Effects of solid waste-based soil conditioner and arbuscular mycorrhizal fungi on crop productivity and heavy metal distribution in foxtail millet (Setaria italica).

Shanxi is a large coal-producing province, and it also produces a lot of solid waste. Solid waste can leach heavy metals, which can harm soil and affect food security at the beginning of the food chain. To investigate the impacts of solid waste-based soil conditioner (SWSC) and arbuscular mycorrhizal fungi (AMF) on millet safety and crop production, a field experiment with foxtail millet (Setaria italica) was conducted in Tunliu. The results of this study demonstrate that SWSC + AMF, SWSC and AMF can increase millet yield by 28.0%, 27.1% and 19.5%, respectively, compared with CK. This is mainly due to increased mycorrhizal infection. Besides, the pollution index (Pi) and the Nemerow-integrated pollution index (PN) of the soil with SWSC and AMF were both below 0.7, indicating safe pollution levels. The application of AMF and SWSC inhibits plants from absorbing heavy metals from the soil and reduces the TFroot/soil of the millet. SWSC + AMF application inhibited the transfer of heavy metals from the roots to the upper part of the ground and reduced the TFshoot/root of the millet. The TFgrain/soil of the millet was below 1. The HQ and HI of the millet grains did not exceed 1, indicating the absence of a potential health risk. Therefore, SWSC combined with AMF is applicable for millet production in Tunliu, and the combined treatment can decrease heavy metal phytoavailability and post-harvest transfer risks. This work provides a way to utilize solid waste while also improving millet yields in dry farming. Based on the review, we suggested future researches to better understand the mechanisms of SWSC + AMF long-term application to promote awareness on its role over time through alterations in its surface chemistry, soil microbial community and environmental implications.

Mammalian STE20-Like Kinase 2 Promotes Lipopolysaccharides-Mediated Cardiomyocyte Inflammation and Apoptosis by Enhancing Mitochondrial Fission.

In this study, we analyzed the role of mammalian STE20-like protein kinase 2 (Mst2), a serine-threonine protein kinase, in Lipopolysaccharides (LPS)-mediated inflammation and apoptosis in the H9C2 cardiomyocytes. Mst2 mRNA and protein levels were significantly upregulated in the LPS-treated H9C2 cardiomyocytes. LPS treatment induced expression of IL-2, IL-8, and MMP9 mRNA and proteins in the H9C2 cardiomyocytes, and this was accompanied by increased caspase-3/9 mediating H9C2 cardiomyocyte apoptosis. LPS treatment also increased mitochondrial reactive oxygen species (ROS) and the levels of antioxidant enzymes, such as GSH, SOD, and GPX, in the H9C2 cardiomyocytes. The LPS-treated H9C2 cardiomyocytes showed lower cellular ATP levels and mitochondrial state-3/4 respiration but increased mitochondrial fragmentation, including upregulation of the mitochondrial fission genes Drp1, Mff, and Fis1. LPS-induced inflammation, mitochondrial ROS, mitochondrial fission, and apoptosis were all significantly suppressed by pre-treating the H9C2 cardiomyocytes with the Mst2 inhibitor, XMU-MP1. However, the beneficial effects of Mst2 inhibition by XMU-MP1 were abolished by carbonyl cyanide-4-(trifluoromethoxy) phenylhydrazone (FCCP), a potent activator of mitochondrial fission. These findings demonstrate that Mst2 mediates LPS-induced cardiomyocyte inflammation and apoptosis by increasing mitochondrial fission.

MST1-Hippo pathway regulates inflammation response following myocardial infarction through inhibiting HO-1 signaling pathway.

Context: Mammalian STE20-like protein kinases 1 (MST1) has been found to be associated with cardiomyocyte damage following acute myocardial infarction.Aim: The aim of our study is to explore the influence of MST1 in inflammation response following myocardial infarction.Methods: Cardiomyocyte cell line was used in vitro with hypoxia treatment to establish myocardial infarction model. ELISA, qPCR, Western blots, and siRNA technology were used to analyze the role of MST1 in inflammation response following myocardial infarction.Results: The transcription and expression of MST1 was significantly elevated following myocardial infarction. Loss of MST1 attenuated the levels of inflammation response and thus contributed to the survival of cardiomyocyte in vitro. Mechanistically, MST1 deletion reversed the activity of heme oxygenase-1 (HO-1) and thus reduced hypoxia-mediated cardiomyocyte death.Conclusions: Altogether, in this study, we found that MST1-Hippo pathway is activated in myocardial infarction and contributes to the inflammation response in cardiomyocytes through inhibiting the HO-1 signaling pathway. This finding would provide a potential target to reverse cardiomyocyte viability and reduce inflammation response in myocardial infarction.

Target Intestinal Microbiota to Alleviate Disease Progression in Amyotrophic Lateral Sclerosis.

PURPOSE: Emerging evidence has demonstrated that gut microbiome plays essential roles in the pathogenesis of human diseases in distal organs. Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease characterized by the progressive loss of motor neurons. Treatment with the only drug approved by the US Food and Drug Administration for use in ALS, riluzole, extends a patient׳s life span by only a few months. Thus, there is an urgent need to develop novel interventions that for alleviate disease progression and improve quality of life in patients with ALS. Here we present evidence that intestinal dysfunction and dysbiosis may actively contribute to ALS pathophysiology. METHODS: We used G93A transgenic mice as a model of human ALS. The G93A mice show abnormal intestinal microbiome and damaged tight junctions before ALS disease onset. The mice were given 2% butyrate, a natural bacterial product, in the drinking water. RESULTS: In mice fed with butyrate, intestinal microbial homeostasis was restored, gut integrity was improved, and life span was prolonged compared with those in control mice. At the cellular level, abnormal Paneth cells-specialized intestinal epithelial cells that regulate the host-bacterial interactions-were significantly decreased in the ALS mice treated with butyrate. In both ALS mice and intestinal epithelial cells cultured from humans, butyrate treatment was associated with decreased aggregation of the G93A superoxide dismutase 1 mutated protein. IMPLICATIONS: The findings from this study highlight the complex role of the gut microbiome and intestinal epithelium in the progression of ALS and present butyrate as a potential therapeutic reagent for restoring ALS-related dysbiosis.

Vitamin D receptor is a novel transcriptional regulator for Axin1.

BACKGROUND: Axin1 is a scaffold protein in the ß-catenin destruction complex, which, if disrupted, contributes to pathogenesis of various human diseases, including colorectal carcinogenesis and inflammatory bowel diseases (IBD). We have previously demonstrated that Salmonella infection promotes the degradation and plasma sequestration of Axin1, leading to bacterial invasiveness and inflammatory responses. Vitamin D and the vitamin D receptor (VDR) appear to be important regulators of IBD and colon cancer. Although VDR and Axin1 are all involved in intestinal inflammation, it remains unclear whether these processes are related or function independently. In the current study, we hypothesize that VDR is an important regulator for the maintenance of physiological level of Axin1. METHODS: Using the intestinal epithelial conditional VDR knockout mouse model (VDRΔIEC) and cultured cell lines, influences of VDR status on the expression of Axin1 was evaluated by Western blots and real-time PCR. Loss- and gain-of-function assays were used to investigate the regulation of VDR on Axin1 at the transcriptional and translational levels. Cells were treated with cycloheximide or actinomycin for molecular mechanistic studies. Candidate genomic VDR binding sites for Axin1 were tested by chromatin immunoprecipitation (ChIP) assay. Physical interactions among VDR, Axin1, and ß-catenin were tested by immunoprecipitation. Cellular localization of Axin1 with different VDR status was determined by fractionation and immunohistochemistry. RESULTS: We found that VDR deletion led to lower protein and mRNA levels of Axin1, whereas knockdown of Axin1 did not change the expression level of VDR protein. Immunoprecipitation data did not support physical interaction between VDR and Axin1. The VDR regulation of Axin1 was through a VDR genomic binding site for Axin1 gene on the regulatory region. Fractionation data showed that cytosolic Axin1 was significantly reduced due to VDR deletion, leaving the nuclear fraction unchanged. In ileum, Axin1 was distributed in the cytosol of apical epithelium and crypts. CONCLUSION: VDR is important for the maintenance of physiological level of Axin1. The discovery of Axin1 as a VDR target gene provides novel and fundamental insights into the interactions between the VDR and ß-catenin signaling pathways.

Lack of Vitamin D Receptor Causes Dysbiosis and Changes the Functions of the Murine Intestinal Microbiome.

PURPOSE: The microbiome modulates numerous aspects of human physiology and is a crucial factor in the development of various human diseases. Vitamin D deficiency and downregulation of the vitamin D receptor (VDR) are also associated with the pathogenesis of diseases such as inflammatory bowel disease, cancers, obesity, diabetes, and asthma. VDR is a nuclear receptor that regulates the expression of antimicrobial peptides and autophagy regulator ATG16L1. Vitamin D may promote a balanced intestinal microbiome and improve glucose homeostasis in diabetes. However, how VDR regulates microbiome is not well known. In the current study, we hypothesize that VDR status regulates the composition and functions of the intestinal bacterial community. METHODS: Fecal and cecal stool samples were harvested from Vdr knockout (Vdr(-/-)) and wild-type mice for bacterial DNA and then sequenced with 454 pyrosequencing. The sequences were denoised and clustered into operational taxonomic units, then queried against the National Center for Biotechnology Information database. Metagenomics were analyzed, and the abundances of genes involved in metabolic pathways were compared by reference to the Kyoto Encyclopedia of Genes and Genomes and Clusters of Orthologous Groups databases. FINDINGS: In the Vdr(-/-) mice, Lactobacillus was depleted in the fecal stool, whereas Clostridium and Bacteroides were enriched. Bacterial taxa along the Sphingobacteria-to-Sphingobacteriaceae lineage were enriched, but no genera reached statistical significance. In the cecal stool, Alistipes and Odoribacter were depleted, and Eggerthella was enriched. Notably, all of the taxa upstream of Eggerthella remained unchanged. A comparison of Vdr(-/-) and wild-type samples revealed 40 (26 enriched, 14 depleted) and 72 (41 enriched, 31 depleted) functional modules that were significantly altered in the cecal and fecal microbiomes, respectively (both, P < 0.05), due to the loss of Vdr. In addition to phylogenetic differences in gut microbiome with different intestinal origins, we identify several important pathways, such as nucleotide-binding oligomerization domain-like receptor, affected by Vdr status, including amino acid, carbohydrate, and fatty acid synthesis and metabolism, detoxification, infections, signal transduction, and cancer and other diseases. IMPLICATIONS: Our study fills knowledge gaps by having investigated the microbial profile affected by VDR. Insights from our findings can be exploited to develop novel strategies to treat or prevent various diseases by restoring VDR function and healthy microbe-host interactions.

Isolation and biological characteristics of chicken adipose-derived progenitor cells.

Adipose-derived stem cells/adipose-derived progenitor cells (ADPCs) are multipotent stem cells that can differentiate in vitro into many cell types. However, the vast majority of experimental materials were obtained from human, mouse, rabbit, and other mammals but rarely from poultry. In this study, ADPCs were isolated from 1-day-old chicks. Primary ADPCs were subcultured to passage 15. The surface markers of ADPCs, CD29, CD44, CD71, and CD73, were detected by immunofluorescence and RT-polymerase chain reaction assays. The growth curves of different passages were all typically sigmoidal. In addition, ADPCs of different passages were successfully induced to differentiate into osteoblasts, adipocytes, and myocardial cells. The results suggest that the ADPCs isolated from chicken possess similar biological characteristics with those derived from other species, and their multilineage differentiation provides many potential applications.

Inhibitory effects of vitamin E on UVB-induced apoptosis of chicken embryonic fibroblasts.

Apoptosis research has been focused on several model species in the past decades, whereas studies concerned with non-mammalian vertebrate, particularly birds, have rarely been involved. In accord with requirements to expand the biodiversity of apoptotic research, a chicken embryonic fibroblasts model involving UVB (ultraviolet B) as the death stimulus was established through primary explantation and serial passage. Myriads of antioxidants can inhibit UVB-induced apoptosis by virtue of scavenging reactive oxygen species. To improve our understanding of the possible anti-apoptotic effects and mechanisms of Vitamin E against UVB-induced apoptosis in chicken embryonic fibroblasts, cells treated with Vitamin E after UVB irradiation were stained with AO/EB and Fluo-3/AM to visualize chromatin distribution and calcium homoeostasis, respectively. They were also analysed by flow cytometry to detect mitochondrial transmembrane potential, and cell cycle progression and apoptotic rates were recorded. RT-PCR was used to analyse the expression of some apoptosis-related genes. Typical apoptotic events, including cell shrinkage, blebbing and nuclear condensation, occurred after radiation. In the presence of Vitamin E following irradiation, apoptotic cells were reduced. Ca2+ release was temporarily prevented, and cell cycle arrest at S/G2 checkpoint had almost completely reverted to normal. fas decreased, while procaspase-3 remained nearly unchanged with and without Vitamin E, and bcl2/bax ratio was up-regulated, indicating possible anti-apoptotic mechanisms through the mitochondrial pathway. This new investigation of an apoptosis model involving chicken embryonic fibroblasts expands the database of knowledge across a wider spectrum of vertebrate species.

Identification of Y chromosome genetic variations in Chinese indigenous horse breeds.

Y chromosome acts as a single nonrecombining unit that is male specific and in effect haploid, thus ensuring the preservation of mutational events as a single haplotype via male lines. In this study, 6 Y chromosome-specific microsatellites (SSR) were tested for the patrilineal genetic variations of 573 male samples from Chinese domestic horse (30 breeds), Przewalski's horse, and donkey. All the 6 loci appeared as a haplotype block in Przewalski's horse and the domestic donkey. There were notable differences, however, at Y chromosome markers between horse and donkey. There were 2 haplotypes of Eca.YA16 in the domestic horse breeds, Haplotype A (Allele A: 156 bp) and Haplotype B (Allele B: 152 bp). Allele A was the common allele among 30 horse breeds, and Allele B was found in 11 horse breeds. This is the first description of a Y chromosome variant for horses. The 2 haplotypes of Y chromosome discovered in the domestic horse breeds in China could be helpful in unveiling their intricate genetic genealogy.

Establishment and characterization of a fibroblast line from landrace.

Up to 32 Landrace ear marginal tissue samples were used for establishing a fibroblast cell bank by the means of primary explantation and cryopreservation. Biological analysis suggested that the Population Doubling Time (PDT) of the cell line was approximately 24h. The diploid accounted for 97.2% of the whole population; isozyme analysis of Lactic Dehydrogenase (LDH) and Malic Dehydrogenase(MDH) disproved cross-contamination from other cell lines. The results of bacterium, fungus, virus and mycoplasma tests were all negative. The transfection rates of three fluorescent proteins were high, indicating that the exogenous genes could be effectively expressed in the cells. It had not only preserved the precious germplasm resource of the Landrace on the cell level but also provided valuable material for the researches of genomics, postgenomics, somatic cloning and so on.