3 research outputs found
A Self-Contained Chemiluminescent Lateral Flow Assay for Point-of-Care Testing
Immunoassays whose
readouts rely on chemiluminescence are increasingly
useful for a broad range of analytical applications, but they are
rarely made into point-of-care (POC) format because of the complex
reagents required (some reagents have to be stored in low temperatures,
and some reagents have to be freshly made right before the assay).
This study reports a self-contained chemiluminescent lateral flow
assay (CLFA), which prestores all necessary reagents. This CLFA contains
three parts: the normal lateral flow assay (LFA) strip, the chemiluminescence
substrate pad, and the polycarbonate (PC) holder. On the LFA strip,
we simultaneously labeled horseradish peroxidase (HRP) and antibody
on the gold nanoparticles (AuNPs) for the conjugate pad. For the substrate
pad, we used sodium perborate as the oxidant and lyophilized the chemiluminescence
substrate on the glass fiber, which allows long-term storage. After
the transfer of substrate from the substrate pad to the nitrocellulose
(NC) membrane, we captured the chemiluminescence signal for the quantification
of the targets. The HRP on the AuNPs can amplify the chemiluminescence
signal efficiently. We used this CLFA system to detect both macromolecules
and small molecules successfully. This self-contained and easily processable
device is exceedingly appropriate for rapid detection and is a convenient
platform for POC testing
Sensitive Interrogation of Enhancer Activity in Living Cells on a Nanoelectroporation-Probing Platform
Enhancers
involved in the upregulation of multiple oncogenes play
a fundamental role in tumorigenesis and immortalization. Exploring
the activity of enhancers in living cells has emerged as a critical
path to a deep understanding of cancer properties, further providing
important clues to targeted therapy. However, identifying enhancer
activity in living cells is challenging due to the double biological
barriers of a cell cytoplasmic membrane and a nuclear membrane, limiting
the sensitivity and responsiveness of conventional probing methods.
In this work, we developed a nanoelectroporation-probing (NP) platform,
which enables intranuclear probe delivery for sensitive interrogation
of enhancer activity in living cells. The nanoelectroporation biochip
achieved highly focused perforation of the cell cytoplasmic membrane
and brought about additional driving force to expedite the delivery
of probes into the nucleus. The probes targeting enhancer activity
(named āPH probeā) are programmed with a cyclic amplification
strategy and enable an increase in the fluorescence signals over 100-fold
within 1 h. The platform was leveraged to detect the activity of CCAT1 enhancers (CCAT1, colon cancer-associated
transcript-1, a long noncoding RNA that functions in tumor invasion
and metastasis) in cell samples from clinical lung cancer patients,
as well as reveal the heterogeneity of enhancers among different patients.
The observations may extend the linkages between enhancers and cancer
cells while validating the robustness and reliability of the platform
for the assay of enhancer activity. This platform will be a promising
toolbox with wide applicable potential for the intranuclear study
of living cells
Video_1_Danhong Injection Reversed Cardiac Abnormality in BraināHeart Syndrome via Local and Remote Ī²-Adrenergic Receptor Signaling.mp4
<p>Ischemic brain injury impacts cardiac dysfunction depending on the part of the brain affected, with a manifestation of irregular blood pressure, arrhythmia, and heart failure. Generally called braināheart syndrome in traditional Chinese medicine, few mechanistic understanding and treatment options are available at present. We hypothesize that considering the established efficacy for both ischemic stroke and myocardial infarction (MI), Danhong injection (DHI), a multicomponent Chinese patent medicine, may have a dual pharmacological potential for treating the braināheart syndrome caused by cerebral ischemic stroke through its multi-targeted mechanisms. We investigated the role of DHI in the setting of braināheart syndrome and determined the mechanism by which it regulates this process. We induced Ischemia/Reperfusion in Wistar rats and administered intravenous dose of DHI twice daily for 14 days. We assessed the neurological state, infarct volume, CT scan, arterial blood pressure, heart rhythm, and the hemodynamics. We harvested the brain and heart tissues for immunohistochemistry and western blot analyses. Our data show that DHI exerts potent anti-stroke effects (infarct volume reduction: <sup>āā</sup>p < 0.01 and <sup>āāā</sup>p < 0.001 vs. vehicle. Neurological deficit correction: <sup>ā</sup>p < 0.05 and <sup>āāā</sup>p < 0.001 vs. vehicle), and effectively reversed the abnormal arterial pressure (<sup>ā</sup>p < 0.05 vs. vehicle) and heart rhythm (<sup>āā</sup>p < 0.01 vs. vehicle). The phenotype of this braināheart syndrome is strikingly similar to those of MI model. Quantitative assessment of hemodynamic in cardiac functionality revealed a positive uniformity in the PV-loop after administration with DHI and valsartan in the latter. Immunohistochemistry and western blot results showed the inhibitory effect of DHI on the Ī²-adrenergic pathway as well as protein kinase C epsilon (PKCĪµ) (<sup>āā</sup>p < 0.01 vs. model). Our data showed the underlying mechanisms of the braināheart interaction and offer the first evidence that DHI targets the adrenergic pathway to modulate cardiac function in the setting of braināheart syndrome. This study has made a novel discovery for proper application of the multi-target DHI and could serve as a therapeutic option in the setting of braināheart syndrome.</p