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The separation and detection of circulating tumor cells (CTCs) have a significant impact on clinical diagnosis and treatment by providing a predictive diagnosis of primary tumors and tumor metastasis. But the responsive release and downstream analysis of live CTCs will provide more valuable information about molecular markers and functional properties. To this end, specific capture and controllable release methods, which can achieve the highly efficient enrichment of CTCs with strong viability, are urgently needed. DNA networks create a flexible, semi-wet three-dimensional (3D) microenvironment for cell culture, and have the potential to minimize the loss of cell viability and molecular integrity. More importantly, responsive DNA networks can be reasonably designed as smart sensors and devices to change shape, color, disassemble, and giving back to external stimuli. Here, a strategy for specifically collecting cells using a dual-aptamer DNA network is designed. The proposed strategy enables effective capture, 3D encapsulation, and responsive release of CTCs with strong viability, which can be used for downstream analysis of live cells. The programmability of CRISPR/Cas12a, a powerful toolbox for genome editing, is used to activate the responsive release of captured CTCs from the DNA network. After activation by a specified double-strand DNA (dsDNA) input, CRISPR/Cas12a cleaves the single-stranded DNA regions in the network, resulting in molecular to macroscopic changes in the network. Accompanied by the deconstruction of the DNA network into fragments, controllable cell release is achieved. The viability of released CTCs is well maintained and downstream cell analysis can be performed. This strategy uses the enzymatic properties of CRISPR/Cas12a to design a platform to improve the programmability and versatility of the DNA network, providing a powerful and effective method for capturing and releasing CTCs from complex physiological samples.

Herein, we demonstrate that MnO2 nanosheets are an excellent carrier of CRISPR/Cas12a due to the two important roles played by them.

Methods to detect and quantify disease biomarkers with high specificity and sensitivity in biological fluids play a key role in enabling clinical diagnosis, including point-of-care testing. Myeloperoxidase (MPO) is an emerging biomarker for the detection of inflammation, neurodegenerative diseases, and cardiovascular disease, where excess MPO can lead to oxidative damage to biomolecules in homeostatic systems. While numerous methods have been developed for MPO analysis, most techniques are challenging in clinical applications due to the lack of amplification methods, high cost, or other practical drawbacks. Enzyme-linked immunosorbent assays are currently used for the quantification of MPO in clinical practice, which is often limited by the availability of antibodies with high affinity and specificity and the significant nonspecific binding of antibodies to the analytical surface. In contrast, nucleic acid-based biosensors are of interest because of their simplicity, fast response time, low cost, high sensitivity, and low background signal, but detection targets are limited to nucleic acids and non-nucleic acid biomarkers are rare. Recent studies reveal that the modification of a genome in the form of phosphorothioate is specifically sensitive to the oxidative effects of the MPO/H2O2/Cl- system. We developed an oxidative cleavage-based three-dimensional DNA biosensor for rapid, ratiometric detection of HOCl and MPO in a "one-pot" method, which is simple, stable, sensitive, specific, and time-saving and does not require a complex reaction process, such as PCR and enzyme involvement. The constructed biosensor has also been successfully used for MPO detection in complex samples. This strategy is therefore of great value in disease diagnosis and biomedical research.

The trans-cleavage activity of CRISPR/Cas12a has been widely used in biosensing applications. However, the lack of exploration on the fundamental properties of CRISPR/Cas12a not only discourages further in-depth studies of the CRISPR/Cas12a system but also limits the design space of CRISPR/Cas12a-based applications. Herein, a "RESET" effect (random extending sequences enhance trans-cleavage activity) is discovered for the activation of CRISPR/Cas12a trans-cleavage activity. That is, a single-stranded DNA, which is too short to work as the activator, can efficiently activate CRISPR/Cas12a after being extended a random sequence from its 3'-end, even when the random sequence folds into secondary structures. The finding of the "RESET" effect enriches the CRISPR/Cas12a-based sensing strategies. Based on this effect, two CRISPR/Cas12a-based biosensors are designed for the sensitive and specific detection of two biologically important enzymes.

To overcome the undesirable immune response and adverse side effects, and improve the therapeutic effect, developing a nanocarrier with good biocompatibility and high biosecurity to achieve highly efficient loading of drugs is still urgently needed. For the carriers of photodynamic therapy (PDT) photosensitizers, one of the special requirements is that the photosensitizers are better loaded on the carrier surface. In this paper, we give a paradigm of applying a pure DNA assembly as a biocompatible nanocarrier for highly efficient photosensitizer loading. Via simple DNA hybridization reactions, a pure DNA assembly (termed as a DNA nanolantern) with abundant modifiable sites was prepared and developed as a good nanocarrier for the loading of a porphyrin/G-quadruplex composite photosensitizer on the surface. The as-prepared DNA-based nanocomposite photosensitizer (DNA-NCPS) was characterized by various techniques and demonstrated to hold the charming features of nanoscale size, good monodispersity, robust biostability, strong light absorption in the biological transparent window, high tumor accumulation and cell internalization efficiency, excellent biological safety, low dark toxicity and high phototoxicity, thus showing great promise for PDT applications. Its PDT potential was verified by both in vitro cell and in vivo animal experiments. This study exhibits the great potential of pure DNA nanoassemblies in drug-loading, demonstrating their promising applications in therapy, diagnosis and theranostics.

The precise control over high-order DNA architecture assembly might be challenging due to complicated circuit design and functional unit synthesis. Here, we show an enzyme-free, catalytic assembly to construct nanometer and micrometer architectures in a bottom-up manner and applied them in nanoparticles and cell assembly.

Accurate and specific analysis of adenosine triphosphate (ATP) expression levels in living cells can provide valuable information for understanding cell metabolism, physiological activities and pathologic mechanisms. Herein, DNA nanolantern-based split aptamer nanoprobes are prepared and demonstrated to work well for in situ analysis of ATP expression in living cells. The nanoprobes, which carry multiple split aptamer units on the surface, are easily and inexpensively prepared by a “one-pot” assembly reaction of four short oligonucleotide strands. A series of characterization experiments verify that the nanoprobes have good monodispersity, strong biostability, high cell internalization efficiency, and fluorescence resonance energy transfer (FRET)-based ratiometric response to ATP in the concentration range covering the entire intracellular ATP expression level. By changing the intracellular ATP level via different treatments, the nanoprobes are demonstrated to show excellent performance in intracellular ATP expression analysis, giving a highly ATP concentration-dependent ratiometric fluorescence signal output. ATP-induced formation of large-sized DNA aggregates not only amplifies the FRET signal output, but also makes in situ ATP-imaging analysis in living cells possible. In situ responsive crosslinking of nanoprobes also makes them capable of lighting up the mitochondria of living cells. By simply changing the split aptamer sequence, the proposed DNA nanolantern-based split aptamer strategy might be easily extended to other targets.

In recent years, DNA has been widely noted as a kind of material that can be used to construct building blocks for biosensing, in vivo imaging, drug development, and disease therapy because of its advantages of good biocompatibility and programmable properties. However, traditional DNA-based sensing processes are mostly achieved by random diffusion of free DNA probes, which were restricted by limited dynamics and relatively low efficiency. Moreover, in the application of biosystems, single-stranded DNA probes face challenges such as being difficult to internalize into cells and being easily decomposed in the cellular microenvironment. To overcome the above limitations, DNA nanostructure-based probes have attracted intense attention. This kind of probe showed a series of advantages compared to the conventional ones, including increased biostability, enhanced cell internalization efficiency, accelerated reaction rate, and amplified signal output, and thus improved in vitro and in vivo applications. Therefore, reviewing and summarizing the important roles of DNA nanostructures in improving biosensor design is very necessary for the development of DNA nanotechnology and its applications in biology and pharmacology. In this perspective, DNA nanostructure-based probes are reviewed and summarized from several aspects: probe classification according to the dimensions of DNA nanostructures (one, two, and three-dimensional nanostructures), the common connection modes between nucleic acid probes and DNA nanostructures, and the most important advantages of DNA self-assembled nanostructures in the applications of biosensing, imaging analysis, cell assembly, cell capture, and theranostics. Finally, the challenges and prospects for the future development of DNA nanostructure-based nucleic acid probes are also discussed., In recent years, DNA has been widely noted as a kind of material that can be used to construct building blocks for biosensing, in vivo imaging, drug development, and disease therapy because of its advantages of good biocompatibility and programmable properties.

Molecular self-assembly is widely used in the fields of biosensors, molecular devices, efficient catalytic materials, and medical biomaterials. As the carrier of genetic information, DNA is a kind of biomacromolecule composed of deoxyribonucleotide units. DNA nanotechnology extends DNA of its original properties as a molecule that stores and transmits genetic information from its biological environment by taking advantage of its unique base pairing and inherent biocompatibility to produce structurally-defined supramolecular structures. With the continuously development of DNA technology, the assembly method of DNA nanostructures is not only limited on the basis of DNA hybridization but also other biochemical interactions. In this review, we summarize the latest methods used to construct higher-order DNA structures. The problems of DNA nanostructures are discussed and the future directions in this field are provided.

Besides gene-editing, the CRISPR/Cas12a system has also been widely used in

The introduction of nanotechnology can overcome some inherent drawbacks of traditional DNA probes, thus promoting their applications in living cells. Herein, a three-dimensional DNA nanostructure, a DNA nanolantern, was prepared via simple nucleotide hybridization of four short-stranded oligonucleotides and successfully applied to the construction of a novel DNA probe and signal amplifier. Compared to most reported DNA nanostructures, a DNA nanolantern shows the distinct advantages of low cost, easy design and preparation, more and arbitrary adjusted probe numbers, and high fluorescence resonance energy transfer (FRET) signal readout. Compared to traditional DNA probes, the constructed nanolantern-based one has improved cell internalization efficiency, enhanced biostability, accelerated reaction kinetics, excellent biocompatibility, and greatly reduced false-positive output and was demonstrated to work well for probing the expression level of tumor-related mRNA and microRNA in living cells. The DNA nanolantern can also be easily integrated with some reported signal amplification strategies, e.g., isothermal hybridization chain reaction (HCR), and the obtained signal amplifier combines the advantages of the DNA nanolantern and the HCR, enabling sensitive imaging detection of ultralow abundance targets in living cells. This work demonstrated that this simple DNA nanostructure can not only improve the performance of traditional DNA probes but can also be easily integrated with reported DNA-based strategy and technology, thus showing a broad application prospect.

As an important step in learning from nature, artificial molecular machines have attracted more and more public attention. Due to the distinct advantages of well-defined Watson-Crick base pairing rule, programmable sequence, diverse and controllable secondary structures, DNA are considered as promising building blocks for artificial molecular machines. In this review, recent research progress on DNA walker, an important member in artificial molecular machine family, is introduced on the basis of difference in driving forces, and the future development of this kind of molecular machines is prospected.

CRISPR (clustered regularly interspaced short palindromic repeats)/Cas (CRISPR-associated) proteins are powerful gene-editing tools because of their ability to accurately recognize and manipulate nucleic acids. Besides gene-editing function, they also show great promise in biosensing applications due to the superiority of easy design and precise targeting. To improve the performance of CRISPR/Cas-based biosensing systems, various nucleic acid-based signal amplification techniques are elaborately incorporated. The incorporation of these amplification techniques not only greatly increases the detection sensitivity and specificity, but also extends the detectable target range, as well as makes the use of various signal output modes possible. Therefore, summarizing the use of signal amplification techniques in sensing systems and elucidating their roles in improving sensing performance are very necessary for the development of more superior CRISPR/Cas-based biosensors for various applications. In this review, CRISPR/Cas-based biosensors are summarized from two aspects: the incorporation of signal amplification techniques in three kinds of CRISPR/Cas-based biosensing systems (Cas9, Cas12 and Cas13-based ones) and the signal output modes used by these biosensors. The challenges and prospects for the future development of CRISPR/Cas-based biosensors are also discussed.

Accurate and specific analysis of adenosine triphosphate (ATP) expression levels in living cells can provide valuable information for understanding cell metabolism, physiological activities and pathologic mechanisms. Herein, DNA nanolantern-based split aptamer nanoprobes are prepared and demonstrated to work well for in situ analysis of ATP expression in living cells. The nanoprobes, which carry multiple split aptamer units on the surface, are easily and inexpensively prepared by a "one-pot" assembly reaction of four short oligonucleotide strands. A series of characterization experiments verify that the nanoprobes have good monodispersity, strong biostability, high cell internalization efficiency, and fluorescence resonance energy transfer (FRET)-based ratiometric response to ATP in the concentration range covering the entire intracellular ATP expression level. By changing the intracellular ATP level via different treatments, the nanoprobes are demonstrated to show excellent performance in intracellular ATP expression analysis, giving a highly ATP concentration-dependent ratiometric fluorescence signal output. ATP-induced formation of large-sized DNA aggregates not only amplifies the FRET signal output, but also makes in situ ATP-imaging analysis in living cells possible. In situ responsive crosslinking of nanoprobes also makes them capable of lighting up the mitochondria of living cells. By simply changing the split aptamer sequence, the proposed DNA nanolantern-based split aptamer strategy might be easily extended to other targets.

We reported a CRISPR/Cas-based dual amplified sensing strategy for rapid, sensitive and selective detection of polynucleotide kinase/phosphatase (PNKP), a DNA damage repair-related biological enzyme. In this strategy, a PNKP-triggered nicking enzyme-mediated strand displacement amplification reaction was introduced to enrich the activator DNA strands for CRISPR/Cas. Such an isothermal DNA amplification step, together with subsequent activated CRISPR/Cas-catalyzed cleavage of fluorescent-labeled short-stranded DNA probes, enable synergetic signal amplification for sensitive PNKP detection. The proposed strategy showed a wide linear detection range (more than 3 orders of magnitude ranging from 1× 10−5 to 2.5 × 10−2 U/mL T4 PNKP) and a detection limit as low as 3.3 × 10−6 U/mL. It was successfully used for the PNKP activity detection in cell extracts with high fidelity and displayed great potential for enzyme inhibitor screening and inhibitory capability evaluation. This work broadens the applications of CRISPR/Cas12a-based sensors to biological enzymes and provides a way to improve the sensitivity by introducing an isothermal signal amplification step. Such an isothermal DNA amplification-CRISPR/Cas-combined biosensor design concept might expand CRISPR/Cas-based sensing systems and promote their applications in various fields such as disease diagnosis and drug screening.

Rapid, reliable and highly selective detection of mercury species, including Hg2+ ions and organomercury, is of significant importance for environmental protection and human health. Herein, a new fluorescent dye 1,1,2,2-tetrakis[4-(3-methyl-1H-benzimidazol-1-yl)phenyl ethylene tetraiodide (Tmbipe) with aggregation-induced emission (AIE) potential was prepared and characterized. The presence of four positively charged methylated benzimidazole groups endows Tmbipe with excellent water solubility and almost undetectable background fluorescence. However, it can coordinate with two Hg2+ ions or two organomercury molecules (e.g. methylmercury and phenylmercury) to form a planar dinuclear HgII tetracarbene complex, which can then self-aggregate to turn on AIE fluorescence. Such a fluorescence turn-on process can be completed in 3 min. In addition, synergic rigidification of the tetraphenylethylene-bridged Tmbipe molecule by mercury-mediated chelate ring formation and subsequent aggregation results in obviously higher fluorescence enhancement than that given by the single aggregation-induced one. Low background, high fluorescence enhancement and rapid response time make Tmbipe a good fluorescent probe for reliable, sensitive and highly selective quantitation of both inorganic and organic mercury species. This probe was also demonstrated to work well for identification of mercury species accumulation in living cells.

Simultaneous detection of multiple microRNAs (miRNAs) with high sensitivity can give accurate and reliable information for clinical applications. By uniformly anchoring hairpin probes on the surface of DNA nanolantern, a three-dimensional DNA nanostructure contains abundant and adjustable modification sites, highly integrated DNA nanoprobes were designed and developed as catalytic hairpin assembly (CHA)-based signal amplifiers for enzyme-free signal amplification detection of target miRNAs. The nanolantern-based CHA (NLC) amplifiers, which were facilely prepared via a simple “one-pot” annealing method, showed enhanced biostability, improved cell internalization efficiency, accelerated CHA reaction kinetics, and increased signal amplification capability compared to the single-stranded DNA hairpin probes used in traditional CHA reaction. By co-assembling multiple hairpin probes on a DNA nanolantern surface, as-prepared NLC amplifiers were demonstrated to work well for highly sensitive and specific imaging, expression level fluctuation analysis of two miRNAs in living cells, and miRNAs-guided tumor imaging in living mice. The proposed DNA nanolantern-based nanoamplifier strategy might provide a feasible way to promote the cellular and in vivo applications of nucleic acid probes.

The value of Sonoclot detection technology to guide the clinical medication of the perioperative anticoagulation and antiplatelet therapy in patients with acute myocardial infarction (AMI) undergoing emergent percutaneous coronary intervention (PCI) was estimated. One hundred and twenty-eight patients were randomly divided into control group and observation group with 64 cases in each group. Control group adopted routine blood coagulation indexes, including prothrombin time, activated partial thromboplastin time, fibrinogen and plasma thrombin time, platelet count and platelet aggregation turbidity analysis; observation group adopted Sonoclot detection technology, including activated clotting time, coagulation rate and platelet function. Anticoagulant therapy selected was of low molecular weight heparin calcium perioperatively, intraoperative unfractionated heparin, and clopidogrel (75 mg) combined with aspirin enteric-coated tablets (100 mg) as antiplatelet drugs. The therapy was administered in accordance with blood coagulation results. The blood coagulation time, postoperative creatine kinase isoenzyme MB, cardiac troponin I and B-type natriuretic peptide levels in the observation group are significantly lower than those in the control group (P0.05). The incidence of recurrent myocardial infarction, microembolism, acute and subacute thrombosis and bleeding events in the observation group are significantly lower than those in the control group (P0.05). Whereas, in the observation group, there is significant difference in coagulation indexes of the patients with thrombosis events or bleeding events or no event (P

A simple and green layer-by-layer assembly strategy is developed for the preparation of a highly bioavailable nanocomposite photosensitizer by assembling near-infrared (NIR) light-sensitive porphyrin/G-quadruplex complexes on the surface of a highly biocompatible nanoparticle that is prepared via Zn

Nonenzymatic nucleic acid amplification techniques (e.g. the hybridization chain reaction, HCR) have shown promising potential for amplified detection of biomarkers., Nonenzymatic nucleic acid amplification techniques (e.g. the hybridization chain reaction, HCR) have shown promising potential for amplified detection of biomarkers. However, the traditional HCR occurs through random diffusion of DNA hairpins, making the kinetics and efficiency quite low. By assembling DNA hairpins at the vertexes of tetrahedral DNA nanostructures (TDNs), the reaction kinetics of the HCR is greatly accelerated due to the synergetic contributions of multiple reaction orientations, increased collision probability and enhanced local concentrations. The proposed quadrivalent TDN (qTDN)-mediated hyperbranched HCR has a ∼70-fold faster reaction rate than the traditional HCR. The approximately 76% fluorescence resonance energy transfer (FRET) efficiency obtained is the highest in the reported DNA-based FRET sensing systems as far as we know. Moreover, qTDNs modified by hairpins can easily load drugs, freely traverse plasma membranes and be rapidly cross-linked via the target-triggered HCR in live cells. The reduced freedom of movement as a result of the large crosslinked structure might constrain the hyperbranched HCR in a confined environment, thus making it a promising candidate for in situ imaging and photodynamic therapy. Hence, we present a paradigm of perfect integration of DNA nanotechnology with nucleic acid amplification, thus paving a promising way to the improved performance of nucleic acid amplification techniques and their wider application.

An ultimate goal of synthetic DNA motor studies is to mimic natural protein motors in biological systems. Here, we rationally designed a highly integrated and biostable DNA motor system with high potential for living body operation, through simple assembly of a Mn2+-dependent DNAzyme-powered DNA motor with a degradable MnO2 nanosheet. The motor system shows outstanding high integration and improved biostability. High integration confers the motor system with the ability to deliver all the core components to the target sites as a whole, thus, enabling precise control of the spatiotemporal distribution of these components and achieving high local concentrations. At the target sites, reduction of the MnO2 nanosheet by intracellular glutathione (GSH) not only releases the DNA motor, which can then be initiated by the intracellular target, but also produces Mn2+ in situ to power the autonomous and progressive operation of the DNA motor. Interestingly, the resultant consumption of GSH in turn protects the DNA m...

Molecular computing holds great promise for intelligent diagnosis and treatment of diseases at molecular level, thus designing molecular logic gates to operate programmably and autonomously for molecular analysis is urgently needed. In this work, controllable assembly/disassembly of DNA tetrahedron, a typical DNA frame structure, was achieved by alternate addition of two stimulating factors including nucleic acid and antisense nucleic acid, Hg2+ and cysteine, or H+ and OH−. Such a DNA frame structure assembly/disassembly behavior had been proved to be reversible and could be cycled for several times, which was clearly reflected by repeated switch-on and switch-off of the fluorescence resonance energy transfer signal. Based on this principle, three binary INHIBIT logic gates were constructed by using different input combinations. Such a principle was also successfully used to develop various sensing systems for the ratiometric detection of microRNA, Hg2+, cysteine and pH. Thanks to the highly efficient internalization of DNA nanostructures by cells, the proposed sensing systems were demonstrated to work well for the ratiometric imaging of microRNA, Hg2+ and pH value in living cells.

A simple and highly sensitive biosensing strategy was reported by cascading terminal deoxynucleotidyl transferase (TdT)-catalyzed substrate extension and CRISPR-Cas12a -catalyzed short-stranded DNA probe cleavage. Such a strategy, which is named as TdT-combined CRISPR-Cas12a amplification, gives excellent signal amplification capability due to the synergy of two amplification steps, and thus shows great promise in the design of various biosensors. Based on this strategy, two representative biosensors were developed by simply adjusting the DNA substrate design. High signal amplification efficiency and nearly zero background endowed the biosensors with extraordinary high sensitivity. By utilizing these two biosensors, ultrasensitive detection of uracil-DNA glycosylase (UDG) and T4 polynucleotide kinase (T4 PNK) was achieved with the detection limit as low as 5 × 10−6 U/mL and 1 × 10−4 U/mL, respectively. The proposed UDG-sensing platform was also demonstrated to work well for the UDG activity detection in cancer cells as well as UDG screening and inhibitory capability evaluation, thus showing a great potential in clinical diagnosis and biomedical research.

Parkinson's disease (PD) is a common neurodegenerative disease characterized by the degeneration of dopaminergic neurons in the substantia nigra and the intraneuronal Lewy bodies in this area. Genetic mutations in PD pathogenesis have been explored and better understood in recent years. GBA variants are now considered to be the single largest risk factor for PD. Gaucher disease (GD) is a lysosomal storage disorder disease and an inherited deficiency of lysosomal glucocerebrosidase (GCase) arising from mutations in the gene GBA. A group of patients with GD exhibit parkinsonian symptoms, meanwhile, GBA mutations are more frequently observed in patients with PD. These lines of evidence suggest a close relationship between GBA mutations and PD. GBA mutations are associated with an earlier onset age and a distinct cognitive decline in PD. GCase loss-of-function caused by GBA mutations interferes with the degradation of α-synuclein, and α-synuclein pathology in turn inhibits normal GCase function in PD, which forms a vicious cycle. However, the exact mechanisms for this bidirectional pathogenic loop have not to be fully elucidated. In this review, we summarize the current understandings on the potential link between GBA mutations and PD pathogenesis, which may show novel insights into PD etiology and therapeutics.

The objective of the study is to calculate the role and underlying the molecular mechanisms of caveolin-1 (Cav-1) in atherosclerosis (AS). Cav-1 was mainly expressed in the endothelial cells of atherosclerotic lesions in both human patients and apolipoprotein E deficient (ApoE–/–) mice. Cav-1 deficiency (Cav-1–/–) attenuated high-fat diet (HFD)-induced atherosclerotic lesions in ApoE–/– mice, supported by the reduced aortic plaques. Cav-1–/– reduced the macrophage content and decreased the release of inflammation-related cytokines or chemokine in serum or abdominal aortas, accompanied with the inactivation of inhibitor κB kinase κ (IKKβ)/p65/IκBα signaling pathway. Also, the activity of mitogen-activated protein kinases 7/c-Jun-N-terminal kinase (MKK7/JNK) signaling was decreased by Cav-1–/–. In addition, oxidative stress induced by HFD in ApoE–/– mice was alleviated by Cav-1–/–. In response to HFD, Cav-1–/– markedly reduced triglyceride (TG), total cholesterol (TC), low-density lipoprotein-cholesterol (LDLC) and very low-density lipoprotein-cholesterol (VLDLC) in serum of HFD-fed ApoE–/– mice, whereas enhanced high-density lipoprotein-cholesterol (HDLC) contents. Consistent with these findings, haematoxylin and eosin (H&E) and Oil Red O staining showed fewer lipid droplets in the liver of Cav-1-deficient mice. Further, real time-quantitative PCR (RT-qPCR) analysis indicated that Cav-1–/– alleviated dyslipidemia both in liver and abdominal aortas of ApoE−/- mice fed with HFD. Cav-1 inhibition-induced attenuation of inflammatory response, oxidative stress and dyslipidemia were confirmed in vitro using mouse vascular smooth muscle cells (VSMCs) treated with ox-LDL. Surprisingly, the processes regulated by Cav-1-knockdown could be abolished through promoting JNK activation in ox-LDL-treated VSMCs. In conclusion, Cav-1 expression could promote HFD-induced AS in a JNK-dependent manner.

This study aims to explore the expression of RORγt, BATF, and IL-17 in Chinese vitiligo patients with 308 nm excimer laser treatment. One hundred and sixty-four vitiligo patients treated with 308 nm excimer laser were enrolled as Case group and 137 health examiners as Control group. Quantitative real-time polymerase chain reaction and immunohistochemistry were conducted to detect the expressions of RORγt, BATF, and IL-17. Expression of RORγt, BATF, IL-17A, and IL-17F were higher in Case group than Control group, with the diagnostic accuracy of 88.04, 87.38, 97.34, and 89.04%, respectively. Pearson correlation analysis showed a positive correlation in RORγt, BATF, IL-17A, and IL-17F mRNAs in vitiligo patients. Moreover, their expressions were higher in active vitiligo patients than stable ones. Besides, the expressions of RORγt, BATF, IL-17A, and IL-17F in vitiligo skin were significantly higher than those in non lesional skin and normal controls. After treatment, their expressions were significantly decreased. Active vitiligo and the high expressions of RORγt, BATF, and IL-17F were the independent risk factors for the ineffectiveness of 308 nm excimer laser treatment. The expressions of RORγt, BATF, IL-17 were significantly enhanced in vitiligo patients, which were correlated with the activity of vitiligo and 308 nm excimer laser therapeutic effects.

The cone is widely used in mechanical design for rotation, centering and fixing. Whether the conicity error can be measured and evaluated accurately will directly influence its assembly accuracy and working performance. According to the new generation geometrical product specification(GPS), the error and its measurement uncertainty should be evaluated together. The mathematical model of the minimum zone conicity error is established and an improved immune evolutionary algorithm(IIEA) is proposed to search for the conicity error. In the IIEA, initial antibodies are firstly generated by using quasi-random sequences and two kinds of affinities are calculated. Then, each antibody clone is generated and they are self-adaptively mutated so as to maintain diversity. Similar antibody is suppressed and new random antibody is generated. Because the mathematical model of conicity error is strongly nonlinear and the input quantities are not independent, it is difficult to use Guide to the expression of uncertainty in the measurement(GUM) method to evaluate measurement uncertainty. Adaptive Monte Carlo method(AMCM) is proposed to estimate measurement uncertainty in which the number of Monte Carlo trials is selected adaptively and the quality of the numerical results is directly controlled. The cone parts was machined on lathe CK6140 and measured on Miracle NC 454 Coordinate Measuring Machine(CMM). The experiment results confirm that the proposed method not only can search for the approximate solution of the minimum zone conicity error(MZCE) rapidly and precisely, but also can evaluate measurement uncertainty and give control variables with an expected numerical tolerance. The conicity errors computed by the proposed method are 20%-40% less than those computed by NC454 CMM software and the evaluation accuracy improves significantly.

Single-microphone speech enhancement algorithms by using nonnegative matrix factorization can only utilize the temporal and spectral diversity of the received signal, making the performance of the noise suppression degrade rapidly in a complex environment. Microphone arrays have spatial selection and high signal gain, so it applies to the adverse noise conditions. In this paper, we present a new algorithm for speech enhancement based on two microphones with nonnegative matrix factorization. The interchannel characteristic of each nonnegative matrix factorization basis can be modeled by the adopted method, such as the amplitude ratios and the phase differences between channels. The results of the experiment confirm that the proposed algorithm is superior to other dual-microphone speech enhancement algorithms.

This article selects HOL theorem proving systems for hardware Trojan detection and gives the symbol and meaning of theorem proving systems, and then introduces the symbol table, item and the meaning of HOL theorem proving systems. In order to solve the theorem proving the application of the system in hardware Trojan detection requirements, this article analyses basic hardware Trojan detection methods which applies for theorem proving systems and introduces the implementation methods and process of theorem proving about hardware Trojan detection.

Due to the complexity and non-rotational symmetry of free-form surface, it is difficult to achieve accurate and efficient inspection method. In order to solve this problem, three types of sampling sequences are proposed to specify a set of measuring points of free-form surface. For comparing the results of different sampling strategies, the profile errors of free-form surface are calculated based on a quasi particle swarm optimization (QPSO) searching the transformation parameters to implement localization and surface subdivision method finding the closest points on the design model corresponding to measured points. In order to obtain effective sampling strategies, four design models are generated by non-uniform rational basis spline (NURBS) and parts are manufactured on two machining centers to obtain surfaces of different roughness and measured on CMMs by selecting different sampling methods and sample sizes. The profile errors of parts are calculated by the proposed method and CMMs software, respectively. The results show that randomized Hammersley sampling sequence and medium sample size are preferred for the profile error inspection of given parts if accuracy and time are all considered. The research provides a method for free-form surface accurate inspection while minimizing the sampling time and cost.

Measurement uncertainty is one of the most important concepts in geometrical product specification (GPS). The “Guide to the expression of uncertainty in measurement (GUM)” is the internationally accepted master document for the evaluation of uncertainty. The GUM method (GUMM) requires the use of a first-order Taylor series expansion for propagating uncertainties. However, when the mathematical model of measurand is strongly non-linear the use of this linear approximation may be inadequate. Supplement 1 to GUM (GUM S1) has recently been proposed based on the basis of probability density functions (PDFs) using the Monte Carlo method (MCM). In order to solve the problem that the number of Monte Carlo trials needs to be selected priori, adaptive Monte Carlo method (AMCM) described in GUM S1 is recommended to control over the quality of the numerical results provided by MCM. The measurement and evaluation of cylindricity errors are essential to ensure proper assembly and good performance. In this paper, the mathematical model of cylindricity error based on the minimum zone condition is established and a quasi particle swarm optimization algorithm (QPSO) is proposed for searching the cylindricity error. Because the model is non-linear, it is necessary to verify whether GUMM is valid for the evaluation of measurement uncertainty of cylindricity error. Then, AMCM and GUMM are developed to estimate the uncertainty. The procedure of AMCM scheme and the validation of GUMM using AMCM are given in detail. Practical example is illustrated and the result shows that GUMM is not completely valid for high-precision evaluation of the measurement uncertainty of cylindricity error if only the first-order terms in the Taylor series approximation are taken into account. Compared with conventional methods, not only the proposed QPSO method can search the minimum zone cylindricity error precisely and rapidly, but also the Monte Carlo simulation is adaptive and AMCM can provide control variables (i.e. expected value, standard uncertainty and lower and higher coverage interval endpoints) with an expected numerical tolerance. The methods can be extended to the evaluation of measurement uncertainty of other form errors such as roundness and sphericity errors.

In today's Web environment, Service Oriented Architecture (SOA) becomes an efficient paradigm to integrate distributed applications. Due to loosely coupled nature of SOA, security is one of the most important issues that must be considered in SOA-based environments. Most of the existing security solutions are proposed only from one certain point of view, and they are difficult to integrate together. In this paper, an integrated framework for SOA are proposed to provides an overall security solution, which contains a three-dimensional security model, a security architecture and related security standards.

Although significant progress has been made in precision machining of free-form surfaces recently, inspection of such surfaces remains a difficult problem. In order to solve the problem that no specific standards for the verification of free-form surface profile are available, the profile parameters of free-form surface are proposed by referring to ISO standards regarding form tolerances and considering its complexity and non-rotational symmetry. Non-uniform rational basis spline(NURBS) for describing free-form surface is formulated. Crucial issues in surface inspection and profile error verification are localization between the design coordinate system(DCS) and measurement coordinate system(MCS) for searching the closest points on the design model corresponding to measured points. A quasi particle swarm optimization(QPSO) is proposed to search the transformation parameters to implement localization between DCS and MCS. Surface subdivide method which does the searching in a recursively reduced range of the parameters u and v of the NURBS design model is developed to find the closest points. In order to verify the effectiveness of the proposed methods, the design model is generated by NURBS and the measurement data of simulation example are generated by transforming the design model to arbitrary position and orientation, and the parts are machined based on the design model and are measured on CMM. The profile errors of simulation example and actual parts are calculated by the proposed method. The results verify that the evaluation precision of freeform surface profile error by the proposed method is higher 10%–22% than that by CMM software. The proposed method deals with the hard problem that it has a lower precision in profile error evaluation of free-form surface.

To master the influence of deterministic system error to navigation system, so can accurately present the precision index of gyroscopes and accelerometers, which are the main components of rate azimuth platform inertial navigation system, and initial calibration precision. With the error equations under static state, the characteristic equation of rate azimuth platform inertial navigation system is set up. Based on the solutions of the error equation, and the system error characteristics caused by the gyroscope drifts are deduced with analytical method.

New generation geometrical product specification (GPS) links the whole course of a geometrical product from the research, development, design, manufacturing and verification to its release, utilization, and maintenance. Measurement process is one of the most important part of verification/inspection in the new generation GPS. With the knowledge-intensive and globalization trend of the economy, unifying the evaluation and verification of form errors will play a vital role in international trade and technical communication. Considering the plane feature is one of the most basic geometric primitives which contribute significantly to fundamental mechanical products such as guide way of machine tool to achieve intended functionalities, the mathematical model of flatness error minimum zone solution is formulated and an improved genetic algorithm (IGA) is proposed to implement flatness error minimum zone evaluation. Then, two evaluation methods of flatness error uncertainty are proposed, which are based on the Guide to the Expression of Uncertainty in Measurement (GUM) and a Monte Carlo Method (MCM). The calculating formula and the propagation coefficients of each element and correlation coefficients based on GUM and the procedures based on MCM are developed. Finally, two examples are listed to prove the effectiveness of the proposed method. An investigation into the source and effects of different uncertainty contributors for practical measurement on CMM is carried out and the uncertainty contributors significant are analyzed for flatness error verification. Compared with conventional methods, the proposed method not only has the advantages of simple algorithm, good flexibility, more efficiency and accuracy, but also guarantees the minimum zone solution specified in the ISO/1101 standard. Furthermore, it accords with the requirement of the new generation GPS standard which the measurement uncertainty characterizing the reliability of the results is given together. And it is also extended to other form errors evaluation and verification.

To lower the cost of passive gravity navigation, a simple and easy passive gravity navigation, which is composed of a rate azimuth inertial platform with a gravity sensor on it, and a digital gravity map, is proposed. Based on the local observability theory, the relationship between the observation of the integrated system and the gravity field’s roughness is deduced by the singular value of the observation matrix. The simulation of the integrated navigation system is carried out with Matlab/Simulink and gravity anomaly map whose resolution is five over one thousand. Simulation results show that, in the area with evident gravity anomaly, the positioning error of the RAPINS/Gravity integrated navigation system, combined of moderate precision accelerometers and gyroscopes, is small, which can meet up with the demand of underwater passive navigation.

To know the influence of deterministic system error to navigation system well, so the precision index of gyroscopes and accelerometers, which are the main components of rate azimuth platform inertial navigation system, and initial calibration precision, can be put forward. Based on the error equations, the characteristic equation of rate azimuth platform inertial system is established, and the solutions of the error equation under static state are obtained with analytical method, and the system error characteristics caused by the accelerometer drifts are analyzed.

An algorithm based on the differential evolutionary (DE) computation is proposed to evaluate circularity error. It is a heuristic evolutionary algorithm based on population optimization .In the meantime, the suggested method is used to solve the minimum zone circularity error. Compared with other methods, the results show the presented method has very strong self-adaptive ability to environment and better global convergence. Examples proves that the proposed method is effective, convergence and robustness in the process of optimization. And this method makes the circularity error evaluation more accurate.

Iterative Learning Control (ILC) has recently emerged as a powerful control strategy that iteratively achieves a higher accuracy for systems with repetitive tasks. The basic idea of ILC is to construct a compensation signal based on the tracking error in each repetition so as to reduce the tracking error in the next repetition. In this paper, particle swarm optimization (PSO) is proposed to optimize the input of iterative learning controller. The experimental results confirm that the proposed method not only has higher tracking accuracy than that of Improved Genetic Algorithm (IGA) and traditional Genetic Algorithm based elisit strategy (EGA), but also has the advantages of simple algorithm and good flexibility. And compared with conventional iterative learning control methods, it is easy to solve the optimal input for non-linear plant models.

High-speed cutting (HSC) is a practical technique of advanced manufacturing technology, leading the development of cutting and machining. This paper discusses the mechanism of HSC and the worldwide status quo, application and prospects of HSC.

To examine the association of 5-lipoxygenase (5-LOX) expression with clinicopathological factors in colorectal cancer.Immunohistochemical stain was used to detect the 5-LOX expression in 52 resected specimens of colorectal cancer. The association between 5-LOX expression and clinicopathological factors was examined.The positive rate of 5-LOX expression in 52 specimens of colorectal carcinoma was 73.1% (38/52). In 41 colorectal cancer specimens with lymph node metastasis, the positive rate of 5-LOX expression was higher than that in the specimens without metastasis (87.8% vs. 18.2%, P0.05). The positive rate of 5-LOX expression in the specimens with deep infiltration (T3 and T4) was higher than that in the specimens with superficial infiltration (T1 and T2) (81.1% vs. 53.3%, P0.05). The positive rate of 5-LOX expression in TNM stage III and IIII cancer was higher than that in stage I and II (79.5% vs. 53.8%, P0.05). The positive rate of 5-LOX expression in cancers of poor differentiation and non-differentiation adenocarcinoma was higher than that of well and moderately differentiated cancer (100% vs. 50.0%, P0.05). There were no significant differences of 5-LOX expression with tumor size,vascular invasion and peritoneal dissemination.5-LOX expression in colorectal carcinoma is closely associated with lymph node metastasis, infiltration depth, differentiation degree and TNM stage.