Programming Fast DNA Amplifier Circuits with Versatile Toehold Exchange Pathway.

DNA amplifier circuits establish powerful tools to dynamically control molecular assembly for computation, sensing, and biological applications. However, the slow reaction speed remains a major barrier to their practical utility. Here, diverse fast DNA amplifier circuits termed toehold exchange polymerization (TEP) and toehold exchange catalysis (TEC) using toehold exchange-mediated assembly as a fundamental mechanism are built. Both TEP and TEC with a duplex and a hairpin can respond within minutes to diverse nucleic acid inputs with high fidelity. In addition, the circuits can amplify live-cell signals for fluorescence imaging target RNA dynamics and discriminating different cell lines. Compared with existing DNA circuits that involve time scales of hours for transducing small signals, TEP and TEC exhibit much faster dynamics, simpler design, and comparable sensitivity. These features make TEP and TEC promising platforms to develop programmable nucleic acid tools and devices and to create fast sensing and processing systems, amenable to wide practical applications.

Analysis of Factors Related to Physical Activity Levels Among Lung Cancer Survivors Who Underwent Nonsurgical Treatment: A Cross-Sectional Study.

AIM(S): The study aimed to investigate the current status of physical activity (PA) levels and associated factors among lung cancer survivors who have undergone nonsurgical treatments. BACKGROUND: PA has been incorporated as a nonpharmacological intervention in the rehabilitation programmes of cancer patients, playing a crucial role in alleviating symptom burden and enhancing the quality of life among lung cancer survivors. Understanding the potential influencing factors of PA levels aids in formulating targeted intervention strategies. DESIGN: A multicentre cross-sectional study. METHODS: Convenience sampling was utilised to survey lung cancer survivors from the respiratory and oncology departments of 12 hospitals across Eastern, Central and Western China, spanning from June 2023 to January 2024. Social demographic characteristics, disease-related features, health behaviour abilities, psychological factors and levels of PA were collected through a combination of clinical case systems used to record patients' treatment and medical conditions and self-reported questionnaires. Additionally, measurements of grip strength and the 6-min walk test were conducted for patients. Descriptive analysis, bivariate analysis and multivariate logistic regression were conducted. RESULTS: Only 109 patients (16.2%) achieved high PA levels. Multivariate logistic regression analysis indicated differences in age, residential location, employment status, religion, lung cancer stage, grip strength, albumin concentration, blood urea, Anderson symptom, depression and health behaviour capacity among lung cancer survivors with varying PA levels. CONCLUSIONS: Significant associations were observed between age ≥ 75 years, residing in urban areas, unemployment, absence of religious beliefs, Stage IV lung cancer, lower grip strength, lower albumin concentration, higher blood urea, higher Anderson symptom scores, lower health behaviour capacity scores, higher depression scores and lower PA levels among lung cancer survivors. These potential factors should be considered when developing PA intervention plans. RELEVANCE TO CLINICAL PRACTICE: This study offers insights for developing subsequent PA intervention programmes. In clinical practice, healthcare professionals should continuously educate patients about the benefits of exercise and help them incorporate PA into their daily lives. Additionally, emphasising multidisciplinary collaboration involving physical therapists, nutritionists and mental health experts is crucial for ensuring safe and effective PA, thereby improving patients' quality of life. REPORTING METHOD: Our study complies with the Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) Checklist: cross-sectional studies. PATIENT OR PUBLIC CONTRIBUTION: At the phase of collecting data, participants were recruited to fill the questionnaires. TRIAL REGISTRATION: Chinese Clinical Trial Registry: ChiCTR2300072609.

Significant lead isotope 'fractionation' in maize records plant lead uptake, transfer, and detoxification mechanisms.

Lead isotope analysis is the main method to trace the sources and cycling of Pb in the biosphere system. The linchpin of such application hinges on the assumption that there is no negligible or biologically mediated isotopic fractionation of Pb occurs in the environment. However, recent measurements by high-precision multi-collector mass spectrometry revealed that biological isotope fractionation of heavy mass elements is a prevalent phenomenon. This study shows that compared with the Pb sources, the maize plant (Zea mays L.) organs exhibit a wider range of Pb isotope compositions and a depletion of radioactive Pb isotopes (206Pb, 207Pb, and 208Pb). Moreover, three independent studies consistently indicate that the 206Pb/207Pb ratio of maize organs varies as root/leaf > stem/grain, reflecting a continuous loss of light Pb isotopes during transportation. The conventional wisdom fails to account for these phenomena, suggesting that maize may undergo Pb isotope fractionation during the absorption and transportation of Pb. However, compared with other non-traditional isotopes, Pb isotope exhibits a more significant fractionation magnitude. We tentatively attribute this fractionation to the Pb tolerance mechanism of maize and its selective absorption of various forms of Pb, which requires further research to validate. Findings from this study mandate caution in future Pb source trace in plants using Pb isotope methods and open up applications in using Pb isotopic fractionation to track Pb uptake and transfer pathways and decipher the associated detoxification mechanisms in plants.

A bibliometric analysis of domestic and international research on hemorrhagic fever with renal syndrome over the past 2 decades.

BACKGROUND: Bibliometrics and statistics were used to determine and analyze the research status of hemorrhagic fever with renal syndrome (HFRS) from 2004 to 2023, and objectively reflect the development and future trend of HFRS at home and abroad through comparison. METHODS: To search the research literature on HFRS in China National Knowledge Network and Web of Science databases from January 2004 to December 2023, CiteSpace and VOSviewer were used to visually analyze the annual publication trends, authors, research institutions, countries, co-cited literature, keywords and other contents of the included literatures. RESULTS: A total of 4460 Chinese literatures and 2372 foreign literatures were included. The number of HFRS published in the Web of Science database showed a trend of positive growth, while the number of HFRS published in China National Knowledge Network showed a trend of decline. Bai Xuefan and Wang Zhiqiang were the most published authors in China, and foreign scholars Vaheri, Antti, Ahlm, Clas. The main research institutions in the domestic literature were Zhejiang Provincial Center for Disease Control and Prevention, Liaoning Provincial Center for Disease Control and Prevention, while foreign research institutions concentrated on the University of Helsinki and Ministry of Health. The top 3 countries in the literature research of Web of Science are the USA, China, and Germany. CONCLUSION: The analysis results of hot spots and trends suggested that we need to develop more reliable tools and methods in the monitoring and spatio-temporal analysis of HFRS epidemic data in the future, so as to provide references for the surveillance and early warning of zoonotic diseases in the field of public health research.

Serum amyloid A1 induced dysfunction of airway macrophages via CD36 pathway in allergic airway inflammation.

Previous studies showed that serum amyloid A (SAA) and macrophages were associated with allergic airway inflammation. However, the interaction between SAA1 and macrophages in allergic airway inflammation remains to be further elucidated. In this study, the levels of SAA1 were measured in nasal tissues from patients with eosinophilic chronic rhinosinusitis with nasal polyps (CRSwNP), house dust mite (HDM)-treated BEAS-2B cells and the tissues of mice of HDM-induced allergic airway inflammation. Human monocytes-derived macrophages and mouse bone marrow-derived macrophages (BMDMs) were exposed to SAA1, and CCL17 and the other M1/M2-related factors were evaluated using RT-PCR and/or ELISA. To test the effects of SAA1-treated BMDMs on chemotaxis and differentiation of CD4+ T cells, number of migrated cells and the levels of Th1 and Th2 were measured using flow cytometry. SAA1 receptors were examined in BMDMs and lung macrophages of model mice. CD36 neutralizing antibody was applied to explore the mechanisms of SAA1 in regulating BMDMs using RT-PCR and/or ELISA. We found that SAA1 was expressed in epithelial cells, and was increased in the nasal tissues of patients with eosinophilic CRSwNP and HDM-treated BEAS-2B- cells as well as the bronchoalveolar lavage fluid and lung tissues of mice exposed to HDM. We also found that the level of CCL17 was increased in M2 macrophages, more CD4+ T cells were recruited and proportion of Th2 was increased after the treatment of SAA1. The treatment of CD36 neutralizing antibody decreased CCL17 level in SAA1-treated M2 BMDMs. In summary, our results showed that SAA1 was increased in allergic airway inflammation, and the administration of SAA1 upregulated the expression of CCL17 in M2 macrophages via CD36 and promoted the chemotaxis of CD4+ T cells and differentiation of Th2. It may provide a new therapeutic strategy that could mediate allergic airway inflammation via suppressing SAA1 to reduce recruitment of CD4+ T cells and activation of Th2.

Dual-Mode MXene-Based Phase-Change Composite Towards Enhanced Photothermal Utilization and Excellent Infrared Stealth.

Solar thermal collectors based on phase change materials (PCMs) are important to promote the civilian use of sustainable energy. However, simultaneously achieving high photothermal efficiency and rapid heat transfer of the PCM carrier typically involves a high proportion of functional materials, contradicting a satisfying energy storage density. In this work, a surface-engineered anisotropic MXene-based aerogel (LMXA) integrated with myristic acid (MA) to produce phase change composites (LMXA-MA) is reported, in which the laser-treated surface composed of the hierarchically-structured TiO2/carbon composites act as a light absorber to improve solar absorption (96.0%), while the vertical through-hole structure allows for fast thermal energy transportation from surface to the whole. As a result, LMXA-MA exhibits outstanding thermal energy storage (192.4 J·g-1) and high photothermal conversion efficiency (93.5%). Meanwhile, benefiting from the intrinsic low emissivity of MXene material, thermal radiation loss can be effectively suppressed by simply flipping LMXA-MA, enabling a long-term temperature control ability (605 s·g-1). The excellent heat storage property and switchable dual-mode also endow it with an infrared stealth function, which maintains camouflage for more than 240 s. This work provides a prospective solution for optimizing photothermal conversion efficiency and long-term thermal energy preservation from surface engineering and structural design.

Investigation of the impact of papain on the volatile and non-volatile metabolites of soybean meal yogurt.

BACKGROUND: Soybean meal yogurt was prepared from soybean meal using papain and Bifidobacterium animalis subsp. lactis. A non-targeted metabolomics approach was employed to analyze the relevance of papain to the differences in volatile and non-volatile metabolites of soybean meal yogurt. RESULTS: The results showed that the main up-regulated metabolites and metabolic pathways after enzymatic digestion were dominated by amino acids and their derivatives. Conversely, the main down-regulated metabolites and pathways were predominantly associated with flavonoid metabolism. Amino acids and their derivatives, as well as flavonoids, were found to be highly correlated with the formation of sweet, umami, astringent, and bitterness. The addition of papain enriched the content of aromatic compounds in soybean meal yogurt. Aromatic components such as 2-heptanone, naphthalene, and p-xylene increased in concentration. Synthetic peptide of aspartate and serine, gramine, geissospermine, N-desmethyl vinblastine, and 3,7-dihydroxyflavone were the major non-volatile differential metabolites distinguishing the soybean meal yogurt. CONCLUSION: This study provided a comprehensive analysis of the metabolic traits of products co-fermented by papain and Bifidobacterium animalis subsp. lactis, offering insights for the application of papain in fermented goods. © 2024 Society of Chemical Industry.

Antepartum versus postpartum amoxicillin oral challenge in pregnant patients with a reported penicillin allergy: A two-center prospective cohort study.

INTRODUCTION: While 10% of pregnant individuals report a penicillin allergy, there is no established best practice for penicillin allergy delabeling in pregnancy. To better understand options for penicillin delabeling, we aimed to evaluate two penicillin allergy delabeling protocols in pregnancy regarding efficacy, adverse events, and patient satisfaction. MATERIAL AND METHODS: From July 2019 to December 2022, we completed a two-center prospective cohort study, where each site recruited pregnant patients over 24 weeks gestational age with a reported penicillin allergy. One center offered antepartum amoxicillin oral challenges, either directly or after negative skin testing (i.e., antepartum oral challenge site). Our other centers completed a two-step approach with antepartum penicillin skin testing only and deferred oral challenges to the postpartum period (i.e., postpartum oral challenge site). Our primary outcome was the rate of penicillin allergy delabeling, defined as tolerating an antibiotic challenge with penicillin or amoxicillin. Univariate analyses were completed using chi-squared, Fisher's exact, and Wilcoxon rank tests. RESULTS: During the study period, 276 pregnant patients were assessed, with 207 in the antepartum oral challenge site and 69 in the postpartum oral challenge site. Among the 204 patients who completed antepartum oral challenges, 201 (98%) passed without reactions. Deferring oral challenges to the postpartum period led to a loss of follow-up for 37/53 (70%) of eligible individuals. Overall, 97% (201/207) of patients at the antepartum oral challenge site were delabeled from their penicillin allergy-compared to 38% (26/69) of patients referred to the postpartum oral challenge site (p < 0.0001). Three antepartum oral challenge reactions were noted, including two mild cutaneous reactions and a case of transient abdominal discomfort. CONCLUSIONS: Antepartum amoxicillin oral challenge is a more effective method to delabel pregnant patients from their penicillin allergy. Deferral of oral challenges to the postpartum period introduces a significant barrier for penicillin allergy delabeling.

Effect of degree of substitution of octenyl succinate on starch micelles for synthesis and stability of selenium nanoparticles: Towards selenium supplements.

To develop a promising selenium supplement that overcomes the instability and poor water dispersibility of selenium nanoparticles (SeNPs), we synthesized a series of amphiphilic octenyl succinic anhydride starch (OSAS) through esterification. As the degree of substitution (DS) increased, the particle size of OSAS micelles and the critical micelle concentration (CMC) decreased. FTIR and XRD analyses confirmed the successful introduction of octenyl succinic anhydride groups onto starch. Subsequently, OSAS micelles were used as carriers to synthesize SeNPs via in situ chemical reduction, forming SeNPs-loaded self-assembled starch nano-micelles (OSAS-SeNPs). The OSAS-SeNPs exhibited spherical dispersion in water with an average diameter of 116.1 ±â€¯2.3 nm, contributed to enhanced hydrophobic interactions. TEM images showed a core-shell structure with SeNPs as the core and OSAS as the shell. FTIR results indicated hydrogen bonding interactions between OSAS and SeNPs. Due to the negatively charged OSAS shell and hydrogen bonding (OH⋯Se), OSAS-SeNPs remained non-aggregated for one month at room temperature, demonstrating remarkable stability. This study suggests that using OSAS can address the synthesis and stability issues of SeNPs, making it a potential selenium supplement candidate for further evaluation as an anticancer agent.

Synergistic effect of oxygen defects and calabash-like hollow carbon matrix enables VO2 as high-performance cathode for zinc ion battery.

Vanadium dioxide (VO2) materials exhibit significant theoretical specific capacity, which is ascribed to multi-electron transfer reactions and unique tunneled structures. However, the low electronic conductivity and sluggish reaction kinetics of VO2 have impeded its further development. Hence, in this study, we employed a synergistic strategy of defect engineering and compositing with a calabash carbon matrix to reduce Zn2+ diffusion barriers and accelerate electron transfer. The VO2 cathode provided a high specific capacity at a low rate of 303 mA h g-1 at 0.1 A g-1 after 191 cycles, along with good rate performance (168 mA h g-1 at 10 A g-1) and satisfactory long-term stability (170 mA h g-1 at 1 A g-1 after 1100 cycles). The exhaustive structural analyses indicated that oxygen vacancies accelerated the Zn2+ diffusion rate, while a uniform calabash-like hollow carbon matrix improved electronic conductivity during cycling. Moreover, ex-situ measurements demonstrated that during discharge, the composite cathode transformed to layered Zn3+x(OH)2V2O7·2H2O, which then facilitated the subsequent intercalation of Zn2+. This cooperative strategy advances the practical application of aqueous zinc ion batteries by leveraging vanadium-based electrodes.

Weakening the Space Charge Layer Effect Through Tethered Anion Electrolyte and Piezoelectric Effect Toward Ultra-Stable Zinc Anode.

The space charge layer (SCL) dilemma, caused by mobile anion concentration gradient and the rapid consumption of cations, is the fundamental reason for the generation of zinc dendrites, especially under high-rate discharge conditions. To address the issue, a physical (PbTiO3)/chemical (AMPS-Zn) barrier is designed to construct stable zinc ion flow and disrupt the gradient of anion concentration by coupling the ferroelectric effect with tethered anion electrolyte. The ferroelectric materials PbTiO3 with extreme-high piezoelectric constant can spontaneously generate an internal electric field to accelerate the movement of zinc ions, and the polyanionic polymer AMPS-Zn can repel mobile anions and disrupt the anions concentration gradient by tethering anions. Through numerical simulations and analyses, it is discovered that a high Zn2+ transference number can effectively weaken the SCL, thus suppressing the occurrence of zinc dendrites and parasitic side reactions. Consequently, an asymmetric cell using the PbTiO3@Zn demonstrates a reversible plating/stripping performance for 2900 h, and an asymmetric cell reaches a state-of-the-art runtime of 3450 h with a high average Coulombic efficiency of 99.98%. Furthermore, the PbTiO3@Zn/I2 battery demonstrated an impressive capacity retention rate of 84.0% over 65000 cycles by employing a slender Zn anode.

Mediation effect of stroke recurrence in the association between post-stroke lactate dehydrogenase and functional disability.

Background: We aimed to use lactate dehydrogenase (LDH) as a marker of inflammation burden and quantify post-stroke inflammation's direct and indirect effect on functional disability. Methods: We analyzed 5,129 patients with acute ischemic stroke (AIS) admitted to Shenyang First People's Hospital. Stroke recurrence and functional outcome measured by the modified Rankin Scale (mRS) were assessed at 90 days. Functional disability was defined as mRS score > 2. Receiver operating characteristic curve and restricted cubic spline (RCS) analysis were conducted to illustrate the associations between LDH levels and 90-day functional outcomes in patients with AIS. Mediation analyses were performed to examine the potential causal chain in which stroke recurrence may mediate the relationship between LDH and functional outcome. Positive correlation between LDH and hs-CRP was found and mediation effects of stroke recurrence in the association between LDH or hs-CRP and functional disability were both less than 20%. Sensitivity analyses in different subgroups showed comparable results. Results: Among 5,129 included AIS patients, the median (IQR) level of LDH was 186 (161-204.4) U/L. Functional disability was seen in 1200 (23.4%) patients and recurrence was observed in 371(7.2%) patients at 90-day follow-up. Each standard deviation increase in the concentration of LDH was linked to an increased risk of functional disability (adjusted odds ratio[aOR], 1.07; 95%CI,1.04-1.09) and stroke recurrence (aOR,1.02; 95%CI, 1.01-1.04) within 90 days. The highest quartile of LDH (>204.2 U/L) had an elevated risk of suffering functional disability (aOR, 1.21; 95%CI, 1.00-1.47) and recurrence (aOR, 1.21; 95%CI,1.00-1.47) compared with the lowest quartile of LDH (<161 U/L). Stroke recurrence during follow-up explained 12.90% (95%CI, 6.22-21.16%) of the relationship between LDH and functional disability. Positive correlation between LDH and hs-CRP was found and mediation effects of recurrence in the association between LDH or hs-CRP and functional disability were both less than 20%. Sensitivity analyses in different subgroups showed comparable results. Conclusion: The relationship between LDH and functional disability at 90 days among AIS patients is partially mediated by stroke recurrence, accounting for less than 20%. LDH deserves equal attention as hs-CRP in predicting recurrence and functional outcome. In addition to traditional secondary prevention measures, innovative anti-inflammatory strategies warrant further investigation.

Increased circulating LOX-1+ neutrophils activate T cells and demonstrate a pro-inflammatory role in allergic rhinitis.

Background: Low-density neutrophils are heterogeneous immune cells with immunosuppressive (such as polymorphonuclear myeloid-derived suppressor cells [PMN-MDSC]) or pro-inflammatory (such as low-density granulocytes [LDG]) properties that have been well described in multiple cancers and immune diseases. However, its role in allergic rhinitis (AR) is still unclear. Methods: In the present study, we defined low-density neutrophils as CD14-CD11B+CD15+LOX-1+ (LOX-1+ neutrophils), and their levels in the peripheral blood (PB) were evaluated and compared between patients with AR and healthy donors using flow cytometric analysis. LOX-1 expression on polymorphonuclear neutrophils was identified. Carboxyfluorescein succinimidyl ester (CFSE)-stained CD3+ T cells were cultured alone or with LOX-1+ neutrophils, T cell proliferation was assessed using flow cytometry, and pro-inflammatory cytokines in the supernatants were detected using enzyme-linked immunosorbent assay (ELISA). Clinicopathological analyses were performed to gain a thorough understanding of LOX-1+ neutrophils. Results: We determined that LOX-1+ neutrophils were significantly increased in the PB of patients with AR, and LOX-1 expression in neutrophils from patients with AR was elevated. Interestingly, LOX-1+ neutrophils derived from patients with AR, unlike PMN-MDSC, promoted T cell proliferation and pro-inflammatory cytokine production. Moreover, clinicopathological analysis revealed that there was no any relation between circulating LOX-1+ neutrophil levels and the levels of IgE, age and sex. Conclusion: These findings indicate that elevated circulating LOX-1+ neutrophils play a pro-inflammatory role in AR.

Programming DNA Nanoassemblies into Polyvalent Lysosomal Degraders for Potent Degradation of Pathogenic Membrane Proteins.

Lysosome-targeting chimera (LYTAC) shows great promise for protein-based therapeutics by targeted degradation of disease-associated membrane or extracellular proteins, yet its efficiency is constrained by the limited binding affinity between LYTAC reagents and designated proteins. Here, we established a programmable and multivalent LYTAC system by tandem assembly of DNA into a high-affinity protein degrader, a heterodimer aptamer nanostructure targeting both pathogenic membrane protein and lysosome-targeting receptor (insulin-like growth factor 2 receptor, IGF2R) with adjustable spatial distribution or organization pattern. The DNA-based multivalent LYTACs showed enhanced efficacy in removing immune-checkpoint protein programmable death-ligand 1 (PD-L1) and vascular endothelial growth factor receptor 2 (VEGFR2) in tumor cell membrane that respectively motivated a significant increase in T cell activity and a potent effect on cancer cell growth inhibition. With high programmability and versatility, this multivalent LYTAC system holds considerable promise for realizing protein therapeutics with enhanced activity.

Scalable crystal structure relaxation using an iteration-free deep generative model with uncertainty quantification.

In computational molecular and materials science, determining equilibrium structures is the crucial first step for accurate subsequent property calculations. However, the recent discovery of millions of new crystals and super large twisted structures has challenged traditional computational methods, both ab initio and machine-learning-based, due to their computationally intensive iterative processes. To address these scalability issues, here we introduce DeepRelax, a deep generative model capable of performing geometric crystal structure relaxation rapidly and without iterations. DeepRelax learns the equilibrium structural distribution, enabling it to predict relaxed structures directly from their unrelaxed ones. The ability to perform structural relaxation at the millisecond level per structure, combined with the scalability of parallel processing, makes DeepRelax particularly useful for large-scale virtual screening. We demonstrate DeepRelax's reliability and robustness by applying it to five diverse databases, including oxides, Materials Project, two-dimensional materials, van der Waals crystals, and crystals with point defects. DeepRelax consistently shows high accuracy and efficiency, validated by density functional theory calculations. Finally, we enhance its trustworthiness by integrating uncertainty quantification. This work significantly accelerates computational workflows, offering a robust and trustworthy machine-learning method for material discovery and advancing the application of AI for science.

A Methyl-Engineered DNAzyme for Endogenous Alkyltransferase Monitoring and Self-Sufficient Gene Regulation.

The on-demand gene regulation is crucial for extensively exploring specific gene functions and developing personalized gene therapeutics, which shows great promise in precision medicines. Although some nucleic acid-based gene regulatory tools (antisense oligonucleotides and small interfering RNAs) are devised for achieving on-demand activation, the introduction of chemical modifications may cause undesired side effects, thereby impairing the gene regulatory efficacy. Herein, a methyl-engineered DNAzyme (MeDz) is developed for the visualization of endogenous alkyltransferase (AGT) and the simultaneous self-sufficiently on-demand gene regulation. The catalytic activity of DNAzyme can be efficiently blocked by O6-methylguanine (O6MeG) modification and specifically restored via the AGT-mediated DNA-repairing pathway. This simply designed MeDz is demonstrated to reveal AGT of varying expression levels in different cells, opening the possibility to explore the AGT-related biological processes. Moreover, the AGT-guided MeDz exhibits cell-selective regulation on the human early growth response-1 (EGR-1) gene, with efficient gene repression in breast cancer cells and low effectiveness in normal cells. The proposed MeDz offers an attractive strategy for on-demand gene regulation, displaying great potential in biomedical applications.

On-Site Multiply Stimulated Self-Confinement of an Integrated DNA Cascade Circuit for Highly Reliable Intracellular Imaging of miRNA and In Situ Interrogation of the Relevant Regulatory Pathway.

Artificial DNA circuits represent a versatile yet promising toolbox for in situ monitoring and concomitant regulation of diverse biological events within live cells. Nonetheless, their performance is significantly impeded by the diffusion-dominated slow reaction kinetics and the uncontrollable off-target activation. Herein, a self-localized cascade (SLC) circuit is reported for the robust and efficient microRNA (miRNA) analysis in living cells. The SLC circuit consists of the cell-specific localization module and the analyte-specific signal amplification module. By integrating the reaction probes of these two modules, the complexity of the system is reduced to realize the responsive co-localization of circuitry probes and the simultaneous cascade signal amplification. Taking advantage of the specifically activated, self-localized, and cascade design, the SLC circuit successfully achieves the robust miRNA-21 (miR-21) imaging and the accurate cells differentiation. Moreover, the reverse regulation mechanism is successfully explored between messenger RNA (mRNA) and miRNA through the engineered SLC circuit and further elucidates the underlying signaling pathways between them. Therefore, the SLC circuit provides a powerful tool for the sensitive detection of intracellular biomolecules and the study of the corresponding cell regulatory mechanisms.

Clinical and pathogen features of COVID-19-associated infections during an Omicron strain outbreak in Guangzhou, China.

Although the Omicron variant has been associated with greater transmissibility and tropism of the upper respiratory tract, the clinical and pathogenic features of patients infected with the Omicron variant during an outbreak in China have been unclear. Adults with COVID-19 were retrospectively enrolled from seven medical centers in Guangzhou, China, and clinical information and specimens ( BALF, sputum, and throat swabs) from participants were collected. Conventional detection methods, metagenomics next-generation sequencing (mNGS), and other methods were used to detect pathogens in lower respiratory tract samples. From December 2022 to January 2023, we enrolled 836 patients with COVID-19, among which 56.7% patients had severe/critical illness. About 91.4% of patients were infected with the Omicron strain (BA.5.2). The detection rate of possible co-infection pathogens was 53.4% by mNGS, including Klebsiella pneumoniae (16.3%), Aspergillus fumigatus (12.2%), and Pseudomonas aeruginosa (11.8%). The co-infection rate was 19.5%, with common pathogens being Streptococcus pneumoniae (11.5%), Haemophilus influenzae (9.2%), and Adenovirus (6.9%). The superinfection rate was 75.4%, with common pathogens such as Klebsiella pneumoniae (26.1%) and Pseudomonas aeruginosa (19.4%). Klebsiella pneumoniae (27.1%% vs 6.1%, P < 0.001), Aspergillus fumigatus (19.6% vs 5.3%, P = 0.001), Acinetobacter baumannii (18.7% vs 4.4%, P = 0.001), Pseudomonas aeruginosa (16.8% vs 7.0%, P = 0.024), Staphylococcus aureus (14.0% vs 5.3%, P = 0.027), and Streptococcus pneumoniae (0.9% vs 10.5%, P = 0.002) were more common in severe cases. Co-infection and superinfection of bacteria and fungi are common in patients with severe pneumonia associated with Omicron variant infection. Sequencing methods may aid in the diagnosis and differential diagnosis of pathogens. IMPORTANCE: Our study has analyzed the clinical characteristics and pathogen spectrum of the lower respiratory tract associated with co-infection or superinfection in Guangzhou during the outbreak of the Omicron strain, particularly after the relaxation of the epidemic prevention and control strategy in China. This study will likely prompt further research into the specific issue, which will benefit clinical practice.

Isolation and characterization of genetic variants of Orthohantavirus hantanense from clinical cases of HFRS in Jiangxi Province, China.

BACKGROUND: Hemorrhagic fever with renal syndrome (HFRS) is a severe public health problem in Jiangxi province, China. Previous studies reported genetic variants of Orthohantavirus hantanense (Hantaan virus, HTNV) in rodents in this area. However, the relationship between HTNV variants and human infection needs to be confirmed. This study aimed to identify the HTNV variants in patients and to understand the clinical characteristics of HFRS caused by these variants. METHODS: Samples were collected from hospitalized suspected cases of HFRS during the acute phase. HFRS cases were confirmed using quantitative real-time RT-PCR. Peripheral blood mononuclear cells (PBMC) from patients with HFRS were inoculated into Vero-E6 cells for viral isolation. The genomic sequences of HTNV from patients were obtained by amplicon-based next-generation sequencing. A retrospective analysis was conducted on the clinical characteristics of the patients. RESULTS: HTNV RNA was detected in 53 of 183 suspected HFRS patients. Thirteen HTNVs were isolated from 32 PBMCs of HFRS cases. Whole genome sequences of 14 HTNVs were obtained, including 13 isolates in cell culture from 13 patients, and one from plasma of the fatal case which was not isolated successfully in cell culture. Genetic analysis revealed that the HTNV sequence from the 14 patients showed significant variations in nucleotide and amino acid to the HTNV strains found in other areas. Fever (100%, 53/53), thrombocytopenia (100%, 53/53), increased serum aspartate aminotransferase (100%, 53/53), and increased lactate dehydrogenase (96.2%, 51/53) were the most common characteristics. Severe acute kidney injury was observed in 13.2% (7/53) of cases. Clinical symptoms, such as pain, petechiae, and gastrointestinal or respiratory symptoms were uncommon. CONCLUSION: The HTNV genetic variants cause human infections in Jiangxi. The clinical symptoms of HFRS caused by the HTNV genetic variant during the acute phase are atypical. In addition to renal dysfunction, attention should be paid to the common liver injuries caused by these genetic variants.