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The binding reaction between vitamin B12 (B12, cyanocobalamin) and lysozyme (Lys) has been investigated by fluorescence, synchronous fluorescence, ultraviolet–vis (UV) absorbance, and three-dimensional fluorescence. The intrinsic fluorescence of Lys was strongly quenched by the addition of B12 in different pH buffer solutions (pH 3.4, 7.4, and 9.0) and the spectroscopic observations are mainly rationalized in terms of a static quenching process at lower concentration of B12 ( C B12 / C Lys C B12 / C Lys > 5). The structural characteristics of B12 and Lys were probed, and their binding affinities were determined under different pH conditions (pH 3.4, 7.4, and 9.0). The effect of B12 on the conformation of Lys was analyzed using UV, synchronous fluorescence and three-dimensional fluorescence under different pH conditions. These results indicate that the binding of B12 to Lys causes apparent change in the secondary or tertiary structures of Lys. Furthermore, the effect of Zn 2+ on the binding constant of B12 with Lys under various pH conditions (pH 3.4, 7.4, and 9.0) was also studied.

We have designed a novel nerve guidance conduit (NGC) made from silk fibroin and poly(lactic-co-glycolic acid) through electrospinning and weaving (ESP-NGCs). Several physical and biological properties of the ESP-NGCs were assessed in order to evaluate their biocompatibility. The physical properties, including thickness, tensile stiffness, infrared spectroscopy, porosity, and water absorption were determined in vitro. To assess the biological properties, Schwann cells were cultured in ESP-NGC extracts and were assessed by morphological observation, the MTT assay, and immunohistochemistry. In addition, ESP-NGCs were subcutaneously implanted in the backs of rabbits to evaluate their biocompatibility in vivo. The results showed that ESP-NGCs have high porosity, strong hydrophilicity, and strong tensile stiffness. Schwann cells cultured in the ESP-NGC extract fluids showed no significant differences compared to control cells in their morphology or viability. Histological evaluation of the ESP-NGCs implanted in vivo indicated a mild inflammatory reaction and high biocompatibility. Together, these data suggest that these novel ESP-NGCs are biocompatible, and may thus provide a reliable scaffold for peripheral nerve repair in clinical application.

The process of Schwann cells (SCs) forming a sheath around axons is termed as myelination, which plays a pivotal role for proper physiological function in the peripheral nervous system (PNS). The molecular mechanisms regulating SC myelination in the PNS remain to be elucidated. Here, we show that AMP-activated protein kinase (AMPK) in sciatic nerves was gradually decreased during the PNS myelination process. Pharmacological interventions showed that activation of AMPK by AICAR attenuated myelin gene expression in SCs, whereas inhibition of AMPK by Compound C (ComC) or AMPKα1 knockdown stimulated myelin gene expression. Following experiments revealed that c-Jun, a negative modulator of PNS myelination, was activated by AMPK in SCs. The application of ComC in newborn rats markedly downregulated c-Jun expression in sciatic nerves. The lipid and protein synthesis in sciatic nerves was greatly potentiated by ComC. As a consequence, myelin gene expression in sciatic nerves, as well as myelin sheath thickness, were promoted in the ComC-treated rats. All together, our data identify that AMPK is an important negative regulator of Schwann cell myelination in the PNS, and this regulation role may rely on c-Jun activation.

Incorporation of nerve growth factor (NGF) into a nerve conduit can improve peripheral nerve regeneration. Here, genipin, a natural and low toxic agent, was used to crosslink chitosan, a natural polysaccharide, and concurrently to immobilize NGF onto modified chitosan, followed by fabrication of chitosan (CS)-genipin (GP)-NGF nerve conduits. MTT test showed that the cell viability of Schwann cells cultured in the conduit extract was not significantly different from that in plain medium. The neurite outgrowth measurement and immunocytochemistry with anti-growth-associated protein-43 and anti-neurofilament indicated that NGF released from CS-GP-NGF nerve conduits retained the bioactivity of stimulating neuronal differentiation of PC12 cells. Fracture strength measurements and vitamin B12 release analysis confirmed that CS-GP-NGF nerve conduits possessed good mechanical properties and adequate permeability. We also investigated the in vitro release kinetics of NGF from CS-GP-NGF nerve conduits by ELISA. The continuous release profile of NGF, within a 60-day time span, consisted of an initial burst that was controlled by a concentration gradient-driven diffusion, followed by a zero-order release that was controlled by a degradation of chitosan matrix. Collectively, CS-GP-NGF nerve conduits had an integrated system for continuous release of NGF, thus holding promise for peripheral nerve repair applications.

Background. Tissue-engineered nerve grafts (TENGs) constitute a promising alternative to nerve autografts that are recognized as the gold standard for surgical repair of peripheral nerve gaps. Objective. To investigate the feasibility of using TENGs for bridging extra large peripheral nerve gaps in large animals. Methods. TENGs were constructed by incorporating autologous bone marrow mesenchymal stem cells (MSCs) into a neural scaffold that consisted of a chitosan conduit inserted with poly(lactic-co-glycolic acid) (PLGA) fibers. A 60-mm-long sciatic nerve gap in dogs was bridged by TENGs, chitosan/PLGA scaffolds, or nerve autografts. At 12 months postsurgery, behavioral analysis, electrophysiology, retrograde fluorogold tracing, and histological examination were performed. Results. The outcomes of TENGs were similar to those of autografts and better than those of scaffolds alone. Conclusion. Introduction of autologous MSCs to a chitosan/PLGA scaffold improved the repair and rehabilitation of a large gap after peripheral nerve injury in dogs. Autologous MSCs may be a source of support cells for neural tissue engineering.

Tissue-engineered nerve grafts (TENGs), typically consisting of a neural scaffold included with support cells and/or growth factors, represent a promising alternative to autologous nerve grafts for surgical repair of large peripheral nerve gaps. Here, we developed a new design of TENGs by introducing bone marrow mesenchymal stem cells (MSCs) of rats, as support cells, into a silk fibroin (SF)-based scaffold, which was composed of an SF nerve guidance conduit and oriented SF filaments as the conduit lumen filler. The biomaterial SF had been tested to possess good biocompatibility and noncytoxicity with MSCs before the TENG was implanted to bridge a 10-mm-long gap in rat sciatic nerve. Functional and histological assessments showed that at 12 weeks after nerve grafting, TENGs yielded an improved outcome of nerve regeneration and functional recovery, which was better than that achieved by SF scaffolds and close to that by autologous nerve grafts. During 1-4 weeks after nerve grafting, MSCs contained in the TENG significantly accelerated axonal growth, displaying a positive reaction to S-100 (a Schwann cell marker). During 1-3 weeks after nerve grafting, MSCs contained in the TENG led to gene expression upregulation of S100 and several growth factors (brain-derived neurotrophic factor, ciliary neurotrophic factor, and basic fibroblast growth factor). These results suggest that the cell behaviors and neurotrophic functions of MSCs might be responsible for their promoting effects on peripheral nerve regeneration.

Neuroprotective agents have been in the focus of attention in the treatment of ischemic stroke. Salidroside, a phenylpropanoid glycoside isolated from Rhodiola rosea L., possessed a wide range of biological activities, especially neuroprotection. In an attempt to improve neuroprotective effects of new salidroside analogs for ischemic stroke, a series of novel aralkyl alcoholic 2-acetamido-2-deoxy-β-d-pyranosides were synthesized and their protective activities against the hypoglycemia and serum limitation induced cell death in rat pheochromocytoma cells (PC12 cells) were studied. Most compounds showed strong neuroprotective effects, especially for 4g and 4h, which exhibited a great potency superior to salidroside. MTT assay, Hoechst 33342 staining, and flow cytometry with annexin V/PI staining collectively showed that pretreatment with 4g and 4h attenuated cell viability loss and apoptotic cell death in cultured PC12 cells. Caspase-3 colorimetric assay and Rhodamine 123 staining revealed the changes in expression levels of caspase-3 and mitochondrial membrane potential in PC12 cells on exposure to hypoglycemia and serum limitation with and without 4g and 4h pretreatment, respectively. All the results suggested that 4g and 4h protects the PC12 cells against hypoglycemia and serum limitation induced apoptosis possibly by modulation of apoptosis-related gene expression and restoration of the mitochondrial membrane potential. Therefore, these novel findings may provided a new framework for the design of new aralkyl alcoholic 2-acetamido-2-deoxy-β-d-pyranosides as neuroprotective agents for treating cerebral ischemic stroke and neurodegenerative diseases.

In this study, a chitosan conduit loaded with bone marrow stromal cells (BMSCs) was developed to bridge the gap in the transected spinal cord of adult rats, and the nerve repair outcomes were evaluated by functional and histological techniques at 12 weeks after implantation. As compared to chitosan conduits alone, incorporation of BMSCs within chitosan conduits yielded additional improving effects on nerve regeneration and function restoration. The measurements with the Basso, Beattie and Bresnahan locomotor rating scale or of motor evoked potentials indicated that motor functional recovery was enhanced; retrograde tracing confirmed that the ascending tract was regenerated and the neural pathway was established; and histological analyses revealed that axon growth and remyelination in the regenerated nerve was promoted. The three-dimensional reconstruction showed that the chitosan conduit loaded with BMSCs significantly reduced the spinal cord cavity volume at the injured site. Taken together, the results collectively suggest that implantation with BMSCs-loaded chitosan conduits may become a promising approach to the repair of spinal cord injury.

Chitooligosaccharides (COSs), the biodegradation product of chitosan, possess a wide range of biological activities. In this study, we investigated the influences of COSs on primary cultured cortical neurons exposed to glucose deprivation (GD). The cell viability assessment by MTT assay, in couple with cell apoptosis analysis by Hoechst 33342 and TUNEL staining, indicated that GD-induced cell apoptosis in cultured cortical neurons was attenuated by COSs co-treatment in a dose-dependent manner. Light micrography following tetramethylrhodamine methyl ester staining revealed that COSs protected cultured cortical neurons from GD insult through the stabilization of mitochondrial membrane potentials. COSs co-treatment also led to the increase in Bcl-2/Bax protein ratio and the inhibition of caspase-3 activation in cultured cortical neurons exposed to GD insult. We further found that COSs were able to transiently cause the activation of Akt and ERK1/2 proteins, and anti-apoptotic effects of COSs could be blocked by chemical inhibition of PI3K and MEK. Taken together, the results suggest that COSs hold a promise to serve as a potential neuroprotective agent for treating cerebral ischemic stroke and neurodegenerative diseases.

The aim of this study was to investigate the changes in the molecular structure and physiological activities of silk fibroin induced by three different sterilization methods (steam, gamma radiation and ethylene oxide) with different dose or time period of sterilization by means of Fourier transform infrared (FT-IR) spec-troscopy, X-ray diffraction, mechanical properties and assessment of molecular weight. The results showed that the steam sterilization darkened the color of silk fibroin and obviously affected the mechanical property; gamma irradiation slightly degraded the molecular weight of silk fibroin and the speed of degradation increased with increasing irradiation dose; and ethylene oxide almost had no influence on silk fibroin expect for some slight hydrolysis on mo-lecular weight. Because ethylene oxide sterilization had the smallest influence on the quality of silk fi-broin with compared to other sterilization methods, it could be used as an efficient method to make fibroin more suitable for the development of functional foods and cosmetics.

In this study, electrospinning was used to fabricate silk-fibroin (SF)-based mats, which served as substrates for the culturing of rat Schwann cells. Microscopic observation and physical parameter measurements revealed that the electrospun SF mats had a nanofibrous structure with favorable physical properties. Fourier transform infrared analysis provided chemical characterization of the molecular confirmation of the SF proteins in the mats. The morphology and immunocytochemistry showed that the mats supported the survival and growth of the cultured Schwann cells, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis indicated that the electrospun SF mat extract had no cytotoxic effects on Schwann cell proliferation. Collectively, all of the results suggest that the electrospun SF mats might become a candidate scaffold for tissue-engineered nerve grafts to promote peripheral nerve regeneration. © 2010 Wiley Periodicals, Inc. J Appl Polym Sci, 2011

Complexity pursuit is an extension of projection pursuit to time series data and the method is closely related to blind separation of time-dependent source signals and independent component analysis. The goal is to find projections of time series that have interesting structure, defined using criteria related to Kolmogoroff complexity or coding length. In this paper, we first derive a simple approximation of coding length for unifying model that takes into account nongaussianity of sources, their autocorrelations and their smoothly changing nonstationary variances. Next, a fixed-point algorithm is proposed by using approximate Newton method. Finally, simulations verify the fixed-point algorithm converges faster than the existing gradient algorithm and it is more simple to implement due to it does not need any learning rate.

We have developed a new design of nerve guidance conduits (NGCs) made up of silk fibroin (SF), referred to as SF-NGCs, by a well-established method. The present study aimed to comprehensively investigate the degradation behaviors of SF-NGCs versus SF fibers. After they were allowed to incubate in the protease XIV solution or to be subcutaneously implanted in rabbits, SF-NGCs and SF fibers were subjected to degradation level measurement, mass loss assessment and gel electrophoresis, or light and electron microscopy and mass loss assessment for testing the dynamic course of in vitro or in vivo degradation. The results collectively indicated that SF-NGCs were able to degrade at a significantly increasing rate as compared to SF fibers, thus meeting the requirements of peripheral nerve regeneration. Furthermore, based on the possible involvement pathway in the in vivo degradation of SF-NGCs, the time-dependent changes in the mRNA level of lysosome-related genes (Hip1R, cathepsin D, and tPA) in subcutaneous implantation site within 24-week period post-implantation was determined by real-time RT-PCR, and the resulting data might contribute to our understanding of the molecular aspects that affect in vivo degradation and absorption of SF-NGCs.

Chitosan membranes loaded with bovine serum albumin (BSA; BSA/chitosan membranes) were prepared by either oven-drying or lyophilization. The encapsulation of BSA was mostly >80%. Controlled release of BSA from the membranes into phosphate-buffered saline (PBS; pH 7.4) was determined with Coomassie brilliant blue at different time periods. The highest cumulative amount of released BSA was 17.30% after 20 days. The optimized chitosan concentration was found to range from 10 to 20 g/l. The most favorable drying temperature for the membrane preparation was 55°C. And membranes made by lyophilization were more porous and uniform than those made by oven-drying.

Chitooligosaccharides (COSs), the biodegradation product of chitosan, have shown many biological functions. In this study, we examined the possible benefits of treatment with COSs (M.W. 800) on regeneration of rat crushed sciatic nerves. The rats with sciatic nerve crush injury were administered intraperitoneally daily with 3 or 6 mg/kg body weight of COSs over a 3-week period. During and at the end of COSs treatment, a series of functional and histological examinations, including the measurement of withdrawal reflex latency (WRL) values, walking track analysis, electrophysiological assessments, morphometric analysis of gastrocnemius muscle, as well as immunohistochemistry and electromicroscopy to regenerated sciatic nerves, were performed to evaluate the therapeutic outcomes of COSs. The experimental data demonstrated that COSs promoted peripheral nerve regeneration with the desired functional recovery in the rat sciatic nerve crush injury model. This study raises a possibility of developing COSs as a potential neuroprotective agent for peripheral nerve repair applications.

The natural biomaterial chitosan has been widely used as a promising nerve guidance conduit material for peripheral nerve repair. This study aimed to investigate in vitro biocompatibility of chitosan to primarily cultured hippocampal neurons, one type of central nervous system (CNS) cells. The substrate made up of chitosan fibers or membranes was found to support the survival and growth of the attached hippocampal neurons by using light and electron microscopy as well as immunocytochemistry for neurofilament 200, growth-associated protein-43, microtubule-associated protein 2, beta-tubulin III and synaptophysin. MTT assay indicated that the cell viability of hippocampal neurons in chitosan fiber or membrane extract was not significantly different from that in hydroxyapatite extract or plain neuronal medium, but significantly higher than that in organotin extract after culture for different times. Western analysis revealed that no significant difference in the protein level of growth-associated protein-43 and beta-tubulin III was detected between hippocampal neurons cultured in chitosan extract and in plain neuronal culture medium. The results collectively demonstrate that chitosan is biocompatible to primary culture of hippocampal neurons without cytotoxic effects on cell phenotype and functions, raising a potential possibility of using chitosan for CNS therapy.

Chitooligosaccharides (COSs), the biodegradation product of chitosan, have demonstrated a diverse array of biological activities. Here we report the protective effect of COSs (M.W. 800) against glutamate-induced neurotoxicity in cultured hippocampal neurons. The cell viability assessments, together with Hoechst 33342 staining and flow cytometry for cell apoptosis analysis, indicated that glutamate (125 microM)-induced cell apoptosis in cultured hippocampal neurons was attenuated in a concentration-dependent manner by COSs pretreatment. After measurement with Fluo 4-AM, COSs were found to depress glutamate-induced elevation in intracellular calcium concentration ([Ca(2+)](c)). The enzymatic assay indicated that COSs antagonized glutamate-evoked activation of caspase-3. These results collectively suggest that COSs prevent cultured hippocampal neurons from glutamate-induced cell damage by interfering with an increase in [Ca(2+)](c) and inhibiting caspase-3 activity.

A unifying model that combines three properties is proposed by Hyvarinen, and a gradient ascent algorithm for independent component analysis (ICA) is performed by maximum likelihood estimation. In this paper, we consider the estimation of the data model of ICA when Gaussian noise is present and the independent components are time dependent. Firstly, according to the useful property of Gaussian moments, we introduce Gaussian moments algorithm to estimation of the noisy unifying model when the noise covariance matrix is known. Next, when the noise covariance is unknown, a new Gaussian moments algorithm is developed. Finally, the validity and performance of our algorithms are demonstrated by computer simulations.

Silk fibroin (SF), derived from natural silk long used as a textile material, has recently become an important biomaterial for tissue engineering applications. We have previously reported on good in vitro biocompatibility of SF fibers with peripheral nerve tissues and cells. In the present study, we developed a novel biomimetic design of the SF-based nerve graft (SF graft) which was composed of a SF-nerve guidance conduit (NGC) inserted with oriented SF filaments. The SF-NGC prepared via well-established procedures exhibits an eggshell-like microstructure that is responsible for its superior mechanical and permeable properties beneficial to nerve regeneration. The SF graft was used for bridge implantation across a 10-mm long sciatic nerve defect in rats, and the outcome of peripheral nerve repair at 6 months post-implantation was evaluated by a combination of electrophysiological assessment, FluoroGold retrograde tracing and histological investigation. The examined functional and morphological parameters show that SF grafts could promote peripheral nerve regeneration with effects approaching those elicited by nerve autografts which are generally considered as the gold standard for treating large peripheral nerve defects, thus raising a potential possibility of using these newly developed nerve grafts as a promising alternative to nerve autografts.

A partially purified potato polyphenol oxidase (PPO) was immobilized in a cross-linked chitosan–SiO2 gel and used to treat phenol solutions. Under optimized conditions (formaldehyde 20 mg/ml, PPO 4 mg/ml and pH 7.0), the activity of immobilized PPO was 1370 U/g and its K m value for catechol was 12 mm at 25°C. The highest activity of immobilized enzyme was at pH 7.4. Immobilization stabilized the enzyme with 73 and 58% retention of activity after 10 and 20 days, respectively, at 30°C whereas most of the free enzyme was inactive after 7 days. The efficiency of removing phenol (10 mg phenol/l) by the immobilized PPO was 86%, and about 60% removal efficiency was retained after five recycles. The immobilized PPO may thus be a useful for removing phenolic compounds from industrial waste-waters.

The aim of this study was to investigate the effects induced by three different sterilization methods [steam, γ-radiation, and ethylene oxide (EO)] with different dose or time period of sterilization by means of Fourier transform infrared spectroscopy, X-ray diffraction, and assessments of molecular weight and degree of deacetylation. The experiment results clearly showed that the steam sterilization greatly darkened the color of chitosan powder; γ-irradiation strongly depolymerized the chitosan especially at irradiation dose above 10 kGy; and exposure to EO led to minor change in crystallinity and network structure of chitosan powder. Therefore, these three methods have their respective advantages and disadvantages, suggesting that the selection of the sterilization method of chitosan should depend upon the specific requirement of the final application. © 2007 Wiley Periodicals, Inc. J Appl Polym Sci 104: 1968–1972, 2007

Cervical erosion is one of the common diseases of women. The loop electrosurgical excisional procedure (LEEP) has been used widely in the treatment of the cervical diseases. However, there are no effective wound dressings for the postoperative care to protect the wound area from further infection, leading to increased secretion and longer healing time. Iodine is a widely used inorganic antibacterial agent with many advantages. However, the carrier for stable iodine complex antibacterial agents is lack. In the present study, a novel iodine carrier, Carboxymethyl chitosan-g-(poly(sodium acrylate)-co-polyvinylpyrrolidone) (CMCTS-g-(PAANa-co-PVP), was prepared by graft copolymerization of sodium acrylate (AANa) and N-vinylpyrrolidone (NVP) to a carboxymethyl chitosan (CMCTS) skeleton. The obtained structure could combine prominent property of poly(sodium acrylate) (PAANa) anionic polyelectrolyte segment and good complex property of polyvinylpyrrolidone (PVP) segment to iodine. The bioactivity of CMCTS could also be kept. The properties of the complex, CMCTS-g-(PAANa-co-PVP)-I2, were studied. The in vitro experiment shows that it has broad-spectrum bactericidal effects to virus, fungus, gram-positive bacteria and gram-negative bacteria. A CMCTS-g-(PAANa-co-PVP)-I2 complex contained cervical antibacterial biomembrane (CABM) was prepared. The iodine release from the CABM is pH-dependent. The clinic trial results indicate that CABM has better treatment effectiveness than the conventional treatment in the postoperative care of the LEEP operation.

Schwann cell (SC) myelination is pivotal for the proper physiological functioning of the nervous system, but the underlying molecular mechanism remains less well understood. Here, we showed that the expression of bone morphogenetic protein 7 (BMP7) inversely correlates with myelin gene expression during peripheral myelination, which suggests that BMP7 is likely a negative regulator for myelin gene expression. Our experiments further showed that the application of BMP7 attenuates the cAMP induced myelin gene expression in SCs. Downstream pathway analysis suggested that both p38 MAPK and SMAD are activated by exogenous BMP7 in SCs. The pharmacological intervention and gene silence studies revealed that p38 MAPK, not SMAD, is responsible for BMP7-mediated suppression of myelin gene expression. In addition, c-Jun, a potential negative regulator for peripheral myelination, was up-regulated by BMP7. In vivo experiments showed that BMP7 treatment greatly impaired peripheral myelination in newborn rats. Together, our results established that BMP7 is a negative regulator for peripheral myelin gene expression and that p38 MAPK/c-Jun axis might be the main downstream target of BMP7 in this process.

The binding of isorhamnetin to bovine liver catalase (BLC) was first investigated at 302, 310 and 318 K at pH 7.4 using spectroscopic methods including fluorescence spectra, circular dichroism (CD) and UV-vis absorption. Spectrophotometric observations are rationalized mainly in terms of a static quenching process. The binding constants and binding sites were evaluated by fluorescence quenching methods. Enzymatic activity of BLC in the absence and presence of isorhamnetin was measured using a UV/vis spectrophotometer. The result revealed that the binding of isorhamnetin to BLC led to a reduction in the activity of BLC. The positive entropy change and enthalpy change indicated that the interaction of isorhamnetin with BLC was mainly driven by hydrophobic forces. The distance r between the donor (BLC) and acceptor (isorhamnetin) was estimated to be 2.99 nm according to fluorescence resonance energy transfer. Fluorescence, synchronous fluorescence, and CD spectra showed no obvious change in the conformation of BLC upon the binding of isorhamnetin. In addition, the influence of pH on the binding of isorhamnetin to BLC was investigated and the binding ability of the drug to BLC deceased under other pH conditions (pH 9.0, 6.5, 5.0, 3.5, or 2.0) as compared with that at pH 7.4. Copyright © 2016 John WileySons, Ltd.

One major obstacle in the application of drug delivery systems for cancer chemotherapy is their poor penetration in tumor tissues. Conjugating active ligand moieties to the surface of nanoparticles may be a promising approach for enhancing the tumor accumulation and penetration of nanoparticles. Herein, the cell-penetrating peptide twin-arginine translocation (Tat)-conjugated epirubicin-loaded poly(lactic-glycolic acid) nanoparticles were prepared to achieve deep tumor penetration. The morphology and size of nanoparticles were characterized by scanning electron microscopy and dynamic light scattering, and the biological behaviors of nanoparticles were evaluated. It is demonstrated that Tat-conjugated nanoparticles have a significant improvement in antitumor activity and biodistribution compared with nonconjugated nanoparticles. Importantly, Tat conjugated on the surface of nanoparticles could facilitate the encapsulated drug penetration into deeper tumor tissue. Additionally, Tat-conjugated nanoparticles have good biocompatibility, as demonstrated by hemolytic tests, in vitro cytotoxicity, and histology study. These results suggested that the Tat-conjugated nanoparticles, as a powerful delivery system for chemotherapeutic drug, would have a promising application in human cancer therapy.

Reptiles are the most morphologically and physiologically diverse tetrapods, and have undergone 300 million years of adaptive evolution. Within the reptilian tetrapods, geckos possess several interesting features, including the ability to regenerate autotomized tails and to climb on smooth surfaces. Here we sequence the genome of Gekko japonicus (Schlegel's Japanese Gecko) and investigate genetic elements related to its physiology. We obtain a draft G. japonicus genome sequence of 2.55 Gb and annotated 22,487 genes. Comparative genomic analysis reveals specific gene family expansions or reductions that are associated with the formation of adhesive setae, nocturnal vision and tail regeneration, as well as the diversification of olfactory sensation. The obtained genomic data provide robust genetic evidence of adaptive evolution in reptiles., Geckos are small, agile reptiles with nocturnal habits. Here, the authors sequence the genome of the Schlegel's Japanese Gecko and reveal gene family expansions and reductions associated with formation of adhesive setae, nocturnal vision, tail regeneration, and diversification of olfactory sensation.

The spacecraft telemetry data is the only way for operation managers to understand the spacecraft's real working state in space. In face with the increasing amount of telemetry data growing day by day, it is hard to overcome the long data time span, large data volume, dirty data and etc. It is very necessary to explore some effective analytical methods to analysis the big telemetry data. In this paper, the K Charts analysis method is referenced and used to deal with the telemetry big data for safety analysis. Using the transformation and improvement, particle size analysis technique, the fault-tolerant K Charts is proposed and established for spacecraft telemetry data change analysis. K Charts cycle through the transforming of variables, different time granularity, formation, K Charts combination, to realize mass spacecraft telemetry sampling data association analysis of structure the shape and the state. The results of application show that, combinations of different sizes of K Charts can reflect the law of the data intrinsic change more accurately than ordinary timing diagrams.

Despite an intrinsic regenerative capacity of the peripheral nervous system, in most cases of peripheral nerve injury, especially those involving substantial nerve gaps, therapeutic interventions are always required to support peripheral nerve regeneration and functional recovery. With the advancement of tissue engineering and translational medicine, extensive research has focused on the development of tissue-engineered nerve grafts, as a promising alternative to autologous nerve grafts, for bridging nerve gaps in peripheral nerve repair. The typical design of tissue-engineered nerve grafts is composed of a nerve scaffold combined with cellular and/or molecular components. This chapter separately describes a diverse array of physical and biochemical components of tissue-engineered nerve grafts with regard to their features, functions, structure or composition, and fabrication or procurement, and further discusses many representative examples of tissue-engineered nerve grafts and their pros and cons. We also highlight the current clinical applications of tissue-engineered nerve grafts and outline the future prospects for tissue engineering in peripheral nerve regeneration.

The biotransformation of digoxigenin by Colletotrichum lini AS3. 4486 was investigated. The conversion reaction was carried out in a 72h process, and the sole products was isolated by column chromatography and elucidated as digoxigenone by HR-ESI-MS, H-NMR , C-NMR and single–crystal X-ray diffraction. The crystal of digoxigenone belongs to triclinic, space group P1 with 7.4017 (15), 7.7450 (15), 10.215 (2) A , α = 99.51(3), β = 94.70 (3), γ = 114.97 (3)°, Z = 1. This study povides a new method for the synthesis of digoxigenone.

Retinoid signaling is essential for development of vertebrate embryos, and its action is mainly through retinoic acid (RA) binding to its RA receptors and retinoid-X receptors, while the critical concentration and localization of RA in embryos are determined by the presence and activity of retinal dehydrogenases (for RA synthesis) and cytochrome P450 RAs (Cyp26s) (for degradation of RA). Previously, we identified a novel cyp26 gene (cyp26d1) in zebrafish that is expressed in hindbrain during early development. Using reverse-phase HPLC analyses, we show here that zebrafish Cyp26D1 expressed in 293T cells could metabolize all-trans RA, 9-cis RA, and 13-cis RA, but could not metabolize retinol or retinal. The metabolites of all-trans RA produced by Cyp26D1 were the same as that produced by Cyp26A1, which are mainly 4-hydroxy-all-trans-RA and 4-oxo-all-trans-RA. Performing mRNA microinjection into zebrafish embryos, we demonstrated that overexpression of Cyp26D1 in embryos not only caused the distance between rhombomere 5 and the first somite of the injected embryos to be shorter than control embryos but also resulted in left-right asymmetry of somitogenesis in the injected embryos. These alterations were similar to those caused by the overexpression of cyp26a1 in zebrafish embryos and to that which resulted from treating embryos with 1 μm 4-diethylamino-benzaldehyde (retinal dehydrogenase inhibitor), implying that cyp26d1 can antagonize RA activity in vivo. Together, our in vitro and in vivo results provided direct evidence that zebrafish Cyp26D1 is involved in RA metabolism.

Two chitooligosaccharides, Ch1100 (average molecule weight: 1,100; average degree of polymer: 7) and Ch500 (average molecule weight: 500; average degree of polymer: 3), were obtained by partial hydrolysis of chitosan. Their antioxidant activities were investigated with scavenging activity of chitooligosaccharide against superoxide radical using phenazin methosulphate (PMS)–β-nicotinamide adenine dinucleotide (NADH)–nitroblue tetrazolium (NBT) system, scavenging activity of chitooligosaccharide against carcinogen-induced active oxygen species and scavenging activity of chitooligosaccharide against hydrogen peroxide released from polymorphonuclear leukocytes stimulated by phorbol-12-myristate-13-acetate (PMA). By the aforementioned three measurements, the radical scavenging activity (Inhibition%) of Ch1100 was determined to 54.8, 35.3 and 38.9%, respectively, at the concentration of 100 μmol/L, while that of Ch500 was 40.1, 13.9 and 18.2%, respectively, at the same concentration. These results clearly suggest that Ch1100 is a potent radical scavenger, which has the high radical scavenging activity.

Prestress losses have a direct impact on concrete stress development and deflection behavior of highway bridge members. A poor estimate of prestress losses can result in a structure in which allowable stresses are exceeded or camber and deflection behavior is poorly predicted, such that the serviceability of a structure may be adversely affected. This paper reports the prestress losses observed throughout fabrication, shipment, erection, and the first 2 years of service for the first high-performance superstructure concrete bridge in Missouri. The prestress losses investigated included prerelease losses, elastic shortening losses, relaxation losses, creep losses, and shrinkage losses. Results from the study were compared with eight commonly used loss estimate models for total prestress losses, including AASHTO and Prestressed Concrete Institute methods. Recommendations were proposed by the authors for the most appropriate methodology to use to predict prestress losses in high-strength concrete girders accurately.

A porous, biodegradable, natural chitin/chitosan nerve conduit was constructed. Scanning electron microscopy confirmed that it was homogeneous and highly porous. FT-IR spectra showed that there were no residues arising from the preparation process in the conduit. Addition of chitin to the chitosan solution increased the mechanical strength and maximum tensile strength from 7.2 to 9.6 MPa. Preliminary animal tests indicated that porous chitin/chitosan conduits did not swell in vivo and were compatible with surrounding tissue.

The complete mitochondrial genome sequence of the Hylopetes alboniger was sequenced and reported for the first time using muscle tissue. The mitochondrial genome is 16,584 bp in size, including 22 tRNA genes, 13 protein-coding genes, 12S rRNA, 16S rRNA, and one control region. Phylogenetic analysis was performed using whole mitogenome sequences with other 15 closely related taxa to assess their phylogenetic relationship. This mitochondrial genome sequence will provide a better understanding for H. alboniger evolution in the future.

A new adsorbent for heavy metal ions, cys-chitin, was produced by modifying chitin in THF with L-cysteine in the presence of sulfuric acid as a catalyst. Its structure was confirmed by elemental analysis and FTIR spectra analysis. The adsorption properties of cys-chitin for CuII, CdII, PbII, ZnII, and NiII were investigated. The effect of pH value on adsorption and adsorption kinetics was examined. The results indicate that the cys-chitin has much better adsorption for CdII, PbII, and ZnII than chitin itself. The adsorption capacities of cys-chitin for CdII, PbII, and ZnII were 214.6, 351.5, and 107.0 mg/g, respectively. It also had good adsorption properties for heavy metal ions. The adsorption capacities were also affected by the acidity of medium. The adsorbed CdII, PbII could be eluted by diluted nitric acid. © 2003 Wiley Periodicals, Inc. J Appl Polym Sci 88: 2575–2579, 2003

For its significant economical savings and greater design flexibility, high-performance concrete (HPC) is becoming more widely used in highway bridge structures. High-performance bridges with HPC and large-diameter prestressed strands are becoming attractive to designers. Bridge A6130 is the first fully HPC superstructure bridge in Missouri. The bridge has HPC cast-in-place deck and high-strength concrete girders reinforced with 15.2-mm (0.6-in.) diameter strands. The bridge was instrumented with embedded strain gauges and thermocouples to monitor the early-age and later-age behavior of the structures from construction through service. To investigate the overall behavior of the bridge under live load, a static live-load test was developed and carried out. During the live-load test, 64 embedded vibrating wire strain gauges and 14 embedded electrical-resistance strain gauges were used to acquire the changing strain rate in the bridge caused by the varying live-load conditions. Girder deflections and rotations were also recorded with external sensors and a data acquisition system. Based on the test results, the load distribution to the girders was studied. The AASHTO specifications live-load distribution factor recommended for design was compared with the measured value and found to be overly conservative. The AASHTO load and resistance factor design live-load distribution factors recommended for design were found to be comparable to measured values. Two finite element models were developed with ANSYS and compared with measured values to investigate the continuity level of the Missouri Department of Transportation interior bent detail.

Medicinal oligoglucosamine was prepared by oxidative degradation of chitosan with neutral hydrogen peroxide under microwave irradiation. The effects of concentration of H2O2, volume of solution, amount of chitosan and irradiation time were investigated by the orthogonal test, and the optimal conditions of degradation are described. The structure of the product was confirmed by FT-IR spectrum analysis. The number average molecular weight of oligoglucosamine was determined by the method of end group analysis. The experimental results show that oligoglucosamine can be conveniently and effectively obtained by oxidative degradation of chitosan under microwave irradiation. The number average molecular weight of chitosan decreased to 900–1000. The changes in the yield of oligoglucosamine are strongly dependent on the reaction time and the concentration of H2O2.

Salidroside is a phenylpropanoid glycoside isolated from Rhodiola rosea L., a traditional Chinese medicinal plant, and has displayed a broad spectrum of pharmacological properties. In this paper, two analogues were prepared with the glucosamine and N-acetylglucosamine as glycosyl donor, 2-(4-hydroxyphenyl)ethanol as glycosyl acceptor. The effects of them over PC12 cell model exposed to hypoglycemia and serum limitation were assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, Flow Cytometry and Western blot analysis.

Natural polysaccharides are biomaterials widely used for constructing scaffolds in tissue engineering. While natural polysaccharides have been shown to robustly promote tissue regeneration, the underlying molecular mechanism remains largely unknown. Here, we show that chitooligosaccharides (COS), the intermediate products of chitosan degradation, stimulate peripheral nerve regeneration in rats. Our experiment also shows that COS stimulate the proliferation of Schwann cells (SCs) during nerve regeneration. By analyzing the transcriptome and gene regulatory network, we identified the miR-27a/FOXO1 axis as the main signaling pathway for mediating the proliferative effects of COS on SCs. COS increase the expression level of miR-27a and cause a reduction of FOXO1, which subsequently accelerates the cell cycle and stimulates SC proliferation to stimulate nerve regeneration. These findings define a basic pathway for oligosaccharides-mediated cell proliferation and reveal a novel aspect of polysaccharide biomaterials in tissue engineering.

Chitosan is found to promote the regeneration of peripheral nerve system in our previous studies, whereas the regeneration speed is not satisfied with clinical request. Micropatterning could promote cell orientation and growth, however, the effect of porous chitosan micropatterning on nerve regeneration is rarely reported. In this study, the porous chitosan micropatterning with surface ridge/groove and inner porosity structure was fabricated using a combination of micromodeling and lyophilization method. The morphology and stability of the prepared chitosan micropatterning were evaluated, the regulation of Schwann cells behavior by chitosan micropatterning was evaluated. The results showed that the chitosan micropatterning displayed stripe-like structure with a clear and complete edge. The micropatterning with 30/30 μm was more stable than 20/20 μm sample. Schwann cells on chitosan micropatterning showed orientation adhesion and began to grow along a certain direction after culture for 2 h, and displayed the minimal orientation angle and the largest length/width ratio on 30/30 μm micropatterning after further culture for 3 d and 5 d, indicating the most obvious cell orientation. Moreover, the secretion of nerve growth factor (NGF) demonstrated that the micropatterned chitosan had no negative influence on the physiological function of Schwann cells. Thus, the results indicate that the porous chitosan micropatterning can regulate Schwann cell growth well, which may have potential application in nerve regeneration. The study provides an important basis for constructing porous nerve conduit with micropatterning structure in the inner wall.

The advantages of high-performance concrete (HPC) have been well documented in recent years. These include enhanced design flexibility and improved durability performance resulting in reduced maintenance costs and an increased service life. In spite of these obvious benefits, implementation of HPC has been very slow. This can be attributed to several factors including the uncertainty related to current design codes and a lack of familiarity of designers and contractors with proper design and construction practices and requirements for HPC structures. This paper introduces and discusses some fundamental issues that affect the implementatation of HPC and impact the practitioner. These include those related to quality control/quality assurance, specifications, material performance, and structural behavior. Within the scope of this discussion, the fundamental differences and similarities between HPC and conventional concrete are discussed. The aim is to provide the practicing engineer with a conceptual understanding of the practical issues that affect the design and use of HPC for highway structures with the desire to further stimulate the implementation of HPC.

Salidroside is proven to be a neuroprotective agent of natural origin, and its analog, 2-(4-Methoxyphenyl)ethyl-2-acetamido-2-deoxy-β-d-pyranoside (named SalA-4g), has been synthesized in our lab. In this study, we showed that SalA-4g promoted neuronal survival and inhibited neuronal apoptosis in primary hippocampal neurons exposed to oxygen and glucose deprivation (OGD) and in rats subjected to ischemia by transient middle cerebral artery occlusion (MCAO), respectively, and that SalA-4g was more neuroprotective than salidroside. We further found that SalA-4g elevated glucose uptake in OGD-injured primary hippocampal neurons and increased the expression and recruitment of glucose transporter 3 (GLUT3) in ischemic brain. Signaling analysis revealed that SalA-4g triggered the phosphorylation of CREB, and increased the expression of PKA RII in primary hippocampal neurons exposed to OGD injury, while inhibition of PKA/CREB by H-89 alleviated the elevation in glucose uptake and GLUT3 expression, and blocked the protective effects of SalA-4g. Moreover, SalA-4g was noted to inhibit intracellular Ca(2+) influx and calpain1 activation in OGD-injured primary hippocampal neurons. Our results suggest that SalA-4g neuroprotection might be mediated by increased glucose uptake and elevated GLUT3 expression through calpain1/PKA/CREB pathway.

Complexity pursuit is a recently developed algorithm using the gradient descent for separating interesting components from time series. It is an extension of projection pursuit to time series data and the method is closely related to blind separation of time-dependent source signals and independent component analysis. The goal is to find projections of time series that have interesting structure, defined using criteria related to Kolmogoroff complexity or coding length. In this paper, we derived a simple approximation of coding length that takes into account the nongaussianity, the autocorrelations and the variance nonstationary of the time series. We give a simple algorithm for its approximative optimization.