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AIM: To test our hypothesis that activation of protein kinase A (PKA) signal pathway by β-adrenergic agonist plays an important role in the protecting of cultured retinal pigment epithelial (RPE) cells against the hydroxychloroquine (HCQ) toxicity. METHODS: Cultured human RPE cells were treated with 1) HCQ, 2) HCQ with salbutamol (a β2-adrenergic receptor agonist), and 3) HCQ with salbutamol and a PKA inhibitor, and compared these to 4) untreated cells (controls). After treated for 24h, cell vacuolation, cells viability, PKA and PKA kinase activity levels were determined by the measurement of the size of vacuoles using Image J software, the cell counting with a dye-exclusion testing, Western blot and PKA kinase detection, respectively. RESULTS: Cell vacuolation and cell death of cultured RPE cells were significantly increased by the treatment of HCQ. Salbutamol significantly elevated PKA and PKA activity levels and this was associated with the inhibition of the vacuolation and cell death. The PKA inhibitor significantly decreased the PKA levels and eliminated the protective effects of salbutamol on HCQ-treated RPE cells. CONCLUSION: The PKA pathway plays an important role in the protective effects of β2-adrenergic agonist on the RPE cells against HCQ toxicity. These findings reveal a novel potential strategy against HCQ retinopathy by treatment with PKA activating medications.

As a disease characterized by severe liver necrosis and hemorrhage, duck viral hepatitis (DVH) is mainly caused by duck hepatitis A virus (DHAV). The positive-strand RNA genome of DHAV type 1 (DHAV-1) contains an internal ribosome entry site (IRES) element within the 5&prime, untranslated region (UTR), structured sequence elements within the 3&prime, UTR, and a poly(A) tail at the 3&prime, terminus. In this study, we first examined that insulin-like growth factor-2 mRNA-binding protein-1 (IGF2BP1) specifically interacted with the DHAV-1 3&prime, UTR by RNA pull-down assay. The interaction between IGF2BP1 and DHAV-1 3&prime, UTR strongly enhanced IRES-mediated translation efficiency but failed to regulate DHAV-1 replication in a duck embryo epithelial (DEE) cell line. The viral propagation of DHAV-1 strongly enhanced IGF2BP1 expression level, and viral protein accumulation was identified as the key point to this increment. Collectively, our data demonstrated the positive role of IGF2BP1 in DHAV-1 viral proteins translation and provided data support for the replication mechanism of DHAV-1.

Background Reverse genetics systems enable the manipulation of viral genomes and therefore serve as robust reverse genetic tools to study RNA viruses. A DNA-launched rescue system initiates the transcription of viral genomic cDNA from eukaryotic promoter in transfected cells, generating homogenous RNA transcripts in vitro and thus enhancing virus rescue efficiency. As one of the hazardous pathogens to ducklings, the current knowledge of the pathogenesis of duck astrovirus type 1 (DAstV-1) is limited. The construction of a DNA-launched rescue system can help to accelerate the study of the virus pathogenesis. However, there is no report of such a system for DAstV-1. Methods In this study, a DNA-launched infectious clone of DAstV-1 was constructed from a cDNA plasmid, which contains a viral cDNA sequence flanked by hammerhead ribozyme (HamRz) and a hepatitis delta virus ribozyme (HdvRz) sequence at both terminals of the viral genome. A silent nucleotide mutation creating a Bgl II site in the ORF2 gene was made to distinguish the rescued virus (rDAstV-1) from the parental virus (pDAstV-1). Immunofluorescence assay (IFA) and western blot were conducted for rescued virus identification in duck embryo fibroblast (DEF) cells pre-treated with trypsin. The growth characteristics of rDAstV-1 and pDAstV-1 in DEF cells and the tissue tropism in 2-day-old ducklings of rDAstV-1 and pDAstV-1 were determined. Results The infectious DAstV-1 was successfully rescued from baby hamster kidney (BHK-21) cells and could propagate in DEF cells pre-treated with 1 μg/ml trypsin. Upon infection of DEF cells pre-treated with trypsin, DAstV-1 mRNA copies were identified after serial passaging, and the result showed that rDAstV-1 and pDAstV-1 shared similar replication kinetics. Animal experiment showed that the rDAstV-1 had an extensive tissue tropism, and the virus was capable of invading both the central and the peripheral immune organs in infected ducklings. Conclusions An improved DNA-launched reverse genetics system for DAstV-1 was firstly constructed. Infectious virus recovered from BHK-21 cells could propagate in DEF cells pre-treated with trypsin. This is the first report of the successful in vitro cultivation of DAstV-1. We believe this valuable experimental system will contribute to the further study of DAstV-1 genome function and pathogenesis.

Upon entering the body, nerve agents can bind active amino acid residues to form phosphonylated adducts. Tabun derivatives (O-alkyl-N,N-dialkyl phosphoroamidocyanidates) have strikingly different structural features from other G-series nerve agents, such as sarin and soman. Here, we investigate the binding mechanism for the phosphonylated adducts of nerve agents of tabun derivatives. Binding sites for three tabun derivatives, O-ethyl-N,N- dimethyl phosphoramidocyanidate (GA), O-ethyl-N,N-ethyl(methyl) phosphoramidocyanidate, and O-ethyl-N,N-diethylphosphoramidocyanidate were studied. Quadrupole-orbitrap mass spectrometry (Q-Orbitrap-MS) coupled to proteomics was used to screen adducts between tabun derivatives and albumin, immunoglobulin, and hemoglobin. The results reveal that all three tabun derivatives exhibit robust selectivity to lysine residues, rather than other amino acid residue types. A set of 10 lysine residues on human serum albumin are labeled by tabun derivatives in vitro, with K525 (K*QTALVELVK) and K199 (LK*CASLQK) peptides displaying the most reactivity. Tabun derivatives formed stable adducts on K525 and K414 (K*VPQVSTPTLVEVSR) for at least 7 days and on K351 (LAK*TYETTLEK) for at least 5 days in a rabbit model. Three of these peptides-K525, K414, and K351-have the highest homology with human serum albumin of all 5 lysine residues that bound to examined rabbit blood proteins in vivo. Molecular simulation of the tabun-albumin interaction using structural analysis and molecular docking provided theoretical evidence supporting lysine residue reactivity to phosphonylation by tabun derivatives. K525 has the lowest free binding energy and the strongest hydrogen bonding to human albumin. In summary, these findings identify unique binding properties for tabun derivatives to blood proteins.

Induced eddy current in the secondary conductor plate of long primary double-sided linear induction motor (LPDSLIM) is asymmetrically distributed since the dynamic longitudinal end effect caused by open-end structure in secondary. This is one of the most influential reasons of the thrust fluctuation. In this paper, a thrust fluctuation reduction method based on the secondary of a longitudinal skew end has been proposed. The thrust fluctuation reduced effect of three different proposed secondary structures, i.e. parallelogram shape, swallow-tail shape and shuttle shape has been firstly comparatively analyzed based on the 3D finite element method (FEM). Then the influence of end skew distance on the thrust performance of LPDSLIM with shuttle structure secondary has also been studied. It has been found that shuttle type secondary can effectively reduce thrust fluctuation without sacrifice of average thrust. This effect is independent on motor slip frequency.

Using 3D FEM to achieve optimal design for long stator Ironless permanent magnet linear synchronous motor (PMLSM) with many design parameters is computationally an expensive task. To reduce design time without loss of accuracy, the end winding inductance model is established in this paper. Then the end winding inductance model and 2D FEM(Semi-Analytical) were combined to replace the 3D FEM model. Finally, the optimal design set of PMLSM can be obtained quickly and accurately based on Semi-Analytical by Genetic-Particle Swarm Optimization Algorithm (GAPOS).

The medium-speed maglev traffic with permanent magnet linear synchronous motor (PMLSM) based on single-sided ironless coil is a new traffic system which has the advantages of light weight, small normal force, little thrust fluctuation and low cost. Analytical model of medium-speed maglev traffic traction system considering the influence of the feeder cable and the switching of primary segment is established in this paper. On this basis, a simulation study is carried out, and the results obtained are of great significance to the design and the optimization of the traction system for medium-speed maglev traffic.

The long primary permanent magnet linear synchronous motor (PMLSM) used for high speed maglev train needs to adopt flux weakening (FW) control to increase its speed due to the limitation of the traction inverter output voltage. In this paper, a subsection FW scheme in the linear modulation region based on the direct traction control (DTC) is proposed. By controlling the reference primary flux directly in different working states, the target speed is achieved faster compared with SFOC-FW control, and the demagnetization current is saved. The primary flux and traction fluctuation are reduced in the low speed working condition by leading the space vector modulation (SVM) algorithm into the proposed DTC-FW scheme. The simulation analysis is carried out and the effectiveness of the proposed scheme is verified.

In order to accelerate the maglev train to the high or ultra-high speed range, a double-secondary permanent magnet linear synchronous motor (DS-PMLSM) with N-S configuration is proposed which has the merits of high thrust and high power density. The two secondaries of the motor share a long primary and the armature is ironless. In this paper, a layered model of DS-PMLSM is established, and the expressions of the air gap magnetic density and propulsion in Fourier form are derived. A virtual N-N configuration motor compensation method is proposed in order to reflect the longitudinal end effect of the secondaries. The expression of the attraction between the two secondaries is derived based on the equivalent surface current method. Finally, the consistency and correctness of the analytical derivation are verified by the finite element (FEM) simulation. The analytical derivations in this paper provide the theoretical support for the design of the motor structure parameters.

4-Hydroxyphenylacetate 3-hydroxylase (EcHpaB) from Escherichia coli is capable of efficient ortho-hydroxylation of a wide range of phenolic compounds and demonstrates great potential for broad chemoenzymatic applications. To understand the structural and mechanistic basis of its catalytic versatility, we elucidated the crystal structure of EcHpaB by X-ray crystallography, which revealed a unique loop structure covering the active site. We further performed mutagenesis studies of this loop to probe its role in substrate specificity and catalytic activity. Our results not only showed the loop has great plasticity and strong tolerance towards extensive mutagenesis, but also suggested a flexible loop that enables the entrance and stable binding of substrates into the active site is the key factor to the enzyme catalytic versatility. These findings lay the groundwork for editing the loop sequence and structure for generation of EcHpaB mutants with improved performance for broader laboratory and industrial use.

A major challenge in organophosphate compound (OP) and OP nerve agent (OPNA) research has been in the identification and utilization of reliable biomarkers for rapid, sensitive, and efficient detection of OP exposure. Albumin has been widely studied as a biomarker for retrospective verification of exposure to OPNAs, including soman (GD), by detecting the phosphonylation of specific amino acid residues. The aim of the present study was to identify binding sites between GD and rabbit serum albumin in vitro and in vivo. A nano-liquid chromatography coupled with a quadrupole-orbitrap mass spectrometry (nLC-Q-Orbitrap-MS) was used to examine the GD-modified adducts of rabbit albumin. A total of 11 GD-modified sites were found in rabbit serum albumin across three experimental models. The following five GD-modified rabbit albumin sites, which were all lysine residues, were established in vivo: K188, K329, K162, K233, and K525. Two of these five lysine residues, K188 in peptide EK*ALISAAQER and K162 in peptide YK*AILTECCEAADK, were stable for at least 7 days in vivo. Molecular simulation of the GD–albumin interaction provided theoretical evidence for reactivity of the identified lysine residues. The findings suggest that these modifiable lysine residues are potential biomarkers of GD exposure for retrospective analysis by Q-Orbitrap-MS.

Antisense RNA (asRNA) technology is an important tool for downregulating gene expression. When applying this strategy, the asRNA interference efficiency is determined by several elements including scaffold design, loop size, and relative abundance. Here, we take the Escherichia coli gene fabD encoding malonyl-CoA-[acyl-carrier-protein] transacylase as an example to describe the asRNA design with reliable and controllable interference efficiency. Real-time PCR and fluorescence assay methods are introduced to detect the interference efficiency at RNA level and protein level, respectively.

Accumulating evidence suggests granulocyte macrophage-colony stimulating factor (GM-CSF) can function as an inflammatory mediator, but whether GM-CSF-producing CD4

The aged kidney is susceptible to acute injury due presumably to its decreased ability to handle additional challenges, such as endoplasmic reticulum (ER) stress. This was tested by giving tunicamycin, an ER stress inducer, to either old or young mice. Injection of high dose caused renal failure in old mice, not in young mice. Moreover, injection of low dose resulted in severe renal damage in old mice, confirming the increased susceptibility of aged kidney to ER stress. There existed an abnormality in ER stress response kinetics in aged kidney, characterized by a loss of XBP-1 splicing and decreased PERK-eIF2α phosphorylation at late time point. The presence of excessive oxidative stress in aged kidney may play a role since high levels of oxidation increased ER stress-induced cell death and decreased IRE1 levels and XBP-1 splicing. Importantly, treatment with antioxidants protected old mice from kidney injury and normalized IRE1 and XBP-1 responses. Furthermore, older mice (6 months old) transgenic with antioxidative stress AGER1 were protected from ER stress-induced kidney injury. In conclusion, the decreased ability to handle ER stress, partly due to the presence of excessive oxidative stress, may contribute to increased susceptibility of the aging kidney to acute injury.

Accumulating evidence suggests granulocyte macrophage-colony stimulating factor (GM-CSF) can function as an inflammatory mediator, but whether GM-CSF-producing CD4+ T cells (TH-GM-CSF) are a distinct T helper cell subset is lacking. Herein we demonstrate that interleukin (IL)-1b exclusively drives differentiation of naive CD4+ T cells into TH-GM-CSF cells via inducing ubiquitination of IL-1 receptor-associated kinase 1 (IRAK1) and subsequent activation of the transcription factor NF-kappaB (NF-κB), independent of RAR-related orphan receptor gamma (RORt) required for TH17 differentiation. In vivo, TH-GM-CSF cells are present in murine Citrobacter Rodentium infections and mediate colitis following adoptive transfer of CD4+ T cells into Rag1-/- mice via GM-CSF-induced macrophage activation. The TH-GM-CSF cell phenotype is stable and distinct from the TH17 genetic program, but IL-1b can convert pre-formed TH17 cells into TH-GM-CSF cells, thereby accounting for previously reported associations between IL-17 and GM-CSF. Together, our results newly identify IL-1b/NFκB-dependent TH-GM-CSF cells as a unique T helper cell subset and highlight the importance of CD4+ T cell-derived GM-CSF induced macrophage activation as a previously undescribed T cell effector mechanism. Funding Statement: Huabao Xiong was supported by NIH funding RO1AI104688. Peter. S Heeger was supported by RO1AI132405 and RO1AI071185. Declaration of Interests: The authors declare that they have no competing interests. Ethics Approval Statement: The animal study protocols were approved by the Institutional Animal Care and Use Committees of Mount Sinai School of Medicine.

In this paper, a method for designing a sparse 2-D (two-dimensional) FIR notch filter based on BP neural network and iterative reweighted orthogonal matching pursuit algorithm is proposed. At each iteration, the iteratively reweighted orthogonal matching pursuit algorithm is employed to generate a sparse filter whose response is the best approximation to the desired filter. The sparse filter obtained by the iteration is then modified through the BP neural network. The iteration stops when the modified filter meets the design specifications. A sparse 2-D FIR notch filter contains a large number of zero coefficients, so it can save the multiplier and adder corresponding to the zero coefficients during the circuit implementation, which will reduce the cost and power of the hardware implementation. Moreover, the BP neural networks are simple in structure and avoid complicated computation of matrix inversion. Simulation results demonstrate that the proposed scheme can obtain sparse solutions and is effective in image processing.

Thorax disease classification is a challenging task due to complex pathologies and subtle texture changes, etc. It has been extensively studied for years largely because of its wide application in computer-aided diagnosis. Most existing methods directly learn global feature representations from whole Chest X-ray (CXR) images, without considering in depth the richer visual cues lying around informative local regions. Thus, these methods often produce sub-optimal thorax disease classification performance because they ignore the very informative pathological changes around organs. In this paper, we propose a novel Part-Aware Mask-Guided Attention Network (PMGAN) that learns complementary global and local feature representations from all-organ region and multiple single-organ regions simultaneously for thorax disease classification. Specifically, multiple innovative soft attention modules are designed to progressively guide feature learning toward the global informative regions of whole CXR image. A mask-guided attention module is designed to further search for informative regions and visual cues within the all-organ or single-organ images, where attention is elegantly regularized by automatically generated organ masks and without introducing computation during the inference stage. In addition, a multi-task learning strategy is designed, which effectively maximizes the learning of complementary local and global representations. The proposed PMGAN has been evaluated on the ChestX-ray14 dataset and the experimental results demonstrate its superior thorax disease classification performance against the state-of-the-art methods.

Objective In some cases of single focus epilepsy, conventional video electroencephalography (EEG) cannot reveal the epileptogenic focus even when intracranial electrodes are used. Here, we tested whether analyzing high frequency oscillations (HFOs) can be used to determine the ictal onset zone in suspected bitemporal epilepsy and improve seizure outcome. Methods We prospectively studied 13 patients with refractory temporal seizures who were treated over a 4-year period and underwent bilateral placement of intracranial electrodes. Subdural strips were used in all cases, and depth electrodes were implanted into mesial temporal lobes in 10 patients. The mean patient age was 30.92 years, and 30.7% of patients were male. Patients were monitored by conventional and wide-band frequency amplifiers. Results Conventional invasive EEG monitoring of interictal periods showed bilateral epileptiform abnormalities in 12 patients (92.3%) and unilateral epileptiform abnormalities in one (7.7%), and monitoring of ictal periods revealed unilateral seizure origins in nine patients (69.2%) and bilateral origins in four (30.8%). In contrast, high frequency invasive EEG monitoring of interictal periods showed bilateral HFOs in seven patients (53.8%) and unilateral HFOs in six (46.2%), and monitoring of ictal periods revealed unilateral HFOs in all 10 patients who were tested. Three patients were not monitored during ictal periods because of time limitations. All 13 patients subsequently underwent a standard unilateral temporal lobectomy and have been followed-up for a minimum of 12 months. Eleven (84%) had a Class I outcome, one (8%) a Class II outcome, and one a Class III outcome. Significance Bilateral placement of subdural strip and depth electrodes for seizure monitoring in patients with suspected bitemporal epilepsy is both safe and effective. Monitoring high frequency oscillations can help determine the laterality of the onset zone when localization using conventional EEG or brain MRI fails.

A near-infrared-driven photoelectrochemical aptasensor was developed as a new method for the detection of the breast cancer cell MCF-7. The upconversion nanoparticles and TiO2/CdTe heterostructure were combined to prepare the film electrode, and the high-affinity aptamer AS1411 was conjugated to the electrode to recognize MCF-7 cells. In this fabrication, the upconversion nanoparticles transferred the near-infrared light to visible light, which could excite the semiconductor to enhance the current response. As a result, the aptasensor revealed good sensitivity and specificity with MCF-7 cell concentrations ranging from 1 × 103 to 1 × 105 cells/mL. The results presented a favorable determination of MCF-7 cells, which was achieved with the help of the upconversion nanoparticles and the photoelectrochemical interface.

Dendritic cells (DCs) play a pivotal role in the development of effective immune defense while avoiding detrimental inflammation and autoimmunity by regulating the balance of adaptive immunity and immune tolerance. However, the mechanisms that govern the effector and regulatory functions of DCs are incompletely understood. Here, we show that DC-derived nitric oxide (NO) controls the balance of effector and regulatory DC differentiation. Mice deficient in the NO-producing enzyme inducible nitric oxide synthase (iNOS) harbored increased effector DCs that produced interleukin-12, tumor necrosis factor (TNF) and IL-6 but normal numbers of regulatory DCs that expressed IL-10 and programmed cell death-1 (PD-1). Furthermore, an iNOS-specific inhibitor selectively enhanced effector DC differentiation, mimicking the effect of iNOS deficiency in mice. Conversely, an NO donor significantly suppressed effector DC development. Furthermore, iNOS−/− DCs supported enhanced T cell activation and proliferation. Finally iNOS−/− mice infected with the enteric pathogen Citrobacter rodentium suffered more severe intestinal inflammation with concomitant expansion of effector DCs in colon and spleen. Collectively, our results demonstrate that DC-derived iNOS restrains effector DC development, and offer the basis of therapeutic targeting of iNOS in DCs to treat autoimmune and inflammatory diseases.

We have developed synthetic gene networks that enable engineered cells to selectively program surface chemistry. E. coli were engineered to upregulate biotin synthase, and therefore biotin synthesis, upon biochemical induction. Additionally, two different functionalized surfaces were developed that utilized binding between biotin and streptavidin to regulate enzyme assembly on programmable surfaces. When combined, the interactions between engineered cells and surfaces demonstrated that synthetic biology can be used to engineer cells that selectively control and modify molecular assembly by exploiting surface chemistry. Our system is highly modular and has the potential to influence fields ranging from tissue engineering to drug development and delivery.

// Juan Wang 1,2 , Liang Peng 2 , Ruihua Zhang 2 , Zihan Zheng 2,3 , Chun Chen 4,5 , Ka Lung Cheung 6 , Miao Cui 7 , Guanglin Bian 2 , Feihong Xu 2 , David Chiang 2 , Yuan Hu 2 , Ye Chen 1 , Geming Lu 2 , Jianjun Yang 2 , Hui Zhang 2 , Jianfei Yang 8 , Hongfa Zhu 7 , Shu-hsia Chen 2 , Kebin Liu 9 , Ming-Ming Zhou 6 , Andrew G. Sikora 2 , Liwu Li 5 , Bo Jiang 1 and Huabao Xiong 2 1 Guangdong Provincial Key Laboratory of Gastroenterology, Department of Gastroenterology, Nanfang Hospital, Southern Medical University, Guangzhou, China 2 Department of Medicine, Immunology Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA 3 Department of Biology, College of Arts and Sciences, University of North Carolina at Chapel Hill, NC, USA 4 Amgen Inc., Thousand Oaks, CA, USA 5 Department of Biological Sciences, Virginia Tech, Blacksburg, VA, USA 6 Department of Structural and Chemical Biology, Icahn School of Medicine at Mount Sinai, New York, NY, USA 7 Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, NY, USA 8 Tempero Pharmaceuticals, GlasxoSmithKline, Cambridge, MA, USA 9 Department of Biochemistry and Molecular Biology, Medical College of Georgia, Georgia Regents University, Augusta, GA, USA Correspondence to: Huabao Xiong, email: // Bo Jiang, email: // Keywords : 5-FU, TH17, TS, Immunology and Microbiology Section, Immune response, Immunity Received : November 24, 2015 Accepted : March 07, 2016 Published : March 24, 2016 Abstract While it is well established that treatment of cancer patients with 5-Fluorouracil (5-FU) can result in immune suppression, the exact function of 5-FU in the modulation of immune cells has not been fully established. We found that low dose 5-FU selectively suppresses T H 17 and T H 1 cell differentiation without apparent effect on Treg, T H 2, and significantly suppresses thymidylate synthase (TS) expression in T H 17 and T H 1 cells but has a lesser effect in tumor cells and macrophages. Interestingly, the basal expression of TS varies significantly between T helper phenotypes and knockdown of TS significantly impairs T H 17 and T H 1 cell differentiation without affecting the differentiation of either Treg or T H 2 cells. Finally, low dose 5-FU is effective in ameliorating colitis development by suppressing T H 17 and T H 1 cell development in a T cell transfer colitis model. Taken together, the results highlight the importance of the anti-inflammatory functions of low dose 5-FU by selectively suppressing T H 17 and T H 1 immune responses.

Clustering and multi-hop routing algorithms substantially prolong the lifetime of wireless sensor networks (WSNs). However, they also result in the energy hole and network partition problems. In order to balance the load between multiple cluster heads, save the energy consumption of the inter-cluster routing, in this paper, we propose an energy-efficient routing algorithm based on Unequal Clustering Theory and Connected Graph Theory for WSN. The new algorithm optimizes and innovates in two aspects: cluster head election and clusters routing. In cluster head election, we take into consideration the vote-based measure and the transmission power of sensor nodes when to sectionalize these nodes into different unequal clusters. Then we introduce the connected graph theory for inter-cluster data communication in clusters routing. Eventually, a connected graph is constituted by the based station and all cluster heads. Simulation results show that, this new algorithm balances the energy consumption among sensor nodes, relieves the influence of energy-hole problem, improve the link quality, achieves a substantial improvement on reliability and efficiency of data transmission, and significantly prolongs the network lifetime.

Glucokinase (GCK) is an important enzyme critical for glucose metabolism, and has been targeted as such in the pursuit of a cure for diabetes mellitus. We show that streptozotocin (STZ)-induced diabetic murine model exhibits low GCK expression with high blood glucose levels; moreover, aggravated glomerulonephritis is observed in the model when there is IL10 deficiency. Although T cells infiltrate into the liver and pancreas in STZ-induced diabetes mice, T helper 1 (Th1) and T helper 17 (Th17) cells decrease significantly with STZ addition in in vitro polarization. Using a mutant GCK gene (GCK 262) with a knocked out cytosine at position 2643 results in lower protein expression and more ubiquitination-led protein degradation compared with wild-type GCK (GCK 261). We further observed that hsa-mir-1302 can bind to 3'-untranslated region of mutant GCK, which can decrease GCK mRNA translation. Finally, delivery of mutant GCK by subcutaneous injection is more effective at decreasing blood glucose in the STZ-treated (STZ) murine diabetes model than insulin treatment alone. Similarly, mutant GCK consistently and moderately decreases blood glucose levels in GK rats over a period of 12 and 70 days without inducing hypoglycemia, whereas insulin is only effective over 12 h. These results suggest that mutant GCK may be a future cure for diabetes.

Colistin has been used as a growth promotant in livestock feed for many years. In China, mcr-1-positive Escherichia coli strains have been isolated from humans, chickens, and pigs. To date, there are few reports about the prevalence and molecular characteristics of fecal E. coli bearing mcr-1 in the meat ducks. In this study, the prevalence of mcr-1 gene was investigated among 120 fecal E. coli strains isolated from healthy meat ducks in Shandong province of China between October 2017 and February 2018. A total of nine mcr-1-containing E. coli strains were identified and two were identified as extra-intestinal pathogenic E. coli (ExPEC) among them. The clonal relationship of the nine E. coli strains was determined by multilocus sequencing typing (MLST) and pulsed field gel electrophoresis (PFGE), and the results indicated that all mcr-1-carrying isolates were clonally unrelated. Two different genetic contexts of mcr-1 were identified among these isolates. Colistin-resistant phenotype of all the isolates was successfully transferred to the recipient strains by conjugation experiments and seven transconjugants carried a single plasmid. The mcr-1 was located on three replicon plasmids: IncI2 (n = 4), IncFII (n = 2) and IncN (n = 1). Complete sequence analysis of a representative plasmid pTA9 revealed that it was strikingly similar with plasmid pMCR1-IncI2 of E. coli, plasmid pHNSHP45 of E. coli, and plasmid pWF-5-19C of Cronobacter sakazakii, implying that pTA9-like plasmids may be epidemic plasmids that mediate the spread of mcr-1 among Enterobacteriaceae. These results highlight that healthy meat duck is a potential reservoir for multidrug resistant mcr-1-containing E. coli strains.

The fracture degree of propellant charge on the projectile base before it is ignited is the key to study the launch safety of propellant charge and simulate the breech-blow phenomenon of the gun. In this paper, by solving the difficult problem of calculating the gas generation law of fractured propellant charge, an interior ballistic two-phase flow calculation method with mixed-charge considering the fractured propellant charge is established. Through the test method for mechanical situation of propellant charge, the time history of compression stress of propellant charge near the projectile base is acquired, which is loaded into the propellant bed model based on discrete element method(DEM). The compression and fracture process of propellant charge in the 30 mm area near the projectile base at low temperature is simulated. Then the corresponding fractured propellant charge is obtained, and next setting it as the initial condition of the interior ballistic two-phase flow calculation; moreover, the simulation of interior ballistic two-phase flow with mixed-charge considering fractured propellant charge is carried out. And the simulation results are in good agreement with the ballistic performance test results. The research in this paper provides a precise calculation method for the abnormal pressure in chamber caused by fractured propellant charge, which can be used to predict the breech-blow phenomenon of the gun and to guide the design of gun charge structure.

Summary α-Synuclein (α-syn) overload is strongly associated with Parkinson disease (PD), and reduction of the α-syn level by targeting the peptide-based system through the autophagy-lysosomal pathway (ALP) is a promising strategy to delay PD progression. However, if the ALP is comprised, targeting the peptide-based proteasomal degradation system would be a good alternative. In this study, we designed a fusion peptide containing an α-syn-binding domain and a short strong proteasome-targeting motif. Our results reveal that this peptide could specifically bind to α-syn, and direct it to the proteasomes for degradation in a recombinant expression system. Furthermore, by adding a membrane-penetrating motif to this fusion peptide, we demonstrated that it could penetrate into cells and consequently suppress the cellular α-syn level through proteasome degradation in a dose- and time-dependent manner. Functionally, these effects rescued the mitochondrial dysfunction and cellular defects caused by α-syn overexpression in the cultured cells and primary neurons.

In this research, multilayered steel (MLS), which is composed of middle-carbon martensite steel, high-carbon martensite steel, and a pure Ni thin layer was obtained by the accumulative roll-bonding method. The microstructure and mechanical properties of the MLS were investigated by scanning electron microscopy (SEM), Vickers microhardness, tensile, and bending tests. In-situ SEM tensile tests were used to observe the crack initiation and propagation processes during the tensile loading. The results show that the ultimate tensile strength and bending strength of the MLS can reach 946 MPa and 3153 MPa, and the maximum elongation can reach 18%, which is related to the better combined quality of the interface. The middle and larger martensite layer (ML) becomes the weakest link of tensile fracture and interfacial delamination of the MLS during the tensile processes, because there are lots of large hard blocks Cr23C6 phases distributed in the middle thicker ML layer. Besides, the MLS can withstand larger bending deformation. When the MLS was bent to 180 degrees, neither macro-cracks in the outer side of the bending parts nor interfacial delamination can be found.

Lung cancer is a common malignant disease, nearly 2.09 million new patients occurred last year. Approximately 85% of the patients are classified as non-small-cell lung cancer (NSCLC). It is therefore important to identify new diagnostic and prognostic biomarkers for the early detection of this disease. The presented study identifies biomarkers in the serum of NSCLC patients. The expression of 274 cytokines was measured by a novel antibody array methodology and ELISA was applied to validate the array results. The levels of MIP-1 α, IL-8, MIP-1 β, Resistin, GDF-15, HGF, CA125, FLRG, VCAM-1, DKK-3, sTNF-R1, CTACK, Acrp30, CXCL-16 and LYVE-1 were significantly higher in serum from NSCLC patients, while the level of TIMP-2 and IGFBP-6 were lower. More importantly, the validation supported the result of the antibody array. The result of the antibody array indicates that these cytokines might be novel auxiliary biomarkers in the diagnosis and prognosis of NSCLC.

Medium and low speed maglev trains adopt ironless permanent magnet linear synchronous motor (IPMLSM) as their propulsion system. This kind of motor can reduce the normal attractive force between the mover and the stator, and it can also eliminate the cogging force, in the meantime, the problem caused by its flux leakage has drawn people's attention. In this paper, the leakage magnetic fields around the IPMLSM are calculated using 2-D and 3-D finite element methods (FEM). The calculation results include the magnetic fields distribution of the IPMLSM, and the range of flux leakage density equal to 5Gs, which indicates the safety distance people should keep from the motor. Some suggestions on the way to reduce the flux leakage are given as well, which offers the reference for the practical application of this motor.

Permanent magnet linear synchronous motor (PMLSM) is a nonlinear, strongly coupled high order system. It is susceptible to uncertain factors such as internal parameters change, external load disturbance and so on. In view of this problem, an improved backstepping controller(BSC) is proposed in this paper. According to the Lyapunov stability theory, appropriate Lyapunov function and intermediate virtual control quantity are defined to realize the ideal tracking of speed of PMLSM. Experiment results validate that the backstepping controller can achieve fast response and stable tracking of speed. Compared with the traditional PI controller, the overshoot decreases obviously, and the dynamic and static characteristics are better.

Iron sleepers which are laid under the long stator of the ironless permanent magnet linear synchronous machine (IPMLSM) to support it are critical parts to the whole maglev system. It is important to analyze the iron sleepers' effect on the force characteristics of IPMLSM and to reduce this effect by decreasing the bias amount of power angle characteristics and the force fluctuation. In this paper, the model of the IPMLSM is built up based on the finite element method (FEM), and different structure parameters of the iron sleepers are researched to find out the relation between structure parameters and the effect caused by iron sleepers. This paper also analyzes the way to reduce this effect. As the iron sleepers are important in the construction of maglev system, especially the experimental line, and may bring inevitable problems, the simulation results about iron sleepers could provide a guidance for the construction of maglev experimental line.

In this paper, single current regulator flux-weakening control based on the d-q current cross-coupling effect for permanent magnet linear synchronous motor (PMLSM) is improved and a stable control process is implemented. When the speed exceeds a certain value, the control strategy is switched from the conventional double closed-loop control method to the flux-weakening control method. The method of taking the average value of multi-point sampling is used for mode switching judgment. And the improved switching method of the control mode achieves a stable switching process.

The follicular helper T cell (T(FH)) are established regulators of germinal center (GC) B cells, whether T(FH) have pathogenic potential independent of B cells is unknown. Based on in vitro T(FH) cell differentiation, in vivo T cell transfer animal colitis model, and intestinal tissues of inflammatory bowel disease (IBD) patients, T(FH) and its functions in colitis development were analyzed by FACS, ChIP, ChIP-sequencing, WB, ELISA and PCR. Herein we demonstrate that intestinal tissues of patients and colon tissues obtained from Rag1(−/−) recipients of naïve CD4(+) T cells with colitis, each over-express T(FH)associated gene products. Adoptive transfer of naïve Bcl6(−/−) CD4(+) T cells into Rag1(−/−) recipient mice abrogated development of colitis and limited T(FH) differentiation in vivo, demonstrating a mechanistic link. In contrast, T cell deficiency of interferon regulatory factor 8 (IRF8) resulted in augmentation of T(FH) induction in vitro and in vivo. Functional studies showed that adoptive transfer of IRF8 deficient CD4(+) T cells into Rag1(−/−) recipients exacerbated colitis development associated with increased gut T(FH)-related gene expression, while Irf8(−/−)/Bcl6(−/−) CD4(+) T cells abrogated colitis, together indicating that IRF8-regulated T(FH) can directly cause colon inflammation. Molecular analyses revealed that IRF8 suppresses T(FH) differentiation by inhibiting transcription and transactivation of the TF IRF4, which is also known to be essential for T(FH) induction. Our documentation showed that IRF8-regulated T(FH) can function as B-cell-independent, pathogenic, mediators of colitis suggests that targeting T(FH) could be effective for treatment of IBD.

To investigate the expression of kisspeptin in gastric adenocarcinoma and its effect on the proliferation and migration of gastric cancer cells.The level of kisspeptin in 50 gastric cancer tissues and their paracancerous tissues were detected by quantitative real-time PCR and Western blotting. Kisspeptin-siRNA,control-siRNA,p EGFP-N1-kisspeptin,p EGFP-N1 were separately transfected into MKN-45 gastric cancer cells. After 48-hour culture,the levels of matrix metalloproteinase-2( MMP-2),β-catenin,C-myc,MMP-9,kisspeptin were detected by Western blotting; MTT assay was used to detect the proliferation of MKN-45 cells; wound healing assay was performed to assess the migration ability of MKN-45 cells. After MKN-45 cells were treated with Wnt / β-catenin signal pathway inhibitor FH535,MTT assay and flow cytometry were used to evaluate cell proliferation ability and cell apoptosis,respectively.Kisspeptin expression in gastric adenocarcinoma tissues was significantly lower than that of the adjacent normal tissues. The cell survival rate and migration rate of p EGFP-N1-kisspeptin group were significantly lower than those of p EGFP-N1 group. Cell survival rate and migration rate of kisspeptin-siRNA group were significantly higher than those of control siRNA group. The levels of MMP-9,MMP-2,β-catenin and C-myc in p EGFP-N1-kisspeptin group were significantly lower than those in p EGFP-N1 group. The levels of MMP-9, MMP-2, β-catenin and C-myc in kisspeptin-siRNA group were significantly higher than those in control-siRNA group. The proliferation and migration trend of gastric cancer cells treated with FH535 was similar to that of the p EGFP-N1-kisspeptin group.The expression of kisspeptin decreases in gastric cancer tissues,and kisspeptin can interact with Wnt / β-catenin signaling pathway to inhibit the proliferation and migration of gastric cancer cells.

Sox7, Sox17 and Sox18 are the members of the Sry-related high-mobility group box family (SoxF) of transcription factors. SoxF factors regulate endothelial cell fate as well as development and differentiation of blood cells and lymphatic vessels. There is very less information about the functions of these genes in fish. We obtained the full-length cDNA sequence of SoxF genes including Sox7, Sox17 and Sox18 in Cyprinus carpio, where Sox7 and Sox18 had two copies. The construction of a phylogenetic tree showed that these genes were homologous to the genes in other species. Chromosome synteny analysis indicated that the gene order of Sox7 and Sox18 was highly conserved in fish. However, immense change in genomic sequences around Sox17 had taken place. Numerous putative transcription factor binding sites were identified in the 5_ flanking regions of SoxF genes which may be involved in the regulation of the nervous system, vascular epidermal differentiation and embryonic development. The expression levels of SoxF genes were highest in gastrula, and was abundantly expressed in the adult brain.We investigated the expression levels of SoxF genes in five specific parts of the brain. The expression levels of Sox7 and Sox18 were highest in the mesencephalon, while the expression level of Sox17 was highest in the epencephalon. In carp, the expression patterns of SoxF genes indicated a potential function of these genes in neurogenesis and in vascular development. These results provide new information for further studies on the potential functions of SoxF genes in carp.

Generally, duck hepatitis A virus type 1 (DHAV-1) only infects young ducklings. Since December 2016, severe outbreaks of duck viral infection with egg drop, feed consumption decline, and ovary-oviduct disease have occurred in some laying duck flocks in Shandong Province of China. DHAV-1 isolated from the affected ducks was confirmed as the causative pathogen of the egg drop. Compared with other DHAV-1 strains, the novel isolate has three special amino acid mutation points in the most variable regions at the C-terminus of VP1. The experimental infection in laying ducks indicated that successful immunization with DHAV-1 vaccine could protect laying duck from infection. To the best of our knowledge, this is the first reported incidence of a severe duck disease outbreak involving egg drop syndrome caused by DHAV-1.

This work investigated the impact of high intensity ultrasound (HIUS) pretreatment on the functional properties and structural characteristics of micellar casein concentrate (MCC). Microfiltered casein protein retentates were treated with HIUS for 0.5, 1, 2, and 5 min prior to spray drying. The results showed that conductivity, solubility, emulsifying, gelling increased significantly as the ultrasonic time prolonged, but the change of pH value were insignificant. In addition, the structural characteristics of MCC for all samples were studied. There was an increase in surface hydrophobicity (Ho) and a reduction in particle size compared with the control (without HIUS pretreatment). The secondary structure of HIUS pretreated MCC samples changed significantly with an increase in β-sheets and random coils and a reduction in α-helix and β-turn. It can be speculated that HIUS pretreatment facilitate the change of functional properties of MCC and these changes would promote its application in food industry.

The GNSS is vulnerable so that it often faces the risk of jamming, and blanket jamming is one of the interference. Power inversion algorithm is a commonly method used to solve this problem, however, the receiver will lose lock when GNSS satellites are close to RFI sources. To improve the receiver’s anti-jamming performance in this harsh scene, a method of vector tracking loop (VTL) based on despread-respread (DR) algorithm is present. Firstly, this paper suppress oppressive interference. Secondly, local reference signal is reconstructed according to predicted tracking loop parameters. Thirdly, beamforming algorithm is present. Theoretical analysis and simulation experiments verify that the VTL based on DR algorithm can reduce phase discriminator output errors, improve receiver’s positioning performance, and has superior anti-jamming performance in harsh scene. The method proposed in this paper can be applied to arbitrary array without prior information of satellites’ directions and is not sensitive to the error of array. Copared to VTL based on EIG, VTL based on DR algorithm has lower complexity.

The dynamic response characteristics of current loop in vector control system is closely related to the realization of vector control strategy. In the system model of multiple motors connected in series (which consists of coupled section, uncoupled section and feeder cable), the inductance and resistance of uncoupled section and feeder cable are taken into consideration in this paper. Due to the ignoring of current coupling problem of d-q axis in traditional PI current regulator, the dynamic performance of the system is poor. To solve this problem, a modified PI regulator based on voltage feed-forward decoupling is applied to build the control system of permanent magnet linear synchronous motor (PMLSM). The model of traditional PI current regulator and the model of modified PI current regulator are built respectively and compared with each other in Matlab/Simulink. The simulation results verify the correctness and effectiveness of PI current regulator with voltage feed-forward decoupling, and indicate that this improved control system has better dynamic and static characteristic than traditional control system.

Objective The objectives of this study were, first, to explore differences in brain activity between normal people and idiopathic restless legs syndrome (RLS) patients during asymptomatic periods; and, second, to determine whether administering repetitive transcranial magnetic stimulation (rTMS) to specific cortical regions would reverse any observed differences in brain activity and alleviate patient symptoms. Methods Fifteen idiopathic RLS patients (nine drug-naive patients) and 14 gender- and age-matched healthy controls were enrolled. Resting-state functional magnetic resonance imaging was used to measure the amplitude of low-frequency fluctuations (ALFF) in spontaneous brain activity during asymptomatic periods. Seven patients received high-frequency (5 Hz) rTMS directed toward the leg area of the primary motor cortex. Scores on the International Restless Legs Syndrome Study Group (IRLSSG) Rating Scale and ALFF values were measured before and after treatment. Results Compared with healthy controls, RLS patients showed lower ALFF in the sensorimotor and visual processing regions, and higher ALFF in the insula, parahippocampal and hippocampal gyri, left posterior parietal areas, and brainstem. These results were largely conserved when only drug-naive patients were considered. After rTMS treatment, ALFF in several sensorimotor and visual regions were significantly elevated and IRLSSG Rating Scale scores decreased, indicating improved RLS symptoms. Conclusions High-frequency rTMS delivered to the leg area of the primary motor cortex may raise functional activity in the sensorimotor and occipital regions, leading to improve symptoms in RLS patients. These results provide novel insight into RLS pathophysiology and suggest a potential mechanism for rTMS therapy in idiopathic RLS patients.

The prevalence of chronic kidney disease (CKD) increases with age, and anemia is known to affect the outcome of subjects with CKD. However, little is known with regard to the associations between metabolic complications and stages of CKD among elderly patients. Thus, the aim of the present study was to investigate the prevalence of CKD in elderly Chinese patients, as well as the associations between stages of CKD and clinically important complications of anemia. In total, 2,258 individuals with CKD, divided into younger (n=989) and elderly (n=1,269) groups, were enrolled in the study between June 2009 and December 2011. The glomerular filtration rate (GFR) was assessed using a 99mTc-DTPA renal dynamic imaging method (modified Gate's method). The levels of serum creatinine (SCr) and hemoglobin (Hb), and the hematocrit (HCT) were measured simultaneously per manufacturer's instructions. In the elderly group, the levels of SCr and proportional ratios were higher, while the GFR, Hb level, HCT and proportional ratios were lower when compared with the values in the younger group. Statistically significant differences were observed in the Hb concentrations when comparing individuals classified with different stages of CKD in the younger and elderly groups. In the younger group, there was no significant difference in the Hb concentrations between the stage 3a and 3b CKD patients. However, in the elderly group, the Hb concentrations were significantly higher in patients classified with stage 3a CKD when compared with those with stage 3b, whose GFR cutoff point was

During hematopoiesis, hematopoietic stem cells constantly differentiate into granulocytes and macrophages via a distinct differentiation program that is tightly controlled by myeloid lineage-specific transcription factors. Mice with a null mutation of IFN regulatory factor 8 (IRF8) accumulate CD11b+Gr1+ myeloid cells that phenotypically and functionally resemble tumor-induced myeloid-derived suppressor cells (MDSCs), indicating an essential role of IRF8 in myeloid cell lineage differentiation. However, IRF8 is expressed in various types of immune cells, and whether IRF8 functions intrinsically or extrinsically in regulation of myeloid cell lineage differentiation is not fully understood. In this study, we report an intriguing finding that, although IRF8-deficient mice exhibit deregulated myeloid cell differentiation and resultant accumulation of CD11b+Gr1+ MDSCs, surprisingly, mice with IRF8 deficiency only in myeloid cells exhibit no abnormal myeloid cell lineage differentiation. Instead, mice with IRF8 deficiency only in T cells exhibited deregulated myeloid cell differentiation and MDSC accumulation. We further demonstrated that IRF8-deficient T cells exhibit elevated GM-CSF expression and secretion. Treatment of mice with GM-CSF increased MDSC accumulation, and adoptive transfer of IRF8-deficient T cells, but not GM-CSF–deficient T cells, increased MDSC accumulation in the recipient chimeric mice. Moreover, overexpression of IRF8 decreased GM-CSF expression in T cells. Our data determine that, in addition to its intrinsic function as an apoptosis regulator in myeloid cells, IRF8 also acts extrinsically to repress GM-CSF expression in T cells to control myeloid cell lineage differentiation, revealing a novel mechanism that the adaptive immune component of the immune system regulates the innate immune cell myelopoiesis in vivo.

This study reported the development of a p–n heterojunction photoelectrochemical biosensor using a G-quadruplex as the recognition system. The n-type ZnO nanorods and p-type BiOI nanoflakes were combined to form the heterostructure by in situ growth of BiOI attributed to a simple impregnating-hydroxylation method. This simple p–n heterojunction fabrication method could facilitate the development of rapid and sensitive visible light activated PEC sensors. In this study, the ZnO/BiOI photoelectrode showed a rapid photocurrent decrease in the presence of Pb2+ due to the electron withdrawing effect of hemin which bound with the G-quadruplex/Pb2+ complex. This PEC biosensor exhibited a good response following the concentration of Pb2+ in the range from 10 to 100 μM.