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Depression is associated with abnormal lipid metabolism, and omega (n)-3 polyunsaturated fatty acids (PUFAs) can effectively treat depression. However, mechanism of lipid metabolism involved in the depressive attenuation remains poorly understood. Olfactory bulbectomy (OB)-induced changes in animal behavior and physiological functions are similar to those observed in depressed patients. Therefore, the present study used wild type (WT) and Fat-1 mice with or without OB to explore whether endogenous n-3 PUFA treatment of depression was through rectifying lipid metabolism, and to discover the possible lipid metabolic pathways. In WT mice, OB enhanced locomotor activity associated with up-regulation of lipid metabolites in the serum, such as phosphatidylcholines, L-a-glutamyl-L-Lysine and coproporphyrinogen III (Cop), which were involved in anti-inflammatory lipid metabolic pathways. OB also increased microglia activation marker CD11b and pro-inflammatory cytokines in the hippocampus. In one of the lipid pathways, increased Cop was significantly correlated with the hyper-activity of the OB mice. These OB-induced changes were markedly attenuated by endogenous n-3 PUFAs in Fat-1 mice. Additionally, increased expressions of anti-inflammatory lipid genes, such as fatty acid desaturase (Fads) and phospholipase A2 group VI (Pla2g6), were found in the hippocampus of Fat-1 mice compared with WT mice. Furthermore, Cop administration increased the production of pro-inflammatory cytokines and nitric oxide in a microglial cell line BV2. In conclusion, endogenous n-3 PUFAs in Fat-1 mice attenuated abnormal behavior in the depression model through restoration of lipid metabolism and suppression of inflammatory response.

The development of a sensing platform with high sensitivity and specificity, especially programmability and universal applicability, for the detection of clinically relevant molecules is highly valuable for disease monitoring and confirmation but remains a challenge. Here, for the first time, we introduce the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system into an immobilization-free electrochemical biosensing platform for sensitively and specifically detecting the disease-related nucleic acids and small molecules. In this strategy, a modular rolling circle amplification (RCA) is designed to transform and amplify the target recognition event into the universal trigger DNA strand that is used as the trigger to activate the deoxyribonuclease activity of CRISPR/Cas12a for further signal amplification. The cleavage of the target-activated blocker probe allows the methylene blue-labeled reporter probes to be captured by the reduced graphene oxide-modified electrode, leading to an obviously increased electrochemical signal. We only need to simply tune the sequence for target recognition in RCA components, and this strategy can be flexibly applied to the highly sensitive and specific detection of microRNAs, Parvovirus B19 DNA, and adenosine-5'-triphosphate and the calculated limit of detection is 0.83 aM, 0.52 aM, and 0.46 pM, respectively. In addition, we construct DNA logic circuits (YES, NOT, OR, AND) of DNA inputs to experimentally demonstrate the modularity and programmability of the stimuli-responsive RCA-CRISPR/Cas12a system. This work broadens the application of the CRISPR/Cas12a system to the immobilization-free electrochemical biosensing platform and provides a new thinking for developing a robust tool for clinical diagnosis.

Objective: The iridoid glycosides were extracted and separated from Osmanthus fragrans seeds, and the potential protective effect of Osmanthus fragrans seed extract on concanavalin A-induced immune liver injury in mice was studied.Methods:Osmanthus fragrans seeds were extracted by 95% ethanol reflux. Then, the iridoid glycosides were enriched by extraction refined through petroleum ether (60°C–90°C), ethyl acetate, and water-saturated n-butanol in sequence, so as to purify the n-butanol part (Osmanthus fragrans seed’s n-butanol extraction, OFSN) by macroporous resin. Specnuezhenide and Nuezhenoside G13 were used as the reference substances to determine the concentration of iridoid glycosides by HPLC. On this basis, a mouse immune liver injury model was established by tail intravenous concanavalin A (20 mg/kg); the contents of serum ALT, AST, IFN-γ, and TNF-α and the contents of liver tissue MDA and SOD were determined; the pathological changes of the liver by HE staining were observed; and the expression levels of p38MAPK and p-p38mapk in liver tissue were detected by WB.Results: The linearity, precision, repeatability, recovery, and stability of HPLC all met the requirements by validating with the methodology. The contents of Specnuezhenide and Nuezhenoside G13 in the n-butanol extracts were 39.20% and 39.88%, respectively. Actually, their contents can reach up to 82.56% and 87.9% after being purified by macroporous resin. The results of animal experiments show that OFSN could significantly reduce the liver and spleen index, reduce the ALT and AST contents in plasma and the MDA content in liver tissue, and then increase the SOD content. Besides, OFSN could also reduce the plasma IFN-γ and TNF-α levels. The HE staining result indicates that the pathological changes in the liver tissues of mice treated with OFSN are alleviated to different degrees while the WB result suggests that OFSN could significantly inhibit the expression of p-p38mapk.Conclusion:Osmanthus fragrans seeds are rich in iridoid glycosides, which has a good protective effect on mouse immune liver injury caused by concanavalin A. The mechanism may be related to inhibiting the phosphorylation of p38MAPK, inhibiting the release of inflammatory mediators, improving the antioxidant capacity of liver cells, and weakening the occurrence of lipid peroxidation.

The optical sensor with ratiometric and turn-off dual modes is constructed to detect H2O2 and glucose based on blue fluorescent carbon dots (CDs) and MnO2 nanosheets with great ability of fluorescence quenching and scattering. Employing CDs@MnO2 nanosheets nanocomposite as the probe, H2O2 is detected by simultaneously collecting first-order scattering (FOS, 353.5 nm), fluorescence (440 nm), and second-order scattering (SOS, 710 nm) under the excitation of 350 nm. H2O2 with strong oxidation property can etch the lamellar structure of MnO2 nanosheets into nano-fragments, which made the fluorescence of CDs in the system recover and the scattering intensity (FOS and SOS) of the system decrease significantly. Therefore, the optical sensor combined FOS and fluorescence signals in ratiometric mode, and SOS signal in turn-off mode to realize sensitive determination of H2O2. The linear ranges of ratiometric mode and turn-off mode for H2O2 detection were 0.2–40 and 0.2–15 μM, respectively. And the limits of detection (LODs) of two modes were 73 and 104 nM, respectively. Furthermore, the sensor was also successfully applied to the detection of glucose which can react to produce H2O2. Satisfactorily, the LODs of this sensor for glucose detection were 95 and 113 nM for ratiometric mode and turn-off mode, respectively. This work not only provides a new method for the accurate detection of H2O2 and glucose, but also extends a new idea for the study of the combination of scattering and fluorescence.

DNA methyltransferase (MTase), modulating the level of genomic DNA methylation, harbors both a pharmacological target for clinical therapy and a potential biomarker for genetic disorders and tumorigenesis. Typical homogeneous electrochemical approaches, employing solution phase probes, have been considered simple, efficient, and economical method, yet these architectures usually require electroactive molecules labeling, rely on weak electrostatic adsorption interaction, and possess low sensitivity. For circumventing the above drawbacks, herein, we devise a ‘plug and play’ microelectrode featuring microminiaturization, rapid response time and enhanced mass transport to quantify MTase activity through monitoring the variation of diffusion current of methylene blue (MB) induced by the less-mobile G-quadruplex framework. By coupling the unique signal-transduction approach with Y-motif-mediated primer-free cyclic signal amplification (YPCSA), the miniaturized biosensor possesses low detection limit (down to 2.5 × 10−4 U mL−1), high specificity, good stability and satisfying reusability, and has been successfully applied to the screening of MTase inhibitors, holding great potential in clinical diagnosis and pharmacological research.

Herein, an innovative photocathodic enzymatic biosensor is proposed with poly {4,8-bis[5-(2-ethylhexyl)thiophen-2-yl]-benzo[1,2-b:4,5-b']dithiophene-2,6-diyl-alt-3-fluoro-2-[(2-ethylhexyl)carbonyl]thieno[3,4-b]thiophene-4,6-diyl} (PTB7–Th) as donor–acceptor–type photoactive material and three–dimensional (3D) polyaniline hydrogels (PAniHs) as both electron transfer layer and biomolecule carrier. Based on the enhancement effect of PAniHs on the charge separation and electron transfer of PTB7–Th and the competitive consumption of dissolved oxygen (O2) between the xanthine oxidase (XOD)–guanine catalytic reaction and O2–sensitive PTB7–Th/PAniHs, the proposed photocathodic enzymatic biosensor has been demonstrated to detect guanine with the advantages of low limit of detection (0.02 μM), wide linear range (from 0.1 to 80 μM), simple and convenient preparation process, satisfactory stability, and photochemical signal amplification independent of any exogenous electron donor/acceptor or sensitizer. Remarkably, the proposed photocathodic enzymatic biosensor can not only be extended to other aerobic enzymatic bioanalyses, but also pave a horizon for the application of environmentally friendly conductive hydrogel materials in photoelectrochemical bioanalysis.

Herein, for the first time, we engineered click chemistry reaction to trigger a 3D DNA walking machine for ultrasensitive electrochemical detection of copper ion (Cu2+), which provided a convenient access to overcome the shortcomings of poor selectivity and limited amplification efficiency in traditional determination of Cu2+. Click chemistry reaction drove azido-S2 to bind with alkynyl-S1 for the formation of a walker probe on aminated magnetic polystyrene microsphere@gold nanoparticles (PSC@Au), which opened the hairpin-locked DNAzyme. In the presence of magnesium ion (Mg2+), the unlocked DNAzyme was activated to cleave the self-strand at the facing ribonucleotide site, accompanied by the release of product DNA (S3) and the walker probe. Therefore, the walker probe was able to open the adjacent hairpin-locked DNAzyme strand and then be released by DNAzyme cleavage along the PSC@Au-DNAzyme track. Eventually, the liberated single-strand S3 induced catalytic hairpin assembly (CHA) recycling, resulting in t...

Herein a rigid three-dimensional (3D) DNA "nanosafe-box" (DNB) for encapsulation and release of a recognition probe (N3) is designed to construct an electrochemical biosensor with the use of electroactive two-dimensional metal-organic framework (2D MOF) nanosheets as signal tags for ultrasensitive detection of mercury ion (Hg2+). Initially, N3 is locked in the 3D cavity of DNB by blocker DNA. After addition of target Hg2+, exonuclease III (Exo-III) digestion is initiated to the liberate DNA "key" (K); thereby, the free K triggers a strand displacement reaction for exposing the prelocked N3 to successfully ligate dibenzocyclooctyne (DBCO)-tagged anchor via metal-catalyst-free click chemistry, in which amounts of 2D MOF nanosheets containing Co(II) as electron mediator are introduced accompanied by significant electrochemical response. Compared with traditional linear or stem-loop DNA nanostructure, the well-designed 3D DNB possesses remarkably enhanced mechanical rigidity and structural stability, resulting in improved accessibility of probes and increased loading amounts of signal tags. More importantly, by this way of encapsulation and release of recognition probes, the background signal is decreased dramatically, leading to increased sensitivity of the proposed biosensor. Consequently, this electrochemical biosensor exhibits outstanding analytical performance for Hg2+ detection with a low detection limit of 33 fM and dynamic linear range of 0.1 pM to 10 nM. This strategy offers an ingenious method for detection of metal ions and biomarkers, possessing potential applications in environmental tests and clinical diagnosis.

In this work, on the basis of a new 2D DNA nanoprobe (DNP) and an enzyme-free-target-recycling amplification, an electrochemical biosensor is developed for the ultrasensitive detection of microRNA-21 (miRNA-21). Herein, two ferrocene-labeled bipedal DNPs, which show small steric hindrance and strong stability, are prepared on the basis of the mechanism of the proximity-ligation assay (PLA), improving the space utilization. In the presence of the target, miRNA-21, and a hairpin DNA strand, the DNP will collapse, and then two ferrocene-labeled DNA strands and the miRNA-21 will be simultaneously released from the electrode surface through toehold-mediated strand-displacement reactions (TSDRs), leading to a decrease in the electrochemical signal and realization of enzyme-free target recycling. As a result, the one input target, miRNA-21, could release 2 N ferrocene-labeled DNA strands, achieving a dramatic decrease in the electrochemical signal. Combining DNPs and enzyme-free target recycling, this proposed biosensor showed a linear dependence with miRNA-21 concentration, ranging from 1.0 fM to 10 nM with a detection limit of 0.31 fM. In addition, it is worth mentioning that this biosensor can be regenerated through incubating with three assistant-DNA strands, realizing the reuse of raw materials. Surprisingly, the elaborated biosensor provides a novel strategy for building controllable DNA nanoprobes for the sensitive detection of various biomarkers.

Electrochemical aptasensors have been wildly used in the detection and quantification of protein biomarkers, but their practical applications are limited by inefficient signal amplification and laborious and time-consuming probe surface-immobilization procedure. Here, we combine the homogeneous electrochemical aptasensor with the CRISPR/Cas12a strategy to overcome such limitations. The binding-induced DNA strand displacement (BIDSD) strategy is designed to transform thrombin-aptamer interaction into nucleic acid output, which further triggers the rolling circle amplification (RCA) to regulate the deoxyribonuclease activity of CRISPR/Cas12a. Based on the difference in affinity of graphene to single-stranded DNA and double-stranded DNA, this electrochemical aptasensor is free of probe surface-immobilization procedure, which greatly simplifies operation steps and facilitates its flexibility. By integrating BIDSD with two amplification strategies of RCA and CRISPR/Cas12a, this electrochemical aptasensor achieves specific and sensitive detection of thrombin with a calculated limit of detection of 1.26 fM, which provides a paradigm for the sensitive detection of protein biomarker.

Herein, we construct an ingenious spatially localized amplification reaction (SLAR) by colocalizing the entropy-driven reaction (EDR) in a nanometer space, which greatly accelerates target conversion and realizes the sensitive detection of microRNA-21 (miRNA-21). A large number of EDR complex are hybridized with the prefabricated DNA scaffold via a DNA self-assembly strategy to form the SLAR nanoprobe. Target miRNA-21 triggers interval EDR along the long DNA scaffold, resulting in fluorescence recovery with high signal gain because of the fast release of reporter. Compared with reactions with diffusible components, spatial arrangement of all components of EDR on a nanoscale scaffold can increase the local concentration of reactants, accelerating the interaction between adjacent components, and can also avoid the influence of stochastic diffusion to reduce the unintentional binding interaction between further separated components. Therefore, this SLAR assay displayed an excellent analytical performance for miRNA-21 detection with a detection limit of 6 pM and showed good specificity in distinguishing miRNA-21 from similar miRNAs. In addition, the proposed assay has been experimentally demonstrated for estimation of miRNA-21 in MCF-7 and HeLa cells lysates, which exhibited great promise in the sensitive detection of biomarkers in early diagnosis.

Objectives Olfactory bulbectomy (OB) induced behaviors, hypercortisolism, inflammation and neurotrophin dysfunctions are similar to those observed in depressed patients. Omega (n)-3 polyunsaturated fatty acids (PUFAs) can effectively treat depression via anti-inflammatory and neuroprotective effects. However, n-3 PUFA purities, caloric contents, and ratios in different diets often cause contradictive results. This study used Fat-1 mice, which can convert n-6 to n-3 PUFAs in the brain, to study the effect of n-3 PUFAs on OB-induced behaviors and related changes. Methods Fat-1 and wild-type littermates were fed safflower oil for 3 months. Behaviors were tested on day 21 after surgery. Monoamine neurotransmitters were measured by HPLC. Macrophage activity was measured by MTT assay. Astrocyte phenotypes A1 S100β, A2 BDNF and cholesterol level were measured by ELISA and total cholesterol assay kits respectively. PUFA profile and membrane fluidity were detected by GC and DPH fluorescence probe respectively. Results OB significantly induced animal hyperactivity and spatial memory impairment, while decreased sucrose consumption and social contact with decreased 5-HT turnover, increased the macrophage activity and S100β/BDNF ratio. Meanwhile, n-3/n-6 PUFAs ratio and total cholesterol level were reduced in OB mice. Whereas, OB-induced behavioral changes were attenuated, which were associated with increasing 5-HT turnover, decrease macrophage activity, restored S100β/BDNF and n-3/n-6 PUFAs ratios, and total cholesterol concentrations in Fat-1 mice. Conclusion The present study for the first time demonstrated that endogenous n-3 PUFAs attenuated OB-induced depression-like behaviors and spatial memory impairment through modulating serotonergic and immune function, balancing the astrocyte A1/A2 phenotypes, and normalizing PUFAs profile and membrane function.

DNAzyme and G-quadruplex are purposed as universal recognition elements to achieve multi-functional signal amplification and readout, showing great potential in the design of biosensors and nanodevices. Herein, the single-stranded DNA (ssDNA) opens the dumbbell-shaped hairpin probes, the generated double-stranded DNA (dsDNA) is constructed as a nanobridge, and the released G-rich sequence is folded into a G-quadruplex village. The fluorescence of crystal violet/G-quadruplex complex villages is enhanced by the dsDNA nanobridge connections. Under the excitation wavelength of 580 nm, the fluorescence emission intensity at 624 nm exhibits a good linearity with the concentration of crystal violet (CV) in the range of 10–250 nM, which can be used for sensitive detection of CV. Additionally, combined with Pb2+-dependent DNAzyme, a platform for quantitative detection of Pb2+ is established. In the concentration range of 5 nM – 25 μM, the fluorescence signal increases with the increase of Pb2+ concentration. The quantitative detection limit of Pb2+ by this method is 5 nM, and it possesses good selectivity and recovery for Pb2+ in a variety of water samples. More importantly, the platform can sensitively detect various metal ions by changing the DNAzyme sequence. It has potential application in the field of environmental monitoring.

Herein, a p-n quenching photoelectrochemical (PEC) biosensor is constructed for the ultrasensitive detection of miRNA-21 which the SnO2/CdCO3/CdS ternary composite nanomaterial is used as a signal indicator and the Y-type DNA structure stabilized p-type CuS quantum dot (QD) is employed as a signal quencher. Constructing the cascade energy level structure by using three photosensitive materials, in which stannic oxide (SnO2) is used as the photoactive material and cadmium sulfide (CdS) and cadmium carbonate (CdCO3) are served as the sensitizers. The prepared SnO2/CdCO3/CdS cascade co-sensitization structure with significantly improved photocurrent signal can inhibit the recombination of photogenerated electrons and holes, thereby dramatically enhancing the initial photocurrent response. Amounts of single strands DNA (assistant DNA) are output via the exponential strand displacement amplification (E-SDA) when miRNA-21 is present. The output assistant DNA can stabilize the CuS QD by forming stable Y-type DNA nanostructure on the surface of SnO2/CdCO3/CdS modified electrode. Therefore, the PEC signal of SnO2/CdCO3/CdS ternary composite nanomaterial is greatly quenched, because of the competitive consumption of exciting light energy and electron donors, as well as the steric effect of Y-type DNA structure. The linear range of the biosensor is from 10 fM to 1 μM with the detection limit down to 3.25 fM.

Based on the 7-nitro-1,2,3-benzoxadiazole (NBD) unit, many fluorescent probes have been developed for the differential detection of biothiols. Among them, however, the use of fluorescent probes for rapid simultaneous identification of biothiols in vivo has been rarely reported. In this work, a simple fluorescent probe (NC-NBD) is rationally designed and developed according to the different reactivity of biothiols, which can simultaneously distinguish homocysteine (Hcy), cysteine (Cys), and glutathione (GSH) by adjusting two different fluorescence signals (green and yellow). The probe NC-NBD can discriminatively detect biothiols using two fluorescence channels with fast response (120–360 s), which is suitable for real-time monitoring of biothiols in biological systems. As expected, the probe NC-NBD with excellent performance is able to determine the levels of Cys/Hcy and GSH in human serum samples, and therefore has potential applications in biological specimens. The robust probe shows low toxicity and high sensitivity when applied to living cells, and successfully achieves rapid sequence distinguishing of biothiols and intracellular imaging by using two different fluorescence signals. The probe can provide an effective tool for the real-time monitoring of intracellular biothiols.

Both organic and inorganic phosphates play important roles in biological activities. Considering the conventional methods are not very useful for identifying them from their analogues, the sensor array strategy has emerged in recent years, which is able to distinguish chemical species with similar structure or chemical properties. We propose a fluorescence sensor array based on two 3d-4f metal-organic frameworks (MOFs) by recording their direct one-step fluorescence changes with the addition of various phosphates. With the introduction of d-metal ions in the sensor unit synthesis process, two 3d-4f MOFs display higher sensitivity and noticeable fluorescence differences to five phosphates than the homometallic MOFs. Phosphates show diverse affinities to sensor units, forming the basis of differential sensing. The signals are much easier to collect, enabling the sensor array to simply accomplish the identification. Through statistical analysis, the sensor array is proved to be excellent in discriminating individual phosphate and mixture. Discrimination of five phosphates is successfully performed in serum samples to illustrate the practicability of the sensor array. This work presents an efficient and reliable approach for simultaneous detection of different phosphates, which can be extended to devise other array-based sensing methods.

Ionic liquid (IL) has attracted wide attention as a pretreating solvent for efficient utilization of lignocellulosic biomass. However, IL is known to severely inactivate the cellulases. In this study, the relative activity of Paenibacillus sp. LLZ1 cellulases (PCs) was analyzed in the presence of 1-ethyl-3-methylimidazolium diethylphosphate ([Emim]DEP) with respect to carboxymethylcellulose (CMC), filter paper (FP), and microcrystalline cellulose (MCC). With increasing [Emim]DEP concentration, PCs were differentially inhibited depending on the substrate. Subsequently, the kinetic parameters Km and Vmax were analyzed. A significant increase in the Km indicated that [Emim]DEP exerted a negative impact on the affinity of the enzyme on the substrate MCC. Comparatively, the Vmax was influenced only slightly. Further study on the relative MCCase activity with different substrate content showed that PCs were less inhibited at higher substrate concentration. Finally, sodium dodecyl sulfate polyacrylamide gel elec...

Adipocyte dysfunction is associated with many metabolic diseases such as obesity, insulin resistance and diabetes. Previous studies found that phloretin promotes 3T3-L1 cells differentiation, but the underlying mechanisms for phloretin's effects on adipogenesis remain unclear. In this study, we demonstrated that phloretin enhanced the lipid accumulation in porcine primary adipocytes in a time-dependent manner. Furthermore, phloretin increased the utilization of glucose and nonesterified fatty acid, while it decreased the lactate output. Microarray analysis revealed that genes associated with peroxisome proliferator-activated receptor-γ (PPARγ), mitogen-activated protein kinase and insulin signaling pathways were altered in response to phloretin. We further confirmed that phloretin enhanced expression of PPARγ, CAAT enhancer binding protein-α (C/EBPα) and adipose-related genes, such as fatty acids translocase and fatty acid synthase. In addition, phloretin activated the Akt (Thr308) and extracellular signal-regulated kinase, and therefore, inactivated Akt targets protein. Wortmannin effectively blocked the effect of phloretin on Akt activity and the protein levels of PPARγ, C/EBPα and fatty acid binding protein-4 (FABP4/aP2). Oral administration of 5 or 10 mg/kg phloretin to C57BL BKS-DB mice significantly decreased the serum glucose level and improved glucose tolerance. In conclusion, phloretin promotes the adipogenesis of porcine primary preadipocytes through Akt-associated signaling pathway. These findings suggested that phloretin might be able to increase insulin sensitivity and alleviate the metabolic diseases.

Bone marrow mesenchymal stem cells (BMSCs) could differentiate into various cell types including adipocytes and myocytes, which had important scientific significance not only in the field of tissue regeneration, but also in the field of agricultural science. In an attempt to exhibit the characterization and differentiation into adipocytes and myocytes of porcine BMSCs, we isolated and purified porcine BMSCs by red blood cell lysis method and percoll gradient centrifugation. The purified cells presented a stretched fibroblast-like phenotype when adhered to the culture plate. The results of flow cytometry analysis and immunofluorescence staining demonstrated that the isolated cells were positive for mesenchymal surface markers CD29, CD44 and negative for hematopoietic markers CD45 and the adhesion molecules CD31. Cells were induced to differentiate into adipocytes with adipogenic medium containing insulin, dexamethasone, oleate and octanoate. Oil Red O staining demonstrated that the porcine BMSCs successfully differentiated to adipocytes. Moreover, the findings of real-time PCR and Western blotting indicated that the induced cells expressed adipogenic marker genes (PPAR-γ, C/EBP-α, perilipin, aP2) mRNA or proteins (PPAR-γ, perilipin, aP2). On the other hand, porcine BMSCs were induced into myoctyes with myogenic medium supplemented with 5-azacytidine, basic fibroblast growth factor, chick embryo extract and horse serum. Morphological observation by hochest 33342 staining showed that the induced cells presented as multi-nucleus muscular tube structure. And myogenic marker genes (Myf5, desmin) mRNA or proteins (Myf5, MyoD, myogenin, desmin) were found in the induced cells. In addition, the results of immunofluorescence staining revealed that myogenic marker (Myf5, MyoD, myogenin, desmin, S-MyHC) proteins was positive in the induced cells. Above all, these results suggested that the isolated porcine BMSCs were not only consistent with the characterization of mesenchymal stem cells, but also exhibited the multipotential capacity to form adipocytes and myocytes, which provided the basis to investigate the regulation mechanism involved in the selective differentiation of porcine BMSCs.

Many researchers have shown a great deal of interest in the effects that magnetic fields have when applied in chemical reactions, crystallization, magnetic separation of materials, magnetic levitation, materials processing, and wastewater treatment. However, surprisingly little research has been done on the effects of magnetic fields on the vapor-liquid equilibrium and the thermodynamic model for vapor-liquid phase equilibrium. The influence of magnetic fields on vapor-liquid equilibrium of binary heterogeneous azeotrope was investigated with ethanol-water in this paper. It was found that the vapor-liquid equilibrium of an ethanol-water system is influenced by the external magnetic field, but that the azeotropic point of the ethanol-water system is not changed by the magnetic field when the magnetic intensity reaches 0.8 T. Rather, the exerted magnetic field reduces the equilibrium temperature and shortens the distance between T-x curve and T-y curve in T-x-y diagram of the vapor-liquid equilibrium of the ethanol-water system. A thermodynamic model for vapor-liquid phase equilibrium in the exerted magnetic field was derived theoretically, based on the fundamental thermodynamic theory. The results show that the logarithm value of the ratio of the composition of the certain component in a magnetic field to that without the magnetic field is proportional to the magnetic susceptibility of the solution, and to the square of magnetic field intensity. This template explains and demonstrates how to prepare your camera-ready paper for Trans Tech Publications. The best is to read these instructions and follow the outline of this text.

The transparent water-insoluble silk fibroin(SF) films were casted from the mixture solution of silk fibroin and xylitol/mannitol. The structure, surface morphology, solubility, mechanical properties and light transmittance of the blend films were measured. FT-IR, X-ray diffraction retuslts indicated that the films were mainly composed of Silk I structure. SEM showed the blend films with xylitol were miscible, whereas the blend films with mannitol had phase-separated structure. There were lots of nanopores in the blend films in the wet state. The insoluble SF /xylitol films had excellent mechanical properties while the SF / mannitol films were brittle. The mechanical property of SF/alcohol blend films were consistent with the human cornea in wet state.When the contents of xylitol were 10% and 20%, the blend films had high light transmittance which were similar to human cornea. In summary, the SF /xylitol film containing 10% xylitol provides a great potential to act as repairing materials for cornea.

The aim of the present work was to study the effect of properties of ion exchange membrane on separation performance of electrodeionization (EDI) process for removal of nickel ions from dilute heavy metal solutions. It was shown that the properties of ion exchange membrane had a significant effect on removal of nickel ions. With feed water containing nickel ions at the concentration of 55mg•L-1, the nickel ions concentration of dilute stream was lower than 0.5mg•L-1and the nickel rejection was greater than 99% when using EDI membranes, while that was in the range of 2.48-3.55mg•L-1with conventional heterogeneous ion exchange membranes. With optimum ion exchange material and operating conditions, EDI would be a very interesting and innovative technology for the treatment of dilute heavy metal solutions.

Dengue virus (DENV) is the most prevalent mosquito-borne viral pathogen in humans. Neither vaccine nor antiviral therapy is currently available for DENV. We report here that N -sulfonylanthranilic acid derivatives are allosteric inhibitors of DENV RNA-dependent RNA polymerase (RdRp). The inhibitor was identified through high-throughput screening of one million compounds using a primer extension-based RdRp assay [substrate poly(C)/oligo(G) 20 ]. Chemical modification of the initial “hit” improved the compound potency to an IC 50 (that is, a concentration that inhibits 50% RdRp activity) of 0.7 μM. In addition to suppressing the primer extension-based RNA elongation, the compound also inhibited de novo RNA synthesis using a DENV subgenomic RNA, but at a lower potency (IC 50 of 5 μM). Remarkably, the observed anti-polymerase activity is specific to DENV RdRp; the compound did not inhibit WNV RdRp and exhibited IC 50 s of >100 μM against hepatitis C virus RdRp and human DNA polymerase α and β. UV cross-linking and mass spectrometric analysis showed that a photoreactive inhibitor could be cross-linked to Met343 within the RdRp domain of DENV NS5. On the crystal structure of DENV RdRp, Met343 is located at the entrance of RNA template tunnel. Biochemical experiments showed that the order of addition of RNA template and inhibitor during the assembly of RdRp reaction affected compound potency. Collectively, the results indicate that the compound inhibits RdRp through blocking the RNA tunnel. This study has provided direct evidence to support the hypothesis that allosteric pockets from flavivirus RdRp could be targeted for antiviral development.

Although flaviviruses cause significant human diseases, no antiviral therapy is currently available for clinical treatment of these pathogens. To identify flavivirus inhibitors, we performed a high-throughput screening of compound libraries using cells containing luciferase-reporting replicon of West Nile viruses (WNV). Five novel small molecular inhibitors of WNV were identified from libraries containing 96,958 compounds. The inhibitors suppress epidemic strain of WNV in cell culture, with EC(50) (50% effective concentration) values of10microM and TI (therapeutic index) values of10. Viral titer reduction assays, using various flaviviruses and nonflaviviruses, showed that the compounds have distinct antiviral spectra. Mode-of-action analysis showed that the inhibitors block distinct steps of WNV replication: four compounds inhibit viral RNA syntheses, while the other compound suppresses both viral translation and RNA syntheses. Biochemical enzyme assays showed that two compounds selectively inhibit viral RNA-dependent RNA polymerase (RdRp), while another compound specifically inhibits both RdRp and methyltransferase. The identified compounds could potentially be developed for treatment of flavivirus infections.

Twenty-four compounds including eight steroids (1–8), nine triterpenoids (9–16, 24), three flavonoids (20–22), and four benzenecarboxylic derivatives (17–19, 23) were isolated and identified from stems and twigs of medicinal mangrove plant Sonneratia caseolaris. The structures of the isolated compounds were determined by extensive analysis of their spectroscopic data. Among these metabolites, compounds 1, 4–20 and 22–24 were isolated and identified for the first time from S. caseolaris. In the in vitro cytotoxic assay against SMMC-7721 human hepatoma cells, compound 21 (3′,4′,5,7-tetrahydroxyflavone) exhibited significant activity with IC50 2.8 μg/mL, while oleanolic acid (14), 3,3′-di-O-methyl ether ellagic acid (18), and 3,3′,4-O-tri-O-methyl ether ellagic acid (19) showed weak activity. None of these compounds displayed significant antibacterial activites.

Lycorine potently inhibits flaviviruses in cell culture. At 1.2-microM concentration, lycorine reduced viral titers of West Nile (WNV), dengue, and yellow fever viruses by 10(2)- to 10(4)-fold. However, the compound did not inhibit an alphavirus (Western equine encephalitis virus) or a rhabdovirus (vesicular stomatitis virus), indicating a selective antiviral spectrum. The compound exerts its antiviral activity mainly through suppression of viral RNA replication. A Val-->Met substitution at the 9th amino acid position of the viral 2K peptide (spanning the endoplasmic reticulum membrane between NS4A and NS4B proteins) confers WNV resistance to lycorine, through enhancement of viral RNA replication. Initial chemistry synthesis demonstrated that modifications of the two hydroxyl groups of lycorine can increase the compound's potency, while reducing its cytotoxicity. Taken together, the results have established lycorine as a flavivirus inhibitor for antiviral development. The lycorine-resistance results demonstrate a direct role of the 2K peptide in flavivirus RNA synthesis.

The crystal structure of the layered compound Na2Al2B2O7 is re-examined. It is found that stacking faults can be introduced easily in the sequence of [Al2B2O7]∞2− lamellae. Both rotation and reflection twinning can form in the crystal by introducing appropriate stacking faults. A domain of the Sr2Be2B2O7-type structure might occur at the twin boundary. Single crystal refinement is performed successfully with the twinning model. A similar analysis and discussion is extended to the structures of Sr2Be2B2O7 and AEAl2B2O7 (AE=Alkaline earth elements Ca, Sr and Ba).

A new compound, LiCaBO3, has been identified in the system Li2O–CaO–B2O3 and high-quality single crystals were obtained by the flux method. Its structure has been solved and refined from single crystal and powder X-ray diffraction data, respectively. This compound crystallizes in an orthorhombic unit cell (space group Pbca) with lattice parameters a=13.227(13) A, b=6.1675(6) A, and c=6.0620(6) A. There are 8 formulas per unit cell and 6 unique atoms sites in the unit. Its structure is stacked alternately with [LiBO] and [CaO] layers along the [100] direction. In the [LiBO] layers, the isolated [BO3]3− anionic groups, distributed along two directions, [011] and [0 1 1] , are almost perpendicular to each other. The differences among the structures of the four compounds, LiCaBO3, LiMgBO3, LiSrBO3 and LiBaBO3 have been discussed, and the infrared spectra of LiCaBO3 has also been researched, which is consistent with the crystallographic study.

The subsolidus phase relations of the ternary system, Li2O–CaO–B2O3, are investigated by the powder X-ray diffraction method. The system can be divided into 19 three-phase regions and three two-phase regions based on the phase identifications of 54 ternary samples. There exist 12 binary compounds and two ternary compounds. Two new compounds with composition close to LiCaBO3 and Li6CaB3O8.5 are identified. They both have an orthorhombic structure with different lattice parameters: a=13.2537(6) A, b=6.1799(3) A, c=6.0746(3) A for LiCaBO3, and a=9.2440(6) A, b=8.0999(4) A, c=3.4839(3) A for Li6CaB3O8.5. Moreover, a compound with a composition close to 0.83LiO0.5·0.005CaO·0.165BO1.5 has been observed or confirmed in this system. We believe it is the high temperature phase of Li3BO3.

New aromatic diamines substituted with a trifluoromethyl group in the side chain, 2,4-diamino-3′-trifluoromethylazobenzene, 2,4-diamino-1-[(4′-trifluoromethylphenoxy) phenyl] aniline, and 3,5-diamino-1-[(4′-trifluoromethyl phenoxy) phenyl] benzamide were synthesized and characterized and used to prepare polyimides by a one-step high-temperature polycondensation method. Experimental results indicated that the prepared polyimides possess good solubility in strong organic solvents such as N-methyl-2-pyrrolidinone, N,N′-dimethylformamide, and N,N′-dimethylacetamide. Homogeneous solutions with solid contents as high as 15–20% can be prepared, which are stable for storing longer than 2 weeks at room temperature. The polyimides exhibited glass-transition temperatures of 249–292 °C and good thermal stability. The PI-Ic and PI-IIIc films prepared by casting the fully imidized polymer solutions showed good transparency with cutoff wavelengths at 320–330 nm. © 2002 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 40: 1572–1582, 2002

The ternary system Li2O–Al2O3–B2O3 is reinvestigated with solid-state reaction and X-ray powder diffraction technique to clarify some long-standing uncertainties. The phase relations are constructed based on the phase identifications of 51 ternary samples. Six ternary compounds, Li2AlB5O10, LiAlB2O5, Li3AlB2O6, Li2AlBO4, LiAl7B4O17 and a compound with a composition close to 0.66Li2O·0.06Al2O3·0.28B2O3, are observed or confirmed in this system, and the thermal stability of these ternary compounds is also discussed on the basis of DTA experimental results.

Two new diterpenoids, Maoecrystal U and epi-maoecrystal P, together with nine known diterpenoids and eight other known compounds were isolated from Isodon eriocalyx ( ). The structures of two new diterpenoids were elucidated as 6β-acetoxy-15β-hydroxy-3α,20-epoxy-ent-kaur-16-ene-1,7-dione and 16(S)-methoxymethy-6β,7β-dihydroxy-7α,20-epoxy-ent-kaur-2,3-ethenylene-1,15-dione on the basis of spectroscopic evidence, especially by 2D NMR techniques.

We study theoretically and numerically the acceleration of protons by a combination of laser radiation pressure acceleration and Coulomb repulsion of carbon ions in a multi-ion thin foil made of carbon and hydrogen. The carbon layer helps to delay the proton layer from disruption due to the Rayleigh–Taylor instability, to maintain the quasi-monoenergetic proton layer and to accelerate it by the electron-shielded Coulomb repulsion for much longer duration than the acceleration time using single-ion hydrogen foils. Particle-in-cell simulations with a normalized peak laser amplitude of a_0 = 5 show a resulting quasi-monoenergetic proton energy of about 70 MeV with the foil made of 90% carbon and 10% hydrogen, in contrast to 10 MeV using a single-ion hydrogen foil. An analytical model is presented to explain quantitatively the proton energy evolution; this model is in agreement with the simulation results. The energy dependence of the quasi-monoenergetic proton beam on the concentration of carbon and hydrogen is also studied.

In2O3 nanowires were successfully fabricated through a simple gas-reaction route in argon atmosphere. These nanowires have diameters ranging from 20 nm to 50 nm and lengths up to tens of micrometers. High-resolution transmission electron microscopy observations and the electron-diffraction (ED) pattern reveal that the In2O3 nanowires are formed by the stacking of (\(\bar\Box2\Box\)2\(\bar\Box2\Box\)) planes along the [\(\bar\Box1\Box\)1\(\bar\Box1\Box\)] direction, which is parallel to the wire axis. A strong and wide ultraviolet (UV) emission band centered at around 392 nm is observed for the first time in the room-temperature photoluminescence measurement in addition to the usual blue emission (468 nm). Moreover, five discrete fine peaks (372 nm, 383 nm, 406 nm, 392 nm and 413 nm) are further identified in this broad UV band.

This case—control study explored the relationship between early recurrent spontaneous abortion (RSA) and the expression of two genes: VEGFA, the gene encoding vascular endothelial growth factor (VEGF); and fms-related tyrosine kinase 1 (FLT1), the gene encoding the soluble VEGF receptor-1 (sFlt-1). Women experiencing RSA or undergoing induced abortions in the early stage of normal pregnancy were recruited to the study ( n = 30 per group). There were no significant between-group differences in maternal age or duration of pregnancy. The levels of VEGF and sFlt-1 mRNA in chorionic villus tissue samples were examined by quantitative reverse transcription—polymerase chain reaction. Levels of sFlt-1 and VEGF mRNA in the chorionic villus tissue of women with RSA were significantly higher than levels in the control group. This study demonstrated that there is a relationship between early RSA and VEGF and sFlt-1 levels, suggesting that overexpression of the FLT1 and VEGFA genes may be associated with the pathogenesis of RSA.

We describe a novel translation inhibitor that has anti-dengue virus (DENV) activity in vitro and in vivo . The inhibitor was identified through a high-throughput screening using a DENV infection assay. The compound contains a benzomorphan core structure. Mode-of-action analysis indicated that the compound inhibits protein translation in a viral RNA sequence-independent manner. Analysis of the stereochemistry demonstrated that only one enantiomer of the racemic compound inhibits viral RNA translation. Medicinal chemistry was performed to eliminate a metabolically labile glucuronidation site of the compound to improve its in vivo stability. Pharmacokinetic analysis showed that upon a single subcutaneous dosing of 25 mg/kg of body weight in mice, plasma levels of the compound reached a C max (maximum plasma drug concentration) above the protein-binding-adjusted 90% effective concentration (EC 90 ) value of 0.96 μM. In agreement with the in vivo pharmacokinetic results, treatment of DENV-infected mice with 25 mg/kg of compound once per day reduced peak viremia by about 40-fold. However, mice treated with 75 mg/kg of compound per day exhibited adverse effects. Collectively, our results demonstrate that the benzomorphan compounds inhibit DENV through suppression of RNA translation. The therapeutic window of the current compounds needs to be improved for further development.

Viral replication relies on the host to supply nucleosides. Host enzymes involved in nucleoside biosynthesis are potential targets for antiviral development. Ribavirin (a known antiviral drug) is such an inhibitor that suppresses guanine biosynthesis; depletion of the intracellular GTP pool was shown to be the major mechanism to inhibit flavivirus. Along similar lines, inhibitors of the pyrimidine biosynthesis pathway could be targeted for potential antiviral development. Here we report on a novel antiviral compound (NITD-982) that inhibits host dihydroorotate dehydrogenase (DHODH), an enzyme required for pyrimidine biosynthesis. The inhibitor was identified through screening 1.8 million compounds using a dengue virus (DENV) infection assay. The compound contains an isoxazole-pyrazole core structure, and it inhibited DENV with a 50% effective concentration (EC 50 ) of 2.4 nM and a 50% cytotoxic concentration (CC 50 ) of >5 μM. NITD-982 has a broad antiviral spectrum, inhibiting both flaviviruses and nonflaviviruses with nanomolar EC 90 s. We also show that (i) the compound inhibited the enzymatic activity of recombinant DHODH, (ii) an NITD-982 analogue directly bound to the DHODH protein, (iii) supplementing the culture medium with uridine reversed the compound-mediated antiviral activity, and (iv) DENV type 2 (DENV-2) variants resistant to brequinar (a known DHODH inhibitor) were cross resistant to NITD-982. Collectively, the results demonstrate that the compound inhibits DENV through depleting the intracellular pyrimidine pool. In contrast to the in vitro potency, the compound did not show any efficacy in the DENV-AG129 mouse model. The lack of in vivo efficacy is likely due to the exogenous uptake of pyrimidine from the diet or to a high plasma protein-binding activity of the current compound.

Recent experiments and simulations have demonstrated effective CO2 laser acceleration of quasi-monoenergetic protons from thick gaseous hydrogen target (of thickness tens of laser wavelengths) via hole boring and shock accelerations. We present here an alternative novel acceleration scheme by combining laser radiation pressure acceleration with shock acceleration of protons in a thin gaseous target of thickness several laser wavelengths. The laser pushes the thin gaseous plasma forward while compressing it with protons trapped in it. We demonstrated the combined acceleration with two-dimensional particle-in-cell simulation and obtained quasi-monoenergetic protons ∼44 MeV in a gas target of thickness twice of the laser wavelength irradiated by circularly polarized CO2 laser with normalized laser amplitude a0=10.

The ‘heteropoly-blue’ species, [NBun4]3[Cl2WVWVI17O62], prepared by exposure to UV light, has been structurally determined by X-ray diffraction; the anion contains randomly distributed WV and WVI in a 1 : 17 ratio and is the first reported mixed-valence heterotungstate which contains two chlorine heteroatoms.

The saturation of stimulated Raman scattering （SRS） through Langmuir decay instability （LDI） and electron trapping in laser-plasma interaction process is studied. Analytical expressions for the saturation time of SRS are obtained for both the LDI and electron trapping schemes in 1-D homogeneous plasma irradiated by unrelativistic laser beams. The simple analytical model agrees well with numerical simulation and experiment in respect of the dependence of the saturation time of SRS on the parameters of lasers and plasmas.