Fluorescent DNA tetrahedral probe with catalytic hairpin self-assembly reaction for imaging of miR-21 and miR-155 in living cells.

A CHA-based fluorescent DNA tetrahedral probe (FDTp) has been designed to detect the microRNAs miR-21 and miR-155 sensitively and specifically in living cells. The design consisted of functional elements (H1, H2, and Protector) connected to a DNA tetrahedron modified with two pairs of fluorophores and quenching groups. In the presence of miR-21, the chain displacement effect was triggered and Cy3 fluorescence was emitted. In the presence of miR-155, the signal of the catalytic hairpin assembly (CHA) between H1 and H2 on FDTp was amplified, making the fluorescence of FAM sensitive to miR-155. Using this method, the detection limit for miR-155 was 5 pM. The FDTp successfully imaged miR-21 and miR-155 in living cells and distinguished a variety of cell lines based on their expression levels of miR-21 and miR-155. The detection and imaging of dual targets in this design ensured the accuracy of tumor diagnosis and provided a new method for early tumor diagnosis.

Detection ofmiRNA-378based on a catalytic hairpin self-assembly reaction combined with gold nanoparticle colorimetry.

Recent studies have shown that abnormalmiRNA-378expression is a rule, rather than an exception, in cervical cancer and can be used as a diagnostic and prognostic biomarker to assess tumor initiation. In this study, we developed a general, sensitive strategy for detectingmiRNA-378using catalytic hairpin self-assembly (CHA) combined with gold nanoparticles (AuNP) colorimetry. The presence ofmiRNA-378triggers the repeated self-assembly of two designed hairpin DNAs (H1 and H2) into dsDNA polymers, which leads to changes in the surface plasmon resonance absorption band and the macroscopic color of the AuNP colloids due to the formation of nanoparticle-DNA conjugates. This experimental phenomenon can be observed by ultraviolet-visible spectrometry or even with the naked eye. Using this method,miRNA-378could be quantitatively detected at the picomolar level (as low as 20.7 pM). Compared with traditional methods, such as quantitative polymerase chain reaction and RNA blotting, this strategy has a simple operation, low cost, and high sensitivity and selectivity, and thus, exhibits significant potential for miRNA detection.

Label-Free Ratiometric Homogeneous Electrochemical Strategy Based on Exonuclease III-Aided Signal Amplification for Facile and Rapid Detection of miR-378.

MiR-378 is abnormally expressed in various cancers, such as hepatocellular carcinoma, renal cell carcinoma, and nonsmall cell lung cancer. Here, we developed a label- and immobilization-free ratiometric homogeneous electrochemical strategy based on exonuclease III (Exo III) for the facile and rapid determination of miR-378. Two 3'-protruding hairpin DNA probes (HPs) are designed in this strategy. Doxorubicin (DOX) and potassium ferrocyanide (Fe2+) were used as label-free probes to produce a response signal (IDOX) and a reference signal (IFe2+) in the solution phase. When no target was present in the solution, the HP was stable, most of the DOX was intercalated in the stem of the HP, and the diffusion rate of DOX was significantly reduced, resulting in reduced electrochemical signal response. When miR-378 was present, double-cycle signal amplification triggered by Exo III cleavage was initiated, ultimately disrupting the hairpin structures of HP1 and HP2 and releasing a large amount of DOX into the solution, yielding a stronger electrochemical signal, which was low to 50 pM. This detection possesses excellent selectivity, demonstrating high application potential in biological systems, and offers simple and low-cost electrochemical detection for miR-378.

Real-time in situ fluorescence imaging of telomerase and miR378 in living cells using a two-color DNA tetrahedron nanoprobe combined with molecular beacons.

A biosensor that can detect biomarkers accurately, quickly, and conveniently is important for the diagnosis of various diseases. However, most of the existing detection methods require sample extraction, which makes it difficult to detect and image intracellular molecules or to detect two different types of biomarkers simultaneously. In this study, we constructed a DNA tetrahedral nanoprobe (DTP) capable of detecting both miR378 and telomerase, both of which are tumor markers. In the presence of miR378, FAM on the molecular beacon of DTP fluoresced via Förster resonance energy transfer (FRET), and the limit of detection was 476 pM with excellent specificity. When present, telomerase binds to telomerase substrate (TS) primers, extending the repeat sequence (TTAGGG)n to trigger Cy3 fluorescence. A strong linear relationship existed between the fluorescence intensity of Cy3 and the number of HeLa cells. The limit of detection was 800 HeLa cells. In addition, DTP was less cytotoxic to and biocompatible with HeLa cells and fluoresced only in cancer cells, which can help to sensitively distinguish between normal and cancer cells. In conclusion, DTP can simultaneously detect the content of miR378 and activity of telomerase and realize intracellular imaging, which has broad application prospects in early cancer diagnosis and treatment.

Dietary Potassium and Clinical Outcomes among Patients on Peritoneal Dialysis.

BACKGROUND: The association between dietary potassium and clinical prognosis is unclear in patients with chronic kidney disease (CKD). Here, we explored the association between dietary potassium intake and all-cause and cardiovascular (CV) mortality in peritoneal dialysis (PD) patients. METHODS: Here, we present a retrospective analysis of a prospective study. Patients that began incident PD in our center between 1 October 2002 and 31 August 2014 were screened. We recorded all demographic and clinical data at baseline. Repeated measurements were recorded at regular intervals to calculate time-averaged values. Spline regression analysis and Cox proportional regression models were used to evaluate the relationship between dietary potassium and mortality. RESULTS: We followed 881 PD patients for 45.0 (21.5, 80.0) months; 467 patients died, of which 189 (40.5%) died of CV death and 93 were still on PD treatment. Compared with those who had baseline dietary potassium ≥1200 mg/d, the majority of patients with lower dietary potassium were female, older, or poorly educated. They were prone to have poorer nutritional status, CV disease, and diabetes mellitus (p < 0.05). In the unadjusted analysis, both baseline and time-averaged dietary potassium <1200 mg/d predicted higher all-cause and CV mortality (p < 0.001~0.01). After adjusting for demographic and laboratory data, the association between potassium intake and all-cause and CV mortality weakened, which even disappeared after additional adjustment for dietary fiber, protein, and energy intake. CONCLUSIONS: Dietary potassium in PD patients was not independently associated with all-cause and CV mortality.

Rare combined variations of accessory left hepatic artery and accessory right hepatic artery: a case report and literature review.

Variations in the hepatic artery are commonly described in the literature, which is vital for the success procedure of all hepatobiliary surgery. Usually a variation occurs in either the accessory right hepatic artery (aRHA) or the accessory left hepatic artery (aLHA). However, we report an extremely rare case where the variation occurs in both simultaneously. We over served the aRHA arising from the gastroduodenal artery and branching into the superior pancreatic duodenum artery, while the aLHA arose from the common hepatic artery and branched into right gastric artery. This situation has never been reported in literature. We will discuss the meaning of this hepatic artery variation in a clinical setting.

Circular RNAs: A novel target among non­coding RNAs with potential roles in malignant tumors (Review).

Circular RNAs (circRNAs) are a class of non­coding RNAs that are generated via alternative back­splicing, which connects the terminal 5' and 3'ends. Due to their unique loop structure, circRNAs are resistant to ribonucleases and more stable than linear RNAs. In vivo, they are usually highly conserved and stably expressed in tissue/developmental­stage­specific manners. Generally, circRNAs function as microRNA sponges and splicing regulators, as well as in protein binding and transcription. Some circRNAs contain open reading frames with internal ribosomal entry site elements and can thus encode specific proteins. Previously, circRNAs were thought to be erroneous splicing products or by­products of mRNA splicing. With the development of the next­generation sequencing techniques, it has become increasingly clear that circRNAs are abundantly widespread in eukaryotes and that they play significant roles in malignant tumor progression. The present review briefly introduces the biogenesis and functions of circRNAs, as well as summarizes recent research in several common malignancies. The present review also addresses the prospects of circRNAs in clinical applications.