Construction of a Recombinant Eukaryotic Expression Plasmid Containing Human Calcitonin Gene and Its Expression in NIH3T3 Cells

Aim. To construct a recombinant eukaryotic expression plasmid containing human calcitonin (hCT) gene and express the gene in murine fibroblast NIH3T3 cells. Materials and Methods. A murine Igκ-chain leader sequence and hCT gene were synthesized and cloned into pCDNA3.0 to form the pCDNA3.0-Igκ-hCT eukaryotic expression vector, which was transfected into NIH3T3 cells. The mRNA and protein expressions and secretion of hCT were detected. Primarily cultured osteoclasts were incubated with the supernatant of pCDNA3.0-Igk-hCT-transfected NIH3T3 cells, and their numbers were counted and morphology observed. Results. The expression and secretion of hCT were successfully detected in pCDNA3.0-Igk-hCT-transfected NIH3T3 cells. The number of osteoclasts was decreased and the cells became crumpled when they were incubated with the supernatant of pCDNA3.0-Igk-hCT-transfected NIH3T3 cells. Conclusion. A recombinant eukaryotic expression vector containing hCT gene was successfully constructed and expressed in NIH3T3 cells. The secreted recombinant hCT inhibited the growth and morphology of osteoclasts

Multivariate Relevance Vector Regression based Degradation Modeling and Remaining Useful Life Prediction

Relevance Vector Regression (RVR) is a useful tool for degradation modeling and Remaining Useful Life (RUL) prediction. However, most RVR models are for one-dimensional degradation processes and can only handle univariate observations. This paper proposes a degradation path based RUL prediction framework using a dynamic Multivariate Relevance Vector Regression (MRVR) model. Specifically, a multi-step regression model is established for describing the degradation dynamics and extends the classical RVR into a multivariate one with consideration of the multivariate environment. The paper introduces a matrix Gaussian distribution based RVR approach and then estimates the hyperparameters with Nesterov's accelerated gradient method to avoid the exhausting re-estimation phenomenon in seeking analytical solutions. It further forecasts the degradation path for monitoring the degradation status. Based on the forecasted path, the RUL is predicted by the First Hitting Time (FHT) method. Finally, the proposed methods are illustrated by two case studies, one is presented in the paper and the other in the supplement, both of which investigate the capacitors' performance degradation in the traction systems of high-speed trains

The ectonucleotidases CD39 and CD73 on T cells: The new pillar of hematological malignancy

Hematological malignancy develops and applies various mechanisms to induce immune escape, in part through an immunosuppressive microenvironment. Adenosine is an immunosuppressive metabolite produced at high levels within the tumor microenvironment (TME). Adenosine signaling through the A2A receptor expressed on immune cells, such as T cells, potently dampens immune responses. Extracellular adenosine generated by ectonucleoside triphosphate diphosphohydrolase-1 (CD39) and ecto-5’-nucleotidase (CD73) molecules is a newly recognized ‘immune checkpoint mediator’ and leads to the identification of immunosuppressive adenosine as an essential regulator in hematological malignancies. In this Review, we provide an overview of the detailed distribution and function of CD39 and CD73 ectoenzymes in the TME and the effects of CD39 and CD73 inhibition on preclinical hematological malignancy data, which provides insights into the potential clinical applications for immunotherapy

Whole exome sequencing of insulinoma reveals recurrent T372R mutations in YY1

Functional pancreatic neuroendocrine tumours (PNETs) are mainly represented by insulinoma, which secrete insulin independent of glucose and cause hypoglycaemia. The major genetic alterations in sporadic insulinomas are still unknown. Here we identify recurrent somatic T372R mutations in YY1 by whole exome sequencing of 10 sporadic insulinomas. Further screening in 103 additional insulinomas reveals this hotspot mutation in 30% (34/113) of all tumours. T372R mutation alters the expression of YY1 target genes in insulinomas. Clinically, the T372R mutation is associated with the later onset of tumours. Genotyping of YY1, a target of mTOR inhibitors, may contribute to medical treatment of insulinomas. Our findings highlight the importance of YY1 in pancreatic β-cells and may provide therapeutic targets for PNETs

Cyclization reaction of amines with dialkyl carbonates to yield 1,3-oxazinan-2-ones

A number of six-membered cyclic carbamates (oxazinanones) were synthesized from the reaction of a primary amine or hydrazine with a dicarbonate derivative of 1,3-diols in a one-pot reaction, in good yield, short time span, and in the absence of a solvent. The reaction proceeds in two steps: an intermolecular reaction to give a linear intermediate and an intramolecular cyclization to yield the cyclic carbamate. This is the first example of a carbonate reacting selectively and sequentially, firstly at the carbonyl center to form a linear carbamate and then as a leaving group to yield a cyclic carbamate

FOXO1 Inhibits Tumor Cell Migration via Regulating Cell Surface Morphology in Non-Small Cell Lung Cancer Cells

Background/Aims: Cell surface morphology plays pivotal roles in malignant progression and epithelial-mesenchymal transition (EMT). Previous research demonstrated that microvilli play a key role in cell migration of non-small cell lung cancer (NSCLC). In this study, we report that Forkhead box class O1 (FOXO1) is downregulated in human NSCLC and that silencing of FOXO1 is associated with the invasive stage of tumor progression. Methods: The cell proliferation, migration, and invasion were characterized in vitro, and we tested the expression of the Epithelial-mesenchymal transition (EMT) marker by immunofluorescence staining and also identified the effect of FOXO1 on the microvilli by scanning electron microscopy (SEM). Results: Functional analyses revealed that silencing of FOXO1 resulted in an increase in NSCLC cell proliferation, migration, and invasion; whereas overexpression of FOXO1 significantly inhibited the migration and invasive capability of NSCLC cells in vitro. Furthermore, cell morphology imaging showed that FOXO1 maintained the characteristics of epithelial cells. Immunofluorescence staining and western blotting showed that the E-cadherin level was elevated and Vimentin was reduced by FOXO1 overexpression. Conversely, the E-cadherin level was reduced and Vimentin was elevated in cells silenced for FOXO1. Furthermore, scanning electron microscopy (SEM) showed that FOXO1 overexpression increased the length of the microvilli on the cell surface, whereas FOXO1 silencing significantly reduced their length. Conclusions: FOXO1 is involved in human lung carcinogenesis and may serve as a potential therapeutic target in the migration of human lung cancer

Efficacy and mechanism of Baicao Fuyanqing suppository on mixed vaginitis based on 16S rRNA and metabolomics

BackgroundMixed vaginitis is the infection of the vagina by at least two different pathogens at the same time, both of which contribute to an abnormal vaginal environment leading to signs and symptoms. Baicao Fuyanqing suppository (BCFYQ) is a Miao ethnomedicine, used to treat various vaginitis. The aim of this study was to investigate the efficacy and possible mechanism of BCFYQ in the treatment of mixed vaginitis based on 16S rRNA high-throughput sequencing and metabonomics.MethodsEscherichia coli and Candida albicans were used to establish mixed vaginitis model in SD rats. Three groups of low, medium and high doses (0.18/0.36/0.64 g.kg-1) were established, and administered vaginally once a day for 6 consecutive days. After the last administration, vaginal pH and IL-1β, IL-2, IL-13 and IgA levels were measured, and the vaginal tissue was examined pathologically. In addition, the vaginal flora was characterised by 16S rRNA, and endogenous metabolites in the vaginal tissue were detected by UHPLC-Q-Exactive MS.ResultsCompared with the model group, BCFYQ can reduce the vaginal pH of rats, make it close to the normal group and improve the damaged vaginal epithelial tissue. The results of ELISA showed that BCFYQ decreased the levels of IL-1 β and IL-2 and increased the levels of IL-13 and IgA (P<0.05). In addition, BCFYQ may increase the abundance of vaginal flora, especially Lactobacillus. The differential metabolite enrichment pathway suggests that the therapeutic mechanism of BCFYQ is mainly related to lipid metabolism and amino acid metabolism.ConclusionOur research shows that BCFYQ has a good therapeutic effect on mixed vaginitis. It repairs the damaged vaginal mucosa by regulating the vaginal flora and lipid metabolism disorders to improve the local immune function of the vagina and inhibit the growth and reproduction of pathogens

Atomic-layer-deposited ultrafine MoS2 nanocrystals on cobalt foam for efficient and stable electrochemical oxygen evolution

Ultrafine molybdenum sulfide (MoS2) nanocrystals are grown on a porous cobalt (Co) foam current collector by atomic layer deposition (ALD) using molybdenum hexacarbonyl and hydrogen sulfide as precursors. When used to catalyze the oxygen evolution reaction (OER), the optimal Co@MoS2 electrode, even with a MoS2 loading as small as 0.06 mg cm-2, exhibits a large cathodic shift of ca. 200 mV in the onset potential (the potential at which the current density is 5 mA cm-2), a low overpotential of only 270 mV to attain an anodic current density of 10 mA cm-2, much smaller charge transfer resistance and substantially improved long-term stability at both low and high current densities, with respect to the bare Co foam electrode, showing substantial promise for use as an efficient, low-cost and durable anode in water electrolyzers.L. F. Liu acknowledges the support of the FCT Investigator grant (no. IF/01595/2014) and the Exploratory grant (No. IF/01595/2014/CP1247/CT0001) from the Portuguese Foundation of Science & Technology (FCT). D. H. Xiong and W. Li are thankful for the financial support from Marie Curie Action COFUND fellowships (NanoTrainforGrowth, Grant Agreement no. 600375) under the FP7 framework. D. H. Xiong also acknowledges the financial support from the China Postdoctoral Science Foundation (No. 2015 T80847). This work was partly funded by the European Commission Horizon 2020 project "CritCat" (Grant Agreement No. 686053).info:eu-repo/semantics/publishedVersio