miRNA Expression Analysis of IPEC-J2 Cells Damaged by Soybean 7S Globulin Reveals ssc-miR-221-5p as the Factor Alleviating Cell Damage.

The most significant and sensitive antigen protein that causes diarrhea in weaned pigs is soybean 7S globulin. Therefore, identifying the primary target for minimizing intestinal damage brought on by soybean 7S globulin is crucial. MicroRNA (miRNA) is closely related to intestinal epithelium's homeostasis and integrity. However, the change of miRNAs' expression and the function of miRNAs in Soybean 7S globulin injured-IPEC-J2 cells are still unclear. In this study, the miRNAs' expression profile in soybean 7S globulin-treated IPEC-J2 cells was investigated. Fifteen miRNAs were expressed differently. The differentially expressed miRNA target genes are mainly concentrated in signal release, cell connectivity, transcriptional inhibition, and Hedgehog signaling pathway. Notably, we noticed that the most significantly decreased miRNA was ssc-miR-221-5p after soybean 7S globulin treatment. Therefore, we conducted a preliminary study on the mechanisms of ssc-miR-221-5p in soybean 7S globulin-injured IPEC-J2 cells. Our research indicated that ssc-miR-221-5p may inhibit ROS production to alleviate soybean 7S globulin-induced apoptosis and inflammation in IPEC-J2 cells, thus protecting the cellular mechanical barrier, increasing cell proliferation, and improving cell viability. This study provides a theoretical basis for the prevention and control of diarrhea of weaned piglets.

[Distribution Characteristics and Environmental Driving Factors of Cyanobacteria Community in Impounded Lakes and Reservoirs in Shandong on the East Route of South-to-North Water Diversion Project].

To investigate the distribution characteristics of the cyanobacteria community and the driving factors in impounded lakes and reservoirs in Shandong on the east route of the South-to-North Water Diversion Project, monthly samples of phytoplankton and the aquatic environment from Nansi Lake, Dongping Lake, Datun Reservoir, Donghu Reservoir, and Shuangwangcheng Reservoir were collected from May to November during 2010 to 2019. A total of 44 planktonic cyanobacteria taxa were identified with 23 filamentous cyanobacteria taxa. Pseudanabaena limnetica, Cylindrospermopsis raciborskii, Microcystis aeruginosa, and Microcystis wesenbergii were the dominant harmful cyanobacteria species, with a high detection frequency and abundance in all lakes and reservoirs. By analyzing the distribution characteristics of the cyanobacteria community in impounded lakes and reservoirs, we found that filamentous cyanobacteria had growth advantages in the water with large hydraulic disturbances, which should be the key points of cyanobacteria prevention and control in the future. Pearson correlation analysis and generalized linear fitting curve results showed that total nitrogen, total phosphorus, water temperature, and water depth played a key role in affecting the growth of P. limnetica, C. raciborskii, M. aeruginosa, and M. wesenbergii. The nitrogen and phosphorus nutrients could promote the growth of harmful cyanobacteria. Due to the good temperature adaptability, P. limnetica could still become the dominant species in early summer and late autumn, and C. raciborskii, M. aeruginosa, and M. wesenbergii had growth advantages when the water temperature was higher than 25℃. In addition, shallow water was more conducive to the growth of C. raciborskii. It was suggested that based on strengthening of the control of nitrogen and phosphorus nutrient input in lakes and reservoirs, the key monitoring of P. limnetica in lakes should be conducted in early summer and late autumn, and the growth of C. raciborskii in shallow water areas should be paid close attention in the high temperature period to ensure the safety of water quality.

UPLC-MS/MS simultaneous quantification of urinary circadian rhythm hormones and related metabolites: Application to air traffic controllers.

Civil aviation flight crew and civil aviation air traffic controllers are prone to circadian rhythm abnormalities, which can lead to a slew of other maladies. It could endanger people's health and provide a serious threat to the safety of civil aviation flights if it is not appropriately evaluated and addressed. Early detection of rhythm irregularities and prompt treatment for particular populations that are vulnerable to rhythm disorders are crucial for enhancing civil aviation safety. In general, monitoring of the classical circadian rhythm biomarkers (melatonin or cortisol) in plasma or saliva is an effective way to evaluate the rhythm status. Due to the challenging sample procedure and the trauma of plasma, urine sample testing has received an increasing amount of attention. While, urine circadian rhythm biomarkers have seldom been examined, and the relationship between urinary steroid hormones and melatonin is still poorly understood. In most cases, hormones are determined by immunoassays respectively, mainly enzyme-linked immunosorbent assay (ELISA) or radioimmunoassay (RIA). There are also reports describing the liquid chromatography with tandem mass spectrometry (LC-MS/MS) technique as a method of melatonin or few steroid hormones quantification, however, the simultaneous detection of multiple rhythmic hormones in human urine is rarely reported. For the quantification of the rhythmic hormones in human urine, an accurate approach using ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was devised in this work. Nine endogenous hormones (melatonin, 6-hydroxymelatonin, 6-sulfatoxymelatonin, cortisol, corticosterone, cortisone, testosterone, epitestosterone and androsterone), in human overnight urine, were quantified after solid phase extraction (SPE). A reverse phase HSS C18 column was used for chromatographic separation with a 9-minute gradient elution and deuterated analogues of each analyte were applied as internal standards. This method was successfully applied to the analysis of 596 overnight urine samples (23:00-9:00) collected from 84 air traffic controllers in the Beijing area during shift work. This study's findings showed a clear correlation not only between melatonin and its metabolites; cortisol-related metabolites, but also between melatonin metabolites and endogenous metabolites upstream and downstream of cortisol, implying that these two categories of hormones can be used as potential biological rhythm indicators to provide circadian rhythm data support for future studies on circadian rhythm disorders.

Bacillus toyonensis SAU-19 and SAU-20 Isolated From Ageratina adenophora Alleviates the Intestinal Structure and Integrity Damage Associated With Gut Dysbiosis in Mice Fed High Fat Diet.

This study was performed to identify potential probiotic endophytes from Ageratina adenophora and evaluate their ameliorating effects on gut injury and integrity damage associated with microbiota dysbiosis in mice fed high fat diet. Using morphological and biochemical tests, and 16S rRNA gene sequencing technique, two bacteria endophytes were identified as strains of Bacillus toyonensis and were named Bacillus toyonensis SAU-19 (GenBank No. MW287198) and Bacillus toyonensis SAU-20 (GenBank No. MW287199). Sixty (60) mice were divided into five groups, group 1 was the negative control fed normal diet (NS), group 2 was fed High fat diet (HF), Group 3 was fed High fat diet + 106 Lactobacillus rhamnosus (LGG), group 4 was fed High fat + 106 Bacillus toyonensis SAU-19 and group 5 fed High fat diet + 106 Bacillus toyonensis SAU-20. After 35 days, histological and immunohistochemistry examination were performed in the ileum tissues. Furthermore, DAO and antioxidants activities were measured in serum, mRNA expressions of tight junction proteins (occludin and ZO-1) and inflammation related cytokines (IL-1ß, TFN-α, IL-2, IL-4, and IL-10) in the ileum tissues as well as sIgA levels and total bacteria (Escherichia coli, Salmonella, Staphylococcus, and Lactobacillus) in the small intestine and cecum content. The results showed an increase in the DAO activity, oxidative stress parameter (MDA), pro-inflammation cytokines (IL-1ß, TFN-α, IL-2), reduce immunity (sIgA), and destroyed intestinal structure and integrity (reduce tight junction proteins) in the high fat diet group and this was associated with destruction of the gut microbiota composition (increasing pathogenic bacteria; E. coli, Salmonella, Staphylococcus and reducing beneficial bacteria, Lactobacillus spp.) in mice (P < 0.05). However, the administration of Bacillus toyonensis SAU-19 and SAU-20 reverted these effects. Our findings indicated that, Bacillus toyonensis SAU-19 and SAU-20 isolated from A. adenophora could prevent the excess weight gain from high fat diet feeding, improved antioxidant status and alleviated the intestine integrity damage as well as reduce the population of enteric bacteria such as E. coli, Salmonella, and S. aureus and increasing the population of beneficial bacteria such as Lactobacillus in the gut of mice fed high fat diet, therefore, can serve as a potential probiotics in humans and animals.

An Overview: The Toxicity of Ageratina adenophora on Animals and Its Possible Interventions.

Ageratina adenophora is one of the major invasive weeds that causes instability of the ecosystem. Research has reported that A. adenophora produces allelochemicals that inhibit the growth and development of food crops, and also contain some toxic compounds that cause toxicity to animals that consume it. Over the past decades, studies on the identification of major toxic compounds of A. adenophora and their toxic molecular mechanisms have been reported. In addition, weed control interventions, such as herbicides application, was employed to reduce the spread of A. adenophora. However, the development of therapeutic and prophylactic measures to treat the various A. adenophora-induced toxicities, such as hepatotoxicity, splenotoxicity and other related disorders, have not been established to date. The main toxic pathogenesis of A. adenophora is oxidative stress and inflammation. However, numerous studies have verified that some extracts and secondary metabolites isolated from A. adenophora possess anti-oxidation and anti-inflammation activities, which implies that these extracts can relieve toxicity and aid in the development of drug or feed supplements to treat poisoning-related disorders caused by A. adenophora. Furthermore, beneficial bacteria isolated from rumen microbes and A. adenophora can degrade major toxic compounds in A. adenophora so as to be developed into microbial feed additives to help ameliorate toxicity mediated by A. adenophora. This review presents an overview of the toxic mechanisms of A. adenophora, provides possible therapeutic strategies that are available to mitigate the toxicity of A. adenophora and introduces relevant information on identifying novel prophylactic and therapeutic measures against A. adenophora-induced toxicity.

Ageratina adenophora Disrupts the Intestinal Structure and Immune Barrier Integrity in Rats.

The aim of this study was to investigate the effects of Ageratina adenophora on the intestines morphology and integrity in rat. Rats were randomly divided into two groups and were fed with 10 g/100 g body weight (BW) basal diet and 10 g/100 g BW experimental diet, which was a mixture of A. adenophora powder and basal diet in a 3:7 ratio. The feeding experiment lasted for 60 days. At days 28 and 60 of the experiment, eight rats/group/timepoint were randomly selected, weighed, and sacrificed, then blood and intestinal tissues were collected and stored for further analysis. The results showed that Ageratina adenophora caused pathological changes and injury in the intestine, elevated serum diamine oxidase (DAO), D-lactate (D-LA), and secretory immunoglobulin A (sIgA) levels, reduced occludin levels in intestinal tissues, as well as increased the count of intraepithelial leukocytes (IELs) and lamina propria leukocytes (LPLs) in the intestine (p < 0.05 or p < 0.01). In addition, the mRNA and protein (ELISA) expressions of pro-inflammation cytokines (IL-1ß, IL-2, TNF-α, and IFN-ϒ) were elevated in the Ageratina adenophora treatment groups, whereas anti-inflammatory cytokines such as IL-4 and IL-10 were reduced (p < 0.01 or p < 0.05). Therefore, the results obtained in this study indicated that Ageratina adenophora impaired intestinal function in rats by damaging the intestine structure and integrity, and also triggered an inflammation immune response that led to intestinal immune barrier dysfunction.

Toxic mechanisms and pharmacological properties of euptox A, a toxic monomer from A. adenophora.

A. adenophora (Spreng.) R.M. King & H. Rob. is as invasive plant known to cause toxicity in humans and animals. The plant's toxic activities have been associated with some toxic phytochemicals present in the plant. One of the major phytochemicals that have been reported to induce toxicity in various organs is euptox A (9-oxo-10, 11-dehydroageraphorone). Previous studies have reported that the main target organs of euptox A are the liver and spleen. Although, many studies have reported on euptox A toxicity in rats and mice, the mechanism of action and the beneficial uses of this toxin as well as it potential uses have not been fully established in literatures. Therefore, this review firstly, aims at elaborating on the toxic effects and mechanism of action of euptox A to give basic knowledge to researchers to help in the development of strategies that will reduce its toxicity to the environment. Secondly, this paper will also report on some beneficial uses of euptox A in recent years as well as suggest some future potential applications of this toxin to help in the utilization of this plant resource.

Ageratina adenophora Inhibits Spleen Immune Function in Rats via the Loss of the FRC Network and Th1-Th2 Cell Ratio Elevation.

The objective of this study was to determine the impact of Ageratina adenophora (A. adenophora) on splenic immune function in a rat model. Rats were fed with 10 g/100 g normal feed and an experimental feed, which was composed of 3:7 A. adenophora powder and normal feed for 60 days. On days 14, 28, and 60, subsets of rats (n = 8 rats/group/time point) were selected for blood and spleen tissue sample collection. The results showed that the proportion of CD3+ T cells in the spleen was decreased at day 60 (vs. control). Also, mRNA and protein expression of chemokines CCL21 and CCL19 and functional protein gp38 in spleen decreased significantly versus the control at day 60. In addition, ER-TR7 antigen protein expression was also decreased at day 60. Levels of T-helper (Th)1 cells significantly increased, whereas those of Th2 cells decreased significantly versus the control at day 60 in spleen. The finding revealed that A. adenophora could affect splenic immune function in rats by altering the fibroblast reticulocyte (FRC) network, as well as by causing an imbalance in Th1/Th2 cell ratios. This research provides new insights into potential mechanisms of spleen immunotoxicity due to exposures to A. Adenophora.

Identification of characteristic heroin metabolites in urine based on data-mining technology and multivariate statistics analysis combined with a targeted verification approach for distinguishing heroin abusers.

A common phenomenon shows that ingestion of opium poppy shell-containing drugs can result in a "false-positive" urinalysis test result for mandatory or workplace heroin abuse screening. Owing to the short detection window (8 h in urine) of the characteristic heroin metabolite 6-monoacetylmorphine (6-MAM) confirmation or exclusion of heroin abusers still presents major challenges for toxicologists. In this work, we developed an ultra-performance liquid chromatography-time-of-flight mass spectrometry method (UPLC-TOF-MS) with online data acquisition and multiple post-data-mining technologies combined with a multivariate statistical and batch validation analysis workflow to assess the characteristic urine metabolites of heroin abusers. Based on the proposed methods, 28 characteristic metabolites were structurally identified, and their fragmentation patterns and metabolite pathways were also summarized. Correlation analysis was used to investigate the internal relationship and similarities among the identified metabolites, and seven representative metabolites were selected as "Target-metabolites". Multi-batch urine of samples of heroin abusers were certified based on the UPLC-MS/MS method for further validation of the practicability of using this method for routine analysis. Overall, the target-metabolites can be utilized as assistant "biomarkers" in workplace or mandatory drug screenings. This approach encourages further studies on the development of the "false-positive" identification system.

[Simultaneous determination of 7 benzodiazepines in human plasma by UHPLC-MS/MS].

The study was designed to develop the method for determination of 7 benzodiazepines concentration in human plasma. UHPLC-MS/MS was adopted to analyze plasma with protein precipitated by acetonitrile. Citalopram was used as an internal standard. Plasma samples were separated on CORTECS UHPLC C18 column with the mobile phase of aqueous solution (0.01% formic acid) - methanol (0.01% formic acid) at a flow rate of 0.3 m L·min(-1). Multiple reaction monitoring (MRM) mode was performed in combiation with electrospray ionization source operating in the positive ionization mode. The liner calibration curve of midazolam, nitrazepam, estazolam, clonazepam, lorazepam, triazolam and diazepam were obtained in the concentration range of 1.05-840 (r = 0.999 4), 2.06-824 (r = 0.998 1), 2.02-1 616 (r = 0.994 7), 6.18-2 472 (r = 0.997 9), 6.12-2 448 (r = 0.997 4), 3.02-2 416 (r = 0.990 2), 1.02-816 (r = 0.998 8) ng·m L(-1), respectively. The lowest detection limit were 0.02, 0.52, 0.51, 1.55, 0.77, 0.76, 0.02 ng·m L(-1), respectively. The RSD of inter-day and intra-day were less than 10.81%. The relative recovery was 81.46%-106.53%. The method was successfully applied to clinical analysis of blood samples from patients.

Does copper reduce cadmium uptake by different rice genotypes?

A hydroponics experiment was conducted to investigate the effect of copper (Cu) on cadmium (Cd), calcium (Ca), iron (Fe), and zinc (Zn) uptake by several rice genotypes. The experiment was carried out as a 2x2x4 factorial with four rice genotypes and two levels of Cu and Cd in nutrient solution. Plants were grown in a growth chamber with controlled environment. The results showed a significant difference between the biomass of different rice genotypes (P < 0.001). The Cd and Cu concentration in the solution had no significant effect on the biomass. The addition of Cu significantly decreased Cd uptake by shoots and roots of rice (P < 0.001). The Cd concentration did not significantly influence Ca uptake by plants, whereas the Cu concentration did (P = 0.034). There was a significant influence of Cd on Fe uptake by shoots and roots (P < 0.001, P = 0.003, respectively). Zn uptake decreased significantly as the addition of Cd and Cu increased in shoots. We concluded that Cu had significant influence on Cd uptake. The possible mechanisms were discussed.

[Effects of arbuscular mycorrhizal fungi on the toxicity of chlorothalonil on upland rice (Oryzal sativa L.)].

Upland rice was selected as a host plant in a greenhouse-pot-culture experiment to investigate the effects of chlorothalonil and arbuscular mycorrhizal fungi (AMF) on the growth and oxidative damage under chlorothalonil stress. The plants were grown with three concentrations of chlorothalonil (0, 50 and 100 mg x kg(-1) soil). The results suggested that chlorothalonil inhibited the growth of upland rice and reduced dry weight of upland rice shoot from 2.5 g x pot(-1) to 1.0 g x pot(-1), and the root dry weight was reduced from 0.9 g x pot(-1) to 0.3 g x pot(-1). However, with AMF colonization the dry weight of upland rice was increased to 2.1 g x pot(-1). With 50 mg x kg(-1) chlorothalonil addition, the P uptake was reduced from 3200 microg x pot(-1) to 860 microg x pot(-1), but the AMF colonization could increased the P uptake to 1900 microg x pot(-1). Chlorothalonil induced oxidative stress indicating by the changes in activities of antioxidative enzyme and AMF colonization could alleviate the oxidative stress. These results showed that chlorothalonil induced oxidative stress and inhibited P uptake in upland rice and AMF could decrease the side effects of chlorothalonil by increasing P uptake and decreasing oxidative damages.

[Adverse health effects of cadmium and related factors].

Cd is a very toxic chemical to animal and human beings. Cd contamination has been ubiquitous because of industry and the human activities. Cd not only affects adversely the growth, yield and quality of crops but also effects on the health of animal and human being. The damage of Cd on bone, kidney, liver, immune and procreation systems as well possible factors that influence Cd absorption, accumulation and damage in the body are summarized in this paper.

Exposure to metal mixtures and human health impacts in a contaminated area in Nanning, China.

Cadmium and lead have been identified as very toxic metals, which are widely present in the environment due to natural and anthropogenic emissions. Many studies have shown that the food chain is the main pathway of cadmium and lead transfer from the environment to humans. It is well documented that many factors will affect their transfer through food chains. Previous investigations on heavy metals were mostly concentrated on one contaminant in isolation. However, in real environments, exposure to mixtures of metals is ubiquitous such that cadmium pollution is invariably being associated with lead and zinc, etc. This study focuses on the contamination and health effects of the metal mixtures. For this purpose, a dietary survey was taken for 3 groups in Nanning in October 2002. Samples of soils, plants (vegetables), urine and blood of humans were measured for Cd, Fe, Cu, Zn, Ca and Pb, in addition, the urinary indicators of renal dysfunction Albumin (ALB), N-acetyl-beta-D-glucosaminidase (NAG), Beta-2-microglobulin (beta2-MG) and Retinol-binding protein (RBP) in urine were also measured. Results showed that soil contamination with metal mixtures had caused significant renal dysfunction of the local residents living in the contaminated area, and the dose-response curve was somewhat altered by the mixed contamination of Cd and Pb as well as the intake of other minerals. The importance of mixtures of metal contamination and human health are also discussed in this paper.

[Effect of calcium content in diet on the accumulation and toxicity of cadmium in organisms].

Cadmium (Cd) is one of the most toxic elements with no benefit to plants and human health. Cd is absorbed via gastrointestinal track through food chain and hence, poses subsequent health risks. Data available showed that the interdependence exists between nutritional status of the organism and Cd accumulation and its toxicity. In this paper, interaction of dietary calcium (Ca) and Cd toxicity was reviewed and discussed.

Transfer of metals from soil to vegetables in an area near a smelter in Nanning, China.

A field survey was conducted to investigate the metal contamination in soils and vegetables, and to evaluate the possible health risks to local population through foodchain transfer near a smelter in Nanning, southern China. Contamination levels in soils and vegetables with cadmium (Cd), lead (Pb), zinc (Zn) and copper (Cu) were measured, and transfer factors (TF) from soils to vegetable plants and its health risk (risk index, RI) were calculated accordingly. Results showed that both soils and vegetables from villages 1 and 2 (V1 and V2, 1500 m and 500 m from the smelter) were heavily contaminated, compared to a village 50 km from the smelter. Geometric mean of Cd and Pb concentrations in vegetables for V1 and V2, respectively, were 0.15 and 0.24 mg Cd kg(-1) and 0.45 and 0.38 mg Pb kg(-1) (on fresh weight basis). Oral intake of Cd and Pb through vegetables poses high health risk to local residents. Risk indices for V1 and V2, respectively, were 3.87 and 7.42 for Cd, and 1.44 and 13.5 for Pb. The complexity of metal contamination and their health risks are also discussed.

Ecological footprint of Shandong, China.

Ecological footprint has been given much attention and widely praised as an effective heuristic and pedagogic device for presenting current total human resource use in a way that communicates easily to almost everyone since 1996 when Wackernagel and Rees proposed it as a sustainable development indicator. Ecological footprint has been improving on its calculation and still can be a benchmark to measure sustainable development although there are still ongoing debates about specific methods for calculating the ecological footprint. This paper calculates the ecological footprint of Shandong Province, China with the methodology developed by Wackernagel and analyzes the current situation of sustainable development in Shandong.

Cadmium uptake by different rice genotypes that produce white or dark grains.

A pot experiment was conducted to investigate cadmium (Cd) uptake by different rice cultivars that produce white or dark grains. Four cultivars with white grains (hereafter, white rice) and five cultivars with dark colors (hereafter dark rice) were selected for this experiment. Three levels of soil Cd concentrations, background (0), 5 and 10 mg/kg, were used. After harvest, plant biomass, tissue concentrations of Cd, Ca, Fe, Cu and Zn were analyzed. The results showed that Cd concentrations are significantly different between different genotypes, but when comparing the Cd concentrations for the two groups, no significant difference was found. For other divalent cations, Ca concentrations in dark rice were higher than those in white ones (P < 0.001 for shoots, P = 0.037 for roots); Fe concentrations in dark rice were also higher than those in white ones (P = 0.001 either in shoot or root); Zn concentrations in shoot of dark rice were higher than those in white ones, but no significant difference in roots. The total molar concentrations of divalent cations in dark rice were also significantly higher than in white rice. The potential benefit of higher Ca and Fe concentrations in dark rice and similar Cd concentrations in both groups is also discussed in this paper.