توجه: فایل دانلود شده برای دانلود نرم افزار الیگو 7 (oligo 7.60) به صورت فشرده می باشد برای استخراج این فایل از رمز: bio استفاده نمایید.
فیلم آموزش طراحی پرایمر : آموزش نرم افزار الیگو آنالایزر (oligo analyzer) برای بررسی ویژگی های پرایمرها مانند تغیین میزان Tm درصد CG برای پرایمرها و همچنین بررسی ساختارهای ثانویه برای پرایمرها در این قسمت ارائه شده است.
oligo 7 keygen
Chemical structures of ten oligostilbenes isolated from the seed shells of Paeonia suffruticosa. (1) suffruticosol A, (2) suffruticosol B, (3) suffruticosol C, (4) trans-resveratrol, (5) cis-ε-viniferin, (6) trans-ε-viniferin, (7) cis-suffruticosol D, (8) cis-gnetin H, (9) trans-suffruticosol D, and (10) trans-gnetin H. The number was based on their peak order of HPLC
Effects of ten oligostilbenes on apoptosis of OA chondrocytes. Group of experiments: untreated control group, IL-1β group, the following groups were treated with 10 ng/mL IL-1β in addition to 1 μM the listed drug: diacerein, (1) suffruticosol A, (2) suffruticosol B, (3) suffruticosol C, (4) trans-resveratrol, (5) cis-ε-viniferin, (6) trans-ε-viniferin, (7) cis-suffruticosol D, (8) cis-gnetin H, (9) trans-suffruticosol D, and (10) trans-gnetin H
Small interference RNA targeting hsa_circ_0000735 (si-circ-1 and si-circ-2) and matching control (si-NC), as well as lentivirus-mediated sh-hsa_circ_0000735 (sh-circ) and corresponding control (sh-NC), were achieved from Genepharma (Shanghai, China). MiR-7 mimic and inhibitor (miR-7 and anti-miR-7), as well as their negative controls (miR-NC and anti-NC), were purchased from Ribobio (Guangzhou, China). The oligonucleotides were transfected into DTX-resistant PCa cells through using Lipofectamine 3000 reagent (Life Technologies, Carlsbad, CA, USA).
Some binary systems, such as the lac operon system [15], the tetR-based system [16], the GAL4-based system [17] and the Cre/loxP recombination system [18], have been used to turn gene expression on and off transiently or permanently. Bacterial lac operon-regulated gene expression in mammalian cells was first demonstrated by Hu and Davidson [19]. Cronin et al. [15] also showed that the lac operon system was functional in the mouse and could provide tight and reversible gene expression. The lac operon induction system might be the best option for inducible α-galactosidase expression because α-galactosidase hydrolysis of oligosaccharides or milk can produce α-lactose, which binds to the lac repressor and facilitates positive feedback of α-galactosidase expression.
The short hairpin RNA (shRNA) targeting control and THOC1 sequences were as follows: shNC: TTCTCCGAACGTGTCACGT, shTHOC1: GATACCAAACCTACGAGAA. The palindromic DNA oligo was annealed to form a double-strand oligo and then ligated to the linearized pLKD-CMV-EGFP-2A-Puromycin-U6-shRNA (OBIO, Shanghai, China) vector to generate the circled pLKD-CMV-shRNA-Puromycin. The specific primers for full-length THOC1 were summarized as follows: THOC1-F: GCTCTAGAATGTCTCCGACGCCG; THOC1-R: CGGGATCCCTAACTATTTGTCTCATTGTCATTA. The amplified fragments were finally ligated into the pLV-EF1a-MCS-IRES-Blasticidin (Biosettia Inc. San Diego, California, USA) expression vector. The PLC/PRF/5 and the HepG2 cells were infected with a lentivirus carrying pLKD-CMV-shRNA-Puromycin or pLV-EF1a-THOC1-Blasticidin plasmids, followed by the separate selection using puromycin or blasticidin to generate polyclonal cell populations.
The massive loss of oligodendrocytes caused by various pathological factors is a basic feature of many demyelinating diseases of the central nervous system (CNS). Based on a variety of studies, it is now well established that impairment of oligodendrocyte precursor cells (OPCs) to differentiate and remyelinate axons is a vital event in the failed treatment of demyelinating diseases. Recent evidence suggests that Foxg1 is essential for the proliferation of certain precursors and inhibits premature neurogenesis during brain development. To date, very little attention has been paid to the role of Foxg1 in the proliferation and differentiation of OPCs in demyelinating diseases of the CNS. Here, for the first time, we examined the effects of Foxg1 on demyelination and remyelination in the brain using a cuprizone (CPZ)-induced mouse model. In this work, 7-week-old Foxg1 conditional knockout and wild-type (WT) mice were fed a diet containing 0.2% CPZ w/w for 5 weeks, after which CPZ was withdrawn to enable remyelination. Our results demonstrated that, compared with WT mice, Foxg1-knockout mice exhibited not only alleviated demyelination but also accelerated remyelination of the demyelinated corpus callosum. Furthermore, we found that Foxg1 knockout decreased the proliferation of OPCs and accelerated their differentiation into mature oligodendrocytes both in vivo and in vitro. Wnt signaling plays a critical role in development and in a variety of diseases. GSK-3β, a key regulatory kinase in the Wnt pathway, regulates the ability of β-catenin to enter nuclei, where it activates the expression of Wnt target genes. We then used SB216763, a selective inhibitor of GSK-3β activity, to further demonstrate the regulatory mechanism by which Foxg1 affects OPCs in vitro. The results showed that SB216763 clearly inhibited the expression of GSK-3β, which abolished the effect of the proliferation and differentiation of OPCs caused by the knockdown of Foxg1. These results suggest that Foxg1 is involved in the proliferation and differentiation of OPCs through the Wnt signaling pathway. The present experimental results are some of the first to suggest that Foxg1 is a new therapeutic target for the treatment of demyelinating diseases of the CNS.
In the central nervous system (CNS), oligodendrocytes (OLs) are myelin-forming glial cells that play vital roles in rapid impulse conduction and normal axonal functions. Current studies have demonstrated that inflammatory stimuli or immune attacks can damage the myelin sheath, leading to OL death and myelin sheath loss, such as that observed in multiple sclerosis (MS), the most common demyelinating disease [1]. It has been reported that in early MS lesions, neural stem cells (NSCs) rooted in the region of the lateral ventricle-subventricular zone migrate, proliferate, and differentiate into oligodendrocyte progenitor cells (OPCs) induced by the injury signal of the myelin sheath and later differentiate to myelinating OLs to repair the damaged myelin [2]. However, OPCs are slow-cycling cells, and only long-term nutritional factors can stimulate their rapid proliferation and differentiation [3]. Furthermore, the repair ability of endogenous remyelination is limited, resulting in difficulties in the regeneration/repair of the myelin sheath in demyelinating diseases. Therefore, elucidating the regulatory mechanism of the correct differentiation of OPCs is crucial for understanding the myelination and remyelination processes in the CNS and is also fundamental to the treatment of demyelinating diseases [4].
To investigate the role of Foxg1 in the proliferation and differentiation of oligodendrocyte lineage cells and myelin regeneration, we conditionally knocked out Foxg1 in vivo to assess the loss and repair of myelin during demyelinating injury in mice, and we further explored its regulatory mechanism in vitro. Our findings revealed a previously-unknown function of Foxg1 in oligodendrocyte lineage cells, suggesting that downregulating the expression of Foxg1 in the treatment of demyelinating CNS diseases may alleviate demyelination and promote remyelination, thus providing a new gene therapy target for demyelinating CNS diseases such as MS.
The rat OPC proliferation culture was maintained as previously described [24, 25]. Isolated OPCs were plated at 10,000 cells/cm2 on poly-L-lysine-coated flasks and cultured in DMEM/F12 medium supplemented with 2% B27, 10 ng/mL platelet-derived growth factor AA (PDGF-AA, Gibco, Grand Island, NY, USA), and 10 g/mL basic fibroblast growth factor (bFGF, Gibco) for 3 days; then, the medium was replaced with DMEM/F12 medium without PDGF-AA and bFGF for 1 day to generate preOLs. For oligodendrocyte differentiation, 10% fetal bovine serum (FBS) was added to the preOL medium and cultured for 7 days. The medium was changed every 2 days.
BrdU labeling was used to assess cell proliferation (Fig. 3A) and differentiation (Fig. 4A) in the corpus callosum, and the results showed that the number of BrdU-positive cells was significantly lower in the Foxg1 cKO+CPZ group than the WT+CPZ group (Fig. 3C, D). Furthermore, the increase in percentage of NG2+ cells (Fig. 3E, F) and O4+ cells (Fig. 3J) cells in the WT+CPZ group was prevented by Foxg1-cKO treatment. The number of proliferating OPCs, indicated by NG2+/BrdU+ (Fig. 3G, H) or BrdU+/O4+ (Fig. 3I, K), was also decreased; however, the number of new mature oligodendrocytes labeled with BrdU+/MBP+ (Fig. 4C, D), BrdU+/CNPase+ (Fig. 4E, F), and BrdU+/MAG+ (Fig. 4G, H) differentiated from the OPCs was significantly higher in the Foxg1 cKO+CPZ group than in the WT+CPZ group. These data suggested that Foxg1 cKO decreases the production or proliferation of OPCs but promotes their differentiation.
As described in previous reports, Foxg1 is abundantly expressed in the CNS [31] and plays pivotal roles in organogenesis through the regulation of proliferation and the specification of cell fate [32]. However, the role of Foxg1 in the oligodendroglial lineage and the underlying mechanisms remain unclear. Therefore, the present study was designed to determine the effect of Foxg1 and the mechanisms underlying the regulation of the Wnt signaling pathway following CPZ-induced demyelination. 2ff7e9595c
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