Lysosomes Initiating and DNAzyme-Assisted Intracellular Signal Amplification Strategy for Quantification of Alpha-Fetoprotein in a Single Cell.

Cellular trace proteins are critical for maintaining normal cell functions, with their quantitative analysis in individual cells aiding our understanding of the role of cell proteins in biological processes. This study proposes a strategy for the quantitative analysis of alpha-fetoprotein in single cells, utilizing a lysosome microenvironment initiation and a DNAzyme-assisted intracellular signal amplification technique based on electrophoretic separation. A nanoprobe targeting lysosomes was prepared, facilitating the intracellular signal amplification of alpha-fetoprotein. Following intracellular signal amplification, the levels of alpha-fetoprotein (AFP) in 20 HepG2 hepatoma cells and 20 normal HL-7702 hepatocytes were individually evaluated using microchip electrophoresis with laser-induced fluorescence detection (MCE-LIF). Results demonstrated overexpression of alpha-fetoprotein in hepatocellular carcinoma cells. This strategy represents a novel technique for single-cell protein analysis and holds significant potential as a powerful tool for such analyses.

mRNA-activated DNAzyme nanoprobe for tumor cell precise imaging and gene therapy.

A novel Au-nucleic acid nanoprobe, catalyzed by mRNA, has been developed for live cell imaging and precise treatment of tumor cells. This nanoprobe exhibits the remarkable ability to differentiate between tumor cells and normal cells through live cell mRNA imaging, while selectively inducing apoptosis in tumor cells.

A Framework Nucleic Acid-Mediated Multimodal Tandem Multivariate Signal Amplification Strategy for Simultaneous Absolute Quantification of Three Different MicroRNAs in a Single Cell.

The simultaneous quantification of multiple microRNAs (miRNA) in a single cell can help scientists understand the relationship between different miRNA groups and different types of cancers from an miRNA omics perspective at the single-cell level. However, there currently remains a challenge in developing techniques for the simultaneous absolute quantification of multiple miRNAs in single cells. Herein, we propose a framework nucleic acid (FNA)-mediated multimodal tandem multivariate signal amplification strategy for simultaneous absolute quantification of three different miRNAs in a single cell. In this study, DNA hexahedron FNAs (DHFs) and DNA tetrahedron FNAs (DTFs) were first prepared, multiple DNA hairpins and substrates were then connected to the hexahedron frame nucleic acid as the target recognition units, and three substrates with labeled FAM fluorophores on the tetrahedral frame nucleic acid served as signal output units. After the two types of FNAs entered the cell, they reacted with three different miRNAs (miRNA-155, miRNA-373, and miRNA-21) and multimodal tandem multivariate signal amplification was initiated simultaneously, reducing the detection limit of the three miRNAs to 8 × 10-15, 2 × 10-15, and 1 × 10-15 M, respectively. The detection sensitivity of the three miRNAs was simultaneously increased by six orders of magnitude, reaching the quantitative requirement of trace miRNAs in single cells. Combined with single-cell injection, membrane melting, and intracellular component separation technology on a microchip electrophoresis platform, we achieved the simultaneous absolute quantification of three different miRNAs in a single cell, thereby providing an important novel method that can be used to conduct single-cell research.

Activation-Assisted High-Concentration Phosphorus-Doping to Enhance the Electrochemical Performance of Cobalt Carbonate Hydroxide Hydrate.

P-doping into metal oxides has been demonstrated as a valid avenue to ameliorate electrochemical performance because it can tune the electronic structures and increase the active sites for an electrochemical reaction. However, it usually results in a low P-doping concentration via the commonly used gas phosphorization method. In this work, an activation-assisted P-doping strategy was explored to significantly raise the P-doping concentration in cobalt carbonate hydroxide hydrate (CCHH). The activation treatment increased active sites for electrochemical reaction and endowed the sample with a high P content in the subsequent gas phosphorization process, thereby greatly enhancing the conductivity of the sample. Therefore, the final CCHH-A-P electrode exhibited a high capacitance of 6.62 F cm-2 at 5 mA cm-2 and good cyclic stability. In addition, the CCHH-A-P//CC ASC with CCHH-A-P as the positive electrode and carbon cloth as the negative electrode provided a high energy density of 0.25 mWh cm-2 at 4 mW cm-2 as well as excellent cycling performance with capacitance retention of 91.2% after 20,000 cycles. Our work shows an effective strategy to acquire Co-based materials with high-concentration P-doping that holds great potential in boosting the electrochemical performance of electrode materials via P-doping technology.

"Spongy skin" as a robust strategy to deliver 4-octyl itaconate for conducting dual-regulation against in-stent restenosis.

The valid management of inflammation and precise inhibition of smooth muscle cells (SMCs) is regarded as a promising strategy for regulating vascular responses after stent implantation, yet posing huge challenges to current coating constructions. Herein, we proposed a spongy cardiovascular stent for the protective delivery of 4-octyl itaconate (OI) based on a "spongy skin" approach, and revealed the dual-regulation effects of OI for improving vascular remolding. We first constructed a "spongy skin" onto poly-l-lactic acid (PLLA) substrates, and realized the protective loading of OI with the highest dosage of 47.9 µg/cm2. Then, we verified the remarkable inflammation mediation of OI, and surprisingly revealed that the OI incorporation specifically inhibited SMC proliferation and phenotype switching, which contributed to the competitive growth of endothelial cells (EC/SMC ratio âˆ¼ 5.1). We further demonstrated that OI at a concentration of 25 µg/mL showed significant suppression of the TGF-ß/Smad pathway of SMCs, leading to the promotion of contractile phenotype and reduction of extracellular matrix. In vivo evaluation indicated that the successful delivery of OI fulfilled the inflammation regulation and SMCs inhibition, therefore suppressing the in-stent restenosis. This "spongy skin" based OI eluting system may serve as a new strategy for improving vascular remolding, and provides a potential concept for the treatment of cardiovascular diseases.

Effect of a modified regimen on drug-sensitive retreated pulmonary tuberculosis: A multicenter study in China.

Background and objective: Retreatment pulmonary tuberculosis (PTB) still accounts for a large proportion of tuberculosis, and the treatment outcome is unfavorable. The recurrence of retreatment PTB based on long-term follow-up has not been well demonstrated. This study aimed to evaluate effect of a modified regimen on drug-sensitive retreated pulmonary tuberculosis. Methods: This multicenter cohort study was conducted in 29 hospitals from 23 regions of China from July 1, 2009, to December 31, 2020. Patients were divided into two treatment regimen groups including experimental group [modified regimen (4H-Rt2-E-Z-S(Lfx)/4H-Rt2-E)]and control group [standard regimen (2H-R-E-Z-S/6H-R-E or 3H-R-E-Z/6H-R-E)]. The patients enrolled were followed up of 56 months after successful treatment. We compared the treatment success rate, treatment failure rate, adverse reaction rate, and recurrence rate between two regimens. Multivariate Cox regression model was used to identify the potential risk factors for recurrence after successful treatment with proportional hazards assumptions tested for all variables. Results: A total of 381 patients with retreatment PTB were enrolled, including 244 (64.0%) in the experimental group and 137 (36.0%) in the control group. Overall, the treatment success rate was significant higher in the experimental group than control group (84.0 vs. 74.5%, P = 0.024); no difference was observed in adverse reactions between the two groups (25.8 vs. 21.2%, P > 0.05). A total of 307 patients completed the 56 months of follow-up, including 205 with the modified regimen and 102 with the standard regimen. Among these, 10 cases (3.3%) relapsed, including 3 in the experimental group and 7 in the control group (1.5% vs 6.9%, P = 0.035). Reduced risks of recurrence were observed in patients treated with the modified regimen compared with the standard regimen, and the adjusted hazard ratio was 0.19 (0.04-0.77). Conclusion: The modified retreatment regimen had more favorable treatment effects, including higher treatment success rate and lower recurrence rate in patients with retreated drug-sensitive PTB.

Influence of COVID-19 for delaying the diagnosis and treatment of pulmonary tuberculosis-Tianjin, China.

Background: The COVID-19 pandemic has disrupted the diagnosis, treatment, and care for tuberculosis (TB). Delays in seeking TB care may result in increased community transmission and unfavorable treatment outcomes. We sought to understand the influence of the COVID-19 pandemic on the proportion of patients with TB who delayed seeking the diagnosis and care for TB and explore the reasons for their postponement. Methods: We surveyed a representative sample of outpatients treated for pulmonary TB from June to November 2020 using an anonymous standardized questionnaire. Multivariable logistic regression was used to calculate adjusted odds ratios (aOR) and 95% confidence intervals (CIs) of factors associated with the postponement of TB care. We used routinely collected surveillance data to assess trends of TB reports before and after the emergence of COVID-19 (2017-2019 vs. 2020-2022) in Tianjin, China. Results: Among 358 participants who were diagnosed with pulmonary TB during the COVID-19 response, 61 (17%) postponed seeking TB diagnosis due to COVID-19, with 39 (64%) citing fear as the primary reason. Female sex (aOR:2.0; 95% CI: 1.1-3.7), previous antituberculosis treatment (aOR:3.2; 95%CI: 1.4-7.6), and TB diagnosis during the first-level response (aOR = 3.2, 1.7-6.2) were associated with the postponement. Among all 518 participants receiving antituberculosis treatment, 57 (11%) had postponed their regular healthcare visits due to COVID-19, 175 (34%) received no treatment supervision, and 32 (6%) experienced treatment interruption. Compared to 2017-2019, reported pulmonary TB declined by 36.8% during the first-level response to COVID-19, 23.5% during the second-level response, 14% during the third-level response in 2020, and 4.3% in 2021. Conclusion: The COVID-19 response reduced the number of people who sought and received diagnosis, treatment, and care for TB in Tianjin, China. Integrative programs to ensure access and continuity of TB services should be considered and dual testing for SARS-CoV-2 and M. tuberculosis may facilitate finding cases.

Intracellular Multicomponent Synchronous DNA-Walking Strategy for the Simultaneous Quantification of Tumor-Associated Proteins in a Single Cell.

Single-cell protein analysis is very important for understanding cellular heterogeneity and single-cell biology. However, owing to the extremely low levels of some tumor-associated proteins in individual cells, the absolute quantification of such tumor-associated proteins in a single cell remains a challenge. Herein, an intracellular multicomponent synchronous DNA-walking strategy is proposed for the simultaneous quantification of multiple tumor-associated proteins in a single cell. In this strategy, a nanoprobe based on a DNA walker was designed for the simultaneous signal amplification of nucleolin (NCL) and thymidine kinase 1 (TK1) in a single cell. NCL and TK1 in single cells were simultaneously detected on a microchip platform with detection limits of 1.0 and 0.8 pM, respectively. The results obtained from 20 liver cancer cells (HepG2) and 20 normal hepatocytes (HL-7702) indicated that NCL and TK1 were overexpressed in liver cancer cells. However, the levels of NCL and TK1 in normal hepatocytes are only about one-tenth to one-sixth of those in hepatic carcinoma. Using different nucleic acid aptamers, the proposed strategy can be applied for the analysis of other single-cell proteins and in the early diagnosis of cancer.

Giant photonic spin Hall effect in bilayer borophene metasurfaces.

We investigate theoretically the photonic spin Hall effect (PSHE) in bilayer borophene metasurfaces. Based on the combined effect of the Fabry-Perot resonance of the bilayer system and the resonant interaction of individual meta-atoms in borophene metasurface which lead to the topological transition, it is found that there exist giant PSHE shifts of the transmitted beams which can be flexibly regulated by adjusting the twist angle of metasurface bilayers, incident angle, spacer refractive index and spacer thickness. Near the topological transition of borophene metasurface the magnitude of PHSE shifts in bilayer borophene metasurfaces is generally on the order of tens of wavelengths and even on the order of hundreds of wavelengths near the epsilon-near-zero (ENZ) regions. The manipulation frequency range of the large PSHE shifts can reach hundreds of terahertz or even picohertz through adjusting the ribbon width of borophene metasurface or the electron density for borophene. It is found that in bilayer borophene metasurfaces there exist the ultrahigh sensitivity of the PSHE shifts to spacer refractive index, which can be applied to design the refractive index sensors with high performance.

An ultrasensitive bunge bedstraw herb type DNA machine for absolute quantification of mRNA in single cell.

Messenger ribonucleic acids (mRNAs) comprise a class of small nucleic acids carrying genetic information, which exhibit very important role in medical research and diagnosis. If only the mean mRNA expression levels of the mRNA population are considered in medical research, important information linking mRNA expression and cellular function may be lost. Single-cell analysis provides valuable insights into studying its heterogeneity, signaling, and stochastic gene expression. In this study, a "bunge bedstraw herb"-type DNA machine based on DNAzyme catalyzing coupled clamping hybrid chain reaction (c-HCR) is presented. In the DNA machine, a bunge bedstraw herb-type DNA structure was first formed by hybridizing a core junction scaffold cruciform probe to a hairpin probe that can trigger the c-HCR via a target molecule in four directions. This approach can reduce the detection limit of mRNA to 5 × 10-15 M. Absolute quantification of survivin mRNA in individual cells was achieved using the DNA machine on a microfluidic chip electrophoresis platform. The reported method represents an unprecedented single-cell analysis platform for single-cell biology studies.

Genuine Pores in a Stable Zinc Phosphite for High H2 Adsorption and CO2 Capture.

Invited for the cover of this issue are Chih-Min Wang and co-workers at Academia Sinica of Taiwan and National Taiwan Ocean University. The image depicts an unusual organic-inorganic hybrid zinc phosphite with interesting structural features and gas adsorption properties. Read the full text of the article at 10.1002/chem.202200732.

Genuine Pores in a Stable Zinc Phosphite for High H2 Adsorption and CO2 Capture.

An uncommon example of stable mixed-ligand zinc phosphite with genuine pores has been synthesized by using zinc metal, inorganic phosphite acid, thio-functionalized O-donor (2,5-thiophenedicarboxylate, TPDC), and tetradentate N-donor [1,2,4,5-tetrakis(imidazol-1-ylmethyl)benzene, TIMB] units assembled into one crystalline structure according to a hydro(solvo)thermal method. This is a very rare case of a metal phosphite incorporating both N- and O-donor ligands. The tetradentate TIMB linker bound to zinc atoms of the isolated zincophosphite hexamers to form a 3D open-framework structure by crosslinking structural components of 1D chains and 2D layers. Here, the TPDC ligand acts as a monodentate binding model to functionalize its porous structure with the uncoordinated S atom and COO- group. Interestingly, this compound demonstrates the highest H2 storage capacity among organic-inorganic hybrid metal phosphates (and phosphites), and a good CO2 capture at 298 K compared with the majority of crystalline materials. The possible adsorption sites and selectivity for CO2 over H2 , N2 , and CO at 298 K were calculated by using density functional theory (DFT), the ideal adsorption solution theory (IAST), and fitting experimental pure-component adsorption data.

The substrate stiffness at physiological range significantly modulates vascular cell behavior.

Changes in the stiffness of the cellular microenvironment are involved in many pathological processes of blood vessels. Substrate stiffness has been shown to have extensive effects on vascular endothelial cells (VECs) and vascular smooth muscle cells (VSMCs). However, the material stiffness of most previously reported in-vitro models is ranging from ~100 kPa to the magnitude of MPa, which does not match the mechanical properties of natural vascular tissue (10-100 kPa). Herein, we constructed hydrogel substrates with the stiffness of 18-86 kPa to explore the effect of physiological stiffness on vascular cells. Our findings show that, with the increase of stiffness at the physiological range, the cell adhesion and proliferation behaviors of VECs and VSMCs are significantly enhanced. On the soft substrate, VECs express more nitric oxide (NO), and VSMCs tend to maintain a healthy contraction phenotype. More importantly, we find that the number of differentially expressed genes in cells cultured between 18 kPa and 86 kPa substrates (560 in VECs, 243 in VSMCs) is significantly higher than that between 86 kPa and 333 kPa (137 in VECs, 172 in VSMCs), indicating that a small increase in stiffness within the physiological range have a higher impact on vascular cell behaviors. Overall, our results expanded the exploration of how stiffness affects the behavior of vascular cells at the physiological range.

Nitrogen and sulfur co-doped carbon dot-based ratiometric fluorescent probe for Zn2+ sensing and imaging in living cells.

A near-infrared nitrogen and sulfur co-doped carbon dot (N,S-CD)-based ratiometric fluorescent probe is proposed that is synthesized via hydrothermal approach using glutathione and formamide as precursor for sensing and imaging of Zn2+. The prepared N,S-CDs facilitate binding with Zn2+ owing to N and S atom doping. The ratio (I650/I680) of fluorescence intensity at 650 nm and 680 nm increased with the concentrations of Zn2+ when the excitation wavelength was 415 nm. The linearity range was 0.01 to 1.0 µM Zn2+with a detection limit of 5.0 nM Zn2+. The proposed probe was applied to label-free monitoring of Zn2+ in real samples and fluorescent imaging of Zn2+ in living cells, which confirmed its promising applications.

An ultrasensitive multivariate signal amplification strategy based on microchip platform tailored for simultaneous quantification of multiple microRNAs in single cell.

MicroRNAs (miRNAs) play a very important regulatory role in life activities. Abnormal expression levels of miRNAs in cells are associated with various diseases, especially human cancer. Nevertheless, accurate detection of the copy numbers of various miRNA molecules in single cell is still a great challenge. In this study, an intracellular multivariate signal amplification strategy based on microchip platform was proposed, and an ultrasensitive single-cell analysis method was established for simultaneous quantification of absolute copy numbers of multiple miRNAs in a single cell. Using miRNA-21 and miRNA-141 as the analytical models of miRNAs, the detection limits of 1.0 and 2.0 fM were obtained. Based on the developed method, an analysis of 600 randomly acquired different types of cells was performed. The distribution of absolute copy numbers of miRNA-21 and miRNA-141 in six types of cells was obtained. It was found that the number of copies of miRNA-21 and miRNA-141 in different types of cancer cells showed different expression characteristics. The study results can help us more accurately understand cell-to-cell heterogeneity and the relationship between different miRNAs and different types of cancer at the single cell level.

Schottky-type GaN-based UV photodetector with atomic-layer-deposited TiN thin film as electrodes.

In this work, a GaN-based UV photodetector with an asymmetric electrode structure was fabricated by atomic layer deposition (ALD) of TiN layers. The thickness of the TiN can be monitored in situ by a quartz crystal microbalance (QCM) and precisely controlled through the modulation of deposition cycles. During the ALD process, periodic variation in the QCM frequency was observed and correlated to the physical adsorption, chemical bonding, and the excessive precursor exhaust, which included tetrakis(dimethylamino)titanium (TDMAT) and N sources. The asymmetric TiN/GaN/TiN photodetector showed excellent photosensing performance, with a UV-visible rejection ratio of 173, a responsivity of 4.25 A/W, a detectivity of 1.1×1013 Jones, and fast response speeds (a rise time of 69 µs and a decay time of 560 µs). Moreover, the device exhibits high stability, with an attenuation of only approximately 0.5% after 360 nm light irradiation for 157 min. This result indicates the potential of TiN as a transparent contact electrode for GaN-based optoelectronic devices.

Bioinspired NO release coating enhances endothelial cells and inhibits smooth muscle cells.

Thrombus and restenosis after stent implantation are the major complications because traditional drugs such as rapamycin delay the process of endothelialization. Nitric oxide (NO) is mainly produced by endothelial nitric oxide synthase (eNOS) on the membrane of endothelial cells (ECs) in the cardiovascular system and plays an important role in vasomotor function. It strongly inhibits the proliferation of smooth muscle cells (SMCs) and ameliorates endothelial function when ECs get hurt. Inspired by this, introducing NO to traditional stent coating may alleviate endothelial insufficiency caused by rapamycin. Here, we introduced SNAP as the NO donor, mimicking how NO affects in vivo, into rapamycin coating to alleviate endothelial damage while inhibiting SMC proliferation. Through wicking effects, SNAP was absorbed into a hierarchical coating that had an upper porous layer and a dense polymer layer with rapamycin at the bottom. Cells were cultured on the coatings, and it was observed that the injured ECs were restored while the growth of SMCs further diminished. Genome analysis was conducted to further clarify possible signaling pathways: the effect of cell growth attenuated by NO may cause by affecting cell cycle and enhancing inflammation. These findings supported the idea that introducing NO to traditional drug-eluting stents alleviates incomplete endothelialization and further inhibits the stenosis caused by the proliferation of SMCs.

Treatment Patterns and Outcomes in Patients with Esophageal Cancer: An Analysis of a Multidisciplinary Tumor Board Database.

BACKGROUND: Multidisciplinary management strategies are standard in esophageal cancer. Based on a multidisciplinary tumor board (MTB) database in a high-volume center, we aimed to evaluate real-world treatment patterns and patient outcomes in patients with esophageal cancer. In addition, we determined the impact of MTB discussions on patient prognosis. METHODS: Patients diagnosed with esophageal cancer between 2010 and 2019 were retrospectively reviewed. The pattern of treatment modalities and overall survival (OS) of patients with limited, locally advanced, and advanced/metastatic disease were reported. RESULTS: Data from 1132 patients, including 247 patients with limited esophageal cancer, 606 patients with locally advanced esophageal cancer, and 279 patients with advanced/metastatic esophageal cancer were included. Upfront surgery was the most common (56.3%) treatment modality for patients with limited esophageal cancer, while treatment for locally advanced esophageal cancer included upfront surgery (19.1%), neoadjuvant chemoradiotherapy (44.9%), and definitive chemoradiotherapy (36.0%); however, 27.9% of patients undergoing neoadjuvant chemoradiotherapy did not receive planned esophagectomy. Definitive chemoradiotherapy was mainly used for patients with locally advanced and advanced/metastatic disease, but had an incompletion rate of 22.0% and 33.7%, respectively. Regarding survival, the 5-year OS rates were 56.4%, 26.3%, and 5.1% in patients with limited, locally advanced, and advanced/metastatic disease, respectively. Additionally, patients whose clinical management was discussed in the MTB had a significantly better 5-year OS rate than the other patients (27.3% vs. 20.5%, p < 0.001). CONCLUSIONS: We report the real-world data of treatment patterns and patient outcomes in patients with esophageal cancer with respect to multidisciplinary management, and demonstrate the positive impact of MTB discussions on patient prognosis.

Facile preparation of Cu-doped carbon dots for naked-eye discrimination of phenylenediamine isomers and highly sensitive ratiometric fluorescent detection of H2O2.

Changing a detection analyte into a colored material is a key challenge for visual discrimination of isomers. In this work, a novel fluorescent probe incorporating Cu-doped carbon dots (Cu-CDs), for the first time, was developed for naked-eye discrimination of phenylenediamine isomers and highly sensitive ratiometric fluorescence detection of H2O2. In this strategy, Cu-CDs were synthesized by a facile hydrothermal approach using citric acid, formamide, and CuCl2 as reactants. The prepared Cu-CDs exhibited outstanding peroxidase-like activity and stability. Consequently, a chemosensor platform based on Cu-CDs was constructed to enable naked-eye discrimination of phenylenediamine isomers through the H2O2-mediated oxidation reaction. Moreover, a Cu-CDs-based ratiometric fluorescence sensor was proposed as a means to sensitively detect H2O2 with a detection limit of 5.0 nM. The sensor was further employed for monitoring H2O2 in human serum, indicating its potential applications in other biologically related study.

Enhanced Production of Formic Acid in Electrochemical CO2 Reduction over Pd-Doped BiOCl Nanosheets.

Bismuth oxyhalides (BiOX, X = F, Cl, Br, I) are emerging energy materials because of their remarkable catalytic activity. The BiOX compounds usually have a tetragonal type crystal structure with unique layered morphology consisting of [X-Bi-O-Bi-X] sheets. Although the BiOX nanosheets exposed with {001} facets perform superior photoactivity, there is lack of understanding about their capability in the electrochemical CO2 reduction reaction (CO2RR). Herein, we adopt wet-chemical syntheses to make 2D BiOCl and Pd-doped BiOCl nanosheets for CO2RR. In the results, formic acid is the only one kind of product converted from CO2 along with H2 gas from water reduction over both BiOCl and Pd-doped BiOCl nanosheets. By thorough analyses with ex situ and in situ spectroscopy, the results reflect that (1) metallic Bi0 atoms generated by the applied negative potentials serve as the catalytic sites for the hydrogen evolution reaction (HER) and CO2RR and (2) the existence of doped Pd ions in the BiOCl structure reduces the barrier of charge transfer over the nanosheets, which enhances HER and CO2RR activities. We believe that the observations are important references for making catalysts toward CO2RR performance.