Construction of Multiplexed Assays on Single Anisotropic Particles Using Microfluidics.

Considerable efforts have been made to develop microscale multiplexing strategies. However, challenges remain due to the difficulty in deploying functional objects and decoding high-density signals on anisotropic microcarriers. Here, we report a microfluidic method to fabricate architecture-marked anisotropic particles for performing designable multiplexed assays in a label-free manner.

Linking Metastatic Behavior and Metabolic Heterogeneity of Circulating Tumor Cells at Single-Cell Level Using an Integrative Microfluidic System.

Circulating tumor cells (CTCs) are pivotal biomarkers in tumor metastasis, however, the underlying molecular mechanism of CTCs behavioral heterogeneity during metastasis remains unexplored. Here, an integrative workflow is developed to link behavior characteristics to metabolic profiling within individual CTCs, which simulates the metastatic process on a microfluidic system and combined with single-cell mass spectrometry (MS) detection. Spheroid-derived HCT116 cells are tracked and extracted via a temporary vascular system, revealing various arrest patterns under biomimetic vascular shear flow.

Microfluidic-Enabled Self-Directed Hydrogel Microspheres for Multiplexed MicroRNA Assays.

Multiplexed microRNA (miRNA) detection has proven valuable in disease diagnosis; yet, the development of advanced tools for their analysis remains a subject of broad interest. Here, we propose a novel single-particle method for multiplexed miRNA detection using self-directed hydrogel microspheres, which feature supersegmented compartments for loading analyte probes and an air-encapsulated region that grants the microsphere a unique preferred posture in aqueous solutions. By exploiting microfluidic technology, we can widely adjust the size of the microspheres and the number of compartments can be widely adjusted.

Droplets in open microfluidics: generation, manipulation, and application in cell analysis.

Open droplet microfluidics is an emerging technology that generates, manipulates, and analyzes droplets in open configuration systems. Droplets function as miniaturized reactors for high-throughput analysis due to their compartmentalization and parallelization, while openness enables addressing and accessing the targeted contents. The convergence of two technologies facilitates the localization and intricate manipulation of droplets using external tools, showing great potential in large-scale chemical and biological applications, particularly in cell analysis.

Transcriptome analysis of Vero cells infected with attenuated vaccine strain CDV-QN-1.

To better understand the interaction between attenuated vaccines and host antiviral responses, we used bioinformatics and public transcriptomics data to analyze the immune response mechanisms of host cells after canine distemper virus (CDV) infection in Vero cells and screened for potential key effector factors. In this study, CDV-QN-1 infect with Vero cells at an MOI of 0.5, and total RNA was extracted from the cells 24 h later and reverse transcribed into cDNA.

Chemical Plasma Membrane Perforation Generated by a Microfluidic Probe for Single-Cell Intracellular Protein Delivery.

Efficient and convenient delivery of exogenous molecules into cells is important for cell biology research. However, many intracellular delivery methods require carrier-mediated or physical field assistance, complicating the delivery process. Here, a general, simple, and effective method for in situ single-cell intracellular delivery is reported.

Understanding Macrophage-Tumor Interactions: Insights from Single-Cell Behavior Monitoring in a Sessile Microdroplet System.

Interaction between tumor-associated macrophages and tumor cells is crucial for tumor development, metastasis, and the related immune process. However, the macrophages are highly heterogeneous spanning from anti-tumorigenic to pro-tumorigenic, which needs to be understood at the single-cell level. Herein, a sessile microdroplet system designed for monitoring cellular behavior and analyzing intercellular interaction, demonstrated with macrophage-tumor cell pairs is presented.

Quantitative proteomics based on TMT revealed the response of PK15 cells infected PEDV wild strain.

Porcine epidemic diarrhea (PED), caused by porcine epidemic diarrhea virus (PEDV), is an acute and highly contagious enteric disease with a high mortality rate in suckling piglets. Identification of proteins associated with PEDV infection may provide insights into the pathogenesis of this viral disease. In this study, we employed tandem mass tag (TMT) quantitative protein analysis to investigate proteomic changes in PK15 cells following PEDV infection, and differential protein expression profiles were obtained at 0 h, 24 h, and 48 h post-infection.

Fabrication and Performance of Bubble-Containing Multicompartmental Particles: Novel Self-Orienting Carriers.

In this work, a class of bubble-containing multicompartmental particles with self-orienting capability is developed, where a single bubble is enclosed at the top of the super-segmented architecture. Such bubbles, driven by potential energy minimization, cause the particles to have a bubble-upward preferred orientation in liquid, enabling efficient decoding of their high-density signals in an interference-resistant manner. The particle preparation involves bubble encapsulation via the impact of a multicompartmental droplet on the liquid surface and overall stabilization via rational crosslinking.

RSL3 Inhibits Porcine Epidemic Diarrhea Virus Replication by Activating Ferroptosis.

Porcine epidemic diarrhea virus (PEDV) is a highly contagious coronavirus that induces diarrhea and death in neonatal piglets, resulting in substantial economic losses to the global swine industry. The mechanisms of PEDV infection and the roles of host factors are still under exploration. In this study, we used the ferroptosis pathway downstream target activator (1S,3R)-RSL3 compound as a starting point, combined with the interactions of N-acetylcysteine and deferoxamine, to elucidate the effects of a series of compounds on PEDV proliferation.

Multiplex bacteria detection using one-pot CRISPR/Cas13a-based droplet microfluidics.

High-throughput detection of bacteria at low levels is critical in public health, food safety, and first response. Herein, for the first time, we present a platform based on droplet microfluidics coupling with the recombinase aided amplification (RAA)-assisted one-pot clustered regularly interspaced short palindromic repeats together with CRISPR-associated proteins 13a (CRISPR/Cas13a) assay, and droplet encoding strategy for accurate and sensitive determination of nucleic acids from various foodborne pathogens. The workflow takes full advantage of CRISPR/Cas13a signal amplification and droplet confinement effects, which enhances the detection sensitivity and enables end-point quantitation.

Comparative transcriptomic analysis of PK15 cells infected with a PRV variant and the Bartha-K/61 vaccine strain.

Introduction: Pseudorabies virus (PRV) is a herpesvirus that can infect domestic animals, such as pigs, cattle and sheep, and cause fever, itching (except pigs), and encephalomyelitis. In particular, the emergence of PRV variants in 2011 have resulted in serious economic losses to the Chinese pig industry. However, the signaling pathways mediated by PRV variants and their related mechanisms are not fully understood.

Erastin inhibits porcine epidemic diarrhea virus replication in Vero cells.

Background: Porcine epidemic diarrhea virus (PEDV), an intestinal pathogenic coronavirus, has caused significant economic losses to the swine industry worldwide. At present, there are several treatment methods, but there is still a lack of clinically effective targeted drugs, new antiviral mechanisms and drugs need to be explored.

Methods: In this study, we established a model of erastin versus ferrostatin-1 treatment of Vero cells, and then detected virus proliferation and gene expression by RT-qPCR through PEDV infection experiments.

[Effects of different orthodontic treatments on sputum chemokines CX3CL1, RANKL/OPG levels in patients with malocclusion].

Purpose: To investigate the effects of different orthodontic treatments on gingival crevicular fluid chemokine CX3CL1, nuclear factor κB receptor activating factor ligand/osteoprotegerin(RANKL/OPG) levels in patients with malocclusion.

Methods: Ninety-six patients with malocclusion who were scheduled to undergo orthodontic treatment were randomly divided into four groups. All patients were treated with square wire appliance, and 0, 50, 150, 250 g of far-distal orthodontic force were given respectively.