Effects of Long-Lasting High-Definition Transcranial Direct Current Stimulation in Chronic Disorders of Consciousness: A Pilot Study

Transcranial direct current stimulation (tDCS) recently was shown to benefit rehabilitation of patients with disorders of consciousness (DOC). However, high-Definition tDCS (HD-tDCS) has not been applied in DOC. In this study, we tried to use HD-tDCS protocol (2 mA, 20 min, the precuneus, and sustaining 14 days) to rehabilitate 11 patients with DOC. Electroencephalography (EEG) and Coma Recovery Scale–Revised (CRS-R) scores were recorded at before (T0), after a single session (T1), after 7 days’ (T2), and 14 days’ HD-tDCS (T3) to assess the modulation effects. EEG coherence was measured to evaluate functional connectivity during the experiment. It showed that 9 patients’ scores increased compared with the baseline. The central-parietal coherence significantly decreased in the delta band in patients with DOC. EEG coherence might be useful for assessing the effect of HD-tDCS in patients with DOC. Long-lasting HD-tDCS over the precuneus is promising for the treatment of patients with DOC

Transient Focal Cerebral Ischemia/Reperfusion Induces Early and Chronic Axonal Changes in Rats: Its Importance for the Risk of Alzheimer's Disease

The dementia of Alzheimer's type and brain ischemia are known to increase at comparable rates with age. Recent advances suggest that cerebral ischemia may contribute to the pathogenesis of Alzheimer's disease (AD), however, the neuropathological relationship between these two disorders is largely unclear. It has been demonstrated that axonopathy, mainly manifesting as impairment of axonal transport and swelling of the axon and varicosity, is a prominent feature in AD and may play an important role in the neuropathological mechanisms in AD. In this study, we investigated the early and chronic changes of the axons of neurons in the different brain areas (cortex, hippocampus and striatum) using in vivo tracing technique and grading analysis method in a rat model of transient focal cerebral ischemia/reperfusion (middle cerebral artery occlusion, MCAO). In addition, the relationship between the changes of axons and the expression of β-amyloid 42 (Aβ42) and hyperphosphorylated Tau, which have been considered as the key neuropathological processes of AD, was analyzed by combining tracing technique with immunohistochemistry or western blotting. Subsequently, we found that transient cerebral ischemia/reperfusion produced obvious swelling of the axons and varicosities, from 6 hours after transient cerebral ischemia/reperfusion even up to 4 weeks. We could not observe Aβ plaques or overexpression of Aβ42 in the ischemic brain areas, however, the site-specific hyperphosphorylated Tau could be detected in the ischemic cortex. These results suggest that transient cerebral ischemia/reperfusion induce early and chronic axonal changes, which may be an important mechanism affecting the clinical outcome and possibly contributing to the development of AD after stroke

Hypoxia-associated circDENND2A promotes glioma aggressiveness by sponging miR-625-5p

Abstract Background As a newfound type of non-coding RNA, circular RNAs (circRNAs) are involved in various physiological and pathological processes via regulation of gene expression. Increasing evidence shows that aberrantly expressed circRNAs play a crucial role in the initiation and progression of many tumors. However, the functions of different circRNAs in gliomas remain elusive. Methods The levels of circRNAs, miRNAs, and mRNAs were quantified by qPCR. The interaction between circDENND2A and miR-625-5p was determined by luciferase reporter and pull-down assays. The migratory and invasive capabilities of glioma cells were examined by wound healing and Transwell assays. Immunohistochemistry was performed to analyze the HIF1α level in glioma tissues. Results We predicted circDENND2A (has_circ_0002142) to be a hypoxia-responsive circRNA in glioma via a bioinformatic analysis. We found that hypoxia induced the expression of circDENND2A, which promoted migration and invasion of glioma cells. To understand the behaviors of circDENND2A in glioma, we studied the putative miRNAs targeted by circDENND2A and identified circDENND2A as an efficient sponge of miR-625-5p in glioma cells. Phenotype experiments verified that circDENND2A was required for the hypoxia-induced migration and invasion of glioma cells and that this occurred by sponging of miR-625-5p. Notably, glioma tissues overexpressing HIF1α exhibited a high expression of circDENND2A as well as a low expression of miR-625-5p. circDENND2A was negatively correlated with miR-625-5p. Conclusion circDENND2A is required for the hypoxia-induced malignancy of glioma cells and functions by sponging miR-625-5p

Quality Evaluation of Bioactive Ingredients in Lianhua Qingwen Capsule Based on Quantitative Analysis of Multicomponent by the Single Marker Method and the Chemical Recognition Patterns Method

Based on high performance liquid chromatography with the diode array detector (HPLC-DAD), a new strategy for simultaneous determination of ten bioactive ingredients in Lianhua Qingwen capsule (LHQW) was developed for comprehensive quality assessment of LHQW. In this work, with rhein regarded as the internal reference substance (IRS), the relative correction factors (RCFs) of neochlorogenic acid, amygdalin, chlorogenic acid, forsythoside A, quercitrin, phillyin, glycyrrhizic acid, isoforsythiaside, and (+) pinoresinol-β-D-glucoside were calculated for simultaneous determination of ten bioactive ingredients. More importantly, compared to previous work, the simultaneous determination of the content of ten pharmacologically important active ingredients at one detection wavelength with only one reference substance has been achieved. Based on the contents of ten bioactive ingredients, the quality of the 20 batches of LHQW samples was further analyzed by chemical recognition patterns method. Ten bioactive ingredients showed a good linear relationship in their respective concentration ranges (r ≥ 0.999). The relative standard deviations (RSDs) of precision (≤4.62%), stability (≤4.04%), repeatability (≤3.87%), and the average recovery of ten bioactive components (99.8%∼104.1%) demonstrated the QAMS developed for LHQW which had good durability. The correlation coefficient (P>0.05) showed that no significant difference existed in the results of QAMS and external standard method (ESM). Hierarchical clustering analysis (HCA) divided samples into three main groups. Radar plot analysis and principal component analysis (PCA) found some quality differences existed between the three groups of samples. Orthogonal partial least-squares discrimination analysis (OPLS-DA) showed that forsythoside A could be used as the primary marker responsible for the quality differences. In conclusion, the established QAMS method combined with chemometric analysis can simultaneously determine the content of 10 active components and comprehensively evaluate the quality of different batches of LHQW. It can provide scientific basis and reference of quality consistency evaluation for the formulation manufacturers and drug regulatory authorities

An Optimized Microwave-Assisted Digestion Method to Analyze the Amino Acids Profile of <i>Quisqualis Fructus</i> from Different Planted Origins

This study aims to establish a rapid and convenient microwave-assisted digestion method for sample pretreatment to determine amino acid profiles in natural products. This method was applied to analyze the amino acid profiles of Quisqualis Fructus (QF) from different planted origins. The microwave-assisted digestion conditions were optimized by a response surface methodology (RSM), and 17 amino acids in different planted origins of QF were determined by an automatic amino acid analyzer according to the optimized digestion conditions. The contents of 17 amino acids in QF from different planted origins were further analyzed by fingerprint and chemometric analysis. The temperature of microwave digestion at 167 °C, time of microwave digestion at 24 min, and a solid–liquid ratio of 46.5 g/mL was selected as the optimal digestion conditions. The total content of 17 amino acids in QF from different planted origins ranged from 71.88 to 91.03 mg/g. Amino acid composition and nutritional evaluation indicated that the content of medicinal amino acids was higher than aromatic amino acids. The results of fingerprint analysis reflected that the similarity between the 16 batches of QF ranged from 0.889 to 0.999, while chemometrics analysis indicated amino acid content in QF varied from different planted origins, and six important differential amino acids were screened. Compared with the traditional extraction method, microwave-assisted digestion with response surface optimized has the advantages of rapidity, convenience, and reliability, which could be used to study the amino acid profiles in natural products. The amino acid profile of QF indicated that it has a rich medicinal nutritional value. Different planted origins of QF have a high degree of similarity and could be effectively distinguished by chemometric analysis

Semi-quantitative analysis of the degree of axonal changes in different brain regions in transient cerebral ischemia/reperfusion and control rats from axonal tracing evaluation.

<p>The numbers (1–3) represent the degree of axonal changes (see also in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0033722#s4" target="_blank">method</a>). MCAO: middle cerebral artery occlusion and reperfusion; Psc: primary sensory cortex; Pmc: primary motor cortex; Hi: hippocampus; Cpu: caudoputamen; Per-C: perilesional cortex; ND: not detected.</p

Tracing and immunohistochemical double staining show the tracer-labeled neurons and the expression of hyperphosphorylated Tau induced by transient cerebral ischemia/reperfusion.

<p>B, D and F are the magnification of an area in A, C and E, respectively. Some staining for AT8 (brown color) was found in the ischemic cortex (A) and AT8 labeled neuronal bodies could be noticed (B, arrowheads). The arrows in B indicate the tracer-labeled neurons (dark blue color). Strong staining for Tau-5 (brown color) was found in the ischemic cortex (C) and the extensive Tau-5 labeled neurons (D, arrows) and swollen axons (D, arrowheads) were noticed. Inserted figure in C showed the magnification of an area (arrowhead), demonstrating the tracer-labeled neurons (dark blue color). Only very weak staining could be observed for P-tau (E) and tracer-labeled neurons presented the morphological changes as cell shrinkage occurred. Scale bars: 10 µm for B, D, and F, 100 µm for A and C, 200 µm for E.</p