Expression and Regulatory Network Analysis of BICC1 for Aged Sca-1-Positive Bone Narrow Mesenchymal Stem Cells.

Background: The impaired osteoblastic differentiation of bone marrow mesenchymal stem cells (BMSCs) is a major cause of bone remodeling imbalance and osteoporosis. The bicaudal C homologue 1 (BICC1) gene is a genetic regulator of bone mineral density (BMD) and promotes osteoblast differentiation. The purpose of this study is to explore the probable function of BICC1 in osteoporosis and osteogenic differentiation of aged BMSCs. Methods: We examined the GSE116925 microarray dataset obtained from the Gene Expression Omnibus (GEO) database. The GEO2R algorithm identified differentially expressed genes (DEGs) in Sca-1+ BMSCs from young (3 months old) and old (18 months old) mice. Then, to identify the most crucial genes, we used pathway enrichment analysis and a protein-protein interaction (PPI) network. Furthermore, starBase v2.0 was used to generate the regulatory networks between BICC1 and related competing endogenous RNAs (ceRNAs). NetworkAnalyst was used to construct TF-gene networks and TF-miRNA-gene networks of BICC1 and ceRNA. Furthermore, we investigated the Bicc1 expression in aged Sca-1-positive BMSCs. Result: We detected 923 DEGs and discovered that epidermal growth factor receptor (EGFR) was the top hub gene with a high degree of linkage. According to the findings of the PPI module analysis, EGFR was mostly engaged in cytokine signaling in immune system and inflammation-related signaling pathways. 282 ceRNAs were found to interact with the BICC1 gene. EGFR was not only identified as a hub gene but also as a BICC1-related ceRNA. Then, we predicted 11 common TF-genes and 7 miRNAs between BICC1 and EGFR. Finally, we found that BICC1 mRNA and EGFR mRNA were significantly overexpressed in aged Sca-1-positive BMSCs. Conclusion: As a genetic gene that affects bone mineral density, BICC1 may be a new target for clinical treatment of senile osteoporosis by influencing osteogenic differentiation of BMSCs through EGFR-related signaling. However, the application of the results requires support from more experimental data.

Mycoplasma suis Alpha-Enolase Subunit Vaccine Induces an Immune Response in Experimental Animals.

Recombinant protein technology has emerged as an excellent option for vaccine development. However, prior to our study, the immune induction ability of recombinant Mycoplasma suis alpha-enolase (rMseno) in animals remained unclear. The purpose of this study was to develop a rMseno protein subunit vaccine and to determine its ability to elicit an immunological response. To accomplish this, we cloned the gene into pET-15b, expressed it in BL21 cells, and purified it. Following the establishment of immunity, the immunogenicity and potential for protection of rMseno were evaluated in mice and piglets. The results demonstrate that anti-M. suis serum recognized the pure rMseno protein in both mice and piglets as evidenced by high levels of specific anti-rMseno antibodies, significantly increased levels of IFN-γ and IL-4 cytokines, and significantly increased T lymphocyte proliferation index. Piglets also had significantly increased levels of specific IgG1, IgG2a, CD4+, and CD8+ cells. The rMseno findings demonstrated a robust immunological response in mice and piglets, affording partial clinical protective efficacy in piglets.

A ready-to-use fluorescence probe of Pd2+ in water: novel tricyclic heterocyclic base on 1,3,4-oxadiazole.

A ready-to-use hetero-tricyclic compound, 5,5'-(furan-2,5-diyl) bis (1,3,4- oxadiazol-2-amine) (5), was synthesized with a good yield; it has an suitable fluorescence characteristic and research founded that it can respond to trace Pd2+ in water at a normal pH range. A fluorescence titration revealed the detection limit for Pd2+ was as low as 3.97 × 10-9 M. Density-functional theory calculation using Guassian09 implied that the breakage of conjugation and coplanarity of compound 5 led to fluorescence quenching. Compound 5 could be applied as a chemical probe to detect trace amounts of Pd2+ with good accuracy, fast response time, excellent selectivity, and high sensitivity. FT-IR, NMR, and MS were used to characterize the chemical structure of compound 5.

The effect of miR-93 on GH secretion in pituitary cells of Yanbian yellow cattle.

Yanbian yellow cattle breeding is limited by slow growth. We previously found that the miRNA miR-93 was differentially expressed between the blood exosomes of Yanbian yellow cattle and Han Yan cattle, which differ in growth characteristics. In this experiment, we evaluated the effects of miR-93 on growth hormone (GH) secretion by pituitary cells of Yanbian yellow cattle using qPCR, Western blot, Targetscan and RNA hybrid analysis software and Dual-Luciferase reporter gene system. The results showed that miR-93 targeted 3' UTR of GHRHR(growth hormone releasing hormone receptor); GH mRNA and protein levels in pituitary cells of Yanbian yellow cattle were significantly lower in the miR-93-mi group than in the NC control group (p < 0.01), while GH mRNA and protein levels were higher in the miR-93-in group than in the iNC control group, but the difference was not significant (p > 0.05); GHRHR mRNA and protein levels were significantly lower in the miR-93-mi group than in the NC control group (p < 0.01), while GHRHR protein levels were significantly higher in the miR-93-in group than in the iNC control group (p < 0.05), but there was no significant difference about GHRHR mRNA level between two groups (p > 0.05). These results prove that miR-93 regulates GH secretion in pituitary cells via GHRHR.

The symbioses of endophytic fungi shaped the metabolic profiles in grape leaves of different varieties.

Endophytic fungi produce many novel bioactive metabolites that are directly used as drugs or that function as the precursor structures of other chemicals. The metabolic shaping of endophytes on grape cells was reported previously. However, there are no reports on the interactions and metabolic impact of endophyte symbiosis on in vitro vine leaves, which may be examined under well-controlled conditions that are more representative of the natural situation of endophytes within grapevines. The present study used an in vitro leaf method to establish endophyte symbiosis of grapevines and analyze the effects on the metabolic profiles of grape leaves from two different cultivars, 'Rose honey' (RH) and 'Cabernet sauvignon' (CS). The effects of endophytic fungi on the metabolic profiles of grape leaves exhibited host selectivity and fungal strain specificity. Most of the endophytic fungal strains introduced novel metabolites into the two varieties of grape leaves according to the contents of the detected metabolites and composition of metabolites. Strains RH49 and MDR36, with high or moderate symbiosis rates, triggered an increased response in terms of the detected metabolites, and the strains MDR1 and MDR33 suppressed the detected metabolites in CS and RH leaves despite having strong or moderate symbiosis ability. However, the strain RH12 significantly induced the production of novel metabolites in RH leaves due to its high symbiosis ability and suppression of metabolites in CS leaves.

The effect of miR-10b on growth hormone in pituitary cells of Yanbian yellow cattle by somatostatin receptor 2.

This study aimed to evaluate the effect of miR-10b on growth hormone (GH) in pituitary cells of Yanbian yellow cattle. According to analysis of GH and somatostatin receptor 2 (SSTR2) mRNA and protein expression levels, we found that miR-10b targeted 3'UTR of SSTR2. Compared with the negative control (NC) group, GH mRNA transcription and protein expression in pituitary cells of Yanbian yellow cattle were significantly increased by adding miR-10b mimics (p < .01), while these were significantly decreased by adding miR-10b inhibitor (p < .05); compared with the NC group, SSTR2 mRNA transcription and protein expression were significantly inhibited by the addition of miR-10b mimics (p < .01), while these were significantly increased by the addition of miR-10b inhibitor compared with the iNC group (p < .05). This study suggested that miR-10b could regulate GH level by regulating SSTR2 gene expression in pituitary cells of Yanbian yellow cattle.

Exposure to endophytic fungi quantitatively and compositionally alters anthocyanins in grape cells.

Anthocyanins contribute greatly to the organoleptic and biochemical properties of grapes and wines. Although there are broadly documented factors involved in grape anthocyanin synthesis, the present work focused on fungal endophytes and their possible role in grape coloration. Our results showed that exposure to endophytic fungi within a dual culture system differentially affected total anthocyanin concentrations and PAL activities in grape cells. Grape cells dual cultured with fungal strains XH-2, R2-21 and B2-17 showed significant differences of their anthocyanin concentrations were subjected to further analysis of their anthocyanidin compositions. Compared to the no-fungus controls, grape cells exposed to fungal strains XH-2 and R2-21 exhibited quantitative promotion of their total anthocyanidin concentrations by 74% and 28%, respectively, whereas treatment with the fungus B2-17 reduced the anthocyanidin content by 19%. A total of 14 species of anthocyanidins were detected from the grape cells in these experiments. Most interestingly, exposure to any of these fungal strains differentially modified the compositional patterns of grape cellular anthocyanidins. The obvious upregulation of the transcription of VvMYB in grape cells treated with fungal strains XH-2 and R2-21 implies that the increased anthocyanin levels in these grape cells may be due to the activated transcriptional factors. In addition, the exposure of grape cells to extracts of these fungi initiated similar responses of anthocyanin contents and PAL activities to exposure to the living fungi and appeared obvious dosage effects. The influence of fungal endophytes on the coloration of grape berries was also examined in this study.

Polylactic-co-glycolic acid microspheres containing three neurotrophic factors promote sciatic nerve repair after injury.

A variety of neurotrophic factors have been shown to repair the damaged peripheral nerve. However, in clinical practice, nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor are all peptides or proteins that may be rapidly deactivated at the focal injury site; their local effective concentration time following a single medication cannot meet the required time for spinal axons to regenerate and cross the glial scar. In this study, we produced polymer sustained-release microspheres based on the polylactic-co-glycolic acid copolymer; the microspheres at 300-µm diameter contained nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor. Six microspheres were longitudinally implanted into the sciatic nerve at the anastomosis site, serving as the experimental group; while the sciatic nerve in the control group was subjected to the end-to-end anastomosis using 10/0 suture thread. At 6 weeks after implantation, the lower limb activity, weight of triceps surae muscle, sciatic nerve conduction velocity and the maximum amplitude were obviously better in the experimental group than in the control group. Compared with the control group, more regenerating nerve fibers were observed and distributed in a dense and ordered manner with thicker myelin sheaths in the experimental group. More angiogenesis was also visible. Experimental findings indicate that polylactic-co-glycolic acid composite microspheres containing nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor can promote the restoration of sciatic nerve in rats after injury.

[Comparison of the clinical efficacy between simple vertebral canal decompression and decompression plus laminoplasty].

OBJECTIVE: To observe the clinical efficacy of the simple expansion of the spinal canal decompression, decompression plus hydroxyapatite/polyamide artificial lamina reconstruction, and decompression plus titanium mesh reconstruction in the treatment of spinal canal stenosi. METHODS: A total of 39 patients with lumbar spinal stenosis (with or without disc herniation, spondylolisthesis less than I degree), who received therapy of surgery from January, 2011 to January, 2012, were retrospectively analyzed. All patients were divided into 3 groups: a laminectomy surgery alone group (group A, n=15), a decompression plus hydroxyapatite/polyamide artificial lamina reconstruction group (group B, n=14), and a laminectomy decompression plus reconstruction with titanium mesh group (group C, n=10). Intraoperative situation, the postoperative excellent rate and JOA score were analyzed. RESULTS: The duration and blood loss in surgery in group A was much less than that in the group B and C (P<0.05), but there was no statistical significance between the group B and C. The postoperative excellent rate in three groups were similar in 3 months (P>0.05). Twelve months after the surgery, the group B and C showed advantage over the group A (P<0.05). JOA scores in 3 and 12 months after the surgery were all greater than that before the surgery (P<0.05). There was no difference in excellent rates in 3 groups in 3 months after the operation (P>0.05); the group B and C showed advantage over the group A in 12 months after the operation (P<0.05). No serious complications were related to the surgery in the 3 groups. Imaging changes were not significant difference. CONCLUSION: The decompression plus hydroxyapatite/polyamide artificial lamina reconstruction and the decompression plus titanium mesh reconstruction show advantages in long-term effect over the simple vertebral canal decompression.