The aim of this study was to screen and evaluate lactic acid bacteria strains for their ability to ferment alfalfa, Leymus chinensis and native grass into silage. We isolated 186 lactic acid bacteria strains from alfalfa, native grass and selected eight strains by acid production test and growth rate test. The eight stains were identified using morphological observations, Gram staining, physiological and biochemical tests, an acid tolerance test and 16S rDNA sequencing. All eight strains (LP1, LP2, LC1, LC2, PP1, PP2, EF1 and EF2) grew at pH 4.0 and 6.5%NaCl. Strains LP1 and LP2 were identified as Lactobacillus plantarum, LC1 and LC2 as Lactobacillus casei, PP1 and PP2 as Pediococcus pentosaceus, EF1 as Enterococcus faecalis, and EF2 as Enterococcus faecium based on their 16S rDNA sequences. The eight strains and two commercial inoculants (PS and CL) were added with or without sucrose to native grass, L. chinensis, and alfalfa to produce silage. All eight screened strains decreased the pH and ammonia nitrogen (N) content and increased the lactic acid content of alfalfa silages. No butyric acid was detected in the L. chinensis and alfalfa silages. Addition of sucrose with the screened strains resulted in lower pH and ammonia N content, and higher lactic acid content and total fermentation acids content in all three silages. The eight strains improved the fermentation quality of alfalfa silage. All strains were more effective when supplemented with sucrose. No efficient lactic acid bacteria strain was found for ensiling the native grass, but the high fermentation quality native grass silage could be obtained using the combination of lactic acid bacteria strains and 2% sucrose.

BackgroundDisorders of endocrine substances in epicardial adipose tissue are known causes of coronary artery disease (CAD). Adiponectin is associated with cardiovascular disease. However, expression of adiponectin in epicardial adipose tissue and its function in CAD pathogenesis is unclear. This study investigates adiponectin expression in epicardial adipose tissue in CAD patients.MethodsVessels or adipose tissue samples collected from CAD patients and non-CAD controls were examined after immunochemical staining. Adiponectin, cytokines of interleukin-6 (IL-6) and necrosis factor-α (TNF-α) and toll-like receptor 4 (TLR4) expression level in adipose tissue were measured using real-time quantitative RT-PCR. Adiponectin concentrations in peripheral and coronary sinus vein plasma were measured with enzyme-linked immunosorbent assay. Peripheral vein plasma biochemistries were performed with routine laboratory techniques. Monocytes were collected from blood using lymphocyte separation medium. Expression level of cytokines and transcription factor NF-κB were measured to learn the effect of adiponectin on stearic acid-stimulated monocytes. Percentage of TLR4 positive monocytes was analyzed using flow cytometry.ResultsHistological examination revealed increased macrophage infiltration into epicardial adipose tissue of CAD patients. Decreased adiponectin displayed by real-time quantitative RT-PCR was associated with enhanced cytokines of IL-6 and TNF-α or TLR4 expression level in epicardial adipose tissue, suggesting decreased circulating adiponectin may be useful as a more sensitive predictor for coronary atherosclerosis than routine laboratory examinations. Adiponectin suppressed secretion of IL-6 and TNF-α in stimulated monocytes and TLR4 was expressed on cell surfaces.ConclusionsEndocrine disorders in epicardial adipose tissue are strongly linked to CAD, and adiponectin has a protective effect by inhibiting macrophage-mediated inflammation.

To study the microbial population and fermentation dynamics of large needlegrass (LN) and Chinese leymus (CL) during ensiling and subsequent exposure to air, silages were sampled and analyzed using culture-based techniques and denaturing gradient gel electrophoresis (DGGE). A total of 112 lactic acid bacteria (LAB) strains were isolated and identified using the 16S rRNA sequencing method. Lactic acid was not detected in the first 20 days in LN silage and the pH decreased to 6.13 after 45 days of ensiling. The temperature of the LN silage increased after approximately 30 h of air exposure and the CL silage showed a slight temperature variation. Enterococcus spp. were mainly present in LN silage. The proportion of Lactobacillus brevis in CL silage increased after exposure to air. LN silage with a higher proportion of Enterococcus spp. and propionic acid concentration did not show higher fermentation quality or aerobic stability than CL silage, which had a higher concentration of acetic acid, butyric acid and increased proportion of L. brevis after exposure to air.

The expression of long non-coding RNAs (lncRNAs) is dysregulated in non-small cell lung cancer (NSCLC). However, the functions and contributions of lncRNAs remain largely unknown. Here, we identified a critical role of long intergenic non-protein coding RNA 00858 (LINC00858) in NSCLC. Ectopic expression of LINC00858 in NSCLC cells promoted cell proliferation and induced cell migration and invasion. Moreover, LINC00858 functioned as a competitive endogenous RNA (ceRNA), effectively becoming sponge for miR-422a and thereby modulating the expression of kallikrein-related peptidase 4 (KLK4). In NSCLC patients, high expression of LINC00858 closely correlated with tumor progression. Thus, targeting the ceRNA network involving LINC00858 may be used as a treatment strategy against NSCLC.

Increasing seed yield and quality of key cool‐season perennial grasses is an important component of meeting China's environmental, food security, and urban beautification goals. Determining a suitable seed producing region and developing best management practices for this region with specific, high priority grasses will contribute to the availability of seed for reducing desertification, increasing livestock production, and improving urban living conditions. Previous research in other areas has addressed various agronomic practices on some grasses, but none has evaluated optimal row spacings for producing high yields of high quality seed in conditions similar to the Hexi Corridor, currently a major seed producing area in China for other seed crops. We compared our field trials in this area with those of other regions in China and the world to determine if this area would be suitable as a grass seed production center. Five important perennial, cool‐season grasses {[Elymus kamoji (ohwi) S.L. Chen, slender wheatgrass [Elymus trachycaulus (Link) Gould ex Shinners ssp. Trachycaulus], smooth bromegrass (Bromus inermis Leyss), Siberian wildrye (Elymus sibiricus L.), and Chinese sheepgrass [Leymus chinensis (Trin) Tzvel]} were evaluated from 2010 to 2012 under four row spacing treatments (30, 50, 70, and 90 cm). Results showed all five grasses have potential for high seed production in this region, with yields equivalent to or significantly higher than other areas. Optimal row spacing for Elymus kamoji and smooth bromegrass was 30 cm while a 50 cm row spacing was better for seed production and weed control for slender wheatgrass, Siberian wildrye, and Chinese sheepgrass. This work will allow multinational seed companies, development agencies, and government program directors to better evaluate the potential economic sustainability of grass seed production in this region compared to importing seed from other regions of the world.

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Objective: To investigate the expression of long non-coding RNA ZXF2 in lung adenocarcinoma tissues and its effect on cell proliferation, migration and invasion. Methods: Forty pairs of cancerous and adjacent non-cancerous lung adenocarcinoma specimens were collected for the studies. Quantitative real-time PCR was used to analyze the expression of ZXF2 in tumor tissues and adjacent normal tissues. The expression of ZXF2 was correlated with patients' clinico-pathological data. Molecular pathway controlled by ZXF2 was explored by using small interfering RNA (siRNA) technology. CCK-8 cell proliferation assay, flow cytometry analysis and transwell assays were used to evaluate cell proliferation, migration and invasion. Results: The expression of ZXF2 was 2 fold or higher in 27 out of 40 (67.5%) cases of lung adenocarcinoma specimens than that in non-cancerous tissues (P<0.05). The relative expression level of ZXF2 was positively correlated with tumor lymph node metastasis (χ²=8.485, P<0.05) and poor prognosis of the patients (p=0.0217). In order to explore the molecular mechanisms of ZXF2 mediated tumor progression, ZXF2 expression was inhibited by siRNA in A549 cells, a highly aggressive and metastatic lung adenocarcinoma cell line. We found that siRNA-ZXF2 treatment inhibited cell proliferation (P<0.01) leading to cell cycle arrest (P<0.01). The cell migration and invasion were suppressed by siRNA-ZXF2 treatment (P<0.01). Further biochemical studies revealed that the knockdown of ZXF2 led to down regulation of c-Myc signaling. Conclusion: ZXF2 was overexpressed in lung adenocarcinoma tissues and the high expression of ZXF was closely related to tumor progression through c-Myc related pathway. Given the fact that both ZXF2 and c-Myc are located in the same chromosome 8q24.2 loci, the potential interaction between ZXF2 and c-Myc might be a novel target for treatment of lung adenocarcinoma.

Zinc finger proteins (ZNF) play important roles in various physiological processes. Here we report that ZNF300, a novel zinc finger protein, identified specifically in humans, promotes tumour development by modulating the NF-κB pathway. Inflammatory factors were found to induce ZNF300 expression in HeLa cell line, and ZNF300 expression further enhanced NF-κB signalling by activating TRAF2 and physically interacting with IKKβ. Furthermore, ZNF300 overexpression increased ERK1/2 phosphorylation and the expression of c-myc, IL-6, and IL-8 but decreased the expression of p21waf-1 and p27Kip1; whose down-regulation led to the opposite effect. Most importantly, ZNF300 overexpression stimulated cancer cell proliferation in vitro and significantly enhanced tumour development and metastasis in mouse xenograft model, while knocking down ZNF300 led to the opposite effects. We have identified a novel function for ZNF300 in tumour development that may uniquely link inflammation and NF-κB to tumourigenesis in humans but not in mice.