Twenty-four hours after the last exposure, the rats were sacrificed. Th1- and Th2-related transcription factors and cytokines were assessed in left ventricle of rats. The mRNA expressions of Th1- and Th2-related transcription factors signal transducer and activator of transcriptionl 1 (STAT1), signal transducer and activator of transcriptional 6 (STAT6), GATA-3 and T-bet were assessed in left ventricle of rats using real-time PCR. Meanwhile, the levels of Th1- and Th2-related cytokines IL-4, IL-13 and interferon gamma (IFN-γ) were determined by ELISA kits in cardiac homogenate supernatant of rats. Furthermore, the protein expression of IL-4 and IFN-γ were detected in myocardium by Western blot. The results of cardiac histology demonstrated exacerbated cardiac lesions and histological characterization of inflammation and degeneration in rats after exposure to fine particles. Moreover, fine particles induced significant increase of IL-4 and IL-13 and decrease of IFN-γ in myocardium of rats. The mRNA expression of STAT1, STAT6 and GATA-3 were up-regulated in left ventricle of rats in a dose-dependent manner, whereas T-bet was significantly down-regulated. The variations of these cytokines demonstrated the imbalance of Th1 and Th2 cytokines existed in cardiac injuries induced by fine particle. The imbalance of Th1/Th2 cytokines might be one of the mechanisms of immunotoxicity of cardiovascular system induced by ambient fine particles.

Effects of atorvastatin on fine particle-induced inflammatory response, oxidative stress and endothelial function in human umbilical vein endothelial cells.

The study is to explore the toxicity of organic extracts and water-soluble fraction of fine particles on human umbilical vein endothelial cells (HUVECs). The exposure doses were 100, 200 and 400 µg/ml, respectively, for two kinds of fractions. Moreover, atorvastatin was used for intervention study. HUVECs were stimulated by 400 µg/ml organic and water soluble extracts, respectively, immediately followed by treatment with atorvastatin in concentrations of 0.1 µmol/L, 1 µmol/L and 10 µmol/L, respectively. Cell viability, malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), reactive oxygen species (ROS) and the expression of interleukin-6 beta (IL-6), tumor necrosis factor-α (TNF-α), endothelin-1 and P-selectin were determined in cells. The results showed that MDA and ROS increased in HUVECs after exposed to organic extracts and water-soluble fraction, whereas cell viability, NO and SOD decreased. The mRNA expression of IL-6, TNF-α, endothelin-1 (ET-1) and P-selectin increased after exposed to different fractions. Meanwhile, at the same exposure dose, water-soluble fraction caused more significant increase of MDA, IL-6, TNF-α and P-selectin and decrease of cell viability and NO when compared to organic extracts. Compared to no atorvastatin group, the levels of MDA, ROS and the expression of IL-6, TNF-α, ET-1 and P-selectin decreased in HUVECs in adding atorvastatin group, but cell viability, NO and SOD increased, which indicated that atorvastatin attenuated fine particle-induced inflammatory response, oxidative stress and endothelial damage. The results hinted that the inflammatory response, oxidative stress and endothelial dysfunction might be the mechanisms of cardiovascular injury induced by different fractions of ambient fine particles.

[Effects of airborne fine particulate matter on human respiratory symptoms and pulmonary function].

OBJECTIVE: to explore effects of airborne fine particulate matter exposure on human respiratory symptoms and pulmonary function. METHODS: one hundred and seven field traffic policemen were recruited as airborne fine particulate matter high-exposure group and one hundred and one male residents as common exposure group. The individual sampler was used to measure fine particulate matter exposure levels of the two groups. To obtain personal information, especially respiratory symptoms such as cough, sputum, etc. a questionnaire survey was used. The pulmonary ventilation function was detected: forced expiratory vital capacity (FVC), the first 1 second forced expiratory volume (FEV1.0), FVC/FEV1.0% and peak flow values (PEF), and the difference of fine particulate matter exposure level and respiratory function of the two groups was compared. RESULTS: 24 h individual average fine particulate matter exposure concentration of traffic police and residents were respectively (115.4 ± 46.17) microg/m(3) and (74.94 ± 40.09) microg/m(3), the traffic police PM2.5 exposure levels were significantly higher than the residents. In the incidence of respiratory symptoms, compared with high-exposure group and common exposure group, coughing, expectoration, throat unwell, asthma, short of breath and nose discomfort, traffic police group was higher than residents group (P < 0.05). The abnormal rate of lung ventilation function indexes, such as FVC and FEV1.0 was 25.23% and 12.15% respectively in high-exposure group, 11.88% and 2.97% in common exposure group, there was no statistical difference between two groups. Besides, the abnormal rate of FVC and FEV1.0, showed rising trend in high-exposure group with seniority. CONCLUSION: long-term higher levels of airborne fine particulate matter exposure, may impact respiratory health and impair pulmonary function.

[Effects of airborne fine particulate matters on human immunological indicators].

OBJECTIVE: To explore effects of airborne fine particulate matters on human immunological indicators. METHODS: All subjects were measured the level of PM2. 5 individual exposure with personal sampling devices. Serum immunological indicators, such as WBC, NE, NE%, Ly, Ly%, CD4+, CD8+, CD4+ / CD8+, IgA, IgM, IgG and IgE were detected through enzyme-linked immunosorbent assay. Then, comparing the difference of PM2.5 exposure and immunological indicators between two groups. RESULTS: For the level of PM2.5 daily exposure, traffic policemen (115.4 +/- 46.2) microg/m3) was significantly higher than residents ((74.9 +/- 40.1) +/- g/ m(-3)). And there was also significant difference in some immunological indicators, such as Ly%, CD4+, CD8+, IgM, IgG, IgE, CC 16 and CRP, between two groups. CONCLUSION: Long-term high level of airborne fine particulate matters exposure may change some blood immunological indicators, affect the health of the immune system.

Acute effects of fine particles on cardiovascular system: differences between the spontaneously hypertensive rats and wistar kyoto rats.

The study is to explore the potential mechanisms linking fine particles and cardiovascular toxicity. Spontaneous hypertensive rats (SHR) and age-matched wistar kyoto (WKY) rats were exposed by intratracheal instillation to fine particles with the doses of 0.0, 1.6, 8.0 and 40.0mg/kg b.w., respectively. The exposure was conducted once a day, for three continuous days. Twenty-four hours after the last exposure, the rats were killed and the levels of high sensitive C-reactive proteins (hsCRPs), nitrous oxide (NO), D-dimer and endothelin-1 (ET-1) were measured in blood. Reverse transcriptase polymerase chain reaction (RT-PCR) assay assessed the expression of interleukin-1 beta (IL-1beta), intercellular adhesion molecule-1 (ICAM-1), ET-1, Bax and Bcl-2 in left ventricle of rats. Meanwhile, cardiac histological lesions were assessed. The expression transforming growth factor-beta 1 (TGF-beta(1)) in left ventricle was measured by immunohistochemical staining. The results showed that the levels of hsCRP, D-dimer, ET-1 and the expression of IL-1beta, ICAM-1, ET-1, Bax and TGF-beta(1) increased in a dose-dependent manner, but NO and Bcl-2 decreased. Cardiac histology demonstrated exacerbated cardiac lesions in SHR when compared to WKY rats. Meanwhile, at the same dose exposed, the levels of hsCRP, d-dimer, ET-1 and the expression of IL-1beta, ICAM-1, ET-1, Bax and TGF-beta(1) were higher in SHR than those in WKY rats. The results indicated that ambient fine particles which entered into lungs could influence the cardiovascular system. When exposed to fine particles, SHR exhibited more severe cardiovascular injury in comparison to WKY rats. The results indicated that the inflammation, endothelial dysfunction, coagulation disorders and imbalance of apoptosis/anti-apoptosis might be the mechanisms of cardiovascular injury induced by fine particles. Different response between SHR and WKY rats after exposed to fine particles indicated that SHR was more susceptible than WKY rats to acute cardiac impairments from fine particle exposure.

[Study of genetic damage of human B lymphocyte cell line induced by 14 nm and 280 nm carbon black particles in vitro].

OBJECTIVE: To assess the genetic damage of human B lymphocyte cell line induced by 14 nm and 280 nm carbon black (CB) particles with micronucleus assay (CBMN), comet assay and hprt gene mutation test in vitro. METHODS: The genetic damage of human B lymphocyte cells exposed to 14 nm and 280 nm CB particles at the doses of 0, 128, 256, 384 and 512 microg/ml for 24 h and 48 h was detected using above three genotoxic assays. Micronucleus (MN) assay, comet assay, hprt gene mutation test were used to detect the genetic damage of human B lymphocyte cells induced by CB. Micronucleus rate (MNR), micronucleated cell rate (MCR), nuclear buds (Buds), nucleoplasmic bridges (NPBs), nuclear division index (NDI) and numbers of apoptotic cells served as indexes of CBMN assay; the percentage of DNA in the tail (% tail DNA) and the olive tail moment (OTM) were used as DNA damage indicators of comet assay; the hprt gene mutation frequency (Mf-hprt) served as the index of hprt gene mutation test. RESULTS: The % tail DNA, OTM in 14 nm CB group at the doses of 384 and 512 microg/ml for 48 h were 8.23% +/- 0.19%, 11.23% +/- 0.42% and 3.72 +/- 0.08, 4.90 +/- 0.18, respectively, which were significantly higher than those in control (5.10% +/- 0.08% and 2.22 +/- 0.03) (P < 0.01). The apoptotic cell rates in 14 nm CB group at the doses of 384 and 512 microg/ml for 48 h were 4.67 +/- 0.33 and 5.33 +/- 0.33, respectively, which were significantly higher than in control (0.00 +/- 0.00) (P < 0.05). The results of Mf-hprt were negative. CONCLUSION: The genetic damage of human B lymphocyte cells exposed to 14 nm CB particles for 48 h could be detected. But the similar effects didn't appear in 280 nm CB group.

[Establishment of a murine model of Alternaria alternate induced allergic asthma].

OBJECTIVE: To develop a method of establishment of airway allergic inflammation model of C57BL/6 mice sensitized and challenged with the spores of Alternaria alternate. METHODS: C57BL/6 mice were divided into negative control group, positive control group and model group, twelve mice in each group. Mice of model group were sensitized with spores at day 0, 5, 10 by intraperitoneal injection, and challenged four times by intranasal drip from day 14 to day 17. OVA instead of spores were used in the positive control group, PBS used in the negative control group. Bronchoalveolar lavage fluid (BALF) was taken at 24h after the last time of challenge, and the pathological manifestation of the lung, cell counts and differential count and IL-4, IFN-gamma total protein levels in BALF were assessed. The levels of total IgE and specific IgE in plasma were identified, the airway resistance and lung compliance were determined. RESULTS: There was pulmonary eosinophilic inflammation in the positive control group and model group. Total cells count, differential cells count and IL-4 and total protein levels in BALF were enhanced significantly while IFN-gamma declined remarkably. The levels of total IgE and specific IgE were increased remarkably in plasma in positive control group and model group, the airway resistance was enhanced and lung compliance decreased. CONCLUSION: An airway allergic inflammation model of C57BL/6 mice has been established by sensitizing and challenge with spores of Alternaria alternate.

[Research on the subchronic health effects of patina in rats by gavage].

OBJECTIVE: To study the 90-day subchronic health effects of patina, and its effects on anti-oxidative enzymes activities and lipid peroxidation levels in important organs of SD rats. METHODS: Eighty SD rats (40 males, 40 females) were divided into control group (treated with sodium carboxymetbycellulose) and patina exposure groups of different dose levels (7.3, 24.4 and 73.2 mg/kg bw). Animals were administered once daily by gavage, 6 times a week, continuously for 13 weeks. At the end of the study, clinical signs, body weights, serum biochemistry, urinalysis, organ weights were examined. Pathological examination, the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-N), and the concentrations of malondialdehyde (MDA) were respectively determined in brain, heart, liver, kidney, spleen, stomach. RESULTS: During the test period, there were no abnormal changes in the behavior, dieting, drinking, urine indicator, pathological changes and the body weights increased. At the dose of 7.3 mg/kg, significant increase was found in body Weight of male rats (P < 0.05). At the exposed group, the activities of LDH of serum in the 73.2 mg/kg appeared a significant decrease comparing with control group (P < 0.05), and there were significant increased in activities of SOD in livers, brains, hearts and activities of GSH-Px in hearts (P < 0.05). Meanwhile, the stomach levels of SOD, GSH-Px were significant reduced and MDA were significant increased at the two higher doses (24.4 and 73.2 mg/kg patina), compared with control group (P < 0.05). CONCLUSION: At certain levels, patina may increase body weight, improve the activities of anti-oxidative enxyme in brains, hearts, livers, but overload patina may cause oxidative damage to stomachs of rats.