MicroRNA-34a suppresses neuronal apoptosis and alleviates microglia inflammation by negatively targeting the Notch pathway in spinal cord injury.

OBJECTIVE: The purpose of this study was to investigate role of inhibition of microRNA-34a (miR-34a) in neural damage and repair after spinal cord injury, and to explore the underlying mechanism. MATERIALS AND METHODS: In BV2 microglia, we conducted classical activation using lipopolysaccharide (LPS) and pre-treatment using miR-34a mimics. The expressions of miR-34a, Notch 1, and Jagged 1 were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Moreover, the protein expressions of inflammatory microglia markers were evaluated by Western blotting. In vivo, SCI model was successfully established in rats. Subsequently, the expression levels of miR-34a, Notch 1, and Jagged 1 levels within 1 week were measured by qRT-PCR. Meanwhile, protein expressions of inflammatory mediators were determined by enzyme-linked immunosorbent assay (ELISA) assay. Immunofluorescence was conducted to display the activation degree of microglia and residual neural structure. Furthermore, locomotor function recovery was estimated using BBB rating scale. RESULTS: Compared with the only LPS-activated group, pre-treatment of miR-34a mimics significantly decreased the expressions of Notch 1 and Jagged 1. Similarly, the protein expressions of CD11b and iNOS were significantly down-regulated. In vivo, the levels of Notch 1 and Jagged 1 within 1 week increased significantly, while miR-34a was negatively regulated following spinal cord injury (SCI). Furthermore, the contents of interleukin-1 beta (IL-1ß) and IL-6 were reduced with the treatment of miR-34a mimics when compared with SCI group. With the treatment of miR-34a, the number of inflammatory microglia decreased significantly, and the remaining neural structure was similarly improved. In addition, locomotor function recovery of hindlimbs in rats was significantly ameliorated after the administration of miR-34a mimics. CONCLUSIONS: Increase of miR-34a suppresses neuronal apoptosis and alleviates microglia inflammation by negatively targeting the Notch pathway, thereby improving neural recovery and locomotor function.

Glucagon-like Peptide-1 Analogues Inhibit Proliferation and Increase Apoptosis of Human Prostate Cancer Cells in vitro.

Background: Research has shown that the incidence of prostate cancer is increased in patients with type 2 diabetes mellitus (T2DM) 1. Glucagon-like peptide-1 (GLP-1) is a gastrointestinal hormone that enhances glucose-dependent insulin secretion and suppresses glucagon release. Method: Here, we examined the effect of exenatide and liraglutide, 2 types of GLP-1 analogues, on prostate cancer cells growth by CCK-8 assay, Hoechst33258 staining assay, and western blot analysis of apoptosis-related proteins Bax and Bcl-2. Also the kinase pathways maybe involved and the expression of GLP-1 receptor (GLP-1 R) in LNCap cells was detected. Results: In our experiments, exenatide and liraglutide significantly inhibited the proliferation of the LNCap cell lines and induced the cell apoptosis. Exenatide (1-100 nmol/L) increased the ratio of Bax/Bcl-2 in a dose-dependent manner, whereas liraglutide increased Bax/Bcl-2 ratio only at concentrations of 10 nmol/L. And we found that GLP-1 analogues activate p38 but not ERK1/2 or AKT in LNCap cells. And classical GLP-1 receptor was detected in LNCap cells. Conclusion: These data suggest that exenatide and liraglutide attenuate prostate cancer growth through regulating P38 pathway by binding with GLP-1R.

Obesity is an independent risk factor for non-alcoholic fatty liver disease: evidence from a meta-analysis of 21 cohort studies.

BACKGROUND: The association between obesity and nonalcoholic fatty liver disease (NAFLD) has not been fully quantified, and the magnitude of NAFLD risk associated with obesity is still unclear. A meta-analysis of cohort studies was performed to elucidate the NAFLD risk associated with obesity. METHODS: Pubmed, Web of Science and Embase were searched for cohort studies assessing NAFLD risk associated with obesity or increased body mass index (BMI). Relative risks (RRs) with 95% confidence intervals (95%CIs) were pooled using random-effects model of meta-analysis. RESULTS: Twenty-one cohort studies including 13 prospective studies and 8 retrospective studies were finally included. There were a total of 381,655 participants in the meta-analysis. Compared with normal weight, obesity independently led to a 3.5-fold increased risk of developing NAFLD (RR = 3.53, 95%CI 2.48 to 5.03, P < 0.001). Meta-analysis also suggested an obvious dose-dependent relationship between BMI and NAFLD risk (per 1-unit increment in BMI: RR = 1.20, 95%CI 1.14 to 1.26, P < 0.001). Subgroup analyses further identified the robustness of the association above. No obvious risk of publication bias was observed. CONCLUSION: Obese individuals have a 3.5-fold increased risk of developing NAFLD, and there is an obvious dose-dependent relationship between BMI and NAFLD risk. © 2016 World Obesity.

A feedback circuit between miR-133 and the ERK1/2 pathway involving an exquisite mechanism for regulating myoblast proliferation and differentiation.

MiR-133 was found to be specifically expressed in cardiac and skeletal muscle in previous studies. There are two members in the miR-133 family: miR-133a and miR-133b. Although previous studies indicated that miR-133a was related to myogenesis, the signaling pathways regulated by miR-133 were still not very clear. In this study, we showed that both miR-133a and miR-133b were upregulated during myogenesis through Solexa sequencing. We confirmed that miR-133 could promote myoblast differentiation and inhibit cell proliferation through the regulation of the extracellular signal-regulated kinase (ERK) signaling pathway in C2C12 cells. FGFR1 and PP2AC, which both participate in signal transduction of the ERK1/2 pathway, were found to be negatively regulated by miR-133a and miR-133b at the post-transcriptional level. Also, downregulation of ERK1/2 phosphorylation by miR-133 was detected. FGFR1 and PP2AC were also found to repress C2C12 differentiation by specific siRNAs. In addition, we found that inhibition of ERK1/2 pathway activity can inhibit C2C12 cell proliferation and promote the initiation of differentiation but form short and small myotubes. Furthermore, we found that the expression of miR-133 was negatively regulated by ERK1/2 signaling pathway. In summary, we demonstrated the role of miR-133 in myoblast and further revealed a new feedback loop between miR-133 and the ERK1/2 signaling pathway involving an exquisite mechanism for regulating myogenesis.

miR-29 targets Akt3 to reduce proliferation and facilitate differentiation of myoblasts in skeletal muscle development.

MicroRNAs (miRNAs) are a type of endogenous noncoding small RNAs involved in the regulation of multiple biological processes. Recently, miR-29 was found to participate in myogenesis. However, the underlying mechanisms by which miR-29 promotes myogenesis have not been identified. We found here that miR-29 was significantly upregulated with age in postnatal mouse skeletal muscle and during muscle differentiation. Overexpression of miR-29 inhibited mouse C2C12 myoblast proliferation and promoted myotube formation. miR-29 specifically targeted Akt3, a member of the serine/threonine protein kinase family responsive to growth factor cell signaling, to result in its post-transcriptional downregulation. Furthermore, knockdown of Akt3 by siRNA significantly inhibited the proliferation of C2C12 cells, and conversely, overexpression of Akt3 suppressed their differentiation. Collectively and given the inverse endogenous expression pattern of rising miR-29 levels and decreasing Akt3 protein levels with age in mouse skeletal muscle, we propose a novel mechanism in which miR-29 modulates growth and promotes differentiation of skeletal muscle through the post-transcriptional downregulation of Akt3.

Characterization of the major histocompatibility complex class I A alleles in cynomolgus macaques of Vietnamese origin.

Cynomolgus macaques (Macaca fascicularis, Mafa) have emerged as an important animal model for infectious disease and transplantation research. Extensive characterization of their major histocompatibility complex (MHC) polymorphism regions therefore becomes urgently required. In this study, we identified 41 MHC class I A nucleotide sequences in 34 unrelated cynomolgus macaques of Vietnamese origin farmed in Southern China, including eight novel Mafa-A sequences. We found two sequences with perfect identity and six sequences with close similarity to previously defined MHC class I alleles from other populations, especially from Indonesian-origin macaques. We also found three Vietnamese-origin cynomolgus macaque MHC class I sequences for which the predicted protein sequences identical throughout their B and F binding pockets to Mamu-A1*001:01 and Mamu-A3*13:03, respectively. This is important because Mamu-A1*001:01 and Mamu-A3*13:03 are associated with longer survival and lower set-point viral load in simian immunodeficiency virus (SIV)-infected rhesus monkeys. These findings have implications for the evolutionary history of Vietnamese-origin cynomolgus macaque as well as for the use of this model in SIV/SHIV (a virus combining parts of the HIV and SIV genomes) research.

BCL10 as a new candidate gene for immune response in pigs: cloning, expression and association analysis.

BCL10 is an apoptotic regulatory molecule identified through its direct involvement in t(1; 14)(p22; q32) of mucosa-associated lymphoid tissue lymphoma, and was implicated in the pathogenesis of this and several other tumour types. BCL10 was recognized as an antigen receptor-specific regulator of NF-kappaB, which showed close association with immune responses. In this study, we cloned and characterized BCL10 from the porcine spleen and analysed its genomic structure. BCL10 was mapped to SSC4q21-q23 by the IMpRH panels, it is closely linked to the marker S0161 and SW1461. This gene has three exons and two introns. Reverse transcriptase-polymerase chain reaction analyses showed that BCL10 was widely expressed in all the examined tissues. Transient transfection indicated that porcine BCL10 was located in cytoplasm in Pig Kidney Epithelial cells. BCL10 gene displays the opposite expression trend between the two treatments mimic virus and bacteria of polyriboinosinic-polyribocytidylic acid (Poly I:C) and lipopolysaccharide (LPS). The level of the BCL10 mRNA was up-regulated during 12-24 h and peaking at 48 h when treated with LPS, whereas it was down-regulated during 0-48 h and highest at 0 h (cells without treating with Poly I:C) when treated with Poly I:C. One single nucleotide polymorphism (SNP) site was identified in the 3'-untranslated region of porcine BCL10. Association analysis revealed that this SNP was significantly associated with intermediate cell mass (eosinophile granulocyte, basophile granulocyte and histoleucocyte) percentage, absolute intermediate cell mass count and mean red blood cell volume of 0-day-old pigs, and red blood cell count of 17-day-old pigs (P < 0.05), and also had significant associations with red blood cell count and haemoglobin concentration of 32-day-old pigs (P < 0.01).

Current knowledge of microRNA characterization in agricultural animals.

MicroRNA (miRNA) is a class of single-stranded small (19-24nt) regulatory RNA that silences gene expression post-transcriptionally. miRNAs regulate a wide range of biological processes through the recognition of complementary sequences between miRNAs and their target genes. Profiling studies in livestock have revealed that many miRNAs are species- and tissue-specific, indicating that miRNAs play important roles in essential biological processes in livestock, such as muscle and organ development, the immune response and metabolism. The allelic variation of miRNA target sites and possibly in miRNAs themselves are also likely to be contributing factors to many phenotypic differences in livestock. In this review, we summarize the current miRNA studies undertaken in livestock.

Treatment with peginterferon plus ribavirin vs. interferon plus ribavirin for 48 weeks in Chinese patients with chronic hepatitis C.

INTRODUCTION: Virological responses to interferon (IFN) treatment for chronic hepatitis C (CHC) have been found to be correlated with racial differences. Furthermore, higher sustained virological response (SVR) rates were obtained in many randomised clinical trials (RCT) in Caucasian patients with CHC after treating with peg-IFN than those with IFN. However, it is not clear whether this conclusion can be adapted to patients of Chinese descent. METHODS: A search of MEDLINE and China National Knowledge Infrastructure between 1966 and 2008 was performed. Trials comparing the use of peg-IFN plus ribavirin vs. IFN plus ribavirin in treating Chinese patients with CHC were assessed. RESULTS: Of the 94 studies screened, seven RCTs including 398 patients (peg-IFN therapy 232, IFN therapy 166) were analysed. The SVRs obtained in patients treated with peg-IFN plus ribavirin were significantly higher than in patients treated with IFN plus ribavirin [70% vs. 35%, relative risk, 1.76; 95% confidence interval: 1.21-2.56; p (0.01)]. Withdrawal rates were similar between patients treated with peg-IFN plus ribavirin and IFN plus ribavirin. CONCLUSION: Chinese patients with CHC also have a greater likelihood of achieving an SVR with peg-IFN plus ribavirin.

Cloning, chromosomal localization, SNP detection and association analysis of the porcine IRS-1 gene.

Insulin receptor substrate-1(IRS-1) gene is one member of the Insulin receptor substrate (IRS) gene family, which plays an important role in mediating the growth of skeletal muscle and the molecular metabolism of type 2 diabetes. Here, we cloned a 3,573 bp fragment of the partial CDS sequence of porcine IRS-1 gene by in silicon cloning strategy and RT-PCR method. The porcine IRS-1 gene was assigned to SSC15q25 by using IMpRH. Sequencing of PCR products from Duroc and Tibetan pig breeds identified one SNP in exon 1 of porcine IRS-1 gene (C3257A polymorphisms). Association analysis of genotypes with the growth traits, anatomy traits, meat quality traits and physiological biochemical indexes traits showed that different genotypes at locus 3,257 of IRS-1 have significant differences in carcass straight length in pigs (P = 0.0102 \ 0.05).

Six placenta permeability-related genes: molecular characterization and expression analysis in pigs.

The nutrient transportation ability of placenta depends on placental size, vascular density and permeability. Regulation of angiogenesis in the placenta is critical for successful gestation. Placenta vascularity exhibits disparity in different gestation stages and different pig breeds. To investigate the expression of genes related to permeability in the porcine placenta of different gestation stages and breeds, molecular cloning and gene expression analysis of six porcine genes, vascular endothelial growth factor (VEGF), VEGF-R1, VEGF-R2, endothelial nitric oxide synthase (eNOS), vascular endothelial cadherin (CDH5) and ß-arrestin2 (Arrb2), were performed in this study. The results demonstrated that from gestation day 33 to day 90, Landrace exhibited significant increase (P < 0.05) in placental VEGF and Arrb2 mRNA expression. Moreover, expression levels of VEGF, VEGF-R1, VEGF-R2 and eNOS mRNA were higher (P < 0.01) in the placenta of Erhualian than those in Landrace on day 90 of gestation. In contrast, CDH5 placental mRNA expression level exhibited significant decrease (P < 0.05) from day 33 to day 90 gestation in Landrace. Erhualian placental CDH5 and Arrb2 expression levels were significantly lower (P < 0.01) than those in Landrace conceptuses on day 90 of gestation. Our study offered new data on the expression of genes in VEGF signal transduction pathway in porcine placenta.