[Source Analysis and Pollution Assessment of Soil Heavy Metals in Typical Geological High Background Area in Southeastern Chongqing].

In order to understand the pollution characteristics of soil heavy metals in typical geological high background areas in southeastern Chongqing, analyze its pollution sources, and put forward risk management and control suggestions, 211 topsoil samples (0-20 cm) were collected in Xinglong Town and Muye Township in the north of Youyang County, Chongqing City, and the contents of eight heavy metals (As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn) and the soil pH value in the surface soil were analyzed. Additionally, mathematical statistics, pollution load index (P), potential ecological risk index (RI), correlation analysis, and the positive matrix factorization (PMF) model were used to study the content and spatial distribution characteristics of heavy metals in the topsoil, and pollution assessment and source analysis of heavy metals were carried out. The results showed that the average concentrations of heavy metals in the surface soil all exceeded the soil background value in Chongqing, showing different levels of enrichment. Among them, the proportion of Cd exceeding the screening value in the "soil environmental quality risk control standard for soil contamination of agricultural land" (GB 15618-2018) was as high as 83.41%, the proportion of exceeding the control value was 7.58%, and the potential risk was relatively high. The evaluation results of the pollution load index method showed that the study area was slightly polluted as a whole, and the evaluation results of the potential ecological risk index showed that the overall ecological risk of the study area was relatively high. Among them, the contribution rates of heavy metals Hg, Cd, and As were 50.27%, 28.63%, and 11.70%, respectively. The PMF results showed that there were three main sources of heavy metals in the soil in the study area:natural parent material sources, agricultural sources, and mixed sources of industrial activities and traffic emissions, accounting for 40.49%, 29.12%, and 30.39%, respectively. The quantitative source analysis results of soil heavy metals in the study area have important reference significance for soil heavy metal pollution control in geologically high background areas.

[Source Analysis and Ecological Risk Assessment of Heavy Metals in the Arable Soil at the Geological High Background, Based on the Township Scale].

Due to the extensive development of carbonate rocks in southwest China, heavy metals are naturally occurring elements that have high natural background levels in the environment. Therefore, it is important to conduct ecological risk assessments and identify potential sources of heavy metals in the geological high background area. Based on the township scale, a total of 307 surface soil samples were collected in Qinglong Town, Fengjie County, Chongqing. The concentrations of As, Cd, Cr, Cu, Hg, Ni, Pb, and Zn and pH were analyzed and determined. The spatial distribution and source of soil heavy metals were discussed using the geostatistical analysis and an absolute principal component score-multiple linear regression (APCS-MLR) model in the studied area. The results showed that the average values of seven heavy metals (As, Cd, Cr, Cu, Hg, Ni, and Zn) in the arable soil exceeded the background values of Chongqing, and the cumulative effect of Cd and As was obvious. The concentrations of Cd significantly exceededthe screening values in The Risk Control Standard for Soil Environmental Quality and Soil Pollution in Agricultural Land (GB 15618-2018), with the over-standard rates of 52.12%. The spatial characteristics of soil heavy metal contents exhibited a pattern of high in the south and low in the north. PCA and APCS-MLR modeling revealed that the contributions of natural sources to Cr, Cu, Ni, and Zn were 86.62%, 64.34%, 76.44%, and 85.46%, respectively. As, Pb, and Hg mainly derived from industrial activities, which accounted for 74.63%, 61.90%, and 73.49%, respectively, and Cd was affected by both natural sources and industrial activities (accounting for 47.74% and 39.56% of the total Cd content, respectively). The evaluation of the soil by the Nemerow comprehensive index (P) showed that Cd pollution was relatively serious, accounting for 27.04% of soil pollution. The potential ecological hazard index showed that Cd and Hg were the main ecological hazard elements, and the distribution range of RI was 51.77 to 2228, indicating mainly mild and moderate risks, and the moderate and above risk areas in the study area were mainly located around the southern industrial source area. Altogether, our results revealed that in the study area, the heavy metal pollution was mainly caused by industrial activities, and the heavy metal pollution caused by geological background was mainly light to moderate. In conclusion, the medium and above risk areas in the study area were mainly caused by mineral and industrial activities, whereas the heavy metal pollution caused by geological background was mainly light to moderate pollution.

[Availability of Se-rich Arable Land in High Geological Background Area and Suggestions for Zoning].

In order to promote the results of land quality geochemical surveys to serve the development of characteristic land resources and realize the safe utilization of Se-rich resources in high geological background areas, Wuli Town, Qianjiang District, Chongqing City were taken as the research object to evaluate the Se-rich level and heavy metal content of soil crops and carry out research on Se-rich cultivated area division and the availability method system in the high geological background area, in order to improve the utilization efficiency of Se-rich resources and provide ideas for the safe utilization of Se-rich resources in high geological background areas. The results showed that the soil and crops in the study area were rich in Se and had the potential to develop Se-rich resources, but some samples had the problem of excessive Cd. Soil pH was a key factor affecting the content of Se and Cd in crops, and alkaline conditions were conducive to the safe utilization of Se-rich resources. The areas of Class A-E (A:selenium-enriched arable land, B:crops that were not selenium-enriched but were safe, C:crops that were selenium-enriched but unsafe, D:crops that were not selenium-enriched and unsafe, and E:soil selenium content less than the selenium-enriched threshold and soil heavy metals not exceeding the standard)cultivated land were 0.72, 0.75, 0.28, 0.13, and 0.56 km2, respectively. It is recommended to develop safe and Se-rich resources in the distribution area of Class A cultivated land and to carry out soil acidification conditioning and low-accumulation crops in the distribution area of Class C and D cultivated land; other agronomic measures should be taken to reduce the risk of excessive heavy metals in crops and strengthen the protection of cultivated land in the region to prevent the input of new pollutants.

[Zoning and Safe Utilization Method of Heavy Metal Contaminated Cultivated Land at Block Scale].

The safe use of arable land is one of the important measures to ensure food security and to realize the construction of ecological civilization. In order to solve the problem of blindly selecting technical measures in the process of safe use and restoration of pollution-risk cultivated land, 244 surface soil samples and 100 sets of rice-root soil samples were collected in Echi Town, Qianjiang District, Chongqing. Based on the contents of five heavy metals of Cd, Hg, Pb, As, and Cr, as well as soil oxides, organic matter, and pH and other indicators, a prediction model was established using multiple regression and geostatistical analyses, and the plots were assigned values, combined with the soil and soil in the plots. The total amount of heavy metals in the rice, the available content of heavy metals in the soil, and the pH of the soil guided suggestions for the safe use of cultivated land at the corresponding plot scale. The results showed that the soil in the study area was mainly and strongly acidic. The percentages of Cd exceeding the soil pollution risk screening value and control value were 33.61% and 2.05%, respectively. The effective content of Cd accounted for 60%, and the Cd exceeding rate of rice was 28%. There was an obvious ecological risk of Cd in the study area. The available Cd content of the soil was mainly affected by the total amount of soil Cd and pH. The enrichment of Cd in rice was mainly affected by the content of soil organic matter, Mn, and CaO. The zoning results showed that the priority protection category of the soil in the study area accounted for 59.30%; the safe use category accounted for 40.44%, of which safe use (IAa), safe use (IAb), safe use (IBa), safe use (IIAa), safe use (IIAb), and safe use (IIBa) accounted for 19.49%, 8.01%, 1.43%, 7.04%, 1.41%, and 3.06%, respectively; and strict control accounted for 0.26%. This method combined the safety risks of soil and agricultural products and aided formation of specific recommendations for safe use, which provided a method of reference for the safe use of contaminated farmland in accordance with local conditions.

RETRACTED ARTICLE: Experimental study and numerical prediction of HTO and 36Cl- diffusion in radioactive waste at Téguline Clay.

Statement of Retraction: Experimental study and numerical prediction of HTO and 36Cl- diffusion in radioactive waste at Téguline ClayWe, the Editor and Publisher of Environmental Technology, have retracted the following article:Yun-yi Zhang, Yong He, Bing-bing Li & Ke-neng Zhang, Experimental study and numerical prediction of HTO and 36Cl- diffusion in radioactive waste at Téguline Clay, Environmental Technology, Latest Articles, 10.1080/09593330.2021.1939433Following publication, errors have been identified in the experimental work and numerical simulation as follows: 1) The experimental results presented in this paper are too limited to effectively support the numerical analysis.2) Although XRD, MIP, SEM and other microstructural tests were conducted and presented in this paper, the results of these microstructural tests have not been used in the numerical analysis, leading to incorrect results shown in Figure 11.3) There is a serious discrepancy between the simulated results and the real conditions, due to an incorrect selection of diffusion parameters and initial concentration (200 mg/L and 500 mg/L) as well as an incorrect simulation model and boundary condition (Figure 4).4) The simulation software (Geo-studio + MATLAB) appears to be unsuitable for the simulation work in this paper due to physical, mechanical and adsorption properties of the sample not being considered in the simulation.These errors were identified and acknowledged by the authors. The errors are too substantial to be corrected in a corrigendum and will alter the conclusions of the paper. There are no concerns regarding research or publishing conduct, and the authors plan to revisit the relevant mature results of their research and may resubmit the revised paper for consideration by the journal.We have been informed in our decision-making by our policy on publishing ethics and integrity and the COPE guidelines on retractions.The retracted article will remain online to maintain the scholarly record, but it will be digitally watermarked on each page as "Retracted".

A Framework for the Automatic Integration and Diagnosis of Building Energy Consumption Data.

Buildings account for a majority of the primary energy consumption of the human society, therefore, analyses of building energy consumption monitoring data are of significance to the discovery of anomalous energy usage patterns, saving of building utility expenditures, and contribution to the greater environmental protection effort. This paper presents a unified framework for the automatic extraction and integration of building energy consumption data from heterogeneous building management systems, along with building static data from building information models to serve analysis applications. This paper also proposes a diagnosis framework based on density-based clustering and artificial neural network regression using the integrated data to identify anomalous energy usages. The framework and the methods have been implemented and validated from data collected from a multitude of large-scale public buildings across China.

Regulating the balance of Th17/Treg cells in gut-lung axis contributed to the therapeutic effect of Houttuynia cordata polysaccharides on H1N1-induced acute lung injury.

Our previous study had demonstrated that oral administration of Houttuynia cordata polysaccharides (HCP) without in vitro antiviral activity ameliorated gut and lung injuries induced by influenza A virus (IAV) in mice. However, as macromolecules, HCP was hard to be absorbed in gastrointestinal tract and had no effect on lung injury when administrated intravenously. The action mechanism of HCP was thus proposed as regulating the gut mucosal-associated lymphoid tissue (GALT). Actually, HCP treatment restored the balance of Th17/Treg cells firstly in GALT and finally in the lung. HCP reduced the expression of chemokine CCL20 in the lung and regulated the balance of Th17/Treg carrying CCR6+ (the CCL20 receptor), which was associated with specific migration of Th17/Treg cells from GALT to lung. In vitro, HCP inhibited Th17 cell differentiation through the downregulation of phospho-STAT3, whereas it promoted Treg cell differentiation by upregulating phospho-STAT5. Furthermore, its therapeutic effect was abolished in RORγt-/- or Foxp3-/- mice. These findings indicated that oral administration of macromolecular polysaccharides like HCP might ameliorate lung injury in IAV infected mice via directly regulating the balance of Th17/Treg cells in gut-lung axis. Our results provided a potential mechanism underlying the therapeutic effect of polysaccharides on pulmonary infection.

Flavonoids from Houttuynia cordata attenuate H1N1-induced acute lung injury in mice via inhibition of influenza virus and Toll-like receptor signalling.

BACKGROUND: Influenza virus is one of the most important human pathogens, causing substantial seasonal and pandemic morbidity and mortality. Houttuynia cordata is a traditionally used medicinal plant for the treatment of pneumonia. Flavonoids are one of the major bioactive constituents of Houttuynia cordata. PURPOSE: This study was designed to investigate the therapeutic effect and mechanism of flavonoid glycosides from H. cordata on influenza A virus (IAV)-induced acute lung injury (ALI) in mice. METHODS: Flavonoids from H. cordata (HCF) were extracted from H. cordata and identified by high-performance liquid chromatography. Mice were infected intranasally with influenza virus H1N1 (A/FM/1/47). HCF (50, 100, or 200 mg/kg) or Ribavirin (100 mg/kg, the positive control) were administered intragastrically. Survival rates, life spans, weight losses, lung indexes, histological changes, inflammatory infiltration, and inflammatory markers in the lungs were measured. Lung virus titers and neuraminidase (NA) activities were detected. The expression of Toll-like receptors (TLRs) and levels of NF-κB p65 phosphorylation (NF-κB p65(p)) in the lungs were analysed. The effects of HCF on viral replication and TLR signalling were further evaluated in cells. RESULTS: HCF contained 78.5% flavonoid glycosides. The contents of rutin, hyperin, isoquercitrin, and quercitrin in HCF were 8.8%, 26.7%, 9.9% and 31.7%. HCF (50, 100 and 200 mg/kg) increased the survival rate and life span of mice infected with the lethal H1N1 virus. In H1N1-induced ALI, mice treated with HCF (50, 100 and 200 mg/kg) showed lesser weight loss and lower lung index than the model group. The lungs of HCF-treated ALI mice presented more intact lung microstructural morphology, milder inflammatory infiltration, and lower levels of monocyte chemotactic protein 1 (MCP-1), interleukin-8 (IL-8), tumor necrosis factor-α (TNF-α) and malondialdehyde (MDA) than in the model group. Further investigation revealed that HCF exerted antiviral and TLR-inhibitory effects in vivo and in vitro. HCF (50, 100 and 200 mg/kg) reduced lung H1N1 virus titers and inhibited viral NA activity in mice. HCF (100 and 200 mg/kg) elevated the levels of interferon-ß in lungs. HCF also decreased the expression of TLR3/4/7 and level of NF-κB p65(p) in lung tissues. In vitro experiments showed that HCF (50, 100 and 200 µg/ml) significantly inhibited viral proliferation and suppressed NA activity. In RAW 264.7 cells, TLR3, TLR4, and TLR7 agonist-stimulated cytokine secretion, NF-κB p65 phosphorylation, and nuclear translocation were constrained by HCF treatment. Furthermore, among the four major flavonoid glycosides in HCF, hyperin and quercitrin inhibited both viral replication and TLR signalling in cells. CONCLUSION: HCF significantly alleviated H1N1-induced ALI in mice, which were associated with its dual antiviral and anti-inflammatory effects via inhibiting influenzal NA activity and TLR signalling. among the four major flavonoid glycosides in HCF, hyperin and quercitrin played key roles in the therapeutic effect of HCF.

Bupleurum polysaccharides ameliorated renal injury in diabetic mice associated with suppression of HMGB1-TLR4 signaling.

Bupleurum polysaccharides (BPs) is isolated from Bupleurum smithii var. parvifolium, a key traditional Chinese medicine. The study was to investigate the effects of BPs on diabetic kidney injury. After two intraperitoneal injections of streptozotozin (STZ) 100 mg·kg-1, renal injury in diabetic mice was induced and BPs was orally administrated at dosages of 30 and 60 mg·kg-1·d-1. The STZ injected mice developed renal function damage, renal inflammation and fibrosis known as diabetic kidney disease (DKD). BPs significantly reduced serum creatinine level and urinary albumin excretion rate, with the attenuated swelling of kidneys. BPs treatment obviously alleviated the pathological damage of renal tissue. The progression of renal injury in BPs treated mice was inhibited with less expression of type IV collagen (Col IV), fibronectin (FN) and α-smooth muscle actin (α-SMA). The inhibition of inflammation in kidney was associated with the reduced level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6). BPs administration suppressed the over-expression of toll like receptor 4 (TLR4) and high-mobility group box 1 (HMGB1) with lowered activity of nuclear factor kappa B (NF-κB) in renal tissue of diabetic mice. Oral administration of BPs effectively prevented the development ofrenal injury in diabetic mice. This study suggested that the protection provided by BPs might affect through the interruption of HMGB1-TLR4 pathway, leading to the inhibition of renal inflammation and fibrotic process.

[RAM study on general standard of maximum residue limits for pollution-free traditional Chinese medicine based on Chinese Pharmacopoeia formula].

The excessive pesticide residues and heavy metals in traditional Chinese medicine seriously endanger human health and the sustainable development of Chinese medicine industry. In order to improve the quality of traditional Chinese medicine and establish a general standard for maximum residue limits(MRL) of pesticides in pollution-free traditional Chinese medicine and decoction pieces, and to ensure the safety of clinical medication from its origin, MRLs were calculated based on the formula(MRL=A×W/100M) from Chinese Pharmacopeia, comparing it with the current Chinese and international standards as well as literature review, the RAND/UCLA appropriateness method(RAM) was applied to determine the categories and MRLs of pesticides in pollution-free traditional Chinese medicine and decoction pieces. Two questionnaires were drafted for expert panel and appropriateness analysis was carried out with the 9-point Likert scale to determine the general standard for MRLs of pollution-free traditional Chinese medicine and decoction pieces. The results showed that a total of nine experts from different fields scored the necessity of standard-setting and 206 pesticide residue limits respectively. The appropriateness scores of 206 pesticides were greater than 7, and appropriateness rate was 100%, which signifies that the expert panel has reached consensus. In summary, based on the RAM, the general standard for maximum residue limits of pesticides in pollution-free Chinese medicines and decoction pieces has reached an expert consensus. Comparing with the MRLs of medicinal plants and plant-sourced food from CAC, Europe Union, the United States, South Korea, Japan, Australia, New Zealand and Canada, 206 MRLs from this general standard share 88.8% in common, 4.4% of which is higher and 6.8% lower than those international standards. This has provided a basis for standardizing the use of pesticides in pollution-free traditional Chinese medicine.

Factors affecting variations of soil pH in different horizons in hilly regions.

Soil pH is a key factor that controls soil nutrient availability, soil microbial activities, and crop growth and development. However, studies on the soil pH variations of cultivated lands in different horizons at the regional scale remain limited. In this work, 348 soil samples were collected from three soil horizons (A, B, and C) at 120 sites over the hilly region of Chongqing, southwestern China. Six topographic indicators, four climate parameters, and parent material were considered. Classification and regression trees (CARTs) were applied to investigate the relationships between soil pH and the variables in the A, B, and C horizons. Model performances were evaluated by root mean square error (RMSE), relative root mean square error (RRMSE), and coefficient of determination (R2). Results showed that soil pH increased obviously from the A to C horizons. Soil pH was predicted well by the forcing factors with the CART models in all horizons. RMSE, RRMSE, and R2 varied between 0.37 and 0.435, between 5.93 and 7.23%, and between 0.71 and 0.80, respectively. The relative importance of the studied variables to soil pH differed with the horizons. Annual temperature range (ATR), terrain wetness index (TWI), and Melton ruggedness number were critical factors that controlled soil pH variability in the A horizon. Parent material, precipitation of warmest quarter (PWQ), ATR, and TWI were important variables in the B horizon. Parent material, PWQ, ATR, and precipitation were key factors in the C horizon. The results are expected to provide valuable information for designing appropriate measurements for agricultural practices and preventing soil acidification.

Houttuynia cordata polysaccharide alleviated intestinal injury and modulated intestinal microbiota in H1N1 virus infected mice.

Houttuynia cordata polysaccharide (HCP) is extracted from Houttuynia cordata, a key traditional Chinese medicine. The study was to investigate the effects of HCP on intestinal barrier and microbiota in H1N1 virus infected mice. Mice were infected with H1N1 virus and orally administrated HCP at a dosage of 40 mg(kg-1(d-1. H1N1 infection caused pulmonary and intestinal injury and gut microbiota imbalance. HCP significantly suppressed the expression of hypoxia inducible factor-1α and decreased mucosubstances in goblet cells, but restored the level of zonula occludens-1 in intestine. HCP also reversed the composition change of intestinal microbiota caused by H1N1 infection, with significantly reduced relative abundances of Vibrio and Bacillus, the pathogenic bacterial genera. Furthermore, HCP rebalanced the gut microbiota and restored the intestinal homeostasis to some degree. The inhibition of inflammation was associated with the reduced level of Toll-like receptors and interleukin-1ß in intestine, as well as the increased production of interleukin-10. Oral administration of HCP alleviated lung injury and intestinal dysfunction caused by H1N1 infection. HCP may gain systemic treatment by local acting on intestine and microbiota. This study proved the high-value application of HCP.

In vivo effect of quantified flavonoids-enriched extract of Scutellaria baicalensis root on acute lung injury induced by influenza A virus.

BACKGROUND: Scutellaria baicalensis root is traditionally used for the treatment of common cold, fever and influenza. Flavonoids are the major chemical components of S. baicalensis root. PURPOSE: To evaluate the therapeutic effects and action mechanism of flavonoids-enriched extract from S. baicalensis root (FESR) on acute lung injury (ALI) induced by influenza A virus (IAV) in mice. METHODS: The anti-influenza, anti-inflammatory and anti-complementary properties of FESR and the main flavonoids were evaluated in vitro. Mice were challenged intranasally with influenza virus H1N1 (A/FM/1/47) 2  h before treatment. FESR (50, 100 and 200  mg/kg) was administrated intragastrically. Baicalin (BG), the most abundant compound in FESR was given as reference control. Survival rates, life spans and lung indexes of IAV-infected mice were measured. Histopathological changes, virus levels, inflammatory markers and complement deposition in lungs were analyzed. RESULT: Compared with the main compound BG, FESR and lower content aglycones (baicalein, oroxylin A, wogonin and chrysin) in FESR significantly inhibited H1N1 activity in virus-infected Madin-Darby canine kidney (MDCK) cells and markedly decreased nitric oxide (NO) production from lipopolysaccharide (LPS)-stimulated RAW264.7 cells. In vitro assays showed that FESR and BG had no anti-complementary activity whereas baicalein, oroxylin A, wogonin and chrysin exhibited obvious anti-complementary activity. Oral administration of FESR effectively protected the IAV-infected mice, increased the survival rate (FESR: 67%; BG: 33%), decreased the lung index (FESR: 0.90; BG: 1.00) and improved the lung morphology in comparing with BG group. FESR efficiently decreased lung virus titers, reduced haemagglutinin (HA) titers and inhibited neuraminidase (NA) activities in lungs of IAV-infected mice. FESR modulated the inflammatory responses by decreasing the levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1), and increasing the levels of interferon-γ (IFN-γ) and interleukin-10 (IL-10) in lung tissues. Although showing no anti-complementary activity in vitro, FESR obviously reduced complement deposition and decreased complement activation product level in the lung . CONCLUSION: FESR has a great potential for the treatment of ALI induced by IAV and the underlying action mechanism might be closely associated with antiviral, anti-inflammatory and anti-complementary properties. Furthermore, FESR resulted in more potent therapeutic effect than BG in the treatment of IAV-induced ALI.

Polymyxin B as an inhibitor of lipopolysaccharides contamination of herb crude polysaccharides in mononuclear cells.

Lipopolysaccharides (LPS) contamination in herbal crude polysaccharides is inevitable. The present study was performed to explore the effect of polymyxin B on abolishing the influence of LPS contamination in mononuclear cells. LPS was pretreated with polymyxin B sulfate (PB) at different concentrations for 1, 5 or 24 h, and then used to stimulate RAW264.7 and mouse peritoneal macrophages (MPMs). The nitric oxide (NO) and tumor necrosis factor-α (TNF-α) in cell culture supernatant, as the indications of cell response, were assayed. Bupleurum chinensis polysaccharides (BCPs) with trace amount contamination of LPS was treated with PB. 30 µg·mL-1 of PB, treating LPS (10 and 1 000 ng·mL-1 in stimulating RAW264.7 and MPMs respectively) at 37 °C for 24 h, successfully abolished the stimulating effect of LPS on the cells. When the cells were stimulated with LPS, BCPs further promoted NO production. However, pretreated with PB, BCPs showed a suppression of NO production in MPMs and no change in RAW264.7. In the in vitro experiments, LPS contamination in polysaccharide might bring a great interference in assessing the activity of drug. Pretreatment with PB (30 µg·mL-1) at 37 °C for 24 h was sufficient to abolish the effects of LPS contamination (10 and 1 000 ng·mL-1).

Polysaccharides from Arnebia euchroma Ameliorated Endotoxic Fever and Acute Lung Injury in Rats Through Inhibiting Complement System.

Arnebiaeuchroma (Royle) Johnst (Ruanzicao) is a traditional Chinese herbal medicine (TCM). It is extensively used in China and other countries for treatment of inflammatory diseases. It is known that hyper-activated complement system involves in the fever and acute lung injury (ALI) in rats. In our preliminary studies, anti-complementary activity of crude Arnebiaeuchroma polysaccharides (CAEP) had been demonstrated in vitro. This study aimed to investigate the role and mechanism of crude Arnebiaeuchroma polysaccharides (CAEP) using two animal models, which relate with inappropriate activation of complement system. In lipopolysaccharide (LPS)-induced fever model, the body temperature and leukocytes of peripheral blood in rats were significantly increased, while the complement levels of serum were remarkably decreased. CAEP administration alleviated the LPS-induced fever, reduced the number of leukocytes, and improved the levels of complement. Histological assay showed that there were severe damages and complement depositions in lung of the ALI rats. Further detection displayed that the oxidant stress was enhanced, and total hemolytic activity and C3/C4 levels in serum were decreased significantly in the ALI model group. Remarkably, CAEP not only attenuated the morphological injury, edema, and permeability in the lung but also significantly weakened the oxidant stress in bronchoalveolar lavage fluid (BALF) in the ALI rats. The levels of complement and complement depositions were improved by the CAEP treatment. In conclusion, the CAEP treatment ameliorated febrile response induced by LPS and acute lung injury induced by LPS plus ischemia-reperfusion. CAEP exerted beneficial effects on inflammatory disease potentially via inhibiting the inappropriate activation of complement system.

Polysaccharides extracted from the roots of Bupleurum chinense DC modulates macrophage functions.

The present study aimed to investigate the effects of polysaccharides extracted from Bupleurum chinense DC (BCPs) on macrophage functions. In the in vivo experiment, 1 mL of 5% sodium thioglycollate was injected into the abdomen of the mice on Day 0 and macrophages were harvested on Day 4. The macrophages were cultured in plates and treated with different concentrations of BCPs and stimulus. Effects of BCPs on macrophage functions were assessed by chemotaxis assay, phagocytosis assay and Enzyme-Linked Immunosorbent Assay (ELISA). Our results showed the enhanced chemotaxis, phagocytosis and secretion of nitric oxide (NO) and inflammatory cytokines by macrophages when treated with BCPs. However, when chemotaxis and phagocytosis were up-regulated by complement components or opsonized particles, BCPs inhibited these effects. Also, the NO production induced by lipopolysaccharides (LPS) was suppressed by BCPs mildly. Moreover, BCPs had an inhibitory effect on the [Ca2+]i elevation of macrophages. These results suggested that BCPs exerted modulatory effects on macrophage functions, which may contribute to developing novel approaches to treating inflammatory diseases.

[Effect of Bupleurum polysaccharides on splenocytes from lupus like mouse re-challenged with Campylobacter jejuni-S(131) in vitro].

Mice were immunized with Campylobacter jejuni-S(131) (CJ-S(131)) to establish the lupus-like model. Splenocytes from lupus like mice were challenged with CJ-S(131) to induce inflammatory response in vitro. Bupleurum smithii var. parvifolium polysaccharides (BPs) was added in the inflammatory model to observe its underlying mechanisms of action on lupus. BALB/c mice were randomly divided into three groups including normal control group, adjuvant control group and lupus-like model. Mice were immunized on Day 0 and 14 with CJ-S(131) to establish lupus-like syndrome, and sacrificed on Day 19. Splenocytes from each group were collected and divided into blank control group, BPs added group (BPs 5, 10, 20, 40 µg·m L(-1)), CJ-S(131) stimulated group, and CJ-S(131) plus BPs group. The levels of total IgG, anti-ds DNA antibody, interferon-γ, interleukin-10 (IL-10) and IL-17 were quantified by ELISA. The proliferation of splenocytes was determined in the MTT assay. BPs significantly suppressed the high levels of total IgG, anti-ds DNA antibody, IFN-γ and IL-10 stimulated by CJ-S131 and had no significant effects on increased IL-17 secretion and splenocytes proliferation. The results suggest that re-stimulation of splenocytes with CJ-S(131) could establish an inflammatory model in vitro. The effect of BPs on lupus might is related to its inhibition of the production of autoantibodies and associated cytokines.

Houttuyniacordata Thunb. polysaccharides ameliorates lipopolysaccharide-induced acute lung injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Houttuynia cordata (HC) has been used as a folk therapy to treat pulmonary infections. This study aimed to determine the role and mechanism of action of polysaccharides isolated from HC (HCP) in lipopolysaccharide (LPS)-induced ALI in the mice. MATERIALS AND METHODS: LPS was delivered by the intratracheal route to Balb/c mice 2h before HCP (40, 80 and 160mg/kg) administration. RESULTS: The number of total cells, protein and tumor necrosis factor-α (TNF-α) concentrations in bronchoalveolar lavage fluid, the wet/dry weight ratio (w/d) of lungs and pulmonary pathology of each mouse were analyzed, it was found that HCP significantly alleviated ALI induced by LPS. Moreover, in lungs of mice, it was found that the infiltration of inflammatory cells, the expression of Toll-like receptor 4 and complement deposition were significantly decreased by HCP treatment. In vitro assays showed that C5a, a complement activation product, induced significant macrophage migration and treatment with HCP prevented it. The in vitro results also proved that LPS increased nitric oxide and pro-inflammatory cytokines (TNF-α, interleukin-6, and interleukin-1ß) production, and HCP antagonized these effects of LPS. It was also found that HCP alone augmented secretion of some pro-inflammatory cytokines. CONCLUSION: These results indicate that HCP may alleviate LPS induced lung inflammatory injury, which may be associated with its inhibitory effect on the over activation of complement and macrophages. This suggests a potential role to treat ALI.

Comparative subproteome analysis of three representative Leptospira interrogans vaccine strains reveals cross-reactive antigens and novel virulence determinants.

Pathogenic Leptospira spp. causes leptospirosis in China and throughout the world. Here, we have sequenced two L. interrogans moderately virulent vaccine strains JDL03 (serovar Canicola) and JDL10 (serovar Hebdomadis) used in China. We selected a subproteomic approach to identify surface-exposed proteins including OMPs and extracellular proteins of these two strains plus a highly virulent vaccine strain 56601 (serovar Lai). Comparative surface-exposed proteome among the three strains indicated 81 cores, 61 dispensable and 122 unique surface-exposed proteins. Finally, the 10 highly conserved surface-exposed or subsurface proteins included two known cross-reactive antigens (LipL32 and LA_3469) and another two novel antigens (LA_0136 and LA_0505) displaying conserved immunoreactivity among 15 Chinese epidemic serovars. Furthermore, many potential virulence factors were detected in these identified surface-exposed proteins, such as Loa22, LipL32, LenC, LenF and OmpL37. Interestingly, LipL45, ClpA and ClpB, exhibiting obvious amino acid mutations among str.56601, str.JDL03 and JDL10, might contribute to virulence differences observed among these strains. Additionally, specific surface-exposed proteins in virulent str.56601 were considered to be key virulence determinants, such as Zn-dependent protease, cholesterol oxidase precursor, and so on. In all, we had relatively complete surface-exposed subproteomes of L. interrogans, which will enhance our understanding of leptospiral pathogenesis and key virulence determinants. BIOLOGICAL SIGNIFICANCE: The present work demonstrates the use of genomic sequencing and subproteomic studies for the identification of potential vaccine and diagnostic antigen candidates against leptospirosis. The data show the conserved surface-exposed proteins to be novel potentially vaccine/diagnostic candidates. Furthermore, the data also show that LipL45, ClpA, ClpB and a lipoprotein from these three strains plus another highly virulent strain Fiocruz L1-130 contain specific amino acid mutations in strains JDL03 and JDL10. The surface-exposed subproteome of pathogenic L. interrogans could provide valuable information to gain a more complete understanding of leptospiral pathogenesis and virulence determinants.

Anti-complementary constituents of Houttuynia cordata and their targets in complement activation cascade.

Activity-guided fractionation for complement inhibitors led to the isolation of 23 known compounds from Houttuynia cordata Thunb. Seven flavonoids, two alkaloids, one coumarin and two phenols showed anti-complementary activity. Preliminary inhibitory mechanism of four flavonoids, including quercitrin, afzelin, isoquercitrin and quercetin in the complement activation cascade were examined for the first time. The results indicated that the target components of flavonols are different from those of flavonosides, and the glycoside moieties may be necessary to block C3 and C4 components.