Investigating the regulation of HSP70 by miRNAs

The HSP70 family is the most highly conserved of the high molecular weight HSP's in both non-neuronal and neuronal systems. Recent studies have also shown that the overexpression of HSP70 is neuroprotective for stroke and the polyglutamine (PolyQ) disorders. Hence understanding the mechanisms that regulate HSP70 may allow new therapeutic strategies to be developed. MicroRNAs are a subset of small non-coding RNAs that have been shown to mediate an entirely new level of post-transcriptional gene regulation by inhibiting the translation of target mRNAs. MiRNAs bind complementary sequences within the 3 'UTR of specific mRNAs and mediate their translational repression. An aim of this study was to identify the miRNAs that regulate HSP70 and investigate their involvement in regulating HSP70 during a heat shock response. To achieve this I initially used the online algorithms designed to predict miRNA targets. The predictions made by the different algorithms showed considerable variation and we hence also carried out an empirical analysis. MiR-206 and miR-21 were found to down regulate luciferase expression when linked to the HSP70 3 'UTR. In order to study the role these two miRNAs play in controlling HSP70 expression further, a lentivirus expressing miR-206 and miR-21 was used. Transduction of HeLa cells with this virus and an HSP70 3'UTR luciferase reporter found the virus mediated a dose dependent decrease in translational activity. HeLa and C2C12 cells were used to study function and preliminary experiments showed that HSP70 mRNA expression was increased 1-3 hours after the induction of a heat shock (HS). HSP70 protein levels were also increased following a HS. Following heat shock miR-206 levels were found to fall significantly within the first hour after heat shock (in C2C12 cells) and then start to recover twenty-fours later. The fall in miR-206 levels correlated with increased HSP70 levels, though this trend did not reach statistical significance. Transduction with viruses expressing miR-206 and miR-21 were shown to significantly decrease HSP70 protein but not mRNA expression following a HS. Furthermore, miR-206 specific antagornirs mediated a decrease in endogenous miR-206 levels and a trend towards increased basal levels of HSP70 protein was also seen. These results suggest that miR-206 may be one of a number of miRNAs that help ensure conditions within a cell are permissive or non-permissive for the expression of heat shock proteins following a stress or consitutively.EThOS - Electronic Theses Online ServiceGBUnited Kingdo

Simultaneous Detection of Ascorbic Acid, Dopamine, and Uric Acid Using a Novel Electrochemical Sensor Based on Palladium Nanoparticles/Reduced Graphene Oxide Nanocomposite

A fresh strategy based on two-step electrochemical reduction for the fabrication of palladium nanoparticles/reduced oxide nanocomposite-modified glass carbon electrode (PdNPs/rGO/GCE) was established in this study. Field emission scanning electron microscopy (FESEM) images showed that spherical PdNPs were evenly distributed on the surface of rGO-modified electrode (rGO/GCE), and the introduction of PdNPs has no effect on the morphology of rGO. Electrochemical impedance spectroscopy (EIS) studies revealed that the conductivity of PdNPs/rGO/GCE was higher than that of rGO/GCE and bare GCE. The electrochemical performances of PdNPs/rGO/GCE sensor were investigated by cyclic voltammetry (CV), differential pulse voltammetry (DPV), and chronoamperometry using ascorbic acid (AA), dopamine (DA), and uric acid (UA) as analytes. At the optimized conditions, wide linear ranges of 0.5–3.5 mM (R2 = 0.99), 3–15 μM (R2 = 0.99) and 15–42 μM (R2 = 0.99), and 0.3–1.4 mM (R2 = 0.99) towards AA, DA, and UA in ternary mixture were observed, respectively. In addition to superior anti-interference capability, fast response (≤5 s), excellent reproducibility, and good long-term stability were also given by this sensor. These results suggested that PdNPs/rGO/GCE is promising for the simultaneous detection of AA, DA, and UA in practical application

The efficacy and safety of acupuncture and moxibustion combined with western medicine for obsessive-compulsive disorder A protocol for systematic review and meta-analysis

Background Obsessive-compulsive disorder is common, chronic mental disorder, which is characterized by recurrent, unwanted, or intrusive thoughts (obsessions) and repetitive behaviors or mental action (compulsions). Acupuncture and moxibustion, as a popular form of complementary and alternative therapy, have the advantages of low side effects, high safety and low cost. The research showed that acupuncture and moxibustion have a good clinical efficacy on obsessive-compulsive disorder. However, there is no literature to systematically evaluate the efficacy and safety of acupuncture and moxibustion in treating obsessive-compulsive disorder. Thus, this study is aimed to evaluate the efficacy and safety of acupuncture and moxibustion for obsessive-compulsive disorder patients, providing reliable evidence for clinical application. Methods Randomized controlled trials of acupuncture and moxibustion combined with western medicine for the treatment of obsessive-compulsive disorder will be searched in the databases including PubMed, EMBASE, the Cochrane library, Web of science, China National Knowledge Infrastructure(CNKI), WanFang, the Chongqing VIP Chinese Science and Technology Periodical Database, and China biomedical literature database(CBM) from inception to June, 2020. In addition, Baidu, Google Scholar, International Clinical Trials Registry Platform, and Chinese Clinical Trials Registry will be searched to obtain the gray literature and relevant data that have not yet been published. Two qualified researchers will extract data and assess the risk of bias from included studies dependently. Statistical analysis is performed in RevMan 5.3 software. Results The efficacy and safety of acupuncture and moxibustion combined with western medicine for obsessive-compulsive disorder will be assessed based on the total effective rate, Hamilton Anxiety Scale score, Hamilton Rating Scale for Depression score, Clinical Global Impression score, side effects and so on. Conclusions The proposed systematic review and meta-analysis of acupuncture and moxibustion combined with western medicine for treating obsessive-compulsive disorder is expected to provide reliable evidence for clinical application

The Role of Neuroglial Crosstalk and Synaptic Plasticity-Mediated Central Sensitization in Acupuncture Analgesia

Although pain is regarded as a global public health priority, analgesic therapy remains a significant challenge. Pain is a hypersensitivity state caused by peripheral and central sensitization, with the latter considered the culprit for chronic pain. This study summarizes the pathogenesis of central sensitization from the perspective of neuroglial crosstalk and synaptic plasticity and underlines the related analgesic mechanisms of acupuncture. Central sensitization is modulated by the neurotransmitters and neuropeptides involved in the ascending excitatory pathway and the descending pain modulatory system. Acupuncture analgesia is associated with downregulating glutamate in the ascending excitatory pathway and upregulating opioids, -aminobutyric acid, norepinephrine, and 5-hydroxytryptamine in the descending pain modulatory system. Furthermore, it is increasingly appreciated that neurotransmitters, cytokines, and chemokines are implicated in neuroglial crosstalk and associated plasticity, thus contributing to central sensitization. Acupuncture produces its analgesic action by inhibiting cytokines, such as interleukin-1β, interleukin-6, and tumor necrosis factor-α, and upregulating interleukin-10, as well as modulating chemokines and their receptors such as CX3CL1/CX3CR1, CXCL12/CXCR4, CCL2/CCR2, and CXCL1/CXCR2. These factors are regulated by acupuncture through the activation of multiple signaling pathways, including mitogen-activated protein kinase signaling (e.g., the p38, extracellular signal-regulated kinases, and c-Jun-N-terminal kinase pathways), which contribute to the activation of nociceptive neurons. However, the responses of chemokines to acupuncture vary among the types of pain models, acupuncture methods, and stimulation parameters. Thus, the exact mechanisms require future clarification. Taken together, inhibition of central sensitization modulated by neuroglial plasticity is central in acupuncture analgesia, providing a novel insight for the clinical application of acupuncture analgesia

ST36 Acupuncture Alleviates the Inflammation of Adjuvant-Induced Arthritic Rats by Targeting Monocyte/Macrophage Modulation

Background. Rheumatoid arthritis (RA) is a chronic systemic chronic autoimmune disease characterized by the aggregation of immune cells and secretion of cytokines in the joint synovium, causing hyperblastosis and even bone destruction. Acupuncture has been proven effective in RA treatment. This study aimed to investigate the anti-inflammatory action of acupuncture, specifically, in relation to immune cell interactions and key mediators. Methods. Rats with adjuvant-induced arthritics (AIA) were treated with manual acupuncture (MA) at Zusanli (ST36). Joint edema and paw withdrawal latency were monitored to observe the effects on inflammation. The levels of 24 cytokines, chemokines, and growth factors in ankle joints during the treatment (on days 1, 7, 15, and 21) were detected by multiplex immunoassay. A bioinformatics analysis based on a directed weighted mathematical model was used to construct cell communication network diagrams and identify the key cells through calculation. The monocyte/macrophage polarization in inflamed joints was investigated by detecting M1- and M2-phenotypic populations and their related cytokines. Results. ST36 MA alleviated paw edema and upregulated the nociceptive threshold of AIA rats. Several innate and adaptive immune cytokines were dynamically regulated by MA, and MA-treated rats showed a significant improvement in symptoms compared with AIA rats by day 21. The immune cell-cell communication networks were intensified with the development of RA but were significantly reduced after treatment with MA. MA was found to specifically regulate monocytes/macrophages in inflamed ankle joints ST36 MA also inhibited M1-phenotype macrophages accompanied by decreased levels of IL-1β. Conclusions. ST36 MA showed anti-inflammatory and analgesic effects as well as inhibition of immune cell communication networks in inflamed joints of AIA rats. Inhibiting the polarization of macrophages to the M1-phenotype in inflamed joints may be one of the key mechanisms of MA anti-inflammatory action. This research highlighted a systematic research paradigm for investigating mechanisms of acupuncture action

Characteristics of long-term cultured UC-MSCs on WJE-coated plates.

<p>(A) Morphological changes of long-term cultured UC-MSCs on WJE-coated plates by microscopy. Scale bars, 20 µm. (B) Proliferation ability of UC-MSCs cultured on WJE-coated plates by MTT assay. (C) Growth curve of long-term cultured UC-MSCs on WJE-coated plates. (D) Flow cytometric analysis of a representative example of UC-MSC (top). Histograms of a representative UC-MSCs culture on WJE-coated plate stained for lineage negative, and positive cell surface markers. Data are mean ± SD (n = 3) (*<i>P</i>>0.05).</p