Encoding DNA Frameworks for Amplified Multiplexed Imaging of Intracellular microRNAs.

Real-time imaging of multiple low-abundance microRNAs (miRNAs) simultaneously in living cells with high sensitivity is of vital importance for accurate cancer clinical diagnosis and prognosis studies. Maintaining stability of nanoprobes resistant to enzyme degradation and enabling effective signal amplification is highly needed for in vivo imaging studies. Herein, a rationally designed one-pot assembled multicolor tetrahedral DNA frameworks (TDFs) by encoding multicomponent nucleic acid enzymes (MNAzymes) was developed for signal-amplified multiple miRNAs imaging in living cells with high sensitivity and selectivity. TDFs could enter cells via self-delivery with good biocompatibility and stability. Two kinds of MNAzymes specific for miRNA-21 and miRNA-155 with fluorescein labeling were encoded in the structure of TDFs respectively through one-step thermal annealing. In the intracellular environment, the TDFs could be specifically bound with its specific miRNA target and form an active DNAzyme structure. The cleavage of the active site would trigger the release of target miRNA and circular fluorescence signal amplification, which enabled accurate diagnosis on miRNA identifications of different cell lines with high sensitivity. Meanwhile, with the specific AS1411 aptamer targeting for nucleolin overexpressed on the surface of the carcinoma cells, this well-designed TDFs nanoprobe exhibited good discrimination between cancer cells and normal cells. The strategy provides an efficient tool for understanding the biological function of miRNAs in cancer pathogenesis and therapeutic applications.

Sources and health risks of PM2.5-bound polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) in a North China rural area.

Polychlorinated biphenyls (PCBs) and organochlorine pesticides (OCPs) are typical persistent organic pollutants (POPs), which have high toxicity, bioaccumulation and long-distance transfer capability. Daily variation, sources of PCBs and OCPs in PM2.5 are rarely explored in polluted rural area. Here, the sources and health risks of the PCBs and OCPs were evaluated for 48 PM2.5 samples collected in winter 2017 in Wangdu, a heavy polluted rural area in the North China Plain. The average diurnal and nocturnal concentrations of Σ18PCBs and Σ15OCPs were 1.74-24.37 and 1.77-100.49, 11.67-408.81 and 16.89-865.60 pg/m3, respectively. Hexa-CBs and penta-CBs accounted for higher proportions (29.0% and 33.6%) of clean and polluted samples, respectively. Hexachlorobenzene (HCB) was the dominant contributor to OCPs with an average concentration of 116.17 pg/m3. Hexachlorocyclohexane (ΣHCHs) and dichlorodiphenyltrichloroethane (ΣDDTs) were the other two main classes in OCPs with the average concentrations of 4.33 and 15.89 pg/m3, respectively. ß-HCH and p,p'-DDE were the main degradation products of HCHs and DDTs, respectively. The principal component analysis and characteristic ratio method indicated both waste incineration and industrial activities were the main sources of PCBs, contributing 76.8% and 12.7%, respectively. The loadings of OCPs were attributed to their application characteristics and the characteristic ratio method reflected a current or past use of OCPs. Health risk assessment showed that the respiratory exposure quantity of doxin-like PCBs (DL-PCBs) and the lifetime cancer risk from airborne OCPs exposure was negligible, while the other exposure modes may pose a risk to human bodies.

Unexpectedly Increased Particle Emissions from the Steel Industry Determined by Wet/Semidry/Dry Flue Gas Desulfurization Technologies.

"Ultralow-emission" standards have started to be implemented for steel plants in China. Flue gas desulfurization (FGD) systems integrating desulfurization and dedusting, common end-of-pipe technologies before the stacks, have been a key process for controlling the complexity of sintering flue gas to meet ultralow-emission requirements. This study reports comprehensive analysis of the influence of wet/semidry/dry FGD systems on particulate emissions via a field investigation of five typical sinter plants equipped with various FGD devices. The size distribution and mass concentration of particulate matter (PM) are adjusted to different ranges by these FGD systems. Chemical analysis of the PM compositions shows that 20-95% of the mass of inlet PM is removed by FGD systems, while it is estimated that approximately 17, 63, 59, and 71% of the outlet PMs are newly contributed by desulfurizers and their byproducts for the tested wet limestone, wet ammonia, semidry circulating fluidized bed, and activated coke FGD systems, respectively. The newly contributed compositions of PM2.5 emitted from these FGD systems are dominated by CaSO4, (NH4)2SO4, CaSO4 + CaO, and coke carbon, respectively. These results suggest that the deployment of FGD technology should be comprehensively considered to avoid additional negative impacts from byproducts generated in control devices on the atmosphere.

The Efficacy of Paired Opposite Clear Corneal Incisions Correcting Preexisting Low-to-Moderate Astigmatism in Implantable Collamer Lens Surgery.

PURPOSE: To evaluate the efficacy of paired opposite clear corneal incisions (OCCIs) in Implantable Collamer Lens (ICL; STAAR Surgical) implantation surgery for correcting preexisting low-to-moderate astigmatism and observe their influence on corneal aberration. METHODS: This prospective controlled randomized study included 123 eyes: 73 eyes in the ICL surgery group (control group) and 50 eyes in the ICL combined with OCCI group (OCCI group). All patients had corneal astigmatism between 0.50 and 3.00 D. Parameters considered included uncorrected distance visual acuity (UDVA), corrected distance visual acuity (CDVA), refraction, keratometry, slit-lamp biomicroscopy, indirect ophthalmoscopy, corneal topography, and higher order aberrations (HOAs). The Alpins method was used to analyze the correction of astigmatism. The follow-up period lasted for 12 months. RESULTS: Both groups showed a statistically significant improvement in mean UDVA and CDVA after surgery. At 1 and 12 months, the OCCI group had statistically better UDVA than the control group (P = .021 and .01). The OCCI group showed a significant reduction in mean refractive astigmatism from -0.835 ± 0.274 to -0.535 ± 0.353 and -0.450 ± 0.346 D postoperatively (P < .05) after 1 and 12 months. No significant change was observed in the control group (P > .05). The average magnitude of surgically induced astigmatism (SIA) was 0.911 ± 0.442 and 0.811 ± 0.397 D at 1 and 12 months postoperatively, respectively, in the OCCI group, which was slightly lower than the target induced astigmatism. The difference in SIA between the OCCI and control groups was significant by the end of 1 and 12 months postoperatively (P < .0001). The mean correction index was below 1, suggesting an undercorrection effect caused by clear corneal incisions in the opposite direction. HOAs increased in both the control and OCCI groups after surgery (t = 5.668, P < .0001, t = 3.773, P = .0003, respectively), with oblique trefoil only significantly increasing in the OCCI group. CONCLUSIONS: This study demonstrated that OCCIs represent a secure, efficient, reliable, and long-term technique for reducing preexisting low-to-moderate corneal astigmatism during ICL surgery. However, OCCIs did not show any advantageous impact in terms of wavefront aberrations when compared to the traditional single-incision method. The presence of oblique trefoil may be the main contributor to the alteration of HOAs during OCCI surgery. [J Refract Surg. 2024;40(1):e20-e29.].

Photothermal Iron-Based Riboflavin Microneedles for the Treatment of Bacterial Keratitis via Ion Therapy and Immunomodulation.

Bacterial biofilm formation protects bacteria from antibiotics and the immune system, excessive inflammation further complicates treatment. Here, iron-based metal-organic framework (MIL-101)-loaded riboflavin nanoparticles are designed for the therapeutic challenge of biofilm infection and hyperinflammation in bacterial keratitis. Specifically, MIL-101 produces a thermal effect under exogenous near-infrared light irradiation, which synergizes with ferroptosis-like bacterial death induced by iron ions to exert an effective biofilm infection eradication effect. On the other hand, the disintegration of MIL-101 sustains the release of riboflavin, which inhibits the pro-inflammatory response of macrophage over-activation by modulating their phenotypic switch. In addition, to solve the problems of short residence time, poor permeability, and low bioavailability of corneal medication, the MR@MN microneedle patch is further prepared by loading nanoparticles into SilMA hydrogel, which ultimately achieves painless, transepithelial, and highly efficient drug delivery. In vivo and ex vivo experiments demonstrate the effectiveness of this approach in eliminating bacterial infection and promoting corneal healing. Therefore, the MRMN patch, acting as an ocular drug delivery system with the ability of rapid corneal healing, promises a cost-effective solution for the treatment of bacterial keratitis, which may also lead to a new approach for treating bacterial keratitis in clinics.

Size distribution and chemical composition of primary particles emitted during open biomass burning processes: Impacts on cloud condensation nuclei activation.

As a major source of fine particles, open biomass burning can affect climate and the hydrological cycle via the formation of cloud condensation nuclei (CCN) and their deposition. This study investigated the effect of aerosol size and chemical composition (water-soluble compounds, elemental carbon, and organic carbon) on CCN activation during the complete combustion of six commonly used crop (rice, wheat, corn, soybean, and cotton) and leaf residues in a simulation chamber developed for this study. Geometric mean diameters (GMD) of emitted primary particles ranged from 68 to 130 nm and water-soluble compound (WSC) content ranged from 33 to 50%. Although GMDs and WSCs possessed similar temporal tendencies during the combustion process, their average values were dependent on the type of biomass. Number concentration ratios of CCN and particle (CCN/CN) ranged from 1.4-2.1% and 5.8-8.3% at supersaturations of 0.2% and 1.0%, respectively. The CCN/CN value increased during the combustion process when the GMD and WSC content increased. Correlation coefficients for GMD and CCN activation ranged from 0.62 to 0.93, while correlations for WSC and CCN activation ranged from 0.33 to 0.95. Soybean biomass had the highest correlations for both GMD and WSC with CCN activation at various supersaturations. These results suggest that the size and chemical composition of biomass aerosols are important in CCN formation, while the aerosol size has a higher impact.

Cancer-Specific MicroRNA Analysis with a Nonenzymatic Nucleic Acid Circuit.

Sensitive detection of gastric cancer-related biomarkers in human serum provides a promising means for early cancer diagnosis. Herein, we report the design of a nucleic acid circuit for gastric cancer-related microRNA-27a (miRNA-27a) detection based on dual toehold-mediated circular strand displacement amplification (CSDA). In the presence of miRNA-27a, the hybridization between miRNA-27a and probe DNA on magnetic beads through toehold 1 leads to the release of fluorescent DNA and the exposure of a new toehold 2 on linker DNA. After hybridization with catalytic DNA, CSDA is initiated and target miRNA-27a is released to participate in the next cyclic reaction; therefore, a greatly enhanced fluorescence signal is produced. The efficient magnetic separation makes the sensitive detection of miRNA-27a be accomplished within 45 min. With the efficient CSDA, the detection limit of the system (0.8 pM) is ∼100 folds lower than that of the system based on strand displacement without CSDA (79.3 pM). Furthermore, the system also showed good stability and sensitivity to discriminate single-base mismatch, which allows the detection of miRNA-27a in human serum samples. This study provides a novel platform and approach for the rapid quantitative determination of miRNA, which has great potential in clinical diagnosis and disease treatment.

Intracellular MicroRNA Imaging with MoS2-Supported Nonenzymatic Catassembly of DNA Hairpins.

Amplification strategies for low-level microRNA detection in living cells are pivotal for gene diagnosis and many cellular bioprocesses. In this work, we develop an amplification strategy for microRNA-21 (miRNA-21) imaging in living cells with MoS2-supported catassembly of DNA hairpins. The MoS2 nanosheet with low cytotoxicity serves as the nanocarrier and excellent fluorescence quencher, which can transfer fluorescent metastable hairpin DNA into the cells easily in a nondestructive manner and significantly reduce background signals. The three-branched catalyzed hairpin assembly (TB-CHA) probes contain three types of designed DNA molecular beacons with the modification of Cy3 in the terminal. In the presence of miRNA-21, the catalyzed hairpin assembly (CHA) reaction would be triggered and a "Y"-shaped three-branched duplex nanostructure would be formed, which would release from the surface of the MoS2 nanosheet due to the reduced affinity between the DNA duplex and MoS2 nanosheet. The multisite fluorescence modification and the circular reaction of TB-CHA probes allowed a significant fluorescence recovery in a live-cell microenvironment. The ultrasensitive detection of miRNA-21 is achieved with a detection limit of 75.6 aM, which is ∼5 orders of magnitude lower than that of a simple strand displacement-based strategy (detection limit: 8.5 pM). This method offers great opportunities for the ultrasensitive live-cell detection of miRNAs and helps in gaining a deeper understanding of the physiological functions of miRNAs in cancer research and life processes.