Concentrations, seasonal trends, sources, health risk and subchronic toxicity to the respiratory and immune system of PAHs in PM2.5 in Xi'an.

PM2.5-bound polycyclic aromatic hydrocarbons (PAHs) have been proved to be hazardous to health. Previous studies have focused on the distribution and sources of PAHs, whereas there is little knowledge of the damage to organs. Here we sought to investigate the pollution level and seasonal variation characteristics of PAHs in PM2.5 in Xi'an and assess the health risk, to establish a PAHs exposure model, and investigate the toxicological effects of PAHs on the respiratory and immune functions. A sub-chronic exposure model of PAHs was established by inhalation. The pathological changes of lung tissues were observed with a light microscope. Inflammatory reactions in alveolar lavage fluid were determined using the corresponding kit. The levels of interleukin-6 (IL-6) and interleukin-8 (IL-8) were detected with enzyme linked immunosorbent assay (ELISA) kit; the proliferation of lymphocytes in spleen was detected with methyl tetrazolium (MTT); DNA immune damage was determined with DNA gel electrophoresis. The results showed that (1) the total concentration of 16 PAHs ranged from 41.1 to 387 ng/m3, with a mean value of 170 ng/m3, and the concentration of PAHs in PM2.5 was higher in winter than in other seasons. (2) The sources of PAHs in the atmosphere of Xi'an urban area were mainly coal combustion, and the equivalent carcinogenic concentration of PAHs in PM2.5 was 3.9 ng/m3. (3) Foreign body granuloma formation and inflammatory cell damage were observed in the lungs of rats infected with toxin; the levels of reactive oxygen species (ROS) and mobile device assistant (MDA) increased while nitric oxide synthase (NOS) decreased with the increase of dose; the expression levels of IL-6 and IL-8 elevated with the increase of toxin dose, showing an obvious dose-effect relationship; the level of PAHs damage to cells showed a dose-effect relationship. Sub-chronic exposure to PAHs could cause sustained inflammatory injury to the organism. Measures should be taken to counter the problems of PAHs in PM2.5 in Xi'an and relevant health promotion strategies should be developed.

Dual effects of atmospheric pressure plasma jet on skin wound healing of mice.

Cold plasma has become an attractive tool for promoting wound healing and treating skin diseases. This article presents an atmospheric pressure plasma jet (APPJ) generated in argon gas through dielectric barrier discharge, which was applied to superficial skin wounds in BALB/c mice. The mice (n = 50) were assigned randomly into five groups (named A, B, C, D, E) with 10 animals in each group. Natural wound healing was compared with stimulated wound healing treated daily with APPJ for different time spans (10, 20, 30, 40, and 50 seconds) on 14 consecutive days. APPJ emission spectra, morphological changes in animal wounds, and tissue histological parameters were analyzed. Statistical results revealed that wound size changed over the duration of the experimental period and there was a significant interaction between experimental day and group. Differences between group C and other groups at day 7 were statistically significant (p < 0.05). All groups had nearly achieved closure of the untreated control wounds at day 14. The wounds treated with APPJ for 10, 20, 30, and 40 seconds showed significantly enhanced daily improvement compared with the control and almost complete closure at day 12, 10, 7, and 13, respectively. The optimal results of epidermal cell regeneration, granulation tissue hyperplasia, and collagen deposition in histological aspect were observed at day 7. However, the wounds treated for 50 seconds were less well healed at day 14 than those of the control. It was concluded that appropriate doses of cold plasma could inactivate bacteria around the wound, activate fibroblast proliferation in wound tissue, and eventually promote wound healing. Whereas, over doses of plasma suppressed wound healing due to causing cell death by apoptosis or necrosis. Both positive and negative effects may be related to the existence of reactive oxygen and nitrogen species (ROS and RNS) in APPJ.

[Effect of Qufeng Tongluo Recipe on the expressions of connexin 36 protein and gene in rat mesangial cells].

OBJECTIVE: To determine the effect of Qufeng Tongluo Recipe (QFTLR) on the expressions of connexin 36 (Cx36) protein and gene in rat mesangial cells (MCs) and the proliferation of the MCs. METHODS: Serum samples containing Benazepril (Bena) and QFTLR were prepared in line with serum pharmacology methodology. The MCs cultured in vitro were divided into normal control and Lipopolysaccharide (LPS), Bena and QFTLR treated groups. The expressions of Cx36 protein and gene were detected by laser scanning confocal microscope (LSCM), Western blot, immunohistochemical assay and quantitative real time polymerase chain reaction (QRT-PCR) respectively. RESULTS: Compared with the control, higher level of Cx36 protein expression was found in the MCs than treated with LPS (P < 0.01). Both Bena and QFTLR lowered the level of Cx36 protein expression in the MCs treated with LPS significantly (P < 0.01 or P < 0.05). Similar results were found with the expression of Cx36 mRNA. CONCLUSION: QFTLR inhibits the proliferation of rat MCs, possibly through down-regulating the expressions of Cx36 protein and gene.

[Effect of artemisinin on the expressions of GRalpha mRNA, GRbeta mRNA and P300/CBP protein in lupus nephritis mice].

OBJECTIVE: To investigate the effects of Artemisinin on the expressions of GRalpha mRNA, GRbeta mRNA and P300/CBP protein in lupus nephritis mice. METHODS: Forty hybrid female mice were randomly and equally divided into four groups with the method of random number table: control group, model group, prednisone group administrated with 6.45 mg/kg Artemisinin (Art) suspension. A mice model of LN was established by injection with living lymph cell suspension. The expressions of GC receptor alpha (GRalpha) mRNA, GC receptor beta (GRbeta) mRNA in peripheral blood mononuclear cells (PBMCs), and transcriptional coactivator P300/CBP protein in renal tissue were respectively measured by the technique of RT - PCR and immunohistochemical assay. RESULTS: Compared with the model group. The expression of transcriptional coactivator P300/CBP protein in renal tissue and GRa mRNA in PBMCs of treatment groups was increased significantly, GRbeta mRNA expression was significantly decreased (P < 0.05 or P < 0.01). And the Art group had a better effect on the expression of GRalpha mRNA and transcriptional coactivator P300/CBP protein than that of the prednisone group (P < 0.01). CONCLUSION: The underlying therapeutic mechanism may be correlated with the regulation of Art on the expressions of GRalpha mRNA, GRbeta mRNA in PBMC and transcriptional coactivator P300/CBP protein in renal tissue.

[Effects of artemisinin on proliferation, apoptosis and Caspase-3 expression of rat mesangial cell].

OBJECTIVE: To study the effects of artemisinin on proliferation, apoptosis and Caspase-3 active of rat mesangial cell. METHODS: Rat mesangial cells were incubated with different concentrations of artemisinin, the proliferation, apoptosis and Caspase-3 active of rat mesangial cell were measured by MTT assay, fluorescent inverted microscope and enzyme-labeled analytical instruments respectively. RESULTS: Compared with control group, the proliferation and Caspase-3 expression of mesangial cell of three other groups were significantly different (P < 0.01). Compared with dexamethasone group, there were significant difference effects of proliferation and Caspase-3 expression of mesangial cell in other two groups of identical concentration drugs (P < 0. 01), especially in the artemisinin + glucocorticoid (ArtGC) group, and the effects of three different drugs on the mesangial cell Caspase-3 expression, proliferation and apoptosis had the tendency of depend on dosage, and mass mortality of mesangial cell in the mediate-dosage and high-dosage ArtGC group. CONCLUSION: Artemisinin could inhibit the proliferation of mesangial cell, enhance the expression of Caspase-3 and promote the apoptosis in a dose-dependent manner.

[Evaluation the Extent of Health Damage Caused by PM2.5 Particulate in Xi'an City].

The study evaluated the acute health damage effect of PM2.5 exposure in winter in Xi'an City, using the average concentration of PM2.5 (24 h) and the Average Poisson Regression Model for three consecutive years, from 2014 to 2016. The Corrected Human Capital Method was used to evaluate the economic loss from premature death. The cost of illness was used to assess the economic loss of hospitalization, illness and clinic. The results show that the economic loss in the study period (2014, 2015, 2016) caused by PM2.5 was about 33.523 billion yuan (95% CI: 24.961-36.975), 21.105 billion yuan (95% CI: 13.560-26.880), and 37.132 billion yuan (95% CI: 27.246-41.164). The proportions of GDP were as follows: 6.10% (4.54%-6.73%), 3.64% (2.34%-4.63%), and 5.91% (4.34%-6.55%) for each of the three years. The health economic loss was positively correlated with the average PM2.5 concentration in winter. PM2.5 pollutants have a significant impact on the health of residents in Xi'an. The number of reported cases was about 1071338 (95% CI: 646432-1385847), 438273 (95% CI: 246842-599989), and 1019503 (95% CI: 611407-1324547) for each of the three years. The effects on children with asthma were significantly higher than that on adults, while adults with chronic bronchitis were affected more significantly than children. This study can provide a scientific basis for cost-benefit analysis of the PM2.5 air quality standard in Xi'an, and provide a reference for the management of environment quality.

Low-temperature Plasma Promotes Fibroblast Proliferation in Wound Healing by ROS-activated NF-κB Signaling Pathway.

Low-temperature plasma (LTP) has shown great promise in wound healing, although the underlying mechanism remains poorly understood. In the present study, an argon atmospheric pressure plasma jet was employed to treat L929 murine fibroblasts cultured in vitro and skin wounds in BALB/c mice. The in vitro analysis revealed that treatment of fibroblasts with LTP for 15 s resulted in a significant increase in cell proliferation, secretion of epidermal growth factor (EGF) and transforming growth factor-ßi (TGF-ßi), production of intracellular reactive oxygen species (ROS), and the percentage of cells in S phase, protein expression of phosphorylated p65 (P-p65) and cyclinD1, but a noted decrease in the protein expression of inhibitor kappa B (IκB). The in vitro experiments demonstrated that 30-s LTP treatment enhanced the number of fibroblasts and the ability of collagen synthesis, while 50-s treatment led to the opposite outcomes. These results suggested that LTP treatment promotes the fibroblast proliferation in wound healing by inducing the generation of ROS, upregulating the expression of P-p65, downregulating the expression of IκB, and activating the NF-κB signaling pathway and consequently altering cell cycle progression (increased DNA synthesis in S phage).

Low temperature plasma promoting fibroblast proliferation by activating the NF-κB pathway and increasing cyclinD1 expression.

The potential applications of low temperature plasma (LTP) in wound healing have aroused the concern of many researchers. In this study, an argon atmospheric pressure plasma jet was applied to generate LTP for treatment of murine fibroblast cell (L929) cultured in vitro to investigate the effect of NF-κB pathway on fibroblast proliferation. The results showed that, compared with the control, L929 cells treated with plasma for less than 20 s had significant increases of proliferation; the productions of intracellular ROS, O2- and NO increased with prolongation of LTP treatment time; NF-κB pathway was activated by LTP in a proper dose range, and the expression of cyclinD1 in LTP-treated cells increased with the same trend as cell proliferation. After RNA interference to block p65 expression, with the same treatment time, RNAi-treated cells proliferated more slowly and expressed less cyclinD1 than normal cells. Furthermore, pretreatment with N-acetyl-L-cysteine (NAC) markedly prevented the plasma-induced changes in cells. In conclusion, the proliferation of L929 cells induced by LTP was closely related to NF-κB signaling pathway, which might be activated by appropriate level of intracellular ROS. These novel findings can provide some theoretical reference of LTP inducing cell proliferation and promoting wound healing.

[Distribution Characteristics and Source Analysis of Polycyclic Aromatic Hydrocarbons (PAHs) in Surface Dust of Xi'an City, China].

A total of 58 surface dust samples were collected from Xi'an city. The concentrations of 16 priority polycyclic aromatic hydrocarbons (PAHs) were analyzed by gas chromatography with a flame ionization detector (GC-FID). At the same time, the distribution and sources of PAHs in surface dust were studied. The results showed that the concentrations of individual PAH ranged from 14.69 to 6370. 48 microg x kg(-1), and the total concentrations of 16 PAHs (∑16 PAHs) ranged from 5039.67 to 47,738.50 microg x kg(-1), with a mean of 13,845.82 microg x kg(-1). Compared to the Y PAHs levels of other cities, the ∑16 PAHs in surface dust of Xi'an belonged to a relatively higher level. PAHs in surface dust were mainly dominated by high molecular weight PAHs with 4-6 rings and the concentration of 7 carcinogenic PAHs (1 CPAHs) accounted for 46.08% of 3 16PAHs. The mean of ∑16 PAHs in surface dust had the highest concentration at industrial area; followed by the educational area, traffic area, business and traffic area; residential area and parks showed relatively lower concentration. The average content of ∑16 PAHs in surface dust presented an increasing trend along the main urban area--the second ring road--the third ring road. ∑16 PAHs in surface dust had higher concentrations at the industrial areas of the east and west suburbs of Xi'an, the south suburb and the north section of the second ring road; ∑16 PAHs concentrations in the main urban area, north suburb and southeast part of Xi'an were relatively lower. The results of ratio, cluster analysis and principal component analysis showed that PAHs in surface dust were mainly originated from the combustion of fossil fuels and coal combustion. Among them, diesel combustion reached 36.07%, gasoline combustion accounted for 32.31%, and coal combustion was resposbe for 23.40%

[Distribution, Source and Human Exposure of Phthalic Acid Esters (PAEs) in Surface Dust in Urban Area of Xi'an City, China].

A total of 58 surface dust samples were collected in urban area of Xi'an City. High Performance Liquid Chromatography (HPLC) was used to analyze the concentrations of six phthalic acid esters (PAEs) listed as priority pollutants by United States Environmental Protection Agency (U.S. EPA). Composition, distribution, environmental sources and exposure characteristics of PAEs in the surface dust were further studied. All analyzed PAEs were detected in the surface dust. The concentration of individual PAE compounds varied from not detectable to 183.19 mg·kg-1 and their mean concentrations decreased in the order of DEHP>DnBP >>DEP >DMP >BBP >DnOP. The total concentration of six PAEs (∑6PAEs) ranged from 0.87 to 250.30 mg·kg-1 with an average of 40.48 mg·kg-1, and followed the order of parks >traffic area >mixed business and traffic area >residential area >educational area >industrial area. The ∑6PAEs presented the decreasing trend along the main urban area - the second ring road - the third ring road. The results of correlation analysis, principal component analysis and cluster analysis showed that PAEs in surface dust of Xi'an City were related to the application of plasticizers, the emission of cosmetics and personal care products along with building materials and home decoration materials. The dose order of human exposure to PAEs in surface dust was the direct ingestion by hand and mouth >>dermal adsorption >inhalation via mouth and nose. Meanwhile, the intake dose of children was higher than that of adults. However, the intake dose of DnBP, DEHP, DBP and BBP was lower than the tolerable daily intake (TDI) suggested by European Union Scientific Committee for Toxicity, Ecotoxicity and the Environment (EU CSTEE) and the reference doses (RfD) proposed by U.S. EPA.

"Qufeng Tongluo" acupuncture prevents the progression of glomerulonephritis by decreasing renal sympathetic nerve activity.

ETHNOPHARMACOLOGICAL RELEVANCE: Although the exact mechanism(s) underlying acupuncture remain unknown, acupuncture and acupuncture-like somatic nerve stimulation have been used to treat different kidney diseases and several complications related to them.The aim of this preliminary study was to assess the effectiveness of acupuncture on glomerulonephritis (GN) according to the theory of "Wind-hided renal collaterals" previously proposed. MATERIAL AND METHODS: We used a New Zealand white rabbit model of cationized bovine serum albumin (cBSA)-induced glomerulonephritis and then administered them metoprolol, irbesartan or acupuncture to evaluate the effectiveness of acupuncture treatment and preliminarily explore its potential mechanism. RESULTS: After immunization, our results showed that compared to the cBSA+MET and cBSA+IRB medication groups, "Qufeng Tongluo" significantly lowered parameters of renal function and improved podocyte injury in the 3rd, 6th and 8th weeks of treatment. Moreover, acupuncture increased the protein expression of phosphorylated ERK1/2. CONCLUSIONS: Our study suggests that a potential mechanism by which acupuncture has an antihypertensive effect and can significantly halt deteriorating renal function due to cBSA GN might be mediated by inhibiting the Erk1/2 MAPK pathway to reduce renal sympathetic nerve activity (RSNA).

Qufeng Tongluo Prescription () inhibits mesangial cell proliferation and promotes apoptosis through regulating cell cycle progression.

OBJECTIVE: To study the effects and possible underlying mechanism of Qufeng Tongluo Prescription (, QFTL) on the regulation of mesangial cells (MCs) proliferation and apoptosis. METHODS: The MCs used in this experiment have undergone five passages induced by lipopolysaccharide (LPS). Changes in the proliferation, apoptosis, cell cycle regulatory proteins and mRNA expression levels of the MCs after administration of Benazepril or QFTL were measured by methyl thiazolyl tetrazolium (MTT) reduction assay, flow cytometry, Western blot and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. RESULTS: The addition of Benazepril or QFTL serum inhibited LPS-induced MC proliferation after treatment for 24, 48 and 72 h, respectively (P<0.05 or P<0.01). Moreover, the inhibitory effect is more significant in the QFTL group at 48 h (P<0.05). Compared with the control group, LPS-induced cell proliferation decreased the number of cells in G1 phase versus cells in S and G2/M phases, while the addition of QFTL and Benazepril serum increased the ratio of cells at G1 phase (P<0.05 or P<0.01) to cells at S phase (P<0.01), implicating the cell cycle inhibition effect exerted by QFTL. LPS decreased the level of MC apoptosis, compared with the control group (P<0.05), while QFTL and Benazepril serum increased the level of MC apoptosis (P<0.01). Moreover, the difference between the QFTL group and the Benazepril group was statistically significant (P<0.01). Compared with the control group, the protein and mRNA expression levels of cylinD1, cyclin dependent kinase 2 (CDK2) and p21 were significantly increased (P<0.05 or P<0.01), p27 was decreased but with no statistical significance (P>0.05); After being treated with QFTL and Benazepril serum, the protein and mRNA expression levels of cylinD1, CDK2, p21 were decreased and p27 increased significantly (P<0.05 or P<0.01); Compared with the Benazepril group, QFTL show better effects on protein and mRNA expression levels of cylinD1, CDK2 (P<0.05 or P<0.01) and p21 protein expression (P<0.05). CONCLUSION: QFTL inhibits MCs proliferation, promotes MCs apoptosis through an underlying mechanism of down-regulating the protein and mRNA expression levels of cylinD1, CDK2, p21 and up-regulation of the expression level of p27.

[Effect of acupuncture on renal function and pathologic changes of kidney in rabbits with nephritis].

OBJECTIVE: To observe the effect of acupuncture on cationized bovine serum albumin (C-BSA) nephritis model in rabbits and to explore its mechanism. METHODS: Fifty rabbits were randomly divided into a blank group, a model group, a metoprolol group, a irbesartan group and an acupuncture group, 10 rabbits in each group. The model was established by ear vein intravenous injection with C-BSA. The positive control groups were treated by intragastric administrated with metoprolol and irbesartan, respectively. The acupuncture group was treated by acupuncture at "Fengmen" (BL 12) and "Shenshu" (BL 23). No interventions were added on the blank group and the model group. The changes of blood pressure (BP), heart rate (HR), plasma norepinephrine (NE), serum creatinine (Scr), blood urea nitrogen (BUN) and 24 hours urine protein (24 h UP) in rabbits at the time points of 3rd, 6th and 8th week of treatment were observed. RESULTS: After the model was established, the Scr of (194.30 +/- 20.09) micromol/L, BUN of (9.19 +/- 0.66) mmol/L and 24 h UP of (277.70 +/- 20.09) mg/24 h in the model group were all higher than the Scr of (66.03 +/- 4. 76) micromol/L, BUN of (4.11 +/- 0.71) mmol/L and 24 h UP of (14.28 +/- 1. 47) mg/24 h in the blank group (all P < 0.01), and the diffused mesenteria hyperplasia and the increase of intercapillary cells in the model group were showed in the pathological sections. After 3 weeks of treatment. The Scr of (99.82 +/- 9.29) micromol/L, BUN of (6.32 +/- 0.75) mmol/L and 24 h UP of (189.67 +/- 15.45) mg/ 24 h in the acupuncture group were all decreased significantly, furthermore, the decrease of BP, HR, NE were better than the other treatment groups (P < 0.05, P < 0.01). Except the level of 24 h up and HR at 8th week, other results were as same as the 3rd week. CONCLUSION: Acupuncture can improve the function of kidney, decrease the content of 24 h UP and the underlying therapeutic mechanism could be correlated with that acupuncture can lower excitability of sympathetic nerve and alleviate the renal pathological lesion induced by nephritis.

Effect of artemisinin combined with glucocorticoid on the expressions of glucocorticoid receptor α mRNA, glucocorticoid receptor ß mRNA and P300/CBP protein in lupus nephritis mice.

OBJECTIVE: To investigate the therapeutic effects and mechanisms of using artemisinin (Art) combined with glucocorticoid (GC) to treat lupus nephritis (LN) mice. METHODS: Forty hybrid female mice were randomly and equally divided into four groups with the method of random number table: control group, model group, prednisone group administrated with 6.45 mg/(kg·d) prednisone suspension, and Art+prednisone group administrated with 150 mg/(kg·d) Art suspension and 3.225 mg/(kg·d) prednisone suspension. A mice model of LN was established by injection with living lymph cell suspension. The changes of urine protein/24h, the expressions of GC receptor α (GRα) mRNA, GC receptor ß (GRß) mRNA in peripheral blood mononuclear cells (PBMCs), and transcriptional coactivator P300/CBP protein in renal tissue were measured. RESULTS: Compared with the model group, the treatment groups had significant decrease in urine protein/24 h, and renal pathological lesion (P<0.01). In the same groups, the expression of transcriptional coactivator P300/CBP protein in renal tissue and GRα mRNA were significantly increased, and GRß mRNA expression was significantly decreased (P<0.01). And the Art+prednisone group has a better therapeutic effect than the prednisone group (P<0.01). CONCLUSIONS: Art has therapeutic sensitization effects on GC in the LN mice. The underlying mechanism could be correlated with the effect of Art on the increase of the expressions of GRα mRNA and transcriptional coactivator P300 300/CBP protein in renal tissue and on the decrease of the expression of GRß mRNA in PBMC.

[Inactivation of bacterial spores using low-temperature plasma].

OBJECTIVE: To investigate the effect of low-temperature plasma on inactivation of bacterial spores and explore the mechanism. METHODS: Dielectric barrier discharge (DBD) was employed to generate the atmospheric low-temperature plasma for treatment of B.subtilis var. niger spores with the gas spacing of 3, 4 and 5 and treatment time intervals of 5, 10, 15, 20, 25, 30 and 35 s. The survived colonies was counted with plate counting method, and the killing log value (KLV) at different treatment times was calculated. The inactivation effect of electric field on B.subtilis var.niger spores was also investigated and the spores treated with low-temperature plasma were observed with transmission electron microscope. RESULTS: With the gap spacing of 3, 4 and 5 mm, the KLV of low-temperature plasma on B.subtilis var.niger spores within 25, 30 and 35 s of exposure was more than 5. The germicidal effects of the electric field on B. subtilis var.niger spores were rather poor. Transmission electron microscopy demonstrated total destruction of the surface and interior structure of the spores by low-temperature plasma. CONCLUSIONS: Low-temperature plasma is effective for inactivation of the bacterial spores with a time and dose dependence. The penetrating effect of charged particles and oxygenation effect of the reactive oxygen species might play a dominant role in plasma-induced bacterial spore inactivation, while the role of electric field is negligible.

[Experimental study on the in vitro resistance to Toxoplasma gondii by murine lymphocyte].

AIM: To study the activities of in vitro resistance to the tachyzoite of Toxoplasma gondii by murine lymphocytes. METHODS: The rat's splenocyte culture method was used to observe the effects of lymphocytes themselves and lymphocytes together with Mandelic( MA) on the invasion and proliferation of T. gondii in lymphocytes. At the same time acetylspiramycin was used as positive control. RESULTS: As compared with other somatic cells, the lymphocytes invaded by T. gondii still inhibited and killed the toxoplasma organisms in the presence of immunity, the effect safety dose of MA on inhibition of the invasion of T. gondii was not notable while the inhibition of the proliferation of T. gondii in lymphocytes was remarkable. CONCLUSION: Cell-mediated immunity(CMI) was an important factor that host resists the T. gondii infection. So, we should pay attention to improving the organism's CMI and take proper medicine so as to enhance the effect of lymphocyte's resistance to T. gondii.