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Background Diffuse intrinsic pontine glioma is an infiltrative, often high-grade glioma of the brainstem that is not amenable to surgical resection. The current treatment of DIPG by radiation therapy showed dramatically improvement of patient’s condition, however, the tumor recurs rapidly. More and more studies are focused on the genetic and epigenetic drivers of DIPGs, which may provide more and more novel therapy target for DIPG. EZH2 has been proved to be a potential therapeutic target for H3K27M-mutant pediatric gliomas recently. Meanwhile, BET family protein is a hot target in many different types of cancers, including DIPG. In this study, we performed the treatment of both EZH2 and BET inhibitor for DIPG cells. Results The combination of these two inhibitors exhibited better inhibition of the tumor growth both in vitro and in vivo compared to use the inhibitor individually. This inhibition was performed by blocking the proliferation and promoting the cell apoptosis. Meanwhile, combination treatment of these two inhibitors would also affect the epigenetic markers which were abnormal in the tumors of the certain set of genes. Conclusion Thus we provided a novel therapy strategy for clinical treatment of DIPG.

Craniofacial bone marrow mesenchymal stem cells (BMSCs) display some site-specific properties that differ from those of BMSCs derived from the trunk and appendicular skeleton, but the characteristics of craniofacial BMSCs and the mechanisms that underlie their properties are not completely understood. Previous studies indicated that special AT-rich binding protein 2 (SATB2) may be a potential regulator of craniofacial skeletal patterning and site-specific osteogenic capacity. Here, we investigated the stemness, autophagy, and anti-aging capacity of mandible-derived BMSCs (M-BMSCs) and tibia-derived BMSCs (T-BMSCs) and explored the role of SATB2 in regulating these properties. M-BMSCs not only possessed stronger expression of SATB2 and stemness markers (pluripotency genes, such as Nanog, OCT-4, Sox2, and Nestin) but also exhibited stronger autophagy and anti-aging capacities under normal or hypoxia/serum deprivation conditions compared to T-BMSCs. Exogenous expression of SATB2 in T-BMSCs significantly enhanced the expression of pluripotency genes as well as autophagy and anti-aging capacity. Moreover, SATB2 markedly enhanced osteogenic differentiation of BMSCs in vitro, and promoted bone defect regeneration and the survival of BMSCs that were transplanted into mandibles with critical size defects. Mechanistically, SATB2 upregulates pluripotency genes and autophagy-related genes, which in turn activate the mechanistic target of rapamycin signaling pathway. Collectively, our results provide novel evidence that site-specific BMSCs have distinct biological properties and suggest that SATB2 plays a potential role in regulating the stemness, autophagy, and anti-aging properties of craniofacial BMSCs. The application of SATB2 to manipulate stem cells for the reconstruction of bone defects might represent a new approach.

Previous studies have shown that craniofacial bone marrow stromal cells (MSCs) have greater osteogenic potential than appendicular bone MSCs. However, detailed phenotypic characterization of MSCs from bone marrow in the different sites remains unclear. To investigate bone repair and regeneration of craniofacial MSCs and the regulatory mechanisms underlying their unique properties, we compared osteogenesis, cell recruitment, autophagy, and apoptosis resistance of MSCs from the mandible (M-MSCs) to those from tibia (T-MSCs) in vitro and in vivo. Compared with T-MSCs, M-MSCs formed more colonies, possessed stronger proliferation activity, exhibited higher expression of pluripotency genes such as Oct4 and Nanog, and held stronger osteogenic differentiation in osteogenic medium. Moreover, M-MSCs had greater autophagy and anti-apoptotic capacities than T-MSCs under hypoxia and serum deprivation conditions. M-MSCs were found to be more capable of recruiting more MSCs than T-MSCs. When these MSCs were transplanted into mandible critical-sized defects, more bone formed in the M-MSC-treated animals than in their T-MSC counterparts. Collectively, these findings reveal that MSCs have unique characteristics and bone-repairing properties from the mandible as compared with those from tibia, presumably by enhanced osteogenic potential, cell recruitment, autophagy and apoptosis resistance.

The proposed Gibraltar Strait tunnel will cross two zones with breccia consisting of a chaotic mixture of blocks and stones embedded in a clay matrix. The breccia is saturated, has a high porosity and exhibits poor mechanical properties in the range between hard soils and weak rocks. The overburden and high in situ pore pressures in combination with the low strength of the breccia may lead to heavy squeezing. The crossing of the breccia zones thus represents one of the key challenges in the construction of the tunnel. In order to improve our understanding of the mechanical behaviour of the breccias, a series of triaxial compressions tests were carried out. Standard rock mechanics test equipment was not adequate for this purpose, because it does not provide pore pressure control, which is important in the case of saturated porous materials. Pore pressure control is routine in soil mechanics tests, but standard soil mechanics equipment allows only for relatively low nominal loads and pressures. In addition, the low hydraulic conductivity of the breccias demands extremely low loading rates and a long test duration. For these reasons, we re-designed several components of the test apparatus to investigate the mechanical behaviour of the breccia by means of consolidated drained and undrained tests. The tests provided important results concerning the strength, volumetric behaviour, consolidation state and hydraulic conductivity of the breccias. The present paper describes the test equipment and procedures, provides an overview of the test results and discusses features of the mechanical behaviour of the breccias which make them qualitatively different from other weak rocks such as kakirites—a typical squeezing rock in alpine tunnelling. The paper also demonstrates the practical importance of the experimental findings for tunnelling in general. More specifically, it investigates the short-term ground response to tunnel excavation from the perspective of elasto-plastic behaviour with the Mohr–Coulomb yield criterion. The computational results indicate that the breccias will probably experience very large deformations already around the advancing tunnel heading, which can be reduced considerably, however, by advance drainage. The analyses additionally show that plastic dilatancy is favourable with respect to the short-term response, thus highlighting the importance of the constitutive model when it comes to theoretical predictions.

In order to improve primary energy utilization, achieve economical operation of distribution network, comprehensively consider the concentration / compensation needs of various groups under typical load levels, and to gain understanding of characteristics of different types of user loads, the present paper proposes a hierarchical cluster algorithm to enhance the cohesion of a distribution feeder load characteristic clustering algorithm circumstances. This will serve to ultimately provide effective guidance for electricity energy conservation as well as to better realize peak load shifting. By cutting distribution network load time sequence data in longitudinal manner, relevant feature were extracted to achieve user load characteristics classification based on hierarchical clustering algorithm. Such classification will therefore assist to optimize distribution network scheduling. It is therefore an effective way to enhance accuracy and effectiveness of relevant power distribution decision-making.

The small GTPase Rab26 is involved in multiple processes, such as vesicle-mediated secretion and autophagy. However, the mechanisms and functions of Rab26 in the human pulmonary microvascular endothelial cells (HPMVECs) are not clear. In this study, we thoroughly investigated the role and novel mechanism of Rab26 in permeability and apoptosis of HPMVECs using a self-assembled Rab26 siRNA loaded DNA Y-motif nanoparticle (siRab26-DYM) and Rab26 adenovirus. We found that siRab26-DYM could be efficiently transfected into HPMVECs in a time- and dose-dependent manner. Importantly, the siRab26-DYM nanovector markedly aggravated the LPS-induced apoptosis and hyper-permeability of HPMVECs by promoting the nuclear translocation of Foxo1, and subsequent activation of Toll-like receptor 4 (TLR4) signal pathway. Overexpression of Rab26 by Rab26 adenoviruses partially inactivated LPS-induced TLR4 signaling pathway, suppressed the cell apoptosis and attenuated the hyperpermeability of HPMVECs. These results suggest that the permeability and apoptosis of HPMVECs can be modulated by manipulating Rab26 derived TLR4 signaling pathway, and that Rab26 can be potential therapeutic target for the treatment of vascular diseases related to endothelial barrier functions.

// Lei Cheng 1, 2, * , Shuhang Gao 3, * , Xiaobo Song 4 , Weijie Dong 5 , Huimin Zhou 6 , Lifen Zhao 1 , Li Jia 1 1 College of Laboratory Medicine, Dalian Medical University, Dalian 116044, Liaoning Province, China 2 Department of Laparoscopic Surgery, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, Liaoning Province, China 3 Department of Oncology, The First Affiliated Hospital of Dalian Medical University, Dalian 116011, Liaoning Province, China 4 Department of Medical Biology, Faculty of Health Sciences, University of Tromso, Tromso, Norway 5 Department of Biochemistry, Dalian Medical University, Dalian 116044, Liaoning Province, China 6 Department of Microbiology, Dalian Medical University, Dalian 116044, Liaoning Province, China * These authors contributed equally to this work Correspondence to: Li Jia, email: jiali0386@sina.com Keywords: fucosyltransferase (FUT) gene, human hepatocellular carcinoma cell lines, tumor progression, microRNAs Received: February 05, 2016 Accepted: August 09, 2016 Published: August 13, 2016 ABSTRACT Glycosylation has significant effects on cancer progression. Fucosylation is one of the most important glycosylation events involved in hepatocellular carcinoma (HCC). Here, we compared N-glycan profiles of liver tumor tissues and adjacent tissues of 27 HCC patients to reveal the association between fucosylation and HCC progression, as well as verified the potential role of miRNA in regulating fucosylation. Mass spectrometry (MS) analysis showed pronounced differences of the N-glycosylation patterns and fucosylated N-glycans between the adjacent and tumor tissues. Different fucosyltransferase (FUT) genes were also identified in adjacent and tumor tissues, and two HCC cell lines with different metastatic potential. High-level expression of FUT8 was detected in tumor tissues and highly metastatic HCC cells. Altered levels of FUT8 in HCC cell lines significantly linked to the malignant behaviors of proliferation and invasion in vitro . Furthermore, using microRNA array, we identified FUT8 as one of the miR-26a, miR-34a and miR-146a-targeted genes. An inverse correlation was revealed between the expression levels of FUT8 and these miRNAs. Luciferase reporter assay demonstrated these miRNAs specifically interacted with the 3′UTR of FUT8 and subsequently down-regulated FUT8 expression-level. The forced expression of these miRNAs was able to induce a decrease in FUT8 levels and thereby to suppress HCC cells progression. Altogether, our results indicate that fucosylated N -glycan and FUT8 levels can be used as markers for evaluating HCC progression, as well as miRNAs may be involved in inhibition of fucosylation machinery through targeting FUT8.

High-altitude deacclimatization syndrome (HADAS) is emerging as a severe public health issue that threatens the quality of life of individuals who return to lower altitude from high altitude. In this study, we measured serum levels of SOD, MDA, IL-17A, IL-10, TNF-α, and HADAS score in HADAS subjects at baseline and 50th and 100th days and to evaluate the relationship between interleukins, including IL-17A, and HADAS. Our data showed that and the serum IL-17A levels and HADAS score decreased over time in the HADAS group, and serum IL-17A levels were significantly higher in the HADAS group at baseline and 50th day compared with controls (p<0.05). Furthermore, baseline serum levels of MDA and TNF-αwere significantly higher, while SOD and IL-10 levels were lower in HADAS subjects compared with controls (p<0.05). It is interesting that serum levels of IL-17A were clearly interrelated with HADAS incidence and severity (p<0.05). ROC curve analysis showed that combined serum IL-17A and IL-10 levels were a better predictor of HADAS incidence than serum levels of IL-17A or IL-10 alone. These data suggest that serum levels of IL-17A are a novel predictive index of HADAS.

N-doped Cu2O films are deposited by sputtering a CuO target in the mixture of Ar and N2. The structures zand optical properties have been studied for the films deposited at different temperatures. It is found that N-doping can suppress the formation of CuO phase in the films. The films are highly (100) textured at low temperatures and gradually change to be highly (111) textured at the temperature of 500 °C. With the analysis of (111) and (100) grain sizes, the surface free energy and grain size of critical nuclei are suggested to dominate the film texture. The analysis of the atomic force microscopy shows that the film growth can be attributed to the surface-diffusion-dominated growth. The forbidden rule of band gap transition is found disabled in the N-doped Cu2O films, which can be attributed to the occupation of 2p electrons of nitrogen at the top of valence band. The optical band gap energy is determined to be 2.52 ± 0.03 eV for the films deposited at different temperatures.

Pb1-xSrx (Zr0.53Ti0.47) O3 (PSZT) thin films with different x values (x = 0, 0.02, 0.04, 0.06) are prepared by sol-gel method and their properties are studied. All PSZT thin films get a single perovskite phase. The dielectric properties are studied. The leakage current density at an applied electric field of 61.2 kV/cm is 293.0 × 10−9A/cm2 when x = 0.04. The remanent polarization and coercive field values of the PSZT (x = 0) thin film are 8.4 μC/cm2 and 67.5kV/cm, respectively.

Microforce sensors made of PT/PZT/PT thin films were designed in the micro-Newton range. The PT/PZT/PT thin films were deposited on the Pt/Ti/SiO2/Si substrate by the sol–gel process. Structure and the crystallinity of the deposited films were characterized. The sensing properties of the microforce sensors were measured in static state and in quasi-static state. The sensitivities of the microforce sensors with two different sizes were 0.004 and 0.040 mV μ N−1 in the quasi-static state.

Calibration of the micro force sensors is necessary for the measurement of microNewton forces, which are critical to analytical applications of micro force sensor in the analysis of micromanipulators, microrobots and microgrippers. This paper proposes a novel calibration system to calibrate PZT thin film micro force sensor using the bimorph probe as the loading part. Sensitivities of PZT thin film micro force sensors were calibrated by the calibration system composed of the drive system and the testing system. The drive system combines 3-dimensional positioning stage, a bimorph probe and a frequency synthesizer, and the testing system combines an electronic balance for static state or a charge amplifier and a lock-in amplifier for the quasi static state.

A blind source separation based vibration control method using piezoelectric self-sensing actuator is proposed. Two piezoelectric ceramic patches acting as actuators and two current amplifiers consist of the self-sensing bridge circuit. Each piezoelectric current is a mixture of strain rate current and capacitor current. As they are orthogonal and independent with each other, the blind source separation technique is introduced to obtain the pure strain rate signal. An effort is made to solve the problem of BSS uncertainty in separation order. Open loop experimental results on cantilever beam show the feasibility and validity of the proposed method.

PZT thin film devices are very competitive when charge signal and force outputs are demanded. A Pb(Zr0.5Ti0.5)O3 thin film with 24 coating layers is deposited by a sol–gel method. An array of three piezoelectric microcantilevers is fabricated using the wet and dry combined bulk micromachining techniques. Each microcantilever has a two-segment top electrode in different dimensions. The actuating and sensing capability of the six segments on the three microcantilevers are measured. It is proved that the proposed piezoelectric microcantilevers have both actuating and sensing functions; different proportion partition results in the diversity of the actuating and sensing capability. At the same location, the longer the piezoelectric segment is, the stronger actuating and sensing capability it has. The maximum actuating sensitivity and sensing sensitivity before poling treatment are 16.7 µN V−1 and 13.44 pC µm−1, respectively, which become about twice as big after poling. We may choose the appropriate microcantilevers to satisfy our purpose on some specific condition where force feedback and object manipulation are needed simultaneously. A hysteresis phenomenon is encountered in the actuating process while not in the sensing process.

The aberrant sialylation profile on the surface of leukemia cells has been recognized for its potential diagnostic value towards assessing leukemia multidrug resistance (MDR). MicroRNAs as endogenous regulators of gene expression have been implicated in treating MDR. In this study, we describe the differential expressional profiles of α-2, 3-sialyltransferases (ST) and miR-4701-5p in three pairs of chronic myeloid leukemia (CML) cell lines and 48 clinical samples of bone marrow mononuclear cells from CML patients. The altered expression level of ST3GAL1 was found corresponding to the drug-resistant phenotype (with and without adriamycin resistance) of CML cell lines both in vitro and in vivo. Further the results showed that miR-4701-5p directly targeted ST3GAL1 to reduce CML cells resistance to multiple chemotherapeutics in vitro and to convert tumor cells from adriamycin resistant to susceptible in vivo of mice. These results indicate that differential expression of α-2,3 ST is involved in MDR of CML, and that miR-4701-5p regulates the susceptibility of CML cells to multiple drugs, at least in part, through targeting ST3GAL1.

Fucosylation is the final step in the glycosylation machinery, which produces glycans involved in tumor multidrug resistance development. MicroRNAs (miRNAs) are endogenous negative regulators of gene expression and have been implicated in most cellular processes of tumors, including drug resistance. This study was undertaken to determine the roles of fucosylation and miR-224-3p in multidrug resistance of human breast cancer cell lines. Comparative analysis revealed differential modification patterns of fucosylation of the fucosylated N-glycans in drug-resistant T47D/ADR cells and sensitive line T47D cells. The expressional profiles of fucosyltransferase genes in two pairs of parental and chemoresistant human breast cancer cell lines showed that FUT4 was up-regulated highly in MDR cell lines. Altered level of FUT4 affected the drug-resistant phenotype of T47D and T47D/ADR cells both in vitro and in vivo. By bioinformatics analysis, we identified FUT4 as one of the miR-224-3p-targeted genes. Further studies showed an inverse relationship between of FUT4 and miR-224-3p in parental and ADR-resistant breast cancer cells, wherein miR-224-3p was downregulated in resistant cells. 3'-UTR dual-luciferase reporter assay confirmed that miR-224-3p directly targeted 3'-untranslation region (3'-UTR) of FUT4 mRNA. In addition, miR-224-3p overexpression sensitized T47D/ADR cells to chemotherapeutics and reduced the growth rate of breast cancer xenografts in vivo. Our results indicate that FUT4 and miR-224-3p are crucial regulators of cancer response to chemotherapy, and may serve as therapeutic targets to reverse chemotherapy resistance in breast cancer.

Developing an advanced nucleic acid drug delivery system is of great significance in order to achieve optimal gene delivery. Self-assembled nucleic acid nanoparticles are an excellent platform for the delivery of nucleic acids and other small molecular drugs. In this study, we developed the efficient, three-stranded, RNA/DNA hybrid triangular self-assembled nanoparticles, namely, mTOR single-stranded siRNA-loaded triangular DNA nanoparticles (ssRNA-TNP). The ssRNA-TNP is formed by the complementary association of the above mentioned three components and is more stable in complete medium than standard duplex siRNA. It could be efficiently transfected into NCI-H292 cells in a dose- and time-dependent manner, resulting in high transfection efficiency. Furthermore, ssRNA-TNP uptake is dependent on macropinocytosis and clathrin-mediated endocytosis pathways. Interestingly, ssRNA-TNP is more efficient to inhibit the expression of mTOR. This ssRNA-TNP has a simpler structure, better stability, and higher transfection efficiency; therefore it may become a novel nonviral nanosystem for gene delivery.

Alcian blue staining has been widely used to visualize acidic mucins and mucopolysaccharides in supported molecular matrix electrophoresis (SMME) and on membrane transferred from electrophoresis gels. Mucins with low acidic glycan content, however, cannot be stained with Alcian blue, which is one of the major drawbacks of this staining method. On the other hand, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, regardless of the acidic residue content; however, periodic acid-Schiff staining decomposes glycans. Here, we introduce succinylation-Alcian blue staining as an alternative staining method to visualize mucins, regardless of the acidic residue content, and without glycan decomposition.

This paper studied the effect of curvature on the charge sensitivity of a curved PVDF sensor. The sensor was fabricated by attaching a PVDF film along the X-axis on a group of cylindrical silicon rods to form an outline with several bumps. The open output voltage of a curved sensor was simulated by using ANSYS with a rod curvature of 333 m-1, 400 m-1 and 500 m-1. It was found that the sensitivity was the highest when the curvature was 500 m-1, and the lowest when the curvature was 333 m-1. Curved PVDF sensors were fabricated and tested with a PVDF patch of 30 mm long and 28 μm thick and a radius of silicon rod of 3 mm, 2.5 mm and 2 mm, respectively. A shaker and mass block were used to apply the same dynamic force to the curved sensors and an oscilloscope was used to observe the output charge. The experiment results are not ideal but do show the same tendency as the simulation. The charge sensitivity increased as the curvature of substrate increased.

Ag recognition and Ab production in B cells are major components of the humoral immune response. In the current study, we found that the core fucosylation catalyzed by α1,6-fucosyltransferase (Fut8) was required for the Ag recognition of BCR and the subsequent signal transduction. Moreover, compared with the 3-83 B cells, the coalescing of lipid rafts and Ag-BCR endocytosis were substantially reduced in Fut8-knockdown (3-83-KD) cells with p31 stimulation and then completely restored by reintroduction of the Fut8 gene to the 3-83-KD cells. Indeed, Fut8-null (Fut8−/−) mice evoked a low immune response following OVA immunization. Also, the frequency of IgG-producing cells was significantly reduced in the Fut8−/− spleen following OVA immunization. Our results clearly suggest an unexpected mode of BCR function, in which the core fucosylation of IgG-BCR mediates Ag recognition and, concomitantly, cell signal transduction via BCR and Ab production.

Alcian blue staining has been widely used to visualize acidic mucins and mucopolysaccharides in supported molecular matrix electrophoresis (SMME) and on membrane transferred from electrophoresis gels. Mucins with low acidic glycan content, however, cannot be stained with Alcian blue, which is one of the major drawbacks of this staining method. On the other hand, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, regardless of the acidic residue content; however, periodic acid-Schiff staining decomposes glycans. Here, we introduce succinylation-Alcian blue staining as an alternative staining method to visualize mucins, regardless of the acidic residue content, and without glycan decomposition.

A modified RAKE receiver based on novel Recursive Least Squares (RLS) adaptive algorithm is proposed. The receiver uses L-fingered correlators, which are composed of RLS adaptive filters, to enhance the performance of multipath receiving. It can also track the amplitude of the received signal to form a real-time amplitude estimation which is correlated with the power of excess delay bin. The simulation results based on the IEEE UltraWide Band (UWB) channel models (CM1 to CM4) show that the novel RLS algorithm can alter the attenuation estimation with the finger’s power delay profile, and RAKE receiver with few fingers can be employed to get high performance.

A modified sol-gel multiple coating and annealing process was developed to prepare lead zirconate titanate PbZr 0.5 Ti 0.5 O 3 (PZT) thin films with various thickness on Pt(111)/Ti/SiO 2 /Si(100) substrates. The PZT films with thickness 280, 560 and 810 nm underwent one, two and three annealing cycles, respectively, which consisted of 5 coating layers, 10 coating layers and 15 coating layers. Surface morphology and phase structure of the thin films were studied by SEM and XRD, respectively. The relationship between the thickness, namely different annealing cycles, and degree of texture was investigated. The dielectric constant increases with increasing film thickness. The dielectric constant and dielectric loss of PZT films as functions of applied field and frequency had also been examined and discussed.

Cell surface sialylation is emerging as an important feature of cancer cell multidrug resistance (MDR). We have focused on the influence of 2,8-sialyltransferases in key steps of the development of MDR in chronic myeloid leukemia (CML). The expressional profiles of six α-2,8-sialyltransferases were generated in three pairs of CML cell lines and peripheral blood mononuclear cells (PBMC) of CML patients. Cellular MDR phenotype positively correlated with ST8SIA4 and ST8SIA6 levels. Furthermore, ST8SIA4 mediated the activity of phosphoinositide-3 kinase (PI3K)/Akt signal pathway and the expression of P-glycoprotein (P-gp). Targeting the PI3K/Akt pathway by its specific inhibitor LY294002, or by Akt RNA interfering reversed the MDR phenotype of K562/ADR cells. Inhibition of PI3K/Akt pathway also attenuated the effects caused by the overexpression of ST8SIA4 on MDR. Therefore this study indicated that α-2,8-sialyltransferases involved in the development of MDR of CML cells probably through ST8SIA4 regulating the activity of PI3K/Akt signaling and the expression of P-gp.

In engineering practice the response of squeezing rock to tunnel excavation is usually analysed by assuming that the ground behaves as a linearly elastic perfectly plastic material obeying the Mohr Coulomb yield criterion (the "MC" model). The model has some limitations especially if the ground is waterbearing: Due to the assumption of purely elastic behaviour before failure the MC model cannot predict the excess pore pressure caused by pre failure plastic deformations under undrained conditions. This deficiency may be significant considering that pore pressure effects are generally important for squeezing ground. In order to study the influence of this aspect of the material behaviour we performed comparative computations concerning the short term (undrained) behaviour of the ground in tunnelling assuming two alternative elastoplastic strain hardening models the so called Deviatoric Hardening model ("DH" model) and the Modified Hardening Soil model ("MHS" model). Both models are relatively simple needing only common triaxial tests to identify their parameters and they can simulate contractant plastic behaviour (and thus the pre failure generation of positive excess pore pressure) and map the behaviour of weak rock in consolidated undrained triaxial compression tests more successfully than the MC model. Numerical analysis showed that the inclusion of the excess pore pressure in the two alternative models leads to larger and slightly more rapid displacements of the tunnel wall than under the MC model. According to the numerically determined stress field an extended zone of the ground around the opening exhibits positive excess pore pressures which also indicates the importance of contractant behaviour. However the difference is not so dramatic with respect to displacements of the tunnel boundary. This shows that the widely used MC model is adequate for the specific problem under consideration (estimation of the ground response to tunnel excavation) in spite of its simplifications. © 2015 Taylor Francis Group London.

Bone marrow stromal cells (BMSCs) are multipotent progenitor cells with capacities to differentiate into the various cell types and hold great promise in regenerative medicine. The regulatory roles of histone deacetylases (HDACs) in osteoblast differentiation process have been increasingly recognized; however, little is known about the precise roles of HDAC8 in the osteogenic differentiation of BMSCs. Herein we aimed to investigate the roles of HDAC8 in the osteogenic differentiation of rat BMSCs by pharmacological and genetic manipulations in vitro. During osteogenic differentiation of BMSCs, pharmacological inhibition of HDAC8 by HDAC inhibitor valproic acid (VPA) promoted the level of histone H3 lysine 9 acetylation (H3K9Ac) and significantly enhanced the expression of osteogenesis-related genes Runx2, Osterix, osteocalcin (OCN), osteopontin (OPN) and alkaline phosphatase (ALP). Similarly, knockdown of HDAC8 using short interfering RNA triggered H3K9Ac and enhanced osteogenic differentiation of BMSCs, largely phenocopied the effects of VPA-mediated HDAC8 depletion. However, enforced expression of HDAC8 significantly reduced the level of H3K9Ac and inhibited osteogenic differentiation of BMSCs, which can be attenuated by VPA addition. Mechanistically, HDAC8 suppressed osteogenesis-related genes expression by removing the acetylation of histone H3K9, thus leading to transcriptional inhibition during osteogenic differentiation of BMSCs. Importantly, we found that HDAC8 physically associated with Runx2 to repress its transcriptional activity and this association decreased when BMSCs underwent osteogenic differentiation. Taken together, these results indicate that epigenetic regulation of Runx2 by HDAC8-mediated histone H3K9 acetylation is required for the proper osteogenic differentiation of BMSCs.

Craniofacial bone marrow mesenchymal stem cells (BMSCs) display some site-specific properties that differ from those of BMSCs derived from the trunk and appendicular skeleton, but the characteristics of craniofacial BMSCs and the mechanisms that underlie their properties are not completely understood. Previous studies indicated that special AT-rich binding protein 2 (SATB2) may be a potential regulator of craniofacial skeletal patterning and site-specific osteogenic capacity. Here, we investigated the stemness, autophagy, and anti-aging capacity of mandible-derived BMSCs (M-BMSCs) and tibia-derived BMSCs (T-BMSCs) and explored the role of SATB2 in regulating these properties. M-BMSCs not only possessed stronger expression of SATB2 and stemness markers (pluripotency genes, such as Nanog, OCT-4, Sox2, and Nestin) but also exhibited stronger autophagy and anti-aging capacities under normal or hypoxia/serum deprivation conditions compared to T-BMSCs. Exogenous expression of SATB2 in T-BMSCs significantly enhanced the expression of pluripotency genes as well as autophagy and anti-aging capacity. Moreover, SATB2 markedly enhanced osteogenic differentiation of BMSCs in vitro, and promoted bone defect regeneration and the survival of BMSCs that were transplanted into mandibles with critical size defects. Mechanistically, SATB2 upregulates pluripotency genes and autophagy-related genes, which in turn activate the mechanistic target of rapamycin signaling pathway. Collectively, our results provide novel evidence that site-specific BMSCs have distinct biological properties and suggest that SATB2 plays a potential role in regulating the stemness, autophagy, and anti-aging properties of craniofacial BMSCs. The application of SATB2 to manipulate stem cells for the reconstruction of bone defects might represent a new approach.

Under pressure from social criticism and an unclear future, tobacco researchers have begun to seek alternative uses for the product. Here, we present our study on isolating tobacco compounds with fungicidal activity, which could be used as plant-derived pesticides. Using Valsa mali as the target fungus, agar plate tests were conducted to evaluate the fungicidal activity of various tobacco extracts, including tobacco leaves extracts prepared with different solvents, extracts of different tobacco cultivars, and samples from different tobacco organs. Fungal growth morphology was used as the criterion to evaluate the fungicidal activity of tobacco extracts. Correlation analyses between the fungicidal activities and the chemical components of tobacco extracts indicated the major chemical constituents with fungicidal activity. Then, the active compounds were isolated and their effects on the ultra-microstructures of V. mali was analyzed using scanning- and transmission-electron microscopy. The results suggested that tobacco extracts prepared with solvents of weaker polarity had higher fungicidal activity, and the inhibitory activity of tobacco extracts against V. mali was also cultivar dependent. Furthermore, the fungicidal effects of tobacco flower extracts were higher than those of the leaf extracts. Chemical analysis indicated that cembranoids were the main fungicidal substances, which act by destroying the endometrial structure of the fungus. Tobacco cembranoids at 80 μg/mL could completely inhibit the growth of V. mali, with an EC50 value of 13.18 μg/mL. Our study therefore suggests that tobacco leaves and inflorescences are excellent plant resources for the biological control of V. mali.

Sialylation is one of the altered glycosylation patterns associated with cancer progression. In this study, we investigated the N-glycan profiles of breast cancer patients and cell lines to reveal sialylation associated with breast cancer progression, and provided new evidences of miRNA-mediated sialylation. MALDI-TOF MS analysis revealed that N-glycans found in breast cancer tissues and breast cancer cell MDA-MB-231 featured increased levels of sialylation compared with adjacent tissues and normal breast epithelial cell MCF-10A. The expressional profiles of 20 sialyltransferase genes were then analyzed and found significantly different comparing breast cancer samples with adjacent tissues, and two breast cancer cell lines MDA-MB-231 and MCF-7 with different metastatic potential and MCF-10A cells. Tumor tissues and highly metastatic breast cancer cell line MDA-MB-231 exhibited higher levels of ST8SIA4. Knocking down ST8SIA4 in breast cancer cell lines significantly inhibited their malignant behaviors including cell proliferation and invasion in a sialyltransferase-dependent manner. By applying bioinformatic approaches for the prediction of miRNA targeting 3′-UTR of ST8SIA4, we identified ST8SIA4 as one of the miR-26a/26b-targeted genes. Further data analysis revealed the inversely related expression of ST8SIA4 and miR-26a/26b in breast cancer cells, tumor tissues and corresponding adjacent tissues. The ability of miR-26a/26b to interact specifically with and regulate the 3′-UTR of ST8SIA4 was demonstrated via a luciferase reporter assay. The forced expression of miR-26a/26b was able to induce a decrease of ST8SIA4 level and also to affect breast cancer cells progression, while altered expression of ST8SIA4 in breast cancer cells modulated progression upon transfection with miR-26a/26b mimics or inhibiter. Taken together, these results indicate that changes in the glycosylation patterns and sialylation levels may be useful markers of the progression of breast cancer, as well as miR-26a/26b may be widely involved in the regulation of sialylation machinery by targeting ST8SIA4.

Aberrant sialylation is closely associated with malignant phenotypes of tumor cells, including invasiveness and metastasis. This study investigated sialylation with regard to the modification of invasive properties and chemosensitivity in human hepatocellular carcinoma (HCC) cell lines and the association between the sialyltransferase gene family and clinicopathological characteristics in HCC patients. Using mass spectrometry analysis, we found that the composition profiling of sialylated N-glycans differed between MHCC97H and MHCC97L cells with different metastatic potential. The expressional profiles of 20 sialyltransferase genes showed differential expression in two cell lines, transitional and tumor tissues, from the same patients. Two genes, ST6GAL1 and ST8SIA2, were detected as overexpressed in MHCC97H and MHCC97L cells. The altered expression levels of ST6GAL1 and ST8SIA2 corresponded to a changed invasive phenotype and chemosensitivity of MHCC97H and MHCC97L cells both in vitro and in vivo. Further data indicated that manipulation of the expression of the two genes led to altered activity of the phosphoinositide-3 kinase (PI3K)/Akt signaling pathway. Targeting the PI3K/Akt pathway by its specific inhibitor wortmannin or by Akt RNA interference resulted in a reduced capacity for invasion and chemoresistance of MHCC97H cells. Our results imply that sialylation may function as an internal factor, regulating the invasion and chemosensitivity of HCC, probably through ST6GAL1 or ST8SIA2 regulation of the activity of the PI3K/Akt pathway.

To explore the time-dependent changes in the expressions of ferritin (Fn) and ferroportin (FPN) in macrophages infected with Mycobacterium (M.) tuberculosis of highly virulent strain (H37Rv strain) and Bacillus Calmette-Guerin (BCG) strain.RAW264.7 macrophages were respectively infected with M. tuberculosis H37Rv strain and BCG strain. At 1, 6, 12, 18 and 24 h after infection, the contents of Fn and FPN in the infected macrophage culture supernatants of the two groups were detected using ELISA, and meanwhile, the expression of Fn in the infected macrophages of each group was determined using Western blotting.After the macrophages were infected with different strains, the expression of Fn in macrophages increased as time went by; at 1 h after infection, the Fn expression in the control group was significantly higher than that in the H37Rv group and the BCG group; at 6 h after infection, the Fn expression in the two infected groups were higher than that in the control group; at 12 h, the Fn expression in the H37Rv group was higher than that in the control group, the difference was statistically significant (P0.05); at 18 and 24 h, the Fn expression was reduced in the order as H37Rv groupBCG groupcontrol group (P0.05). The expression of FPN decreased gradually as time went by. Compared with the normal control group, the expression in both H37Rv group and BCG group presented a lower level and H37Rv groupBCG groupcontrol group (P0.05).The infection of M.tuberculosis can lead to increased Fn expression and decreased FPN expression in macrophages, and the levels of Fn and FPN in the infected macrophages are both related to the strength of the virulence of M.tuberculosis.

Offshore platform, which is the base of the production and living in the sea, is the most important infrastructure for developing oil and gas resources. At present, there are almost 6500 offshore platforms servicing in the 53 countries' sea areas around the world, creating great wealth for the world. In general, offshore platforms may work for 20 years, however, offshore platforms are expensive, complex, bulky, and so many of them are on extended active duty. Because of offshore platforms servicing in the harsh marine environment for a long time, the marine environment have a great impact on the offshore platforms. Besides, with the impact and erosion of seawater, and material aging, the offshore platform is possible to be in unexpected situations when a badly sudden situation happens. Therefore, it is of great significance to monitor the marine environment and offshore platforms. The self-contained sensor for deep-sea offshore platform with its unique design, can not only effectively extend the working time of the sensor with the capability of converting vibration energy to electrical energy, but also simultaneously collect the data of acceleration, inclination, temperature and humidity of the deep sea, so that we can achieve the purpose of monitoring offshore platforms through analyzing the collected data. The self-contained sensor for monitoring of deep-sea offshore platform includes sensing unit, data collecting and storage unit, the energy supply unit. The sensing unit with multi-variables, consists of an accelerometer LIS344ALH, an inclinometer SCA103T and a temperature and humidity sensor SHT11; the data collecting and storage unit includes the MSP430 low-power MCU, large capacity memory, clock circuit and the communication interface, the communication interface includes USB interface, serial ports and wireless interface; in addition, the energy supply unit, converting vibration to electrical energy to power the overall system, includes the electromagnetic generator, voltage multiplier circuit and a super capacitor which can withstand virtually unlimited number of charge-discharge cycles. When the seawater impacts on offshore platforms to produce vibration, electromagnetic generator converts vibration to electrical energy, its output(~ 1 V 50 Hz AC) is stepped up and rectified by a voltage multiplier circuit, and the energy is stored in a super capacitor. It is controlled by the MSP430 that monitors the voltage level on the super capacitor. The super capacitor charges the Li-ion battery when the voltage on the super capacitor reaches a threshold, then the whole process of energy supply is completed. The self-contained sensor for deep-sea offshore platform has good application prospects and practical value with small size, low power, being easy to install, converting vibration energy to supply power and high detection accuracy.

Supported molecular matrix electrophoresis (SMME), in which a hydrophilic polymer such as PVA serves as a support within a porous PVDF membrane, was recently developed. This method is similar to cellulose acetate membrane electrophoresis but differs in the compatibility to glycan analysis of the separated bands. In this report, we describe the first instance of the application of SMME to human serum fractionation, and demonstrate the differences with serum fractionation by cellulose acetate membrane electrophoresis. The SMME membrane exhibited almost no EOF during electrophoresis, unlike the cellulose acetate membrane, but afforded comparative results for serum fractionation. The visualization of each fraction was achieved by conventional staining with dye such as Direct Blue-71, and objective quantification was obtained by densitometry after inducing membrane transparency with 1-nonene. Immunostaining was also achieved. Moreover, mass spectrometric analysis of both N-linked and O-linked glycans from the separated bands was demonstrated. Serum fractionation and glycan profiling of each fraction using SMME will enable novel insights into the relationships between various glycosylation profiles and disease states.

The characterization of mucins is critically important for gaining insights into the molecular pathology of diseases, including cancers, as well as for the discovery of biomarkers for disease diagnosis. However, no practical method has yet been reported for identifying mucin proteins. Here, we report a technique for immunological identification of mucins separated by supported molecular matrix electrophoresis (SMME), a recently developed membrane electrophoresis method. The technique involves on-membrane deglycosylation of mucins by using mild periodate oxidation/base-catalyzed elimination, followed by immunostaining with an antibody that specifically recognizes the mucin tandem repeat (TR) peptide. We demonstrated the method's feasibility by using MUC1 derived from 2 cancer cell lines, T47D and HPAF-II. The present method shows potential as an alternative approach for the identification of mucins separated by SMME or blotted from conventional gel electrophoresis, on a PVDF membrane.

The tunnel planned for the Gibraltar Strait fixed link will cross - among other formations - two paleo-channels filled with breccias. The breccias exhibit poor mechanical properties in the range between stiff clays and weak rocks. The overburden and high pore pressures in-situ may lead, in combination with the low strength of the breccias, to heavy squeezing during construction of the tunnel. The mechanical properties of the breccias were studied using special triaxial tests. The testing technique is very demanding due to the need for pore water control in combination with high applied loads. Neither standard rock nor soil mechanics testing equipment is adequate for this purpose. In addition, the low hydraulic conductivity of the breccias demands extremely low loading rates, which leads to very long test durations. The paper presents and comments on the test results on the strength, volumetric behaviour, consolidation state and hydraulic conductivity of the breccias.

This article is based on an investigation into grain stores in Chongwen District, Beijing. The authors find that state-run grain stores are in severe debt, and do not know how to survive in the coming market economy. On the basis of their investigation, the authors analyse the problems of the grain store system, predict future trends of consumption, and propose some adjustments and resolutions that could help the grain stores to get out of the crisis.

The isolation and characterization of mucins are critically important for obtaining insight into the molecular pathology of various diseases, including cancers and cystic fibrosis. Recently, we developed a novel membrane electrophoretic method, supported molecular matrix electrophoresis (SMME), which separates mucins on a polyvinylidene difluoride (PVDF) membrane impregnated with a hydrophilic polymer. Alcian blue staining is widely used to visualize mucopolysaccharides and acidic mucins on both blotted membranes and SMME membranes; however, this method cannot be used to stain mucins with a low acidic glycan content. Meanwhile, periodic acid-Schiff staining can selectively visualize glycoproteins, including mucins, but is incompatible with glycan analysis, which is indispensable for mucin characterizations. Here we describe a novel staining method, designated succinylation-Alcian blue staining, for visualizing mucins on a PVDF membrane. This method can visualize mucins regardless of the acidic residue content and shows a sensitivity 2-fold higher than that of Pro-Q Emerald 488, a fluorescent periodate Schiff-base stain. Furthermore, we demonstrate the compatibility of this novel staining procedure with glycan analysis using porcine gastric mucin as a model mucin.

Mucins are a family of heavily glycosylated high molecular mass proteins that have great potential as novel clinical biomarkers for the diagnosis of various malignant tumors. Supported molecular matrix electrophoresis (SMME) is a new type of membrane electrophoresis that can be used to characterize mucins. In SMME, mucins migrate in a molecular matrix supported by membrane materials. Here, we have developed an immunostaining method for the identification of SMME-separated mucins. The novel method involves stably fixing the mucins onto the SMME membrane and optimizing the molecular matrix for the fixation process. We applied this technique for the detection of MUC1 produced from three cancer cell lines (T47D, HPAF-II and BxPC3) and also analyzed their O-linked glycans by mass spectrometry. Our results revealed that properties of the MUC1 molecules from the three cell lines are different in terms of migrating position in SMME and glycan profile. The present method allows simple and rapid characterization of mucins in terms of both glycans and core proteins. The method will be a useful tool for the exploration of mucin alterations associated with various diseases such as cancer.

For the case that the traditional parameter adjustment consumes too much time, a method using the multi-parameter extremum seeking algorithm (m-p ESA) to tune the backstepping controller parameters is presented. The m-p ESA can obtain the optimized value of parameters by minimizing the cost function to achieve the desired performance and avoid the interaction of parameters by the multiple channels design. Therefore the utilization of the m-p ESA can simplify the process of parameter adjustment. In addition, the stability is analyzed by Lyapunov theory The simulation results demonstrate the backstepping controller optimized by the m-p ESA is effective.

For large civil engineering structures and base establishments, for example, bridges, super-high buildings, long-span space structures, offshore platforms and pipe systems of water & gas supply, their lives are up to a few decades or centuries. Damaged by environmental loads, fatigue effects, corrosion effects and material aging, these structures experience inevitably such side effects as damage accumulation, resistance reduction and even accidents. The traditional civil structure is a kind of passive one, whose performance and status are unpredictable to a great extent, but the informatics' introduction breaks a new path to obtain the status of the structure, thus it is an important research direction to evaluate and improve reliability of civil structures by the use of monitoring and health diagnosis technique, and this also assures the security of service for civil engineering structures. Smart material structure, originated from the aerospace sector, has been a research hotspot in civil engineering, medicine, shipping, and so on. For structural health monitoring of civil engineering, the research about high-performance sensing unit of smart material structure is very important, and this will possibly push further the development and application of monitoring and health diagnosis techniques. At present, piezoelectric materials are one of the most widely used sensing materials among the research of smart material structures. As one of the piezoelectric materials, PVDF(Polyvinylidene Fluoride)film is widely considered for the advantages of low cost, good mechanical ability, high sensibility, the ability of being easily placed and resistance of corrosion. However, only a few studies exit about building a mature monitoring system using PVDF. In this paper, for the sake of using PVDF for sensing unit for structural local monitoring of civil engineering, the strain sensing properties of PVDF are studied in detail. Firstly, the operating mechanism of PVDF is analyzed. Secondly, wireless collection system of PVDF is integrated with PVDF film, charge amplifier, wireless transceiver and the corresponding software. Then, with strain gauge as a reference, experiments have also been done to study the quasi-static and dynamic strain response of PVDF, such as sensitivity, linearity and frequency responding, etc. The experimental results show that PVDF is sensitive to the impact response of civil engineering structures, and can provide local monitoring in different frequency response cooperating with a strain gauge. The developed wireless collection system has the characteristics of no lines, saving cost and installation time, and thus further pushes the practical application of PVDF for civil engineering structures.

This work presents a novel vibration piezoelectric energy harvester, which is a micro piezoelectric cantilever with multi-beam. The characteristics of the PZT (Pb(Zr0.53Ti0.47)O3) thin film were measured; XRD (X-ray diffraction) pattern and AFM (Atomic Force Microscope) image of the PZT thin film were measured, and show that the PZT (Pb(Zr0.53Ti0.47)O3) thin film is highly (110) crystal oriented; the leakage current is maintained in nA magnitude, the residual polarisation Pr is 37.037 μC/cm2, the coercive field voltage Ec is 27.083 kV/cm, and the piezoelectric constant d33 is 28 pC/N. In order to test the dynamic performance of the energy harvester, a new measuring system was set up. The maximum output voltage of the single beam of the multi-beam can achieve 80.78 mV under an acceleration of 1 g at 260 Hz of frequency; the maximum output voltage of the single beam of the multi-beam is almost 20 mV at 1400 Hz frequency.

PZT thin film sensors are widely used in microsensor because of its simple structure, high sensitivity and good linearity. The sol-gel method is used for the PZT thin film deposition on the silicon cantilever structure, actuation and sensing electrodes are made on the PZT micro-cantilever by the bulk micromachining technology. AC voltage is applied to the actuation electrode to make the micro-cantilever oscillate at 70 Hz, and the charge on the sensing electrode serves as sensor signal. When contact force is applied to the tip of the micro-cantilever, the amplitude variation of the induced charge is collected by a charge amplifier and a digital lock-in amplifier. Experiments have shown that the PZT thin film sensors with the micro-cantilever structure are able to sense contact force whether the PZT thin film is in the state of stretching stress or compressive stress. The resolution of the microforce sensor is 3 muN at the stretching stress, and 3.75 muN at the compressive stress

The displacement of micro-motion worktable driven by piezoceramic actuator can be measured by self-sensing method in the absence of independent sensor. And a displacement self-sensing control system is made. The free charges on the wafers of piezoceramic actuator contain displacement information. So a displacement self-sensing method of piezoceramic actuator based on integrator is presented. Integrator circuit is designed for gathering free charges on the wafers of actuator. It is convenient to adjust the circuit and easy to acquire sensitive signal using this method. And the impedance mismatching problem met in bridge method is overcome. In order to improve the static and dynamical performance of the piezoceramic micro-motion worktable, the compounding control method by combining feedforward control and feedback control is designed. The self-sensing displacement is the feedback signal of control system of piezoceramic micro-motion worktable. The availability of the self-sensing compounding control is validated by experiment. Comparison experiments among such kind of control, feedforward control, general PID feedback control with independent sensor and compound control with independent sensor are took. The experimental results show that self-sensing compounding control has the same good performance of compounding control with independent sensor.

Different from the conventional piezoelectric microcantilevers with a piezoelectric film sandwiched between two metal electrodes on cantilever structures, the objective of our investigation is to develop a technology platform based on the integration of two piezoelectric PZT thin-film elements on silicon cantilevers for microsensors and microactuators. Two novel piezoelectric microcantilevers with two piezoelectric elements (bimorph or two segments of PZT films) and three electric electrodes were designed. The microcantilevers were successfully fabricated by bulk micromachining. The generated piezoelectric charge and actuation force of the microcantilevers were characterized. As microsensors or microactuators, the bimorph piezoelectric cantilevers can increase the sensing resolution and actuation displacement (force) of the piezoelectric cantilevers. The bimorph and two-segment piezoelectric cantilevers can be served as the versatile devices for sensing and actuation. The multifunctional cantilevers have many potential applications in fields such as micromanipulation systems, microrobots, and vibration controls of flexible manipulators.