Background
Circulating microRNA expression profiles may be promising biomarkers for diagnosis and assessment of the prognosis of cancer patients. Quantitative polymerase chain reaction (qPCR) is a ...sensitive technique for estimating expression levels of circulating microRNAs. However, there is no current consensus on the reference genes for qPCR analysis of circulating microRNAs.
Aims
In this study we tried to identify suitable reference genes for qPCR analysis of serum microRNA in gastric cancer patients and healthy individuals.
Methods
Six microRNAs (let-7a, miR-16, miR-93, miR-103, miR-192, and miR-451) and RNU6B were chosen as candidate reference genes on the basis of the literature. Expression levels of these candidates were analyzed by qPCR in serum samples from 40 gastric cancer patients and 20 healthy volunteers. The geNorm, Normfinder, bestkeeper, and comparative delta-Ct method algorithms were used to select the most suitable reference gene from the seven candidates. This was then validated by normalizing the expression levels of serum miR-21 across all gastric cancer patients and healthy volunteers.
Results
The algorithms revealed miR-16 and miR-93 were the most stably expressed reference genes, with stability values of 1.778 and 2.213, respectively, for serum microRNA analysis across all the patients and healthy controls. The effect of different normalization strategies was compared; when normalized to the serum volume there were no significant differences between patients and controls. However, when the data were normalized to miR-93, miR-16, or miR-93 and miR-16 combined, significant differences were detected.
Conclusions
Our results demonstrated that reference gene choice for qPCR data analysis has a great effect on the study outcome, and that it is necessary to choose a suitable reference for reliable expression data. We recommend miR-16 and miR-93 as suitable reference genes for serum miRNA analysis for gastric cancer patients and healthy controls.
Pancreatic ductal adenocarcinoma (PDA) is the most lethal of common human malignancies, with no truly effective therapies for advanced disease. Preclinical studies have suggested a therapeutic ...benefit of targeting the Hedgehog (Hh) signaling pathway, which is activated throughout the course of PDA progression by expression of Hh ligands in the neoplastic epithelium and paracrine response in the stromal fibroblasts. Clinical trials to test this possibility, however, have yielded disappointing results. To further investigate the role of Hh signaling in the formation of PDA and its precursor lesion, pancreatic intraepithelial neoplasia (PanIN), we examined the effects of genetic or pharmacologic inhibition of Hh pathway activity in three distinct genetically engineered mouse models and found that Hh pathway inhibition accelerates rather than delays progression of oncogenic Kras-driven disease. Notably, pharmacologic inhibition of Hh pathway activity affected the balance between epithelial and stromal elements, suppressing stromal desmoplasia but also causing accelerated growth of the PanIN epithelium. In striking contrast, pathway activation using a small molecule agonist caused stromal hyperplasia and reduced epithelial proliferation. These results indicate that stromal response to Hh signaling is protective against PDA and that pharmacologic activation of pathway response can slow tumorigenesis. Our results provide evidence for a restraining role of stroma in PDA progression, suggesting an explanation for the failure of Hh inhibitors in clinical trials and pointing to the possibility of a novel type of therapeutic intervention.
A novel cationic curcumin-chitosan PBCA nanoparticle synthesized by emulsion polymerization simply, can not only improve the bioavailability of hydrophobic drug curcumin, but also suppress ...hepatocellular carcinoma growth and inhibit tumor angiogenesis efficiently
in vitro and
in vivo.
We have synthesized novel cationic poly(butyl) cyanoacrylate (PBCA) nanoparticles coated with chitosan, formulation of curcumin nanoparticles. The size and zeta potential of prepared curcumin nanoparticles were about 200
nm and +29.11
mV, respectively with 90.04% encapsulation efficiency. The transmission electron microscopy (TEM) study revealed the spherical nature of the prepared nanoparticles along with confirmation of particle size. Curcumin nanoparticles demonstrate comparable
in vitro therapeutic efficacy to free curcumin against a panel of human hepatocellular cancer cell lines, as assessed by cell viability (3-4,5-dimethylthiazol-2-yl2,5-diphenyltetrazolium bromide assay MTT assay) and proapoptotic effects (annexin V/propidium iodide staining).
In vivo, curcumin nanoparticles suppressed hepatocellular carcinoma growth in murine xenograft models and inhibited tumor angiogenesis. The curcumin nanoparticles’ mechanism of action on hepatocellular cancinoma cells is a mirror that of free curcumin.
Compression tests were conducted on a ceramic-fiber-reinforced SiO2 aerogel at high temperature. Anisotropic mechanical property was found. In-plane Young's modulus is more than 10 times higher than ...that of out-of-plane, but fracture strain is much lower by a factor of 100. Out-of-plane Young's modulus decreases with increasing temperature, but the in-plane modulus and fracture stress increase with temperature. The out-of-plane property does not change with loading rates. Viscous flow at high temperature is found to cause in-plane shrinkage, and both in-plane and out-of-plane properties change. Compression induced densification of aerogel matrix was also found by Scanning Electron Microscope analysis.
Small-cell lung cancer (SCLC) has the highest malignancy among all lung cancers, exhibiting aggressive growth and early metastasis to distant sites. For 30 years, treatment options for SCLC have been ...limited to chemotherapy, warranting the need for more effective treatments. Frequent inactivation of TP53 and RB1 as well as histone dysmodifications in SCLC suggest that transcriptional and epigenetic regulations play a major role in SCLC disease evolution. Here we performed a synthetic lethal screen using the BET inhibitor JQ1 and an shRNA library targeting 550 epigenetic genes in treatment-refractory SCLC xenograft models and identified HDAC6 as a synthetic lethal target in combination with JQ1. Combined treatment of human and mouse SCLC cell line-derived xenograft tumors with the HDAC6 inhibitor ricolinostat (ACY-1215) and JQ1 demonstrated significant inhibition of tumor growth; this effect was abolished upon depletion of NK cells, suggesting that these innate immune lymphoid cells play a role in SCLC tumor treatment response. Collectively, these findings suggest a potential new treatment for recurrent SCLC.
These findings identify a novel therapeutic strategy for SCLC using a combination of HDAC6 and BET inhibitors.
.
A high entropy alloy (HEA) with a composition of Ti2ZrMo0.5Nb0.5 was prepared by vacuum induction melting technology. The mechanical properties at room temperature and elevated temperature ...(900 °C–1150 °C), microstructure evolution and hot deformation behavior during hot deformation are studied. The as-cast alloy is composed of equiaxed grains with the yield strength, the apparent plastic strain of 1306 MPa and 44%, respectively. By TEM analysis, the ZrTi2 Laves C15 phase is found corresponding the precipitation strengthening effect accompanied with the solid solution strengthening as a ramification of the high-entropy alloy. Then the hot compression tests were carried out. The deformed alloy still maintains the same single BCC structure as the as-cast state. It is noted that when the temperature is 900 °C, the structure of the alloy mainly exhibits dynamic recovery (DRV) characteristics, and when the temperature reaches 1000 °C, there are clear dynamic recrystallization (DRX) characteristics in the structure, which are analyzed as the features of continuous dynamic recrystallization (CDRX). Keeping the temperature at 1150 °C, when the strain rate is 0.1 s−1, the characteristics of discontinuous dynamic recrystallization (DDRX) appear, and as the strain rate decreases, CDRX becomes the main recrystallization mechanism again.
Carcinoma-associated fibroblasts (CAFs) can remodel the extracellular matrix to promote cancer cell invasion, but the paracrine signaling between CAFs and cancer cells that regulates tumor cell ...migration remains to be identified. To determine how the interaction between CAFs and cancer cells modulates the invasiveness of cancer cells, we developed a 3-dimensional co-culture model composed of breast cancer (BC) MDA-MB-231 cell spheroids embedded in a collagen gel with and without CAFs. We found that the crosstalk between CAFs and cancer cells promotes invasion by stimulating the scattering of MDA-MB-231 cells, which was dependent on RhoA/ROCK/phospho MLC signaling in cancer cells but independent of RhoA in CAFs. The activation of RhoA/ROCK in cancer cells activates MLC and increases migration, while the genetic-down-regulation of RhoA and pharmacological inhibition of ROCK reduced cell scattering and invasion. Two distinct mechanisms induced the activation of the RhoA/ROCK pathway in MDA-MB-231 cells, the secretion of IGF-1 by CAFs and the upregulation of PAI-1 in cancer cells. In an orthotopic model of BC, IGF-1R inhibition decreased the incidence of lung metastasis, while Y27632-inhibition of ROCK enhanced the lung metastasis burden, which was associated with an increased recruitment of CAFs and expression of PAI-1. Thus the crosstalk between CAFs and BC cells increases the secretion of IGF-1 in CAFs and PAI-1 activity in cancer cells. Both IGF1 and PAI-1 activate RhoA/ROCK signaling in cancer cells, which increases cell scattering and invasion.
The noncontact and wide-band characteristics of the ultrasonic signal excited by the laser make it widely used in nondestructive detection. In this paper, the finite element method is used to ...simulate multiple ultrasonic signal modes and the responses of them to metal defects. The simulation shows that the changes of excited Rayleigh-wave (RW) and shear-wave (SW) can be used to research the surface crack and internal defect of the sample respectively. By analyzing the reflected wave of RW and SW at the inspection point, the effects of surface crack and internal defect on the ultrasonic signals of the two modes are quantitatively compared. The results show that the time delay of the Rayleigh echo has a linear relationship to the width and transverse position of the crack, and the peak-valley difference of Rayleigh echo at the inspection point and the crack depth can also be fitted linearly. In addition, the Y component displacement of the bottom reflected SW also decays with the change of the internal defect. This simulation model provides a theoretical basis for further experimental verification basis.
As an important branch of non-destructive testing, laser ultrasonic testing has attracted increasing attention in the field of material testing because of its instantaneity, non-contact and wide ...adaptability. Based on the finite element method, the process of laser-excited ultrasonic signal is numerically simulated, and the influence of angled cracks on the ultrasonic signal is analyzed. In this paper, the effects of the time function, pulse width, and spot radius of a Gaussian light source are analyzed through the transient field. The different modes of the ultrasonic signal are used to fit the crack’s angle, depth, and width to complete the characteristic analysis of the surface angled crack. The results show that the displacement peak-valley difference of the direct Rayleigh wave is negatively correlated with the crack angle. The displacement extremes of the transmitted Rayleigh wave boundary are negatively correlated with crack depth and width, while the transmitted Rayleigh wave is positively correlated. This paper presents a method for the quantitative analysis of surface-angled cracks and provides a theoretical basis for further experimental verification.
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