Expression of MiR-9 promotes proliferation, migration and differentiation of human neural stem cells

Purpose: To investigate the effect of miR-9 on the proliferation, differentiation and migration of human neural stem cells (NSCs).Methods: The expression of miR-9 was investigated by quantitative real-time polymerase chain reaction (RT-PCR). Cell proliferation was assessed by cell counting kit-8 (CCK8) assay, while cell migration was studied by Transwell assay. The effect of miR-9 on differentiation of NSCs was investigated by western blot analysis of key differentiation marker proteins. Protein expression was determined by western blotting.Results: Transfection and over-expression of miR-9 in NSCs significantly enhanced the proliferation of NSCs (p < 0.05) in a time-dependent manner, as was evident from CCK8 assay data. MiR-9 overexpression caused down-regulation of Nestin and SOX-2, and up-regulation of Tuj-1 and MAP-2. The migration of NSCs was 37 % in the cells transfected with empty vector, compared to 68 % in the cells transfected with miR-9. This effect of miR-9 on cell migration was accompanied by up-regulation of matrix metallopeptidase 9 (MMP-9) and matrix metallopeptidase 2 (MMP-2).Conclusion: These results show that miR-9 promotes the proliferation, differentiation and migration of NSCs, and thus may be an important drug target for the generation of NSCs.Keywords: Neural stem cells, MicroRNA, Mir-9, Migration, Differentiation, Proliferatio

Restoration of Noradrenergic Function in Parkinson’s Disease Model Mice

Dysfunction of the central noradrenergic and dopaminergic systems is the primary neurobiological characteristic of Parkinson’s disease (PD). Importantly, neuronal loss in the locus coeruleus (LC) that occurs in early stages of PD may accelerate progressive loss of dopaminergic neurons. Therefore, restoring the activity and function of the deficient noradrenergic system may be an important therapeutic strategy for early PD. In the present study, the lentiviral constructions of transcription factors Phox2a/2b, Hand2 and Gata3, either alone or in combination, were microinjected into the LC region of the PD model VMAT2 Lo mice at 12 and 18 month age. Biochemical analysis showed that microinjection of lentiviral expression cassettes into the LC significantly increased mRNA levels of Phox2a, and Phox2b, which were accompanied by parallel increases of mRNA and proteins of dopamine β-hydroxylase (DBH) and tyrosine hydroxylase (TH) in the LC. Furthermore, there was considerable enhancement of DBH protein levels in the frontal cortex and hippocampus, as well as enhanced TH protein levels in the striatum and substantia nigra. Moreover, these manipulations profoundly increased norepinephrine and dopamine concentrations in the striatum, which was followed by a remarkable improvement of the spatial memory and locomotor behavior. These results reveal that over-expression of these transcription factors in the LC improves noradrenergic and dopaminergic activities and functions in this rodent model of PD. It provides the necessary groundwork for the development of gene therapies of PD, and expands our understanding of the link between the LC-norepinephrine and dopamine systems during the progression of PD

Norepinephrine Upregulates the Expression of Tyrosine Hydroxylase and Protects Dopaminegic Neurons Against 6-Hydrodopamine Toxicity

As a classic neurotransmitter in the brain, norepinephrine (NE) also is an important modulator to other neuronal systems. Using primary cultures from rat ventral mesencephalon (VM) and dopaminergic cell line MN9D, the present study examined the neuroprotective effects of NE and its effects on the expression of tyrosine hydroxylase (TH). The results showed that NE protected both VM cultures and MN9D cells against 6-hydroxydopamine-caused apoptosis, with possible involvement of adrenal receptors. In addition, treatment with NE upregulated TH protein levels in dose- and time-dependent manner. Further experiments to investigate the potential mechanisms underlying this NE-induced upregulation of TH demonstrated a marked increase in protein levels of the brain-derived neurotrophic factor (BDNF) and the phosphorylated extracellular signal-regulated protein kinase 1 and 2 (pERK1/2) in VM cultures treated with NE. In MN9D cells, a significantly increase of TH and pERK1/2 protein levels were observed after their transfection with BDNF cDNA or exposure to BDNF peptides. Treatment of VM cultures with K252a, an antagonist of the tropomyosin-related kinase B, blocked the upregulatory effects of NE on TH, BDNF and pERK1/2. Administration of MEK1 & MEK2 inhibitors also reversed NE-induced upregulation of TH and pERK1/2. Moreover, ChIP assay showed that treatment with NE or BDNF increased H4 acetylation in the TH promoter. These results suggest that the neuroprotection and modulation of NE on dopaminergic neurons are mediated via BDNF and MAPK/ERK pathways, as well as through epigenetic histone modification, which may have implications for the improvement of therapeutic strategies for Parkinson\u27s disease

Critical Role of Oxidatively Damaged DNA in Selective Noradrenergic Vulnerability

An important pathology in Parkinson\u27s disease (PD) is the earlier and more severe degeneration of noradrenergic neurons in the locus coeruleus (LC) than dopaminergic neurons in the substantia nigra. However, the basis of such selective vulnerability to insults remains obscure. Using noradrenergic and dopaminergic cell lines, as well as primary neuronal cultures from rat LC and ventral mesencephalon (VM), the present study compared oxidative DNA damage response markers after exposure of these cells to hydrogen peroxide (H2O2). The results showed that H2O2 treatment resulted in more severe cell death in noradrenergic cell lines SK-N-BE(2)-M17 and PC12 than dopaminergic MN9D cells. Furthermore, there were higher levels of oxidative DNA damage response markers in noradrenergic cells and primary neuronal cultures from the LC than dopaminergic cells and primary cultures from the VM. It included increased tail moments and tail lengths in Comet assay, and increased protein levels of phosphor-p53 and γ-H2AX after treatments with H2O2. Consistent with these measurements, exposure of SK-N-BE(2)-M17 cells to H2O2 resulted in higher levels of reactive oxygen species (ROS). Further experiments showed that exposure of SK-N-BE(2)-M17 cells to H2O2 caused an increased level of noradrenergic transporter, reduced protein levels of copper transporter (Ctr1) and 8-oxoGua DNA glycosylase, as well as amplified levels of Cav1.2 and Cav1.3 expression. Taken together, these experiments indicated that noradrenergic neuronal cells seem to be more vulnerable to oxidative damage than dopaminergic neurons, which may be related to the intrinsic characteristics of noradrenergic neuronal cells

Several problems about strain calculation and analysis and correction of related deviation*

There exists many kinds of calculation models of plane and spherical strain fields, but the results of these models are different. The representative models was analyzed, and got some useful conclusions, in which some models are unbiased, some have deviations that can be corrected, some can only be used to compute strain in a uniform medium and can not be extended, and some can be used in the calculation and analysis of continuous strain field as well. Meanwhile, the correction relationship for spherical difference movement (displacement) computed from strain results was given, and the meaning of the non – differential term in spherical strain model was demonstrated

Effects of the differences between the ITRF2000 and ITRF2005 models in GNSS data processing

Abstract:In comparison with the ITRF2000 model, the ITRF2005 model represents a significant improvement in solution generation, datum definition and realization. However, these improvements cause a frame difference between the ITRF2000 and ITRF2005 models, which may impact GNSS data processing. To quantify this impact, the differences of the GNSS results obtained using the two models, including station coordinates, baseline length and horizontal velocity field, were analyzed. After transformation, the differences in position were at the millimeter level, and the differences in baseline length were less than 1mm. The differences in the horizontal velocity fields decreased with as the study area was reduced. For a large region, the differences in these value were less than 1mm/a, with a systematic difference of approximately 2 degrees in direction, while for a medium-sized region, the differences in value and direction were not significant

Impregnation of 1-n-Butyl-3-methylimidazolium Dicyanide [BMIM][DCA] into ZIF-8 as a Versatile Sorbent for Efficient and Selective Separation of CO2

The efficient capture of carbon dioxide (CO2) has industrial, economic, and environmental significance. Here, CO2 capture performance in terms of CO2/CH4 selectivity for the zeolitic imidazolate framework (ZIF-8) is enhanced by impregnation of 1-nbutyl-3-methylimidazolium dicyanide [BMIM][DCA]. The preparation of ionic liquids (ILs)/ZIF-8 composites, their comprehensive characterization, and the impact of IL impregnation are all investigated. The crystal structure and morphology of ZIF-8 remained intact even after the impregnation of ILs, according to X-ray diffraction and scanning electron microscopy studies. The infrared spectroscopy known as Fourier transform infrared spectroscopy determines the interaction between ILs and ZIF-8 in the composite. The isotherm fitting was also performed by dual site Langmuir models, which showed good agreement with the experimental results. The CO2/CH4 selectivity results demonstrated that the ideal selectivity for the IL-impregnated ZIF-8 composite has five times higher selectivity than pristine ZIF-8 at 0.05 bar which is the highest reported selectivity. The normalized selectivity of this type of IL/ZIF-8 composite is also higher. This study suggested that the IL/MOF composite with a tunable structure have a strong potential for enhanced acid gas separation performance to meet the environmental challenges

The Regulation of Corticosteroid Receptors in Response to Chronic Social Defeat

Our previous studies demonstrated that chronic social defeat (CSD) up-regulated expression of the serotonin transporter (SERT) and norepinephrine transporter (NET) in the brain, which was mediated by corticosteroid receptors. In the present study we first analyzed the alterations of corticosteroid receptors in different brain regions after the CSD paradigm. The results showed that CSD significantly reduced glucocorticoid receptor (GR) protein levels in the CA1 and dentate gyrus of the hippocampus, as well as in central and basolateral nuclei of the amygdala, which was accompanied by the translocation of GR from cytoplasm to nuclei. CSD also markedly reduced GR mRNA levels and MR immunoreactivity in the CA1, CA3 and dentate gyrus areas of the hippocampus. Conversely, CSD pronouncedly enhanced GR mRNA and protein levels in the dorsal raphe nucleus and locus coeruleus relative to the control. As an extension of our previous studies, in situ hybridization and immunohistochemical staining demonstrated that CSD regimen caused a notable increase of SERT mRNA levels in the dorsal raphe nucleus and increased SERT immunoreactivities in CA1 and CA3 of the hippocampus, as well as those in the basolateral nuclei of the amygdala. Likewise, CSD regimen resulted in an evident enhancement of NET immunoreactivity in the CA1 of the hippocampus and in the basolateral nuclei of the amygdala. Our current findings suggest that GR expressional alterations in response to CSD are complex and brain region-specific, which may correspond to their different functions in these regions

Colorectal cancer–associated T cell receptor repertoire abnormalities are linked to gut microbiome shifts and somatic cell mutations

ABSTRACTAs with many diseases, tumor formation in colorectal cancer (CRC) is multifactorial and involves immune, environmental factors and various genetics that contribute to disease development. Accumulating evidence suggests that the gut microbiome is linked to the occurrence and development of CRC, and these microorganisms are important for immune maturation. However, a systematic perspective integrating microbial profiling, T cell receptor (TCR) and somatic mutations in humans with CRC is lacking. Here, we report distinct features of the expressed TCRβ repertoires in the peripheral blood of and CRC patients (n = 107) and healthy donors (n = 30). CRC patients have elevated numbers of large TCRβ clones and they have very low TCR diversity. The metagenomic sequencing data showed that the relative abundance of Fusobacterium nucleatum (F. nucleatum), Escherichia coli and Dasheen mosaic virus were elevated consistently in CRC patients (n = 97) compared to HC individuals (n = 30). The abundance of Faecalibacterium prausnitzii and Roseburia intestinalis was reduced in CRC (n = 97) compared to HC (n = 30). The correlation between somatic mutations of target genes (16 genes, n = 79) and TCR clonality and microbial biomarkers in CRC had been investigated. Importantly, we constructed a random forest classifier (contains 15 features) based on microbiome and TCR repertoires, which can be used as a clinical detection method to screen patients for CRC. We also analysis of F. nucleatum-specific TCR repertoire characteristics. Collectively, our large-cohort multi-omics data aimed to identify novel biomarkers to inform clinical decision-making in the detection and diagnosis of CRC, which is of possible etiological and diagnostic significance