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The use of microfluidic chip-based two-dimensional separation holds great promise in the proteomics field, given its portability, simplicity, speed, efficiency, and throughput. However, inclusion of sodium dodecyl sulfate, reported to be necessary for increasing protein-resolving capability, was also accompanied by the loss of both protein conformation and biological function. Here, we describe separation of native proteins by introducing blue native gel electrophoresis into isoelectric focusing and gel electrophoresis (IEF/CGE)-coupled protein two-dimensional microfluidic chip electrophoresis. After assessing the influence of various experimental conditions, the best separation ability and reproducibility of blue native IEF/CGE (IEF/BN-CGE) chip electrophoresis achieved until now were demonstrated no matter whether with a simple simulated mixture or with a complex mixture of total Escherichia coli proteins. Finally, instead of theoretical calculations, the image analysis technique was also used for the first time to quantitatively evaluate the actual peak capacities of chip electrophoresis. According to the number of features abstracted in the electrophoresis patterns, the superiority of the IEF/BN-CGE two-dimensional microfluidic chip electrophoresis was then exhibited quantitatively. The high native protein separation performance makes this established chip electrophoresis method possible for further application in widely needed drug screening, analysis of bio-molecular function, and assays of protein–protein interactions.

Electron exchange between aqueous Fe(II) and structural Fe(III) of iron minerals has been illustrated for understanding the reduction of nitroaromatic compounds (NAC). However, factors influencing Fe(II)-induced the reduction of NAC still remain elusive. In this paper, p-nitrophenol (1.5mM) was selected to explore the effects of pH, the stabilizing ligands (Cl(-), SO4(2-)) of ferrous ions and the extra addition of iron hydroxide on the reduction of NAC via Fe(II) species. The results indicate that the reduction degree of is much lower in SO4(2-) medium than that in Cl(-) medium at pH 7.6. p-Nitrophenol reduction increased in SO4(2-) medium and slightly decreased in Cl(-) medium when Fe hydroxide was extra added. Cl(-) strength (0.01-0.1 mol L(-1)) has no obvious effect on p-NP reduction. SO4(2-) species and its dosage have markedly inhibitory effect on p-NP reduction due to the selective adsorption of SO4(2-) and the formation of sulphated surface complexes on the fresh Fe hydroxide.

Background Association of de novo human leukocyte antigen (HLA) and major histocompatibility complex class I chain-related gene-A (MICA) antibodies and proteinuria with graft survival 5 years after renal transplantation. De novo presence of HLA and MICA antibodies after renal transplantation is associated with poor graft survival. Proteinuria after transplantation is also considered a risk factor for premature graft loss. In this study, we investigated the association of de novo HLA and MICA antibodies on proteinuria after renal transplantation and the association of proteinuria and de novo antibodies with graft survival. Methods We enrolled 275 patients without preexisting HLA and MICA antibodies followed for >5 years after renal transplantation. All donor organs were from living-related donors or from an organ donation program. HLA and MICA antibodies were detected by the Luminex method. Patients with proteinuria (>150 mg/d) underwent intermittent 24-hour proteinuria examination. Results The frequencies of de novo HLA and MICA antibody 5 years after transplantation were 25.8% and 12%, respectively. In total, 26.5% of patients had proteinuria at the 5-year follow-up. De novo HLA antibody was associated with increased proteinuria after transplantation (relative risk, 3.12). HLA antibody and proteinuria were both associated with poor 5-year graft survival (P = .027 and P = .006, respectively). Conclusion De novo HLA and MICA antibodies and proteinuria after renal transplantation are all associated with poor graft survival. De novo HLA antibody is independent risk factor for posttransplant proteinuria, and proteinuria affects the association of de novo antibodies with decreased graft survival after transplantation.

As a kind of recognition molecule, aptamer has been studied and applied widely in numerous science fields in recent years. Immobilized enzymatic reactor has drawn much attention because of its striking advantages, such as high digestion efficiency and ease in coupling with the separation and detection systems. In this study, a novel microfluidic enzymatic chip, which immobilized trypsin based on aptamer, was prepared and proposed. An online analysis platform, which consisted of an aptamer-based chip and high performance liquid chromatography tandem mass spectrometry, was established by using a 6-port valve and applied to protein analysis. The enzymatic capacity and stability performance of chip reactor were characterized by using mixed protein sample, which consisted of bovine serum albumin (BSA), myoglobin (Mb) and cytochrome c (Cyt. c). The sample digestion time of the chip reactor was about 5.76 s while 1 microL/min of flow rate was adopted; and moreover, 5 ng of Mb was identified successfully with the sequence coverage of 37%. Furthermore, the sequence coverages and the relative standard deviations were 44.3% and 6.5% for BSA, 65.0% and 2.7% for Mb, 62.0% and 5.6% for Cyt. c respectively when 500 ng digest of mixed proteins were analyzed in three runs. According to experimental results, the online analysis platform possesses the ability of high sensitivity and good stability, which can provide a promising tool for rapid and high-throughput proteomics study in the near future.

Supplementation with iron-fortified foods is an effective method for treating iron deficiency diseases. However, traditional iron agents used to treat anemia of inflammation (AI) have little effect. In this study, two types of iron liposomes, heme liposomes (HEME-LIP) and ferric citrate liposomes (FAC-LIP), were prepared by the rotary-evaporated film-ultrasonication method, and the encapsulation efficiencies, microstructures, size distributions and zeta potentials were assessed. Both types of iron liposomes showed stable physical characteristics. When used to treat rat models of AI, FAC-LIP and HEME-LIP could increase serum iron levels by 119% and 54% higher than did ferric citrate (FAC) and heme, respectively. Furthermore, the hepcidin, a key regulator of iron homeostasis was up-regulated by these iron liposomes, especially by HEME-LIP. These results indicate that the absorption of iron liposomes was improved over that of unencapsulated iron agents. Thus, iron liposomes may be used to fortify food in treating iron deficiency diseases, especially AI.

SUMMARYThe spore wall ofNosema bombycisplays an important role in microsporidian pathogenesis. Protein fractions from germinated spore coats were analysed by two-dimensional polyacrylamide gel electrophoresis and MALDI-TOF/TOF mass spectrometry. Three protein spots were identified as the hypothetical spore wall protein NbHSWP12. A BAR-2 domain (e-value: 1.35e-03) was identified in the protein, and an N-terminal protein-heparin interaction motif, a potential N-glycosylation site, and 16 phosphorylation sites primarily activated by protein kinase C were also predicted. The sequence analysis suggested thatNbhswp12and its homologous genes are widely distributed among microsporidia. Additionally,Nbhswp12gene homologues share similar sequence features. An indirect immunofluorescence analysis showed that NbHSWP12 localized to the spore wall, and thus we renamed it spore wall protein 12 (NbSWP12). Moreover, NbSWP12 could adhere to deproteinizedN. bombycischitin coats that were obtained by hot alkaline treatment. This novelN. bombycisspore wall protein may function in a structural capacity to facilitate microsporidial spore maintenance.

The stress-inducible immediate early response gene X-1 (IEX-1) regulates cell proliferation and apoptosis in a cell type and stimulus-dependent manner. The aim of this study was to investigate IEX-1 expression in preeclampsia placenta and human umbilical vein endothelial cells (HUVECs) cultured in the serum isolated from preeclampsia patients, to explore its relationship with the pathogenesis of preeclampsia. Thirty preeclampsia patients with 10 cases in the moderate group, 20 cases in the severe group (PE group) and 20 cases of normal pregnant women (control group) were randomly obtained. Reverse transcriptase-polymerase chain reaction and immunohistochemistry showed that IEX-1 expression was significantly higher in preeclampsia patients than in normal pregnant women (p 0.05). There was also a significant difference between moderate and severe preeclampsia (p 0.05). IEX-1 protein was mainly present in cytoplasm of placental trophoblast cells and decidual cells. Compared with the normal pregnancy group, IEX-1 in the preeclampsia group was colored more obviously, and the color was strengthened in correlation with the severity of the disease using immunocytochemial method. Flow cytometry and MTT revealed that the serum isolated from preeclampsia patients appeared to promote IEX-1 expression in cytoplasm of HUVECs, inhibited the proliferation and promoted the apoptosis of HUVECs when compared to the serum of normal pregnant women. Our study demonstrates a correlation of IEX-1 expression levels with the severity of preeclampsia. Given a well-known function of IEX-1 in regulation of apoptosis, IEX-1 may be important in the pathogenesis of preeclampsia by regulating placental villous trophoblast and decidual cell apoptosis as well as participation in endothelial cell injury process.

The microsporidian Nosema bombycis is the pathogen causing pébrine leading to heavy economic loss in sericulture. Little is known of the proteases of microsporidia that are important for both parasite development and pathogenesis. Here we identified a subtilisin-like serine protease NbSLP1 which contains an inhibitor_I9 and a peptidase_S8 domain. Three dimensional modeling of the catalytic domain of the NbSLP1 exhibited a typical 3-layer sandwich structure with S1 pocket substituted by Y(359). Phylogenetic analysis confirms that subtilisin-like serine proteases of microsporidia fall into two clades: SLP1 and SLP2, suggesting the initial subtilisin gene duplication events preceded microsporidia speciation. Furthermore, transcripts of Nbslp1 were detected in the midgut of Bombyx mori infection by N. bombycis by RT-PCR. Antibodies against NbSLP1 recognized both the precursor and mature enzyme by 2D Western blotting. Besides, indirect immunofluorescence assay revealed that the NbSLP1 is mainly localized at the two poles of spore which make the spore look like "safety pins". Remarkably, the mature protease is only detected in the apical region of the spore after germination. These studies demonstrate that NbSLP1 is a conserved subtilisin protease in microsporidia and suggest that NbSLP1 play a significant role in polar tube extrusion process.

Cardiac complication is a major cause of death in hemodialysis patients. The aim of the study was to determine the relationship between plasma catestatin level and cardiac death in those people. A total of 330 maintenance hemodialysis patients were included. Blood samples were collected. Plasma catestatin level was detected by enzyme-linked immunosorbent assay. Fluid status of each patient was expressed by overhydration to total body weight ratio and daily diuresis. Each patient was followed-up for 36 months, unless some of them died in the follow-up period. In the follow-up period, only one hemodialysis patient was lost, 29 patients were died of cardiovascular diseases, 28 patients were died of other diseases and remaining 272 patients survived. Logistic multivariate regression analysis revealed that patients with plasma catestatin level ≥1.9 ng/ml were associated with increased cardiac death risk (RR 6.13, 95% CI 2.54, 18.45), and survival analysis also showed that cardiac death rate in patients with plasma catestatin level ≥1.9 ng/ml was elevated than that in patients with plasma catestatin level

A novel impact-time-control guidance law based on nonlinear kinematics is presented for an interceptor against a stationary target. Exact closed form expression for the guidance command is derived by introducing an indirect variable, and the performance index of a pseudo-control energy has been minimized to increase the robustness against disturbances appeared at the terminal stage. The implementation of the guidance law involves no estimation of time-to-go procedure but provides for exact time-on-target control. The new guidance law called NITCG(Nonlinear Impact-Time-Control Guidance) can be used in salvo attack where multiple missiles are launched for a simultaneous hit on the target. Numerical simulation results are carried out to verify the performance of the guidance law for different engagement scenarios.

The terminal impact-angle-control guidance problem of a homing missile with a physical constraint on seeker's field of view (FOV) limit is considered. The problem is formulated as an infinite horizon non-linear regulator problem with a state constraint, and solved using an extension of the State Dependent Riccati Equation (SDRE) method: first a baseline SDRE controller is designed without taking into account the state constraint, and then a SDRE regulator is designed augmenting the cost functional to take into account the state constraint and the base-line performance. In our design, the state weighting matrix Q is chosen to be a function of time-to-go to include the distance information between the missile and target, and the input weighting matrix R is designed to be a function of the distance of the constrained state to the allowable boundary. Numerical simulations are carried out for different impact angles and seeker's FOV limits, the results of which verify the effectiveness of the proposed guidance approach.

A novel and simple coating method was developed by coating bovine serum albumin (BSA) onto the inner surface of a fused-silica capillary, to avoid the adsorption of analytes during CE. The advantage presented here was that the coating process is more simple, fast, stable, and reproducible. The coated capillary avoided the adsorption of analytes onto the inner surface of a fused-silica capillary and might be a promising candidate for separation of complex biological samples with further development. Meanwhile, the efficiencies of the coated capillary were evaluated by EOF, chromatographic peak shape, and theoretical plate number (N m−1) of RNase A. The optimal coating conditions were obtained from the results. The pH value of coating buffer PB was 4.2, the standing time was 12 h at 4 °C, and the coating concentration of BSA was 1.5 mg mL−1. The stability of the coating on the inner wall of the capillary and the reproducibility of the coated capillaries were good. The theoretical plate number values of RNase A were over 1.3 × 105 (N m−1) in the coated capillary. After successive electrophoresis for 48 h using the coated capillary, the RSD values of EOF and the theoretical plate number were 4.14 % and 9.14 %, respectively. In addition, the RSD values of EOF and the theoretical plate number (N m−1) in the coated capillaries were 13.19 % and 8.96 %, respectively. Finally, the coated capillary was successfully applied to separate the mixture of four basic proteins (RNase A, lysozyme, trypsin and myoglobin).

A method for screening of monoamine oxidase (MAO) inhibitor was carried out using capillary electrophoresis (CE) based on the interaction of MAO and its substrate kynuramine (Kyn). Bioactive proteoliposome was reconstituted by liposome and MAO and then was applied as the pseudostationary phase (PSP) of CE to mimic the interaction between the enzyme and its substrate. N-prolmrgyl-R-2-heptylamine (R-2-HPA) and rasagiline [N-propargyl-1-(R)-aminoindan], which are two kinds of MAO inhibitors, were added into the running buffers containing proteoliposome. The results showed that the relative migration time ratio (RMTR × 10(-1)) values of Kyn were enhanced from 8.88 to 9.31 with an increase of the concentrations of rasagiline from 10(-6) to 1 mM. However, the RMTR values of Kyn were enhanced from 8.83 to 9.14 with an increase of the concentrations of R-2-HPA from 10(-6) to 1 mM. The RMTR value of Kyn in the presence of rasagiline was larger than that in the presence of R-2-HPA when rasagiline and R-2-HPA were at the same concentration. The results indicated that the interaction between Kyn and MAO was weakened with the increase of the inhibitors. In addition, the results of offline incubation showed that the inhibitions of rasagiline were 100.0, 72.1, 51.8 and 5.4% at the concentration of 1, 10(-2), 10(-4) and 10(-6) mM; moreover, the inhibitions of R-2-HPA were 70.0, 44.9, 4.1 and 0.9% at the concentrations of 1, 10(-2), 10(-4) and 10(-6) mM. The inhibition efficiency of rasagiline was stronger than that of R-2-HPA at the same concentration. Additionally, the interaction between Kyn and liposome was also investigated. This newly developed method might provide a potential tool for screening MAO inhibitor.

In this study, a new strategy for screening of enzyme inhibitors based on the interaction between enzyme and substrate was established. The bioactive proteoliposome reconstituted by monoamine oxidase (MAO) and liposome was added in the running buffers of capillary electrophoresis (CE) as pseudostationary phase to simulate the interaction between MAO and its substrate kynuramine (Kyn). The results showed that the relative migration time ratio (RMTR) of Kyn decreased obviously with increase of the proteoliposome, indicating strong interaction between proteoliposome and Kyn. After adding MAO inhibitors into the running buffers containing proteoliposome, the decline of the RMTR value of Kyn became slower. This occurred because MAO was inhibited to some degree by the inhibitors, weakening the interaction between MAO and Kyn. However, adding compounds which do not inhibit MAO activity into the running buffers did not affect the RMTR of Kyn obviously. The results obtained demonstrate that the method developed could characterize the inhibition of MAO inhibitors. Compared with traditional inhibitor screening methods, it does not need incubation outside the column to complete the enzyme catalytic reaction, so analysis is faster and less sample is consumed. With further development, the method might be chosen as a useful tool for screening of membrane protein inhibitors and identification of proteins associated with various diseases.

In this paper, photoinduced protein immobilizations on quartz glass slides utilizing benzophenone as photoinitiator without any photoactive group derivatizations involved were developed. Three different methods mediated by benzophenone were investigated, including protein photo-attachment onto untreated glass surface, protein attachment onto glass surface by reacting with pre-photografted maleic anhydride, and protein photo-attachment onto alkylamino silane functionalized glass surface, respectively. Protein immobilizations were characterized by fluorescence microscopy and atomic force microscopy. The preservation of biological activity after protein photo-attachments was confirmed by immunoassays. Area-defined protein immobilization was also primarily investigated. Comparative studies demonstrated that in respect of immobilization density, coverage homogeneities and photo-localization, protein photocoupling to amino-terminated quartz glass surfaces remarkably outperformed other photoinduced methods, as well as one kind of 3-(triethoxysilyl) propyl isocyanate chemical protein immobilization. The feasibility of this protein photo-immobilization on silicon-based materials is promising for widespread application because of its simplicity and effectiveness.

Polypyrrole (PPy) and WO3 were prepared by chemical oxidation polymerization and emulsion method, respectively. PPy/WO3 hybrid materials with different PPy mass percent were prepared by mechanical mixing. WO3 and PPy/WO3 hybrids were characterized by XRD, SEM and HRTEM, and the sensitivity studies of PPy, WO3 and PPy/WO3 to toxic gases NH3, H2S, NOx were also carried out. It was found that the particle size of WO3 was about 29 nm, and PPy/WO3 was about 50–100 nm. Gas sensitivity tests showed that when operated at 90 °C, PPy/WO3 hybrids had better reversibility to H2S than PPy higher sensitivity and selectivity to H2S than WO3. PPy/WO3 materials had short response–recovery time to H2S and their sensitivities had linear relationship with the concentration of H2S. The sensitive mechanism of PPy/WO3 materials to H2S may be the co-effect of proton doping process and the n-type semiconductor's effect.

In the present study, a new enzymatic microreactor is developed by immobilizing trypsin on the matrix-HILIC (hydrophilic interaction chromatography). The influences of different types of buffer solutions (borate, Tris·HCl, and phosphate), their concentrations and pH values on the immobilizing rate and the activity of immobilized trypsin were investigated. The optimal condition for off-line immobilization of trypsin on matix-HILIC was Tris·HCl buffer (20 mmol/L, pH 8.0), and then, under the optimal condition, an enzymatic microreactor was prepared by trypsin dynamically immobilized on HILIC matrix which was packed in a Peeksil column. It can be found that the microreactor possessed high hydrolytic activity according to the hydrolysis results of N-α-benzoyl-DL-arginine P-nitroanilide (BAPNA), a substrate of trypsin. Moreover, bovine serum albumin (BSA), cytochrome C and myoglobin as standard proteins were chosen to characterize the enzymolysis of the microreactor demonstrating the high enzymolysis efficiency by this enzymatic microreactor. Besides, the deactivated trypsin can be easily eluted from the matrix-HILIC to re-immobilize trysin, which is highly cost-effective in the experiment. All the results show that the new enzymatic microreactor associated with HPLC-MS techniques can realize online enzymolysis of proteins, and the time to analyze and identify the proteins is greatly shortened.

Monolayer of 3-(triethoxysilyl) propyl isocyanate was prepared on the slide by self-assembled technique. X-ray photoelectron spectroscopy (XPS) was employed to analyze the elementary composition of the film. Contact angle of distilled water was measured to characterize the surface state. It was shown that 3-(triethoxysilyl) propyl isocyanate had been successfully assembled on the slide. The increase of contact angle to 80° demonstrated that the hydrophobicity of the surface of chip was increased significantly. Moreover, further self-assembly of bovine serum albumin (BSA) on 3-(triethoxysilyl) propyl isocyanate was also carried out with the advantages such as simple and convenient preparation. Therefore, the potential of broader applications in the modification of micro-channel in the μ-TAS system, the immobilization of protein or peptide and the surface modification of materials are all expectative.

The biochemical indicators of wistar rat under low oxygen concentration, such as brain water content, necrosis, lactic acid and Na+-K+-ATPase, was detected to evaluate normobaric hypoxia-induced brain damage and to investigate the mechanism of wistar rat brain injury. Histopathological changes in brain tissue induced by hypoxia were investigated via hematoxylin and eosin stain (HE). Hypoxia induced factor-1α (HIF-1α) expression in brain was confirmed using immunohistochemistry. The results showed that the level of lactic acid was positively correlated with the degree of hypoxia, while concentration-dependent decrease in total Na+-K+-ATPase activity was observed. Compared with the control group, hypoxia group had a significant difference on brain water content under severe hypoxic conditions, the rate of brain necrosis increased obviously, followed by the increase of lactic acid level and the decrease of Na+-K+-ATPase activity. Histopathological analysis of brain confirmed that there was neuronal cell death in hippocampal gyrus. HIF-1α expression enhanced the hypoxia adaptation capability of the rat model through regulating the expressions of multiple genes. Lactic acid, Na+-K+-ATPase and HIF- 1α played an important role in brain injury as a possible mechanism.

This paper describes non-gel capillary sieving electrophoresis employing semi-crosslinked polyacrylamide as a high performance and low viscous replaceable separation matrix for separation of non-denatured protein separation. Arising from the fine sieving and dynamic coating ability of this polymer, a mixture of basic proteins lysozyme, cytochrome C, ribonuclease A, and trypsin was resolved with excellent reproducibility. Mixing different semi-crosslinked polyacrylamides together further improves the separation. The separtion mechanism was analyzed. With network structure developed to an intermediate state between crosslinked gel and linear polymer solutions, these semi-crosslinked olyacrylamide polymers demonstrate a promise as a new class of size sieving separation medium, not only in capillary electrophoresis, but also in microfluidic chip separation schemes.

A polyaniline (PAni)/SnO2 hybrid material was prepared by a hydrothermal method and characterized by X-ray diffraction (XRD), Fourier transform infrared (FT-IR), scanning electron microscopy (SEM) and high-resolution transmission electron microscopy (HRTEM). The XRD pattern suggested that PAni did not modify the crystal structure of SnO2, but SnO2 affected the crystallization of PAni to some extent. The gas sensitivity of the PAni/SnO2 hybrid was also studied to ethanol and acetone at operation temperatures of 30, 60 and 90 °C. It was found that the PAni/SnO2 hybrid material had gas sensitivity only when operated at 60 and 90 °C, and it showed a linear relationship between the responses and the concentrations of ethanol and acetone at 90 °C. The sensing mechanism was also discussed.

To investigate the immunostimulatory role of Propionibacterium acnes (P. acnes) in the differentiation of dendritic cell maturation and T helper (Th) subsets, and further explore the potential mechanism of its granuloma-inducing effect.After the mouse bone marrow-derived dendritic cells (BMDCs) were stimulated by P. acnes, the expressions of co-stimulatory molecules CD40, CD80, CD86 and major histocompatibility complex II (MHCII) on BMDCs were detected by flow cytometry, the mRNA levels of inflammatory cytokines IL-12, IL-6 and IL-23 were measured by real-time quantitative PCR (qRT-PCR), and the protein concentrations of corresponding cytokines were analyzed by ELISA. Thereafter, BMDCs were cultured in the presence of P. acnes and CD4(+) T cells, and the percentages of Th1 and Th17 cells were detected by flow cytometry. In addition, murine model of liver granuloma was constructed by injecting heat-killed P. acnes through tail vein, and the mRNA levels of IL-17 and IFN-γ were measured by qRT-PCR.P. acnes up-regulated the expressions of CD40, CD80, CD86 and MHCII on BMDCs, and the mRNA and protein levels of the IL-12, IL-6 and IL-23 cytokines were also higher on P. acnes-treated BMDCs compared with non-stimulated BMDCs. Furthermore, the percentages of Th1 and Th17 cells in the co-culture system increased. In addition, the mRNA levels of IL-17 and IFN-γ were elevated obviously in the liver with abundant granulomas after P. acnes treatment.P. acnes could activate BMDCs, promote the differentiation of Th1 and Th17 cells, and further induce the formation of granulomas.

Objectives: To investigate the mechanism and effect of sevoflurane on learning and memory function in transgenic mice model of Alzheimer’s disease (AD). Methods: A total of 45 Tg2576 mice were used and randomly assigned to control, sham, and sevoflurane group. Spatial learning and memory ability were measured before and after sevoflurane exposure using morris water maze (MWM) and Y-maze behavioral tests. Moreover, TUNEL assay was carried out to determine the cell death in hippocampal cornuammonis (CA) 1, CA3, and dentate gyrus (DG) region. Apoptosis-related protein (caspase-3 and Bcl-xL) expression in the hippocampus was analyzed by Western blotting. Results: The MWM results showed that there were no significant differences in the swimming speed after sevoflurane exposure among the three groups. However, the escape latency, time spent in original quadrant, and the number of correct trials (Y-maze) were significantly lower after sevoflurane anesthesia exposure in the sevoflurane group than the sham group and control group (P < 0.05). Besides, the apoptotic cell numbers of the CA1 and CA3 region in the sevoflurane group were significantly higher than those in the sham group and control group (P < 0.05). Western blotting results showed that the protein expression levels of Bcl-xL were significantly higher, but the caspase-3 levels were significantly lower in the sevoflurane group than those in the control group and sham group (both P < 0.05). Conclusion: Our results indicate that sevoflurane might impair acquisition learning and memory function in AD by induction of hippocampal neuron apoptosis.

Microsporidia are obligate intracellular parasites with rigid spore walls that protect against various environmental pressures. Despite an extensive description of the spore wall, little is known regarding the mechanism by which it is deposited or the role it plays in cell adhesion and infection. In this study, we report the identification and characterization of two novel spore wall proteins, SWP7 and SWP9, in the microsporidian species Nosema bombycis . SWP7 and SWP9 are mainly localized to the exospore and endospore of mature spores and the cytoplasm of sporonts, respectively. In addition, a portion of SWP9 is targeted to the spore wall of sporoblasts earlier than SWP7 is. Both SWP7 and SWP9 are specifically colocalized to the spore wall in mature spores. Furthermore, immunoprecipitation, far-Western blotting, unreduced SDS-PAGE, and yeast two-hybrid data demonstrated that SWP7 interacted with SWP9. The chitin binding assay showed that, within the total spore protein, SWP9 and SWP7 can bind to the deproteinated chitin spore coats (DCSCs) of N. bombycis . However, binding of the recombinant protein rSWP7-His to the DCSCs is dependent on the combination of rSWP9–glutathione S -transferase (GST) with the DCSCs. Finally, rSWP9-GST, anti-SWP9, and anti-SWP7 antibodies decreased spore adhesion and infection of the host cell. In conclusion, SWP7 and SWP9 may have important structural capacities and play significant roles in modulating host cell adherence and infection in vitro . A possible major function of SWP9 is as a scaffolding protein that supports other proteins (such as SWP7) that form the integrated spore wall of N. bombycis .

Polypyrrole (PPy) was prepared by chemical oxidation polymerization, analyzed by FT-IR, elemental analysis and HRTEM, and studied for gas sensitivity. It suggested that PPy had sensitivity to NH 3 , H 2 S and NO x , and showed irreversibility to NO x gas. The organic–inorganic hybrid materials PPy/ZnO with different PPy weight percents were prepared by mechanical mixing, and the sensitivity study of the materials to toxic gases NH 3 , H 2 S, NO x was carried out at different operating temperatures 30, 60, and 90 °C. It was found that PPy/ZnO materials (PPy(1%)/ZnO, PPy(3%)/ZnO, PPy(5%)/ZnO, PPy(10%)/ZnO, PPy(20%)/ZnO) had better selectivity and reversibility to NO x than pure PPy, and much lower working temperature than the reported working temperature of ZnO (about 350–450 °C). Their sensitivity increased with the increasing concentration of NO x at particular working temperature, and among them PPy(10%)/ZnO had the maximum sensitivity to NO x in the same condition. They showed no response to 1000, 1500, 2000 ppm NH 3 or H 2 S. The response mechanism of PPy/ZnO materials to NO x was discussed.

In this work, a H 2 S sensor formed from hybrid of WO 3 and polypyrrole (PPy) by mechanical mixing was produced. The gas-sensing properties of the hybrid materials were studied and compared with those of PPy and WO 3 . It was found that PPy/WO 3 materials can complement the drawbacks of pure PPy and WO 3 to some extent. PPy/WO 3 materials with varies PPy mass percent (1%, 3%, 5%, 10%, 20%) had better reversibility than PPy, and also had higher sensitivity than WO 3 when operated at 90 °C. The sensing mechanism of PPy/WO 3 materials to H 2 S was presumed to be the synergism of PPy and WO 3 or the effect of p–n heterojunctions.

The Au/SnO2 catalysts were prepared by a co-precipitation method. The structure of the sample treated at different temperatures was investigated by means of HRTEM and XPS. The structure of the samples after various treatments and their activity in the CO oxidation were compared. The results showed that the catalytic behavior was related to the particle size of gold and surface oxygen species on the support. Highly dispersed gold particles and adsorbed surface oxygens and hydroxyl groups on the support were responsible for the high catalytic activity of the Au/SnO2 catalyst.

The fact that different gamma-zirconium phosphate (gamma-ZrP) preintercalation method induced varied degree and type of conformational change of the adsorption protein was confirmed by characterization techniques including circular dichroism (CD), fourier transform infrared spectroscopy (FTIR) and X-ray powder diffraction (XRD) analysis. The results indicated that the association of hemoglobin with gamma-ZrP preintercalated using butylamine was correlated with conformational change in the secondary structure of the protein. gamma-ZrP which was preintercalated with tetra (n-butylammonium) hydroxide caused the conformational change of Hemoglobin in both the secondary structure and the tertiary structure. X-ray powder diffraction analysis was used to analyze the crystalline structure of the nanocomposites prepared by relamination. The adsorption isotherms of Hemoglobin on different matrices were set up and fitted with Langmuir and Freundlich equations.

A new capillary coating of alternate γ-zirconium phosphate lysozyme fabricated for open-tubular electrochromatography enantioseparation. The chiral compounds D, L-tryptophan tested were enantiomerically resolved. The influences brought by experimental parameters, such as organic modifier, temperature, applied voltage and pH of running buffer on the performance of the column, were investigated.

This paper presents a fast nonsingular integral terminal sliding mode control (FNITSMC) for a class of nonlinear systems. First, a novel fast nonsingular integral terminal sliding mode surface (FNITSMS) is designed by introducing power integral terms which contain a boundary-like structure. Next, by employing FNITSMS and the equivalent control technique, the FNITSMC is proposed to achieve the finite-time convergence of the system states. Compared with many existing works on terminal sliding mode control (TSMC), the proposed FNITSMC exhibits three attractive features: (i) can avoid the singularity problem without any constraint, and provide faster responses by tuning the parameters in FNITSMS; (ii) for constant or eventually constant disturbances, zero steady-state error can be achieved when a boundary layer is used to alleviate chattering; (iii) the switching gain is only required to be designed greater than the bound of the disturbance. Finally, Simulation results of both the numerical and application examples show the effectiveness of the proposed control method.

An immobilized pH gradient was directly constructed on the inner wall of a microfluidic chip channel by photoimmobilizing focused carrier ampholytes onto the wall. A mixture of carbonic anhydrase, myoglobin, and trypsin inhibitor was successfully isoelectric-focused and separated with good linearity between the pI values of proteins and the location of the focused bands. Furthermore, coating methods for the resistance of protein nonselective adsorption and simultaneously for pH gradient photocoupling were screened. The PEG-silane coating method was found to be better than the cross-linked polyacrylamide coating and aminosilane modification methods. Finally, based on the open tubular column mode of carrier ampholytes' immobilization and effective antiadsorption coating, the immobilized pH gradient was reused and the chip was recycled for the first time. By virtue of its remarkable features including simplicity, convenience, high efficiency of protein enrichment and separation, and potential for coupling site-selective IEF with other analytical or separation techniques, this novel method promises to be useful in several applications related with zwitterionic biomolecules.

In this paper, a robust adaptive sliding mode controller is proposed for a class of uncertain nonlinear multi-input multi-output (MIMO) systems. The upper bounds of the uncertainties are not needed in the procedure of the controller design, and the controller is continuous and guarantees the tracking error can converge to a small residual set. Furthermore, the size of the tracking error at steady state can be specified a prior and guaranteed by choosing certain design parameters. Finally, a simulation study based on a two-link rigid robotic manipulator model is used to illustrate the effectiveness of the proposed controller.

A new type of high-throughput and parallel optical sensing platform with a single-color probe based on microfluidic chip electrophoresis combined with aptamer–carboxyfluorescein/graphene oxide energy transfer is reported here. Label-free protein multi-targets were detected, even in challenging complex samples without any pre-treatment.

A system of the signal acquisition and data processing used for light-emitting diodes induced fluorescence (LED-IF) detection in our laboratory was described in this paper. A high-speed data acquisition module was developed. The function of signal acquisition in real-time and data processing was realized utilizing the graphical programming environment LabVIEW. To reduce the distorted come from the technical limitations of the equipment, low-pass Butterworth filter and wavelet filter were applied in this system. Some FITC-derived amino acids were separated coupled with HPLC. The result had been analyzed and the report was finalized automatically.

As a kind of recognition molecule, aptamer has been studied and applied widely in numerous science fields in recent years. Immobilized enzymatic reactor has drawn much attention because of its striking advantages, such as high digestion efficiency and ease in coupling with the separation and detection systems. In this study, a novel microfluidic enzymatic chip, which immobilized trypsin based on aptamer, was prepared and proposed. An online analysis platform, which consisted of an aptamer-based chip and high performance liquid chromatography tandem mass spectrometry, was established by using a 6-port valve and applied to protein analysis. The enzymatic capacity and stability performance of chip reactor were characterized by using mixed protein sample, which consisted of bovine serum albumin (BSA), myoglobin (Mb) and cytochrome c (Cyt. c). The sample digestion time of the chip reactor was about 5.76 s while 1 microL/min of flow rate was adopted; and moreover, 5 ng of Mb was identified successfully with the sequence coverage of 37%. Furthermore, the sequence coverages and the relative standard deviations were 44.3% and 6.5% for BSA, 65.0% and 2.7% for Mb, 62.0% and 5.6% for Cyt. c respectively when 500 ng digest of mixed proteins were analyzed in three runs. According to experimental results, the online analysis platform possesses the ability of high sensitivity and good stability, which can provide a promising tool for rapid and high-throughput proteomics study in the near future.

T-helper (Th) 22 and Th17 cells are involved in the pathogenesis of autoimmune diseases. However, the role of Th22 and correlation with Th17 cells in the pathophysiology of IgA nephropathy (IgAN) remain unknown. In our study, Th22 and Th17 cells in peripheral blood of IgAN patients, non-IgA mesangial proliferative glomerulonephritis (non-IgA MsPGN) patients, and healthy controls were measured by flow cytometry. The concentration of plasma interleukin-22 (IL-22) was examined by enzyme linked immunosorbent assay (ELISA). The results showed that Th22 cells, Th17 cells, and plasma IL-22 were significantly elevated in IgAN patients compared with non-IgA MsPGN patients and healthy controls. Th22 cells showed a positive correlation with the levels of plasma IL-22 in IgAN patients. Moreover, a significantly positive correlation between Th22 cells and Th17 in IgAN patients was observed. Furthermore, IgAN patients with proteinuria showed a higher percentage of Th22 cells than IgAN patients without proteinuria. Our data demonstrated that IgAN had increased frequencies of peripheral Th22, Th17 cells and plasma IL-22, indicating that Th22 along with Th17 cells are involved in the immune responses of IgAN.

Microsporidia are a group of eukaryotic intracellular parasites that infect almost all vertebrates and invertebrates. The microsporidian invasion process involves the extrusion of a unique polar tube into host cells. Both the spore wall and the polar tube play an important role in microsporidian pathogenesis. So far, five spore wall proteins (SWP1, SWP2, Enp1, Enp2, and EcCDA) from Encephalitozoon intestinalis and Encephalitozoon cuniculi and five spore wall proteins (SWP32, SWP30, SWP26, SWP25, and NbSWP5) from the silkworm pathogen Nosema bombycis have been identified. Here we report the identification and characterization of a spore wall protein (SWP5) with a molecular mass of 20.3 kDa in N. bombycis . This protein has low sequence similarity to other eukaryotic proteins. Immunolocalization analysis showed SWP5 localized to the exospore and the region of the polar tube in mature spores. Immunoprecipitation, mass spectrometry, and immunofluorescence analyses revealed that SWP5 interacts with the polar tube proteins PTP2 and PTP3. Anti-SWP5 serum pretreatment of mature spores significantly decreased their polar tube extrusion rate. Taken together, our results show that SWP5 is a spore wall protein localized to the spore wall and that it interacts with the polar tube, may play an important role in supporting the structural integrity of the spore wall, and potentially modulates the course of infection of N. bombycis .

The stress-inducible immediate early response gene X-1 (IEX-1) regulates cell proliferation and apoptosis in a cell type and stimulus-dependent manner. The aim of this study was to investigate IEX-1 expression and its role in apoptosis of ovarian epithelial tumors for potential use in clinical diagnosis and therapy.IEX-1 expression was examined in paraffin-embedded specimens from 77 patients with epithelial ovarian tumors using immunohistochemistry. Correlation between IEX-1 expression and other clinicopathological parameters was evaluated. Apoptosis of tumor cells was detected by terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL).IEX-1 expression was significantly lower in ovarian cancers compared to cystadenomas and borderline tumors (p.05). The expression was significantly associated with FIGO stage and histological grade (p.05), but not with age, histological type, or residual tumor (p.05). A positive correlation was also observed between IEX-1 expression and apoptotic index (p.01) or survival (p=.005).With the development of epithelial ovarian tumors from benign to malignant, IEX-1 expression is decreased, concomitant with a decreased rate of cell apoptosis. Thus, IEX-1 is pro-apoptotic in the development of epithelial ovarian cancer. The pro-apoptotic activity may take part in restraining tumor growth at the early stage of ovarian epithelial cancer, whereas its decreased expression probably contributes to the abnormal survival advantage for malignant cancer. Altered IEX-1 expression can potentially be a new predictor of the malignant transformation and a prognostic indicator for cancer therapy.

The purpose of this study was to develop a new method to assess the activity and inhibition of the immobilized enzyme trypsin based on the frontal analysis of enzymatic reaction products. This novel technique was performed by on-line monitoring of the absorption at 253 nm of N-benzoyl-L-arginine (BA) from the hydrolysis of N-benzoyl-L-arginine ethyl ester (BAEE). A microreactor was constructed by immobilizing trypsin on an immobilized artificial membrane (IAM)-packed column. Trypsin was non-covalently and dynamically immobilized on-line in the hydrophobic interface of an IAM liquid chromatographic stationary phase. The trypsin-IAM stationary phase was bioactive. Due to the enzymatic reaction, the substrate of BAEE was completely hydrolyzed when the BAEE concentration was below 0.5 mmol/L and partly hydrolyzed when the BAEE concentration ranged from 0.75 to 1.0 mmol/L. By the addition of soybean trypsin inhibitor (STI), phenylmethane sulfonyl fluoride (PMSF), and benzamidine into the substrate solution, the results obtained from the frontal analysis showed that the activity of trypsin on IAM was strongly inhibited not only by STI but by both benzamidine and PMSF.

Background. Renal transplantation is currently the prevalent therapy for most patients with end-stage renal disease. No clinical markers for such rejection have been universally accepted. We aimed to investigate the possibility of use of human leukocyte antigen (HLA) class I (ABC) on peripheral blood CD3+/CD8+ T lymphocytes as a marker of acute rejection. Methods. For recipients undergoing renal transplantation from September 2007 to November 2008, peripheral blood samples were obtained pretransplantation and at days 3 and 7 posttransplantation when the patients were still hospitalized and at weeks 2 and 3 and months 1, 2, 3, and 6 posttransplantation. For patients with fever, lumbodynia, gross hematuria, or oliguria after transplantation, blood samples were collected immediately before and at days 3 and 7 after the administration of anti-inflammatory regents. The level of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes was measured on flow cytometry. Results. For the 79 transplant recipients, the level of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes was consistently elevated during the first 3 weeks after transplantation, declined gradually to pretransplantation levels, then tapered off and remained stable. Patients experiencing acute rejection (AR) or not after transplantation did not differ in level of HLA class I (ABC) up to 6-month follow-up, except at days 14 and 21 after transplantation, when the level was higher for patients experiencing AR (P Conclusions. Upregulation of HLA class I (ABC) on peripheral-blood CD3+/CD8+ T lymphocytes could be used as an accurate and reliable predictor of AR after renal transplantation.