MiR-146a promotes oligodendrocyte progenitor cell differentiation and enhances remyelination in a model of experimental autoimmune encephalomyelitis.

The death of mature oligodendrocytes (OLs) leads to demyelination in the central nervous system (CNS) and subsequently to functional deficits. Remyelination requires the differentiation of oligodendrocyte progenitor cells (OPCs) into myelinating OLs, which in the CNS with neurodegenerative diseases such as multiple sclerosis (MS), is often inhibited. Among the inhibitors of OPC differentiation are toll-like receptor 2 (TLR2) and interleukin-1 receptor-associated kinase 1 (IRAK1) signaling, and both are negatively regulated by microRNA-146a (miR-146a). Therefore, we hypothesized that increase of miR-146a level in the CNS would foster OPC differentiation and remyelination by inhibiting the TLR2/IRAK1 signaling pathway. Here, we tested this hypothesis using exogenous miR-146a mimics and a mouse model of MS, experimental autoimmune encephalomyelitis (EAE) induced by immunization with myelin proteolipid protein peptide (PLP139-151). EAE mice were treated by miR-146a mimics or miR-146a mimic negative controls, respectively, which initiated at day 14 post immunization, once a week for 6 consecutive weeks. Neurological function was monitored daily. Immunofluorescent staining, qRT-PCR and Western blot were used to measure the differentiation of OPCs and myelination, and to investigate the underlying mechanisms of action of miR-146a. Using a fluorescence tracing approach, we found that miR-146a mimics crossed the blood brain barrier and targeted OPCs and microglia/macrophages after systemic administration. MiR-146a mimic treatment substantially improved neurological functional outcome, increased the number of newly generated OLs which may facilitate remyelination in the CNS. The cell number, cytokine level and protein levels of M2 phonotype of microglia/macrophages significantly increased, while cytokine and protein levels of the M1 phenotype significantly decreased after miR-146a mimic treatment. Increased OPC differentiation and remyelination were associated with reduction of TLR2/IRAK1 signaling pathway activity by miR-146a mimic treatment. This study provides insight into the cellular and molecular bases for the therapeutic effects of miR-146a on OPC differentiation and remyelination, and suggests the potential of enhancing miR-146a as a treatment of demyelinating disorders.

Global Sensitivity Analysis To Characterize Operational Limits and Prioritize Performance Goals of Capacitive Deionization Technologies.

Capacitive deionization (CDI) technologies couple electronic and ionic charge storage, enabling improved thermodynamic efficiency of brackish desalination by recovering energy released during discharge. However, insight into CDI has been limited by discrete experimental observations at low desalination depths (Δ c, typically reducing influent salinity by 10 mM or less). In this study, the performance and sensitivity of three common CDI configurations [standard CDI, membrane CDI (MCDI), and flowable electrode CDI (FCDI)] were evaluated across the operational and material design landscape by varying eight common input parameters (electrode thickness, influent concentration, current density, electrode flow rate, specific capacitance, contact resistance, porosity, and fixed charge). All combinations of designs were evaluated for two influent concentrations with a calibrated and validated one-dimensional (1-D) porous electrode model. Sensitivity analyses were carried out via Monte Carlo and Morris methods, focusing on six performance metrics. Across all performance metrics, high sensitivity was observed to input parameters which impact cycle length (current, resistance, and capacitance). Simulations demonstrated the importance of maintaining both charge and round-trip efficiencies, which limit the performance of CDI and FCDI, respectively. Accounting for energy recovery, only MCDI was capable of operating at thermodynamic efficiencies similar to reverse osmosis.

Differential Regulation of BMAL1, CLOCK, and Endothelial Signaling in the Aortic Arch and Ligated Common Carotid Artery.

The circadian clock is rhythmically expressed in blood vessels, but the interaction between the circadian clock and disturbed blood flow remains unclear. We examined the relationships between BMAL1 and CLOCK and 2 regulators of endothelial function, AKT1 and endothelial nitric oxide synthase (eNOS), in vascular regions of altered blood flow. We found that the aortic arch from WT mice exhibited reduced sensitivity to acetylcholine (Ach)-mediated relaxation relative to the thoracic aorta. In Clock-mutant (mut) mice the aorta exhibited a reduced sensitivity to Ach. In WT mice, the phosphorylated forms of eNOS and AKT were decreased in the aortic arch, while BMAL1 and CLOCK expression followed a similar pattern of reduction in the arch. In conditions of surgically induced flow reduction, phosphorylated-eNOS (serine 1177) increased, as did p-AKT in the ipsilateral left common carotid artery (LC) of WT mice. Similarly, BMAL1 and CLOCK exhibited increased expression after 5 days in the remodeled LC. eNOS expression was increased at 8 p.m. versus 8 a.m. in WT mice, and this pattern was abolished in mut and Bmal1-KO mice. These data suggest that the circadian clock may be a biomechanical and temporal sensor that acts to coordinate timing, flow dynamics, and endothelial function.

Fingolimod treatment promotes proliferation and differentiation of oligodendrocyte progenitor cells in mice with experimental autoimmune encephalomyelitis.

Multiple sclerosis (MS) is a major demyelinating disease of the central nervous system (CNS) leading to functional deficits. The remyelination process is mediated by oligodendrocyte progenitor cells (OPCs). In this study, we tested the hypothesis that Fingolimod, a sphingosine 1-phosphate (S1P) receptor modulator, stimulates OPC differentiation into mature oligodendrocytes, in addition to its well-known anti-inflammatory effect. Using an animal model of MS, experimental autoimmune encephalomyelitis (EAE), we performed a dose-response study of Fingolimod (0.15 or 0.3mg/kg bw), which was initiated on the day of EAE onset. The neurological function was tested to determine the optimal dose of Fingolimod. Immunofluorescent staining was performed to measure the profile of OPC proliferation and differentiation. The mechanistic premise underlying the therapeutic effect of Fingolimod, was that Fingolimod stimulates the sonic hedgehog (Shh) pathway, and this pathway promotes OPC differentiation. To test this hypothesis, a loss-of-function study using cyclopamine, an inhibitor of the sonic hedgehog (Shh) pathway, was employed in vivo. Protein levels of the Shh pathway were measured by Western blot analysis. We found that Fingolimod treatment (0.3mg/kg bw) significantly decreased cumulative disease score compared to the EAE control group. Concurrently, OPCs and proliferation of OPCs were significantly increased in the white matter of the brain and spinal cord at day 7 and day 30 after EAE onset, and oligodendrocytes, myelination and differentiation of OPCs were significantly increased at day 30 compared with the EAE control group. EAE mice treated with Fingolimod exhibited substantially elevated levels of Shh, its receptor Smoothened and effector Gli1 in the white matter of the CNS. However, combination treatment of EAE mice with cyclopamine-Fingolimod decreased Fingolimod monotherapy elevated protein levels of Smoothened and Gli1, and abolished the effect of Fingolimod on OPC proliferation and differentiation, as well as on neurological function outcome. Together, these data demonstrate that Fingolimod is effective as a treatment of EAE by promoting OPC proliferation and differentiation, which facilitate remyelination. In addition, the Shh pathway likely contributes to the therapeutic effects of Fingolimod on OPCs.

MiR-133b promotes neural plasticity and functional recovery after treatment of stroke with multipotent mesenchymal stromal cells in rats via transfer of exosome-enriched extracellular particles.

To test, in vivo, the hypothesis that exosomes from multipotent mesenchymal stromal cells (MSCs) mediate microRNA 133b (miR-133b) transfer which promotes neurological recovery from stroke, we used knockin and knockdown technologies to upregulate or downregulate the miR-133b level in MSCs (miR-133b(+) MSCs or miR-133b(-) MSCs) and their corresponding exosomes, respectively. Rats were subjected to middle cerebral artery occlusion (MCAo) and were treated with naïve MSCs, miR-133b(+) MSCs, or miR-133b(-) MSC at 1 day after MCAo. Compared with controls, rats receiving naïve MSC treatment significantly improved functional recovery and exhibited increased axonal plasticity and neurite remodeling in the ischemic boundary zone (IBZ) at day 14 after MCAo. The outcomes were significantly enhanced with miR-133b(+) MSC treatment, and were significantly decreased with miR-133b(-) MSC treatment, compared to naïve MSC treatment. The miR-133b level in exosomes collected from the cerebral spinal fluid was significantly increased after miR-133b(+) MSC treatment, and was significantly decreased after miR-133b(-) MSC treatment at day 14 after MCAo, compared to naïve MSC treatment. Tagging exosomes with green fluorescent protein demonstrated that exosomes-enriched extracellular particles were released from MSCs and transferred to adjacent astrocytes and neurons. The expression of selective targets for miR-133b, connective tissue growth factor and ras homolog gene family member A, was significantly decreased in the IBZ after miR-133b(+) MSC treatment, while their expression remained at similar elevated levels after miR-133b(-) MSC treatment, compared to naïve MSC treatment. Collectively, our data suggest that exosomes from MSCs mediate the miR-133b transfer to astrocytes and neurons, which regulate gene expression, subsequently benefit neurite remodeling and functional recovery after stroke.

Exosome-mediated transfer of miR-133b from multipotent mesenchymal stromal cells to neural cells contributes to neurite outgrowth.

Multipotent mesenchymal stromal cells (MSCs) have potential therapeutic benefit for the treatment of neurological diseases and injury. MSCs interact with and alter brain parenchymal cells by direct cell-cell communication and/or by indirect secretion of factors and thereby promote functional recovery. In this study, we found that MSC treatment of rats subjected to middle cerebral artery occlusion (MCAo) significantly increased microRNA 133b (miR-133b) level in the ipsilateral hemisphere. In vitro, miR-133b levels in MSCs and in their exosomes increased after MSCs were exposed to ipsilateral ischemic tissue extracts from rats subjected to MCAo. miR-133b levels were also increased in primary cultured neurons and astrocytes treated with the exosome-enriched fractions released from these MSCs. Knockdown of miR-133b in MSCs confirmed that the increased miR-133b level in astrocytes is attributed to their transfer from MSCs. Further verification of this exosome-mediated intercellular communication was performed using a cel-miR-67 luciferase reporter system and an MSC-astrocyte coculture model. Cel-miR-67 in MSCs was transferred to astrocytes via exosomes between 50 and 100 nm in diameter. Our data suggest that the cel-miR-67 released from MSCs was primarily contained in exosomes. A gap junction intercellular communication inhibitor arrested the exosomal microRNA communication by inhibiting exosome release. Cultured neurons treated with exosome-enriched fractions from MSCs exposed to 72 hours post-MCAo brain extracts significantly increased the neurite branch number and total neurite length. This study provides the first demonstration that MSCs communicate with brain parenchymal cells and may regulate neurite outgrowth by transfer of miR-133b to neural cells via exosomes.

Combination treatment with resveratrol and sulforaphane induces apoptosis in human U251 glioma cells.

Resveratrol is a naturally occurring polyphenolic compound highly enriched in grapes, peanuts, red wine, and a variety of food sources. Sulforaphane belongs to the family of isothiocyanates and is highly enriched in cruciferous vegetables. Our previous study showed that resveratrol, when used at high concentrations, inhibited cell proliferation, caused the cell cycle arrest and induced apoptotic cell death in glioma cells. In the current study, we tested the effect of combination treatment with resveratrol and sulforaphane, when both were used at low concentrations, on cell proliferation, migration and death in human U251 glioma cells. Our study shows that combination treatment with resveratrol and sulforaphane inhibits cell proliferation and migration, reduces cell viability, induces lactate dehydrogenase release, decreases pro-survival Akt phosphorylation and increases caspase-3 activation. The use of combination of bioactive food components, such as resveratrol and sulforaphane, may be a viable approach for the treatment of glioma.

Resveratrol downregulates PI3K/Akt/mTOR signaling pathways in human U251 glioma cells.

Resveratrol (trans-3,4', 5-trihydroxystilbene) is a naturally occurring polyphenolic compound that has antiinflammatory, antioxidant, neuroprotective properties and acts as a chemopreventive agent. Resveratrol causes cell cycle arrest and induces apoptotic cell death in various types of cancer cells. In the current studies, the effect of resveratrol on phosphoinositide kinase-3 (PI3K)/protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling pathway was examined in human U251 glioma cells. Resveratrol decreased both the expression and phosphorylation of Akt. Inhibitors of PI3K (LY294002) and Akt (SH-6) enhanced resveratrol-induced LDH release and caspase-3 activation. Resveratrol reduced phosphorylation of ribosomal protein S6 and the mTOR inhibitor rapamycin further enhanced resveratrol-induced cell death. These results suggest that the downregulation of PI3K/Akt/mTOR signaling pathways may be an important mediator in resveratrol-induced apoptosis in glioma cells.

[Epidemic and research progress of babesiosis].

Babesiosis is an emerging parasitic disease, distributed globally in Europe, Asia, Africa, North and South America, and Australia, and the United States is still the country with the largest number of babesiosis cases reported. Babesiosis in China is mainly distributed in the northeast, followed by the southwest and other regions. As a new vector-borne infectious disease, babesiosis poses a serious threat to human health, and its research foundation is relatively weak, so it requires more attention and recognition. The research hot spots on babesiosis are screening of diagnostic antigens, and the mechanisms of Babesia and the hosts, co-infections between Babesia and other pathogens. The epidemic distribution, screening of diagnostic antigens, host immune response mechanism and co-infection of babesiosis in our country and abroad are reviewed in this paper.

[Design and implementation of field questionnaire survey system of taeniasis/cysticercosis].

A taeniasis/cysticercosis information management system was designed to achieve the dynamic monitoring of the epidemic situation of taeniasis/cysticercosis and improve the intelligence level of disease information management. The system includes three layer structures (application layer, technical core layer, and data storage layer) and designs a datum transmission and remote communication system of traffic information tube in Browser/Server architecture. The system is believed to promote disease datum collection. Additionally, the system may provide the standardized data for convenience of datum analysis.

MiR-146a promotes remyelination in a cuprizone model of demyelinating injury.

The death of mature oligodendrocytes (OLs) which are the sole myelinating cells of the central nervous system (CNS), leads to demyelination and functional deficits. Currently, there is lack of effective remyelination therapies for patients with demyelinating diseases. MicroRNAs (miRNAs) mediate OL function. We hypothesized that miR-146a, by inactivating interleukin-1 receptor-associated kinase 1 (IRAK1), promotes differentiation of oligodendrocyte progenitor cells (OPCs) and thereby enhances remyelination. To test this hypothesis, a demyelination model induced by a cuprizone (CPZ) diet was employed, in which C57BL/6J mice were fed with a CPZ diet for 5weeks. After termination of CPZ diet, the mice were randomly treated with continuous infusion of miR-146a mimics or mimic controls into the corpus callosum for 7days. Compared to the mimic control, infusion of miR-146a mimics facilitated remyelination assessed by increased myelin basic proteins in the corpus callosum, which was associated with augmentation of newly generated mature OLs. Infusion of miR-146a mimics also substantially elevated miR-146a levels in the corpus callosum and fluorescently tagged miR-146a mimics were mainly detected in OPCs. Western blot and double immmunofluorescent staining analysis showed that the miR-146a treatment considerably reduced IRAK1 protein levels and the number of IRAK1-positive cells, respectively. Collectively, these data indicate that exogenous miR-146a enhances remyelination, possibly by promoting OPCs to differentiate into myelinated OLs via targeting IRAK1.

[Risk assessment of secondary transmission induced by imported malaria in Jiangxi Province].

OBJECTIVE: To assess the risk of secondary transmission induced by imported malaria in Jiangxi Province, so as to provide the evidence for adjustment of malaria surveillance strategies in the key groups and areas. METHODS: The Delphi method was used to establish the secondary transmission risk indicator system and the weight of each index was obtained. The data of malaria prevalence, vector distribution and intervention capacity were collected in 100 counties of Jiangxi Province from 2012 to 2015. The transmission potential index (TPI), intervention capacity index (ICI), and malaria risk index (MRI) were calculated for each county. The risk map was drawn with GIS software. RESULTS: The top ten counties with highly potential risk indicators were Linchuan District (2.131), Xinzhou District (1.609), Jiujiang County (1.404), Zhanggong District (1.365), Fengcheng City (1.225), Qingshanhu District (1.184), Yudu County (1.171), Dingnan County (1.018), Xunyang District (1.015) and Zhushan District (1.006). The high risk areas were mainly distributed in the regions of the capitals of their prefectures and in counties with more floating population. CONCLUSIONS: There are the risk of the secondary transmission induced by imported malaria in Jiangxi Province. The high risk of the secondary transmission is shown in the areas with more floating population and weaker intervention capacity.

[Study on the genetic polymorphisms of human glutathione-S-transferase A1 in Hakka population in South China].

OBJECTIVE: Glutathione-S-transferase A1 (GSTA1) is one of the major phase II detoxification enzymes in the cytosol, which genetic polymorphisms distribution is different in different ethnic, national and regional population. Up to now, GSTA1 genetic polymorphisms has been rarely reported in China. The purpose of the study was to investigate the distribution of genetic polymorphisms of human GSTA1 in Hakka population in South China. METHODS: Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) were used to identify the genotypes of GSTA1 gene and the data were analyzed with SPSS10.0 software. RESULTS: The GSTA1 genetic polymorphisms were detected in 480 samples. The frequency of GSTA1 * A/ * A,GSTA1 * A/ * B and GSTA1 * B/ * B were 77.1%, 21.7% and 1.2% respectively. And the GSTA1 genetic polymorphism distribution was in accordance with the Hardy-Weinberg equilibrium rule. There were no difference in the GSTA1 genetic polymorphisms among the different groups of age or gender. Logistic regression analysis showed that there were no association between the GSTA1 genetic polymorphism and family history of hypertension, coronary heart disease, stroke, lung cancer and nasopharyngeal cancer, et al. CONCLUSION: The GSTA1 gene existed polymorphism among Hakka in South China.

[Establishment of a risk assessment model for laboratory biosafety of Cryptosporidium].

OBJECTIVE: To evaluate the risk assessment model of Cryptosporidium laboratory, so as to provide the basis for laboratory personnel engaging in the operation of Cryptosporidium. METHODS: Firstly, the risk factors of Cryptosporidium infection in laboratory were determined by the literature and Delphi, and then the weights of risk factors were determined by fuzzy analytic hierarchy process. A risk assessment model for laboratory biosafety of Cryptosporidium was established. RESULTS: Compared to the indexes, based on the risk assessment model, stool sample processing was the two steps in the laboratory with high risk of infection and high risk factors, with the combination weights of risk possibility and hazard rating were 0.111 and 0.107, respectively. CONCLUSIONS: The risk assessment model established is feasible. It can be used to make some suggestions for the related laboratory staff.

[Pattern analysis of tempo-spatial distribution of schistosomiasis in marshland epidemic areas in stage of transmission control].

OBJECTIVE: To investigate the tempo-spatial patterns of schistosomiasis in Jiangling County, Hubei Province, so as to identify the risk areas and provide the scientific evidence in following intervention plans for marshland epidemic areas in the stage of transmission control. METHODS: The schistosomiasis epidemiological data in Jiangling County from 2009 to 2013 together with the related geographical information were collected and analyzed. The tempo-spatial distribution patterns were analyzed by the spatial autocorrelation analysis and spatial clustering analysis. RESULTS: The human infection rate was decreased from 2.15% in 2009 to 0.63% in 2013, which was the historically low level. The results of tempo-spatial analysis showed that there were spatial clustering effects in human schistosomiasis infection for each of the years. The values of spatial autocorrelation index Moran's I were statistically significant. Eighteen and thirty-five clusters were detected by using SatScan and FlexScan software, respectively. CONCLUSIONS: From 2009 to 2013, the schistosomiasis endemic situation in Jiangling County presented a decline trend and reached the historical low level. The identified spatial clustering areas should be targeted as the prioritized areas for schistosomiasis control.

[Epidemiological analysis of imported malaria cases in 20 counties at border region of Yunnan Province from 2012 to 2014].

OBJECTIVE: To analyze the epidemiological characteristics of the imported malaria cases in 20 counties at the border region of Yunnan Province from 2012 to 2014, so as to provide the evidence-based proof for adjusting the strategies in the elimination stage. METHODS: The malaria epidemic data of the 20 border counties in Yunnan Province from 2012 to 2014 were collected and analyzed by using Microsoft Excel 2010. RESULTS: From 2012 to 2014, a total of 1 558 malaria cases were reported in the 20 border counties in Yunnan Province, among which, 1 336 were imported cases, accounting for 85.75% (1 336/1 558), and 222 were indigenous cases, accounting for 14.25% (222/1 558). The number of the imported cases in the above years took up 80.00% (544/680), 89.10% (425/477) and 91.52% (367/401) of the total reported cases in the whole year, respectively. Among all the 1 336 imported cases, 1 045 (78.22%) were infected with Plasmodium vivax, 284 (21.26%) were infected with P. falciparum, 3 were infected with P. malariae, 3 were mixed infection and 1 was an unclassified case; 2 patients died. And 95.58% of the cases were mainly infected in Myanmar (1 277 cases). Young and middle-aged adult of 20-40 years who worked overseas were the predominant (802 cases, 60.03%) and most of the cases occurred from April to June of the year (679 cases, 50.82%). Those cases mainly distributed in Tengchong (459 cases), Ruili (366 cases), Yingjiang (191 cases) and Mangshi (78 cases). CONCLUSIONS: The epidemic situation of imported malaria is serious in the border region of Yunnan Province. Therefore, the surveillance system of malaria control needs to be well planned and managed to ensure timely case detection and prompt response at the elimination and post-elimination stage.