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The high incidence of lymphatic metastasis is closely related to poor prognosis and mortality in cancers. Potent inhibitors to prevent pathological lymphangiogenesis and lymphatic spread are urgently needed. The VEGF-C-VEGFR3 pathway plays a vital role in driving lymphangiogenesis and lymph node metastasis. In addition, COX2 in tumor cells and tumor-associated macrophages (TAMs) facilitates lymphangiogenesis. We recently reported that aiphanol, a natural stilbenolignan, attenuates tumor angiogenesis by repressing VEGFR2 and COX2. In this study, we evaluated the antilymphangiogenic and antimetastatic potency of aiphanol using in vitro, ex vivo and in vivo systems. We first demonstrated that aiphanol directly bound to VEGFR3 and blocked its kinase activity with an half-maximal inhibitory concentration (IC50) value of 0.29 μM in an in vitro ADP-GloTMkinase assay. Furthermore, we showed that aiphanol (7.5−30 μM) dose-dependently counteracted VEGF-C-induced proliferation, migration and tubular formation of lymphatic endothelial cells (LECs), which was further verified in vivo. VEGFR3 knockdown markedly mitigated the inhibitory potency of aiphanol on lymphangiogenesis. In 4T1-luc breast tumor-bearing mice, oral administration of aiphanol (5 and 30 mg· kg−1·d−1) dose-dependently decreased lymphatic metastasis and prolonged survival time, which was associated with impaired lymphangiogenesis, angiogenesis and, interestingly, macrophage infiltration. In addition, we found that aiphanol decreased the COX2-dependent secretion of PGE2 and VEGF-C from tumor cells and macrophages. These results demonstrate that aiphanol is an appealing agent for preventing lymphangiogenesis and lymphatic dissemination by synergistically targeting VEGFR3 and inhibiting the COX2-PGE2-VEGF-C signaling axis.

Abnormal expression of ADAM29 has been frequently reported in several cancers, however, its role in clear cell renal cell carcinoma (ccRCC) has not evaluated in detail. Herein, we attempt to determine the biological role and the action mechanism of ADAM29 in ccRCC. Bioinformatics analysis based on the ccRCC RNA-Seq dataset from TCGA database revealed that ADAM29 was up-expressed in ccRCC tissues by comparison with normal tissues. And a significant increase of ADAM29 expression was also observed in 3 ccRCC cell lines (UT33A, Caki-1, and786-O) in comparison with normal cell line. Besides, high level of ADAM29 was found to be connected with the poor prognosis and could be considered as an independent prognosticator for patients with ccRCC. Furthermore, functional experiments in vitrodemonstrated that ADAM29 promoted the growth, invasion and migration of ccRCC cells. Moreover, Western blot assays indicated that ADAM29 was positively correlated with the level of proliferation-related proteins Cyclin D1 and PCNA and motion-related proteins MMP9 and Snail. Our data indicate that ADAM29 acts as an oncogene that increases tumour cells proliferation, invasion and migration partly by regulating the expression of Cyclin D1/PCNA/MMP9/Snail, suggesting that ADAM29 may become a prognosticator and therapeutic candidate for ccRCC.

Objective: To investigate the roles and mechanisms of AKAP150/protein kinase Cα ( PKCα) signaling pathway in aerobic exercise-mediated regulation of L-type Cav1.2 channel (Cav1.2) function in cerebral arteries during hypertension. Methods; 12-week-old male spontaneously hypertensive rats ( SHR) and Wistar-Kyoto (WKY) rats were randomly assigned to sedentary (WKY-SED, SHR-SED) and exercise training (WKY-EX, SHR-EX) groups, respectively. Exercise groups were performed a moderate-intensity treadmill running. After 12 weeks, Western blot, immunofluorescence, patch-clamp whole-cell and single channel recording, qPCR were used to detect AKAP150/PKCα signaling pathway, Cav1.2 whole-cell and single channel currents, mRNA and protein expression of α1c subunits. AKAP150 smooth muscle-specific knockin mice (AKAP150 smKI) were constructed. The blood pressure was monitored in conscious WT and AKAP150 smKI mice using an implantable mouse telemetry system. Results; 1) Aerobic exercise significantly reduced body weight and blood pressure in SHR-EX. 2) The protein expression of AKAP150 in cerebral arterial smooth muscle was significantly up-regulated in SHR-SED, while down-regulated in SHR-EX. The protein expression of PKCα did not change in four groups, however, sarcolemmal PKCα translocation was significantly reduced in SHR-EX as compared with SHR-SED myocytes. 3 ) Aerobic exercise significantly decreased the colocalization rate of AKAP150 and PKCα in cerebral arterial myocytes from SHR-EX. 4) Aerobic exercise significantly inhibited SHR-induced increases in Cav1.2 currents in cerebral arterial myocytes, whereas the mRNA and protein expression of α1c subunits were unchanged. 5) Aerobic exercise significantly attenuated the Cav 1.2 channel nPo of SHR-EX cerebral arterial myocytes. 6) Inhibition of PKCα with Gö6976 decreased the Cav1.2 currents nearly 31%, 35%, 48% and 38% in WKY-SED, WKY-EX, SHR-SED and SHR-EX myocytes. 7) Application of the PKC activator PDBu increased a - 1.5-fold and 3.8 fold in the Cav1.2 channel nPo of WKY-SED and SHR-SED, respectively. Aerobic exercise significantly suppressed PDBu-evoked Cav 1.2 channel nPo in SHR-EX. 8) The systolic blood pressure, diastolic blood pressure and mean arterial pressure were significantly higher in AKAP150 smKI than in WT mice. Conclusions; Aerobic exercise effectively reduces blood pressure of SHR, inhibits sarcolemmal PKCα translocation, LTCC channel activity via suppression of AKAP150 protein expression, and ameliorates cerebral arterial function during hypertension. [ABSTRACT FROM AUTHOR]

CD97 belongs to the adhesion GPCR family characterized by a long ECD linked to the 7TM via a GPCR proteolytic site (GPS) and plays important roles in modulating cell migration and invasion. CD97 (EGF1-5) is a splicing variant of CD97 that recognizes a specific ligand chondroitin sulfate on cell membranes and the extracellular matrix. The aim of this study was to elucidate the extracellular molecular basis of the CD97 EGF1-5 isoform in protein expression, auto-proteolysis and cell adhesion, including epidermal growth factor (EGF)-like domain, GPCR autoproteolysis-inducing (GAIN) domain, as well as GPS mutagenesis and N-glycosylation. Both wild-type (WT) CD97-ECD and its truncated, GPS mutated, PNGase F-deglycosylated, and N-glycosylation site mutated forms were expressed and purified. The auto-proteolysis of the proteins was analyzed with Western blotting and SDS-PAGE. Small angle X-ray scattering (SAXS) and molecular modeling were used to determine a structural profile of the properly expressed receptor. Potential N-glycosylation sites were identified using MS and were modulated with PNGase F digestion and glyco-site mutations. A flow cytometry-based HeLa cell attachment assay was used for all aforementioned CD97 variants to elucidate the molecular basis of CD97-HeLa interactions. A unique concentration-dependent GPS auto-proteolysis was observed in CD97 EGF1-5 isoform with the highest concentration (4 mg/mL) per sample was self-cleaved much faster than the lower concentration (0.1 mg/mL), supporting an intermolecular mechanism of auto-proteolysis that is distinct to the reported intramolecular mechanism for other CD97 isoforms. N-glycosylation affected the auto-proteolysis of CD97 EGF1-5 isoform in a similar way as the other previously reported CD97 isoforms. SAXS data for WT and deglycosylated CD97ECD revealed a spatula-like shape with GAIN and EGF domains constituting the body and handle, respectively. Structural modeling indicated a potential interaction between the GAIN and EGF5 domains accounting for the absence of expression of the GAIN domain itself, although EGF5-GAIN was expressed similarly in the wild-type protein. For HeLa cell adhesion, the GAIN-truncated forms showed dramatically reduced binding affinity. The PNGase F-deglycosylated and GPS mutated forms also exhibited reduced HeLa attachment compared with WT CD97. However, neither N-glycosylation mutagenesis nor auto-proteolysis inhibition caused by N-glycosylation mutagenesis affected CD97-HeLa cell interactions. A comparison of the HeLa binding affinities of PNGase F-digested, GPS-mutated and N-glycosylation-mutated CD97 samples revealed diverse findings, suggesting that the functions of CD97 ECD were complex, and various technologies for function validation should be utilized to avoid single-approach bias when investigating N-glycosylation and auto-proteolysis of CD97. A unique mechanism of concentration-dependent auto-proteolysis of the CD97 EGF1-5 isoform was characterized, suggesting an intermolecular mechanism that is distinct from that of other previously reported CD97 isoforms. The EGF5 and GAIN domains are likely associated with each other as CD97 expression and SAXS data revealed a potential interaction between the two domains. Finally, the GAIN and EGF domains are also important for CD97-HeLa adhesion, whereas N-glycosylation of the CD97 GAIN domain and GPS auto-proteolysis are not required for HeLa cell attachment.

In recent years, Klebsiella pneumoniaehas emerged as a leading cause of nosocomial infection owing to the rising prevalence of multidrug-resistant strains, particularly carbapenem-resistant isolates. In this study, the complete genome sequence of carbapenem-resistant K. pneumoniaeSWU01 was determined.

Fluorescence microscopy of live cells is instrumental in deciphering the molecular details of autophagy. To facilitate the routine examination of yeast Atg proteins under diverse conditions, here we provide a comprehensive tool set, including (1) plasmids for the expression of GFP chimeras at endogenous levels for most Atg proteins, (2) RFP-Atg8 constructs with improved properties as a PAS marker, and (3) plasmids for the complementation of common yeast auxotrophic markers. We hope that the availability of this tool set will further accelerate yeast autophagy research.

A series of fatigue tests is performed for polymethyl methacrylate (PMMA) specimens without scratches and with scratches. The scratch depth is set to be 0.2mm, 0.3mm and 0.4mm, respectively. Based on the experimental results, it can be deduced that contained fatigue limit of specimens with scratches significantly decreases compared to specimens without scratches. Besides, contained fatigue limit decreases gradually with scratch depth increasing from 0.0mm to 0.4mm. Consequently, it can be concluded that fatigue properties of PMMA will decrease when there exists scratch and for scratch depth from 0.0mm to 0.4mm, fatigue property decreases remarkably with deeper scratch.

Aiming at the de-noising of GPR echo signal, a de-noising method based on EEMD and wavelet is presented. First the echo signal data is processed with EEMD and yields IMF components. Then the IMF components which indicate noise are subtracted. Next, the high frequency IMF components of the remaining are subjected to wavelet threshold. Finally, the signal is reconstructed using the de-noising IMF and low frequency IMF to realize signal de-noising. Compared with other commonly used methods, EEMD-wavelet method has improvement on SNR. The experiment results show its effectiveness and feasibility in GPR de-noising.

Static tensile tests of polymethyl methacrylate (PMMA) specimens without scratches and containing a scratch whose depth was set to be 0.2mm, 0.3mm and 0.4mm, respectively, were carried out to explore effect of scratch depths on tensile strength of PMMA. Based on the experimental load-displacement curves, the specimens without scratches deform most severely when the failure comes forth. The failure deformation declines gradually with higher scratch depth, namely higher scratch depth leads to lower plastic deformation. It thus follows that the scratch significantly reduces the tensile strength of PMMA while the variation of the scratch depth from 0.2mm to 0.4mm has negligible influence on the tensile strength of PMMA.

Endothelial and erythropoietic lineages arise from a common developmental progenitor. Etv2 is a master transcriptional regulator required for the development of both lineages. However, the mechanisms through which Etv2 initiates the gene-regulatory networks (GRNs) for endothelial and erythropoietic specification and how the two GRNs diverge downstream of Etv2 remain incompletely understood. Here, by analyzing a hypomorphic Etv2mutant, we demonstrate different threshold requirements for initiation of the downstream GRNs for endothelial and erythropoietic development. We show that Etv2 functions directly in a coherent feedforward transcriptional network for vascular endothelial development, and a low level of Etv2expression is sufficient to induce and sustain the endothelial GRN. In contrast, Etv2 induces the erythropoietic GRN indirectly via activation of Tal1, which requires a significantly higher threshold of Etv2to initiate and sustain erythropoietic development. These results provide important mechanistic insight into the divergence of the endothelial and erythropoietic lineages.

In this paper we construct model in computer forensics by using Benford’s Law, which is one of three famous laws in statistics in linguistics. Especially, we use Benford’s Law to analyze data in file analysis and Anomaly detection. We illustrate that Benford’s Law is able to supply the strategy for data analysis. The future work, Benford’s Law will be the theoretical rule for intelligent compression of mass data.

This paper studies Mobile Payment App from Mobile payment and Application. We analyze the data security risks hidden in this payment model. Give the corresponding solutions. Finally, look forward the research direction of Mobile Payment App security.

Life satisfaction is associated with positive development in adolescents. Understanding a path model of life satisfaction can help healthcare providers design interventions to improve positive development in adolescents with type 1 diabetes.

Through the research of design management on the implementation of Hangzhou Cigarette Factory the 11th five-year reconstruction technical reform project,proposes the method of designing stage management and coordination management, for the same type of tobacco construction project reference.

Beam pipe is a key part of the electron positron collider (EPC). It is very expensive and hard to install due to a limited room and high assembly precision requirement. In order to ensure the beam pipe being installed successfully, an installation scheme is introduced. Key factors affected the deformation of installation mechanism are presented by classical mechanics theory. Different finite element models were analyzed to obtain an optimal geometry size for the protective shaft based on commercial software ANSYS®. The deformation of the determinate installation mechanism satisfies the engineering requirement well.

The cone dynamic penetration test is a kind of test method, which has the features of simple device, easy operation, great adaptability, high efficiency and it also is a continuous in-situ test method. But in the common geotechnical exploration activities, we found that bearing capacity of foundation is always larger than the measured value, by analysis to the blow count of cone dynamic penetration in the standard table. This article set the gravel soil geotechnical exploration of Guangxi province’s city planning exhibition center of China as the example, to start with the theory of cone dynamic penetration, and analysis the reason why there are the abnormal value in the dynamic penetration, furthermore, provide the relative error correction models for it. Finally, establish the rational relationship between dynamic penetration results and foundation soil mechanical parameters.

This paper proposes a novel rotor position estimation method based on line back EMF for brushless DC motor (BLDCM). The relationship between position phase error and the symmetric line back EMF is analyzed. It is noted that line back EMF error can exactly reflect rotor position phase error. If the amplitude of line back EMF at the beginning of commutation is equal to the amplitude of line back EMF at the end of commutation, the waveform of line back EMF should be symmetric. Based on the above analysis, a closed loop control method to correct rotor position phase is presented. Line back EMF error is employed to keep the waveform of line back EMF symmetrical by PI controller. This paper also develops a sensorless BLDCM driver to implement the proposed method based on DSP and the experimental results prove the validity of the proposed method.

We propose a theoretical design for a compact photonic crystal (PC) polarization beam splitter (PBS) based on the multimode interference (MMI) effect. The size of a conventional MMI device designed by the self-imaging principle is not compact enough; therefore, we design a compact PC PBS based on the difference of the interference effect between TE and TM modes. Within the MMI coupler, the dependence of interference of modes on propagation distance is weak for a TE wave and strong for a TM wave; as a result, the length of the MMI section can be only seven lattice constants. Simulation results show that the insertion losses are 0.32 and 0.89 dB, and the extinction ratios are 14.4 and 17.5 dB for Port 1 (TE mode) and Port 2 (TM mode), respectively.

Dry core rot of apple (DCR) and Alternaria black rot of citrus (ABR) have in the past respectively been ascribed to Alternaria alternata and A. citri. In recent years, however, it has been speculated that several other species of Alternaria could also be associated with these diseases. In an attempt to elucidate the identity of these taxa, 25 isolates associated with DCR, and 26 isolates associated with ABR were selected for molecular characterisation. Nucleotide sequences of 1116 sites including the histone gene section and the internal transcribed spacers (ITS 1 and 2) of the rRNA gene were determined for these isolates. The gene trees generated from the individual and combined data sets using maximum parsimony, maximum likelihood and neighbour-joining analysis methods distinguished five clades with strong bootstrap support, namely Alternaria sp., A. arborescens, A. infectoria, A. tenuissima, and a clade containing isolates variable in morphology, referred to as the Alternaria group. In the alignment of the combined ITS and histone data set, unique transition/transversion substitutions, as well as positional insertions and deletions were observed for each of the above clades. In addition, key sequences in the form of serial composing nucleotides in both the ITS and histone sections of the alignment were also discovered for the molecular identification of A. arborescens, A. infectoria and A. tenuissima. The final phylogeny also indicated that no host specificity existed among the species associated with these two post-harvest disease complexes. Contrary to the host specificity observed on leaf diseases of these hosts in the field, it appears that the post-harvest diseases are the result of adverse storage conditions and opportunism of different small-spored Alternaria spp.

ABSTRACTThe use of viral thymidine kinase (TK) gene coupled with the administration of ganciclovir to render cancer cell death has been studied extensively. Many of these experiments utilized retrovirus to transfer the TK gene under the control of a nonspecific promoter. Because nonspecific expression of the viral TK gene may cause death of proliferating cells, other than cancer cells, we explored the use of a liver-specific promoter and a tumor-specific AFP enhancer to achieve regulated viral TK gene expression for treatment of hepatocellular carcinoma. We also used the adeno-associated virus (AAV) as vector for the delivery of the TK gene because this virus is not associated with any pathological consequences in humans. Because it can infect nondividing S-phase cells, AAV can transfer genes into noncycling tumor cells. A recombinant AAV virus was constructed to include the selectable marker neoRgene and the herpes simplex virus (HSV)-TK gene driven by the human AFP enhancer and the albumin promoter. The liver-predominant expression pattern of the TK gene was observed when this construct was tested in transgenic mice. When human hepatocellular carcinoma cell lines displaying different levels of AFP and albumin and nonhepatocyte tumor cell lines were infected with the recombinant AAV virus, ganciclovir treatment caused only AFP and albumin-positive hepatocellular carcinoma cells death, but not nonhepatocyte tumor cells or AFP and albumin-negative hepatic tumor cells. Moreover, the dose required to kill the cancer cells was inversely proportional to the level of AFP expression in the cells.

Most of the studies on the herb Chuanxiong Rhizoma(CR) have focused on the l-arginine-nitric oxide (NO) pathway, but the nitrate-nitrite-NO (NO3−-NO2−-NO) pathway was rarely investigated. Therefore, the aim of this study was to evaluate the effects and mechanisms of action of CR in coronary artery disease (CAD).