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Additional file 1: Fig S1. Amino acid sequence alignment of known KstDs. Fig. S2. Construction process of suicide plasmids pKADel. Fig. S3. SDS-PAGE analyses of soluble KstDs in cell-free extracts of recombinant E. coli cells. Fig. S4. Elimination of by-products 9,24-DHC in conversion of phytosterols of mutants. Table S1. Primers, plasmids, and strains used in this study. Table S2. Summary of bioinformatics analysis and secondary structure prediction of the putative KstDs.

This work presents an in-situ technique to quantify the layer-by-layer roughness of thin films and heterostructures by measuring the spectral profile of the reflection high-energy electron diffraction (RHEED). The characteristic features of the diffraction spot, including the vertical to lateral size ratio c/b and the asymmetrical ratio c1/c2 along the vertical direction, are found to be quantitatively dependent on the surface roughness. The quantitative relationships between them are established and discussed for different incident angles of high-energy electrons. As an example, the surface roughnesses of LaCoO3 films grown at different temperatures are obtained using such an in-situ technique, which are confirmed by the ex-situ atomic force microscopy. Moreover, the in-situ measured layer-by-layer roughness oscillations of two LaCoO3 films are demonstrated, revealing drastically different information from the intensity oscillations. The experiments assisted with the in-situ technique demonstrate an outstanding high resolution down to ∼ 0.1 A. Therefore, the new quantitative RHEED technique with real-time feedbacks significantly escalates the thin film synthesis efficiency, especially for achieving atomically smooth surfaces and interfaces. It opens up new prospects for future generations of thin film growth, such as the artificial intelligence-assisted thin film growth.

Pheretima is a common and valuable animal-derived medication used in traditional Chinese medicine. There are four species of Pheretima specified in the Chinese Pharmacopoeia (2015 edition), i.e. Pheretima aspergillum, P. vulgaris, P. guillelmi, and P. pectinifera. A recent report revealed ~ 55% of Pheretima in the commercial marketplace may be adulterated by other species, contrary to the Pharmacopoeia standard. The safety, efficacy, and authenticity of Pheretima is an important issue. Currently, the availability of specific quality-markers for the various species and effective identification methods are still limited. In this study, label-free quantification proteomics of species from Pheretima and Amynthas was carried out using nanoscale liquid chromatography coupled to tandem mass spectrometry (nano LC-MS/MS), and marker peptides were identified based on their ion intensities using multivariate data analysis (principal component analysis and supervised partial least-squares discriminant analysis). A total of 48,476 peptides with high confidence corresponding to 13,397 proteins were identified from all samples. The marker peptides were validated by comparison with synthetic peptide reference standards using LC-MS/MS operating in a multiple-reaction monitoring mode. A multiple-peptide identification strategy was proposed for the authentication of Pheretima and subsequently applied to samples obtained from retail outlets in various regions of China. The results showed that eight out of the 15 samples tested were deemed authentic Pheretima.

Additional file 2: Fig. S2A: Relations between receptors and expression of PBK in pan-cancers (red is positive correlated and blue is negative correlated). B: PBK expression was positively closely related with CCR6 and CCR10, and was negatively correlated with CCR7 and CXCR1.

Additional file 4: Table S1. Multivariate Cox analysis of immune cells infiltration.

Additional file 3: Fig. S3 A: Relations between chemokines and expression of PBK in pan-cancers (red is positive correlated and blue is negative correlated). B: PBK expression was positively or negatively closely related with chemokines (rho>0 indicate positively, rho

Additional file 1: Fig. S1 The expression of PBK in pan-cancers and HCC. A: PBK mRNA levels in 20 types of human cancers analyzed by Oncomine. Red means increased expression and blue means decreased expression. The numbers indicated the amounts of dataset satisfying the threshold in the colored cell. B: PBK mRNA levels analyzed by TIMER. (***P

With the development of low-wind-speed technology, it becomes a trend that low-wind-speed wind turbine generators (LWTGs) are integrated into a microgrid. However, the frequency stability of the microgrid has thereby been challenged since the increased penetration of wind power lowers the inertia of the microgrid. In order to investigate how LWTGs can effectively participate in suppressing the frequency fluctuation of the microgrid, virtual inertia control, over-speed control, as well as droop control, is applied to LWTG. Moreover, the de-loading ratio of over-speed control, along with the control parameters of virtual inertia control and droop control are all optimised under different wind speeds by virtue of the deep belief network, whereas the problem of over-speed control failure with the scheme of fixed de-loading ratio becomes more pronounced under low-wind speeds, which is defined as a blind area problem. To solve this problem, on the one hand, the strategy of the variable de-loading ratio is adopted under low-wind-speeds. On the other hand, the concepts of the minimum and maximum critical wind speed are deduced through theoretical analysis, which greatly restricts the number of feasible solutions of de-loading ratio under different wind speeds so as to improve the optimisation efficiency about 50%.

Among a series of 3D printing techniques, stereolithography provides a new route to produce ceramic architectures with the advantages of high-precision and short cycle time. However, up to now the stereolithography of non-oxide ceramics still face complex and difficult problems. This work focused on the analysis of rheological and curing ability of Si3N4 photocurable slurries. The effects of monomer type, coarse silicon powder, solid loading and ambient temperature on the rheological behavior were intensively studied. The relationships between powder characteristic (involving refractive index, absorbance and the introduce of coarse silicon powder), monomer type and curing ability were discussed in detail. It is expected that this study may benefit the development of Si3N4 or other non-oxide ceramic slurries for stereolithography.

Lin Liu,1,* Dongyu Xie,2,3,* Haina Xie,4,* Wei Huang,1 Jingxian Zhang,5 Wenye Jin,5 Wenjing Jiang,1 Donghao Xie5,6 1Department of Pharmacy, Dahua Hospital, Shanghai, People’s Republic of China; 2Department of Spleen-Stomach, Zhenjiang Affiliated Hospital of Nanjing University of Chinese Medicine, Zhenjiang, People’s Republic of China; 3Department of Spleen-Stomach, Zhenjiang Hospital of Traditional Chinese Medicine, Zhenjiang, People’s Republic of China; 4School of Basic Medical Sciences, Shanghai University of Traditional Chinese Medicine, Shanghai, People’s Republic of China; 5Department of Pharmacy, Guanghua Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai, People’s Republic of China; 6School of Pharmacy, Jiangsu University, Zhenjiang, People’s Republic of China*These authors contributed equally to this workCorrespondence: Donghao XieDepartment of Pharmacy, Guanghua Hospital of Integrated Traditional Chinese and Western Medicine, Shanghai, People’s Republic of ChinaTel +86- 18221220090Email xdhdh2012@126.comIntroduction: ARHGAP10 belongs to the ARHGAP family, which is downregulated in certain human tumors. However, the detailed function of ARHGAP10 remains unclear in human colon carcinoma (CRC). In the current study, we aimed to explore the role of ARHGAP10 in the growth and metastasis of CRC cells.Methods: ARHGAP10 was induced silencing and overexpression using RNA interference (RNAi) and lentiviral-vector in CRC cells. Quantitative real-time PCR (qRT-PCR) and Western blot were used to quantify the mRNA and protein contents of ARHGAP10. Cell proliferation was determined by using Cell counting kit-8 (CCK-8). Transwell assay was utilized to examine the role of ARHGAP10 in the migration and invasion of CRC cells.Results: Our results indicated that ARHGAP10 was downregulated in human CRC tissues and low expression of ARHGAP10 was associated with poor prognosis of patients with CRC. Moreover, ARHGAP10 overexpression significantly inhibited the proliferation and metastasis of CRC cells. Moreover, a PI3K/AKT inhibitor LY294002 was utilized to examine the connection between ARHGAP10 and AKT. Our findings demonstrated that the AKT inhibitor LY294002 could rescue the function of ARHGAP10 in CRC cells.Discussion: It was the first time to elucidate that AKT involved in the ARHGAP10 signaling pathway and ARHGAP10 negatively mediated the phosphorylation of AKT (p-AKT) and RhoA activity in CRC cells. Interestingly, the Rho/MRTF/SRF inhibitor CCG-1423 significantly inhibited the phosphorylation of AKT in ARHGAP10 siRNA transfected CRC cells. Much importantly, overexpression of ARHGAP10 deeply suppressed the metastasis of CRC cells in the lung in vivo. Taken together, our findings not only enhanced the understanding of the anti-cancer effect of ARHGAP10 in CRC cells but also indicated its underlying pathway in CRC.Keywords: ARHGAP10, AKT, RhoA activity, colon carcinoma

Music history, referring to the records of users' listening or downloading history in online music services, is the primary source for music service providers to analyze users' preferences on music and thus to provide personalized recommendations to users. In order to engage users into the service and to improve user experience, it would be beneficial to provide visual analyses of one user's music history as well as visualized recommendations to that user. In this paper, we take a user-centric approach to the design of such visual analyses. We start by investigating user needs on such visual analyses and recommendations, then propose several different visualization schemes, and perform a pilot study to collect user feedback on the designed schemes. We further conduct user studies to verify the utility of the proposed schemes, and the results not only demonstrate the effectiveness of our proposed visualization, but also provide important insights to guide the visualization design in the future.

The presence of illegally adulterated sildenafil and related compounds in dietary supplements was qualitatively analyzed by employing ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS). Samples were extracted by ultrasonic method using methanol. The analysis was performed on an Agilent Poroshell 120 SB-C18 column (75 mm×3.0 mm, 2.7 μm) using gradient elution with a mixture of aqueous formic acid solution (0.1%, v/v) and acetonitrile as the mobile phase at a flow rate of 0.4 mL/min. Q-TOF-MS equipped with ESI ion source was operated in a positive ionization mode. Qualitative analysis was based on the retention time and the accurate relative molecular mass of elemental compositions of the precursor ion and product ions. The limits of detection (LODs) of sildenafil and other 99 compounds were found to be 0.1-50 mg/kg or 0.1~50 mg/L in 10 different matrices. In this manuscript, the fragmentation behavior of 98 phosphodiesterase-5 inhibitors are categorically summarized, which can provide information for referencing and identification of suspicious compounds. The method was successfully applied to the analysis of actual samples, of which 19 compounds combated the illegal adulteration behavior effectively.

The corrosion properties of plasma-sprayed (PS) Ni-coated WC coatings are studied for both before and after laser remelting of the coatings. Optic microscope (OM), scanning electron microscope (SEM) and X-ray diffraction (XRD) are applied to investigate the microstructure and phase composition of PS Ni-coated WC coatings. The results indicate that coatings with laser remelting could improve their microstructure, such as lower porosity rate, weaker lamellar structure and more uniform distribution of phases. XRD shows that the W2C phase is not identified both in as-sprayed and coatings with laser remelting, which means that the decarburization of WC has not happened because of Ni-coated WC grain. However, the Ni compounds were observed both in the plasma spray and the laser-remelting coatings. The salt spray corrosion (SSC) shows that the laser remelting coating has better corrosion resistance, which is mainly due to its fully dense top layer, low number of defects and uniform distribution of the phase and composition.

An analytical method was developed for the determination of 34 illegally adulterated weight loss compounds in foods and dietary supplements by ultra high performance liquid chromatography-triple quadrupole mass spectrometry (UHPLC-MS/MS). A Waters CORTECS T3 column (100 mm×2.1 mm, 2.7 μm) was used with 0.1% (v/v) formic acid aqueous solution-acetonitrile containing 0.1% (v/v) formic acid as mobile phases by gradient elution. The compounds were detected by electrospray ion source in positive or negative mode with multiple reaction monitoring (MRM) mode. The calibration curves showed good linearity in the range of 0.5-10 μg/L for 29 compounds such as sibutramine, and in the range of 2.5-50 μg/L for five compounds such as chlorothiazide. The correlation coefficients (r) of the standard calibration curves for the 34 analytes were all greater than 0.99. The recoveries of the 29 compounds at spiked levels of 5, 10 and 20 μg/kg were in the range of 49.2%-136.2%, and the RSDs were 0.7%-15.0% (n=6). The recoveries of the five compounds at spiked levels of 25, 50 and 100 μg/kg were in the range of 51.5%-130.9%, and the RSDs were 0.8%-14.0% (n=6). The limits of detection (LODs) and limits of quantification (LOQs) were 5 μg/kg and 10 μg/kg for the 29 compounds, 25 μg/kg and 50 μg/kg for the five compounds, respectively. The method was successfully applied to the analysis of actual samples, and 12 compounds were checked out, which combated the illegal adulteration behavior effectively.

During the process of digital campus construction, there emerged many problems, such as, the rapid rise of the servers in DC (Data Center) and the power consumption, various failure of single point. In order to solve these problems, the author proposes virtual server based on virtual software to improve the efficiency of equipment and reduce the energy consumption and thus reducing operating costs.

The low temperature co-fired ceramics (LTCC) is an ideal packaging method that can be used to design highly reliable and miniature microwave multi-chip modules (MMCM). In this paper, modeling and simulation of the three dimensional vertical interconnection were carried out using three dimensional electromagnetic field analysis software HFSS. The BP neural network was used to forecast the parasitics of the vertical through hole. Good agreement was acquired between the simulated results and experimental results. The goal of this paper is to make the vertical vias as a circuit part, so as to enhance the efficiency of the design and processing of microwave circuits based on the LTCC. Simultaneously, it will also improve the design accuracy of the LTCC microwave circuits, providing the technical support for the design of more complex microwave circuits.

Fibrocytes hold a prominent role in inflammatory and tissue repair processes in various organ systems. In this study, we identified and quantified a reactive fibrocyte population in the vocal fold mucosa postinjury using immunohistochemistry and stereological analysis. These cells, which expressed CD11b on their surface and prolyl-4-hydroxylase β (P4H-β) intracellularly, were largely restricted to the lamina propria, and were morphologically and immunochemically distinguishable from newly recruited epithelial cells. We validated our immunohistochemistry findings using flow cytometry, and additionally characterized a reactive fibrocyte population in circulating peripheral blood using a novel detection panel (CD16(-) CD11b(+) P4H-β(+) ). Fibrocyte recruitment peaked at 3 days postinjury in peripheral blood, and 5 days postinjury in the vocal fold mucosa. These findings suggest that circulating fibrocytes are recruited to sites of tissue injury in the vocal fold mucosa, and may play an important role in vocal fold tissue repair. The results of this study are consistent with published data from other organ systems and strongly suggest the importance of fibrocytes as therapeutic targets. Our newly reported antigen panel facilitating the direct characterization of fibrocytes via flow cytometry is a useful tool with the potential to facilitate improved study of this cell population.

The Arabidopsis (Arabidopsis thaliana) gene that encodes the probable ortholog of the 30-kD subunit of the mammalian cleavage and polyadenylation specificity factor (CPSF) is a complex one, encoding small (approximately 28 kD) and large (approximately 68 kD) polypeptides. The small polypeptide (AtCPSF30) corresponds to CPSF30 and is the focus of this study. Recombinant AtCPSF30 was purified from Escherichia coli and found to possess RNA-binding activity. Mutational analysis indicated that an evolutionarily conserved central core of AtCPSF30 is involved in RNA binding, but that RNA binding also requires a short sequence adjacent to the N terminus of the central core. AtCPSF30 was found to bind calmodulin, and calmodulin inhibited the RNA-binding activity of the protein in a calcium-dependent manner. Mutational analysis showed that a small part of the protein, again adjacent to the N terminus of the conserved core, is responsible for calmodulin binding; point mutations in this region abolished both binding to and inhibition of RNA binding by calmodulin. Interestingly, AtCPSF30 was capable of self-interactions. This property also mapped to the central conserved core of the protein. However, calmodulin had no discernible effect on the self-association. These results show that the central portion of AtCPSF30 is involved in a number of important functions, and they raise interesting possibilities for both the interplay between splicing and polyadenylation and the regulation of these processes by stimuli that act through calmodulin.

Locations, numbers and effects of quantitative trait loci (QTLs) for a given trait depend on genetic backgrounds and the environments in which phenotyping is conducted. Few studies address the issue on the consistency of QTLs for drought resistant traits. To test whether there exist consistent QTLs for root thickness and root penetration across genetic backgrounds, we studied three rice populations including one recombinant inbred (CO39/Moroberekan) and two doubled haploid (IR64/Azucena and CT9993/IR62266) populations. Root thickness and root penetration were chosen because they are an important component of drought avoidance mechanism and difficult to be incorporated into conventional plant breeding programs. To make comparison of QTLs consistency across genetic backgrounds possible, following criteria were used: 1) Genotyping was done with a common set of RFLP probes from Cornell University although other types of markers such as AFLPs and SSRs were also used. 2) Phenotyping was conducted under similar environmental conditions in a greenhouse although in different years (1992 for CO39/Moroberekan population; 1995 for IR64/Azucena population; 1997 for CT9993/IR62266 population). Our results indicate that there exist consistent QTLs for root thickness and root penetration across genetic backgrounds. These consistent QTLs would be most useful in the development of marker-assisted selection for drought resistance in rice and most promising candidates for positional cloning (This research is supported by the Rockefeller Foundation's Rice Biotechnology Program). (Texte intégral)

Ernesto Guccione and colleagues report that the transcription factor PRDM15 regulates naive pluripotency in mouse embryos and embryonic stem cells and in derivation of mouse and human iPSCs. They further show that PRDM15 promotes WNT signaling and inhibits MAPK–ERK signaling by directly regulating the expression of R-spondin1 and Sprouty1, respectively. The transcriptional network acting downstream of LIF, WNT and MAPK–ERK to stabilize mouse embryonic stem cells (ESCs) in their naive state has been extensively characterized. However, the upstream factors regulating these three signaling pathways remain largely uncharted. PR-domain-containing proteins (PRDMs) are zinc-finger sequence-specific chromatin factors that have essential roles in embryonic development and cell fate decisions. Here we characterize the transcriptional regulator PRDM15, which acts independently of PRDM14 to regulate the naive state of mouse ESCs. Mechanistically, PRDM15 modulates WNT and MAPK–ERK signaling by directly promoting the expression of Rspo1 (R-spondin1) and Spry1 (Sprouty1). Consistent with these findings, CRISPR–Cas9-mediated disruption of PRDM15-binding sites in the Rspo1 and Spry1 promoters recapitulates PRDM15 depletion, both in terms of local chromatin organization and the transcriptional modulation of these genes. Collectively, our findings uncover an essential role for PRDM15 as a chromatin factor that modulates the transcription of upstream regulators of WNT and MAPK–ERK signaling to safeguard naive pluripotency.

Mast cells (MCs) are versatile immune cells capable of rapidly responding to a diverse range of extracellular cues. Here, we mapped the genomic and transcriptomic changes in human MCs upon diverse stimuli. Our analyses revealed broad H3K4me3 domains and enhancers associated with activation. Notably, the rise of intracellular calcium concentration upon immunoglobulin E (IgE)-mediated crosslinking of the high-affinity IgE receptor (FcεRI) resulted in genome-wide reorganization of the chromatin landscape and was associated with a specific chromatin signature, which we term Ca2+-dependent open chromatin (COC) domains. Examination of differentially expressed genes revealed potential effectors of MC function, and we provide evidence for fibrinogen-like protein 2 (FGL2) as an MC mediator with potential relevance in chronic spontaneous urticaria. Disease-associated single-nucleotide polymorphisms mapped onto cis-regulatory regions of human MCs suggest that MC function may impact a broad range of pathologies. The datasets presented here constitute a resource for the further study of MC function. Cildir et al. map the genomic and transcriptomic changes in human mast cells upon diverse stimuli, defining chromatin changes associated with activation and Ca2+ flux and revealing potential effectors of mast cell function. Disease-associated SNPs mapped onto cis-regulatory regions of mast cells suggest that mast cell function may impact a broad range of pathologies. Refereed/Peer-reviewed