Muscle stem cells (MuSCs) hold great therapeutic potential for muscle genetic disorders, such as Duchenne muscular dystrophy (DMD). The CRISP/Cas9-based genome editing is a promising technology for correcting genetic alterations in mutant genes. In this study, we used fibrin-gel culture system to selectively expand MuSCs from crude skeletal muscle cells of mdx mice, a mouse model of DMD. By CRISP/Cas9-based genome editing, we corrected the dystrophin mutation in expanded MuSCs and restored the skeletal muscle dystrophin expression upon transplantation in mdx mice. Our studies established a reliable and feasible platform for gene correction in MuSCs by genome editing, thus greatly advancing tissue stem cell-based therapies for DMD and other muscle disorders.

Background. The microglia cell transfer has been shown to play a protective role in ischemic stroke. Microglia cells may play this nerve-protective role via the promotion of angiogenesis. However, the underlying mechanisms are largely unknown and need further investigation. Objectives. The aim of this study was to investigate the pro-angiogenesis effects of unpolarized, interleukin 4 (IL-4)-polarized or lipopolysaccharide (LPS)-polarized BV2 microglia cells both in vivo and in vitro. We also investigated the potential mechanisms of these pro-angiogenesis effects. Material and methods. BV2 cells were polarized using phosphate-buffered saline (PBS), LPS or IL-4, respectively. The gene expression pattern was analyzed with reverse transcription polymerase chain reaction (RT-PCR). The transfer of polarized BV2 cells was performed with an intravenous injection into mice 45 min after the middle cerebral artery (MCA) occlusion. Angiogenin expression was assessed with immunofluorescence. We also examined the angiogenesis effect of polarized BV2 cells and their exosomes through 3-dimensional co-cultures in vitro. Finally, the microRNA (miRNA) profiles of exosomes released by BV2 cells under different polarization conditions were examined using miRNA microarray. Results. The IL-4-polarized BV2 transplantation promoted angiogenin expression in the ischemic brain. Interleukin 4-polarized microglia increased the tube formation of endothelial cells by secreting exosomes. The miRNA profiles of exosomes released by BV2 cells under different polarization conditions were different. Exosomes from IL-4-polarized BV2 cells contained higher amounts of miRNA-26a compared to those from the LPS-polarized and unpolarized BV2 cells. Conclusions. Interleukin 4-polarized microglia cells might ameliorate the damage caused by ischemic stroke by promoting angiogenesis through the secretion of exosomes containing miRNA-26a.

We simulated the fourier transform of the correlation function of the Ising model in two and three dimensions using a single cluster algorithm with improved estimators. The simulations are in agreement with series expansion and the available exact results in d = 2, which shows, that the cluster algorithm can succesfully be applied for correlations. We show as a further result that our data do not support a hypothesis of Fisher that in any d = 2 lattice the fourier transform of the correlation function depends on the lattice generating function only. In d = 3 our simulation are again in agreement with the results from the series expansion, except for the amplitudes f ± , where we find f + /f − = 2.06(1).

Previous studies have demonstrated that strabismus or amblyopia can result in marked brain function and anatomical alterations. However, differences in spontaneous brain activity in strabismus and amblyopia (SA) patients as compared with control individuals remain unclear. The present study aimed to analyze the potential brain activity changes in SA patients and their association with behavioral performance. In total, 16 patients with SA (10 women and 6 men) and 16 healthy controls (HCs; 6 men and 10 women) with matched age and sex were recruited. All subjects were examined with resting-state functional magnetic resonance imaging (rs-fMRI), and changes in the spontaneous brain activity of SA patients were evaluated by the regional homogeneity (ReHo) method. The diagnostic ability of the ReHo method was assessed using receiver operating characteristic (ROC) curve analysis. In addition, the association between the mean ReHo value in different brain regions and the behavioral performance was explored by correlation analysis. It was observed that the ReHo value was significantly increased in SA patients compared with HCs in the following brain regions: left lingual gyrus, right middle occipital gyrus/precuneus, bilateral anterior cingulate, left middle occipital gyrus and bilateral precentral gyrus. By contrast, the ReHo value of the left inferior frontal gyrus was significantly lower than that in HCs. ROC curve analysis indicated that the ReHo method has certain credibility for the diagnosis of SA patients. In addition, no similar changes were detected in other brain regions. These results revealed abnormal spontaneous brain activity in certain parts of the brain in adult patients with SA, which suggests the involvement of the neuropathological or compensatory mechanism in these patients, and may be beneficial for clinical treatment.

ObjectivePrevious studies have demonstrated that strabismus or amblyopia can result in markedly brain function and anatomical alterations. However, the differences in spontaneous brain activities of strabismus with amblyopia (SA) patients still remain unclear. This current study intended to use the amplitude of low-frequency fluctuation (ALFF) technique to investigate the intrinsic brain activity changes in SA subjects.Patients and methodsA total of 16 patients with SA (6 males and 10 females) and 16 healthy controls (HCs; 6 males and 10 females) similarly matched in age, gender, and education status were recruited and examined with the resting-state functional MRI. The spontaneous brain activity changes were investigated using the ALFF technique. The receiver operating characteristic curve was performed to classify the mean ALFF signal values of the SA patients from HCs. The correlations between the ALFF values of distinct brain regions and the clinical manifestations in SA patients were evaluated in terms of the Pearson’s correlation analysis.ResultsCompared with HCs, SA patients had significantly decreased ALFF in the left cerebellum posterior lobe, left middle frontal gyrus, and bilateral thalamus. In contrast, SA patients showed increased ALFF values in the right superior frontal gyrus, right precuneus, left cuneus, and bilateral precentral gyrus. Nonetheless, there was no linear correlation between the mean ALFF values in brain regions and clinical features.ConclusionDiverse brain regions including vision-related and motion-related areas exhibited aberrant intrinsic brain activity patterns, which imply the neuropathologic mechanisms of oculomotor disorder and vision deficit in the SA patients.

Objective Neuroimaging studies have shown that patients with pain‐related conditions have altered neuronal activity and structural functions. The purpose of this study was to investigate whether patients with classical trigeminal neuralgia (CTN) exhibit changes in corresponding neuronal activity via analysis of neuronal activity regional homogeneity (ReHo). Methods A total of 28 patients presenting with sore eyes (12 men and 16 women) were matched with 28 healthy controls (12 men and 16 women). All participants underwent functional magnetic resonance imaging (fMRI). This ReHo method was used to assess the consistency of changes in neural activity in various brain regions. The receiver operating characteristic (ROC) curve was applied to differentiate ReHo values of patients with CTN from ReHo values of healthy controls. Pearson's correlation analysis was applied to evaluate the correlation between ReHo values of different brain regions of patients with CTN and clinical manifestations. Results Compared with healthy controls, patients with CTN were found to have increased ReHo values in the inferior cerebellum bilaterally, right inferior temporal gyrus, right middle occipital gyrus, right fusiform gyrus, right superior frontal gyrus, and right precentral gyrus. ROC curve analysis of each brain region revealed near‐perfect accuracy regarding the area under the curve. However, no correlation between ReHo values and clinical manifestations in patients with CTN was found. Conclusions CTN is associated with altered neuronal networks in different areas of the brain. ReHo values all possess different degrees of change, implying that CTN has a certain impact on cerebral function.

Metabolic profiling in liver and serum of mice was studied for the combined toxic effects of deoxynivalenol (DON) and zearalenone (ZEN), through gas chromatography mass spectrum. The spectrum of serum and liver sample of mice, treated with individual 2 mg/kg DON, 20 mg/kg ZEN, and the combined DON + ZEN with final concentration 2 mg/kg DON and 20 mg/kg ZEN for 21 days, were deconvoluted, aligned and identified with MS DIAL. The data matrix was processed with univariate analysis and multivariate analysis for selection of metabolites with variable importance for the projection (VIP) > 1, t-test p value < 0.05. The metabolic pathway analysis was performed with MetaMapp and drawn by CytoScape. Results show that the combined DON and ZEN treatment has an obvious “antagonistic effect” in serum and liver tissue metabolic profiling of mice. The blood biochemical indexes, like alkaline phosphatase, alanine transaminase, and albumin (ALB)/globulin (GLO), reveal a moderated trend in the combined DON + ZEN treatment group, which is consistent with histopathological examination. The metabolic pathway analysis demonstrated that the combined DON and ZEN treatment could down-regulate the valine, leucine and isoleucine biosynthesis, glycine, serine and threonine metabolism, and O-glycosyl compounds related glucose metabolism in liver tissue. The metabolic profiling in serum confirmed the finding that the combined DON and ZEN treatment has an “antagonistic effect” on liver metabolism of mice.

SummarymiR-302/367 is the most abundant miRNA cluster in human embryonic stem cells (hESCs) and can promote somatic cell reprogramming. However, its role in hESCs remains poorly understood. Here, we studied functional roles of the endogenous miR-302/367 cluster in hESCs by employing specific TALE-based transcriptional repressors. We revealed that miR-302/367 cluster dually regulates hESC cell cycle and apoptosis in dose-dependent manner. Gene profiling and functional studies identified key targets of the miR-302/367 cluster in regulating hESC self-renewal and apoptosis. We demonstrate that in addition to its role in cell cycle regulation, miR-302/367 cluster conquers apoptosis by downregulating BNIP3L/Nix (a BH3-only proapoptotic factor) and upregulating BCL-xL expression. Furthermore, we show that butyrate, a natural compound, upregulates miR-302/367 cluster expression and alleviates hESCs from apoptosis induced by knockdown of miR-302/367 cluster. In summary, our findings provide new insights in molecular mechanisms of how miR-302/367 cluster regulates hESCs.