In Situ Detection of Nucleic Acids in Extracellular Vesicles via Membrane Fusion.

Extracellular vesicles (EVs) carry diverse biomolecules (e. g., nucleic acids, proteins) for intercellular communication, serving as important markers for diseases. Analyzing nucleic acids derived from EVs enables non-invasive disease diagnosis and prognosis evaluation. Membrane fusion, a fundamental cellular process wherein two lipid membranes merge, facilitates cell communication and cargo transport. Building on this natural phenomenon, recent years have witnessed the emergence of membrane fusion-based strategies for the detection of nucleic acids within EVs. These strategies entail the encapsulation of detection probes within either artificial or natural vesicles, followed by the induction of membrane fusion with EVs to deliver probes. This innovative approach not only enables in situ detection of nucleic acids within EVs but also ensures the maintenance of structural integrity of EVs, thus preventing nucleic acid degradation and minimizing the interference from free nucleic acids. This concept categorizes approaches into universal and targeted membrane fusion strategies, and discusses their application potential, and challenges and future prospects.

Encoded Fusion-Mediated MicroRNA Signature Profiling of Tumor-Derived Extracellular Vesicles for Pancreatic Cancer Diagnosis.

MicroRNAs (miRNAs) in tumor-derived extracellular vesicles (tEVs) are important cancer biomarkers for cancer screening and early diagnosis. Multiplex detection of miRNAs in tEVs facilitates accurate diagnosis but remains a challenge. Herein, we propose an encoded fusion strategy to profile the miRNA signature in tEVs for pancreatic cancer diagnosis. A panel of encoded-targeted-fusion beads was fabricated for the selective recognition and fusion of tEVs, with the turn-on fluorescence signals of molecule beacons for miRNA quantification and barcode signals for miRNA identification using readily accessible flow cytometers. Using this strategy, six types of pancreatic-cancer-associated miRNAs can be profiled in tEVs from 2 µL plasma samples (n = 36) in an isolation-free and lysis-free manner with only 2 h of processing, offering a high accuracy (98%) to discriminate pancreatic cancer, pancreatitis, and healthy donors. This encoded fusion strategy exhibits great potential for multiplex profiling of miRNA in tEVs, offering new avenues for cancer diagnosis and screening.

Hierarchical Fluid Interface Enables Spatiotemporal Regulation of Ligand Distribution to Increase Kinetics and Thermodynamics of Interfacial Binding Reaction.

In nature, regulation of the spatiotemporal distribution of interfacial receptors and ligands leads to optimum binding kinetics and thermodynamics of receptor-ligand binding reactions within interfaces. Inspired by this, we report a hierarchical fluid interface (HieFluidFace) to regulate the spatiotemporal distribution of interfacial ligands to increase the rate and thermodynamic favorability of interfacial binding reactions. Each aptamer-functionalized gold nanoparticle, termed spherical aptamer (SAPT), is anchored on a supported lipid bilayer without fluidity, like an "island", and is surrounded by many fluorescent aptamers (FAPTs) with free fluidity, like "rafts". Such ligand "island-rafts" model provides a large reactive cross-section for rapid binding to cellular receptors. The synergistic multivalency of SAPTs and FAPTs improves interfacial affinity for tight capture. Moreover, FAPTs accumulate at binding sites to bind to cellular receptors with clustered fluorescence to "lighten" cells for direct identification. Thus, HieFluidFace in a microfluidic chip achieves high-performance capture and identification of circulating tumor cells from clinical samples, providing a new paradigm to optimize the kinetics and thermodynamics of interfacial binding reactions.

A Fluid Multivalent Magnetic Interface for High-Performance Isolation and Proteomic Profiling of Tumor-Derived Extracellular Vesicles.

Isolation and analysis of tumor-derived extracellular vesicles (T-EVs) are important for clinical cancer management. Here, we develop a fluid multivalent magnetic interface (FluidmagFace) in a microfluidic chip for high-performance isolation, release, and protein profiling of T-EVs. The FluidmagFace increases affinity by 105-fold with fluidity-enhanced multivalent binding to improve isolation efficiency by 13.9 % compared with a non-fluid interface. Its anti-adsorption property and microfluidic hydrodynamic shear minimize contamination, increasing detection sensitivity by two orders of magnitude. Moreover, its reversibility and expandability allow high-throughput recovery of T-EVs for mass spectrometric protein analysis. With the chip, T-EVs were detected in all tested cancer samples with identification of differentially expressed proteins compared with healthy controls. The FluidmagFace opens a new avenue to isolation and release of targets for cancer diagnosis and biomarker discovery.

Tumor Exosomal ENPP1 Hydrolyzes cGAMP to Inhibit cGAS-STING Signaling.

To evade immune surveillance, tumor cells express ectonucleotide pyrophosphatase phosphodiesterase 1 (ENPP1) on the surface of their membrane, which degrades extracellular cyclic GMP-AMP (cGAMP), thereby inhibiting the cyclic GMP-AMP synthase (cGAS) stimulator of interferon gene (STING) DNA-sensing pathway. To fully understand this tumor stealth mechanism, it is essential to determine whether other forms of ENPP1 with hydrolytic cGAMP activity also are present in the tumor microenvironment to regulate this innate immune pathway. Herein, it is reported that various tumor-derived exosomes carry ENPP1, and can hydrolyze synthetic 2'3'-cGAMP and endogenous 2'3'-cGAMP produced by cells to inhibit cGAS-STING pathway in immune cells. Moreover, tumor exosomal ENPP1 also can hydrolyze 2'3'-cGAMP bound to LL-37 (an effective transporter of 2'3'-cGAMP) to inhibit STING signaling. Furthermore, high expression of ENPP1 in exosomes is observed isolated from human breast and lung cancer tissue, and tumor exosomal ENPP1 inhibited the immune infiltration of CD8+ T cells and CD4+ T cells. The results elucidate the essential function of tumor exosomal ENPP1 in the cGAS-STING pathway, furthering understanding of the crosstalk between the tumor cells and immune system.

[Retracted] miR­141­3p suppresses proliferation and promotes apoptosis by targeting GLI2 in osteosarcoma cells.

Following the publication of the above article, a concerned reader drew to the Editor's attention that, in the above paper, they had identified multiple instances of overlapping data panels comparing the TUNEL assay data shown in Fig. 2C and D on p. 750 with that shown in Fig. 4C on p. 752, suggesting that data purportedly showing results obtained under different experimental conditions had been derived from a smaller number of original sources. Upon informing the authors about this matter, they consulted their original data and requested a corrigendum to take account of the overlapping data in Figs. 2 and 4; however, given the number of instances of overlapping data panels that were identified comparing these two figures, the Editor of Oncology Reports has decided that this article should be retracted from the publication owing to a lack of overall confidence in the presented data. Upon informing the authors of this decision, they did not accepted the decision to retract this article. The Editor apologizes to the readership for any inconvenience resulting from the retraction of this article. [Oncology Reports 39: 747­754, 2018; DOI: 10.3892/or.2017.6150].

Research on Intelligent Embedded Fan System Based on YOLOv2 Lightweight Algorithm.

With the development of artificial intelligence, the application of intelligent algorithms to low-power embedded chips has become a new research topic today. Based on this, this study optimizes the YOLOv2 algorithm by tailoring and successfully deploys it on the K210 chip to train the face object detection algorithm model separately. The intelligent fan with YOLOv2 model deployed in K210 chip can detect the target of the character and obtain the position and size of the character in the machine coordinates. Based on the obtained information of character coordinate position and size, the fan's turning Angle and the size of air supply are intelligently perceived. The experimental results show that the intelligent fan design method proposed here is a new embedded chip intelligent method of cutting and improving the YOLOv2 algorithm. It innovatively designed solo tracking, crowd tracking, and intelligent ranging algorithms, which perform well in human perception of solo tracking and crowd tracking and automatic air volume adjustment, improve the accuracy of air delivery and user comfort, and also provide good theoretical and practical support for the combination of AI and embedded in other fields.

Knockdown of miR-222 inhibits inflammation and the apoptosis of LPS-stimulated human intervertebral disc nucleus pulposus cells.

It has been demonstrated that miR­222 is upregulated in human intervertebral disc (IVD) degeneration tissues; however, the underlying mechanisms remain unclear. In this study, we aimed to elucidate the mechanisms of action of miR­222 in IVD tissues. Nucleus pulposus (NP) cells were treated with lipopolysaccharide (LPS) to simulate IVD degeneration. The expression level of miR­222 was detected by reverse transcription­quantitative polymerase chain reaction (RT­qPCR) in cells and tissues. Cell apoptosis was analyzed by flow cytometry. Additionally, western blot analysis was used to determine the levels of Toll­like receptor 4 (TLR4), Iκß­alpha (IκBα) and p65. Interleukin (IL)­1ß, tumor necrosis factor­α (TNF­α) and IL­6 protein expression levels were determined by enzyme­linked immunosorbent assay (ELISA). The target gene of miR­222 was determined by TargetScan7.2 and dual luciferase reporter gene analysis. Western blot analysis and RT­qPCR were used to determine the mRNA and protein levels of tissue inhibitor of metalloproteinase 3 (TIMP3). The mRNA expression level of miR­222 was found to be increased in IVD tissues and in LPS­stimulated cells, and its expression was positively associated with the clinical MRI grade. In vitro, apoptosis was promoted/inhibited by miR­222 mimics/inhibitors. Transfection with miR­222 mimics/inhibitors significantly increased/decreased the production of TNF­α, IL­1ß and IL­6 and suppressed/enhanced collagen II and aggrecan expression. The protein levels of TLR4, p­IκΒα and p­p65 were upregulated/downregulated by transfection with the mimics/inhibitors. In addition, it was demonstrated that TIMP3 was a direct target gene of miR­222, and was negatively regulated by miR­222 in NP cells. The silencing of TIMP3 reversed the inhibitory effects of miR­222 inhibitor on cell apoptosis, which was induced by LPS. Thus, on the whole, the findings of this study demonstrate that miR­222 functions as a promoter of IVD development, partly via the regulation of TIMP3.

Long noncoding RNA DANCR promotes colorectal cancer proliferation and metastasis via miR-577 sponging.

Long non-coding RNAs (lncRNAs) play key roles in various malignant tumors, including colorectal cancer (CRC). Long non-coding RNA differentiation antagonizing non-protein coding RNA (DANCR) is overexpressed in CRC patients, but whether it affects CRC proliferation and metastasis via regulation of heat shock protein 27 (HSP27) remains unclear. In the present study, we found that DANCR was highly expressed and correlated with proliferation and metastasis in CRC. In addition, we demonstrated that DANCR and HSP27 were both targets of microRNA-577 (miR-577) and shared the same binding site. Furthermore, we revealed that DANCR promoted HSP27 expression and its mediation of proliferation/metastasis via miR-577 sponging. Finally, using an in vivo study, we confirmed that overexpression of DANCR promoted CRC tumor growth and liver metastasis. The present study demonstrated the function of DANCR in CRC and might provide a new target in the treatment of CRC.

miR-141-3p suppresses proliferation and promotes apoptosis by targeting GLI2 in osteosarcoma cells.

MicroRNAs (miRNAs) have been reported as key regulators in numerous diseases including osteosarcoma. The function of microRNA-141-3p (miR-141-3p) and whether this function is achieved by regulation of GLI family zinc finger 2 (GLI2) in osteosarcoma remain unclear. In the present study, we found decreased expression of miR-141-3p, but increased expression of GLI2 in osteosarcoma tissues and cell lines. In addition, we demonstrated a negative correlation between miR-141-3p and GLI2. Furthermore, we revealed that elevation of miR-141-3p resulted in a marked inhibition of proliferation and promotion of apoptosis as well as an obviously decrease in GLI2 in osteosarcoma cell lines. Furthermore, we determined that GLI2 is a target of miR-141-3p by a constructed luciferase assay. In addition, we showed that miR-141-3p could negatively regulate GLI2 and its downstream parathyroid hormone-related protein 1 (PTHRP1). Finally, through a series of antisense experiments we confirmed that the effect of miR-141-3p on proliferation and apoptosis was achieved through the GLI2 pathway in osteosarcoma cells. The findings of the present study may provide a new target for treating osteosarcoma.

Using data mining techniques on discovering physician practice patterns regarding to medication prescription - an exploratory study.

In this paper, we propose a data mining method for exploring the decision-making processes of physicians from electronic patient records and test it on the medical records of patients with type-2 diabetes mellitus. This method runs in two modes: general and partitioned. In the general mode, it mines rules from the whole medical records. In the partitioned mode, with a given partition factor, medical records are assigned into categories and a corresponding set of rules will be discovered for each category. Medication prescription predictions can be provided based on these rules. By comparing mined rules and prescription prediction accuracy under different modes, we discover that: 1) both the averaged precision and recall rate of the general mode can reach around 80%; 2) physicians tend to conform to the guideline instead of having their own preferences; 3) the medication decision can be affected by some hidden factors. These findings suggest this method show promise in discovering physician practice patterns and obtaining insights from real medical records.