Vertically-stacked W/W2C heterojunctions with high electrocatalytic capability for the hydrogen evolution reaction in a wide pH range.

The hydrogen evolution reaction (HER) in water splitting is among the foremost methods to produce clean and green hydrogen from renewable sources. The practical use of the HER technology is however hindered by the high price and/or the relatively low efficiency of the currently used catalysts. Herein, we report a heterostructured W/W2C electrocatalyst featuring vertically stacked interfaces and embedded in N-doped porous graphitic carbon (denoted as W/W2C@N-PGC) as a high-performance electrocatalyst for the HER in a wide pH range. The catalyst synthesis, accomplished through a straightforward one-pot method, is both facile and highly efficient, involving freeze-drying a suspension of the starting materials followed by pyrolyzing the obtained dry gel. Density functional theory calculations revealed the crucial role of the W/W2C heterojunction in promoting the two key steps of the HER, viz. HOH bond scission and H2 emission. Electrochemical data confirmed the excellent electrocatalytic capability of W/W2C@N-PGC toward the HER process in a wide pH range including alkaline, acidic, and neutral electrolytes. In 1.0 M KOH, we measured a low overpotential of 102 mV to drive a current density of 10 mA cm-2; a long-term stability (up to 24 h) was also realized. The data presented in this work highlight the importance of electrocatalysts with heterojunctions for the HER and the methodology presented in this work may be extended to other contemporary energy-related technologies such as CO2 reduction, oxygen evolution, and oxygen reduction reactions.

Methotrexate polyglutamates.

INTRODUCTION: Methotrexate polyglutamates (MTXPGs) are intracellular metabolites of methotrexate (MTX) that play a critical role in the drug's activity, influencing both its efficacy and toxicity. As the exact implications of MTXPGs in these processes remain a subject of debate, a comprehensive review of MTXPGs could provide valuable insights for clinicians and pharmacists, potentially minimizing adverse reactions and enhancing therapeutic outcomes. AREAS COVERED: A comprehensive search of relevant literature was conducted in PubMed and Web of Science databases, including studies from their inception to April 2024. Eligible studies included reviews, clinical trials, and real-world analyses. Additional manual searches and citation reviews were also performed. This review aims to explore MTXPGs with a primary focus on their pharmacokinetics, analytical methods, determinants of drug exposure, and their correlation with MTX efficacy and toxicity. EXPERT OPINION: MTXPGs have not yet garnered significant attention in clinical practice. However, multiple studies have demonstrated a relationship between MTXPGs and the efficacy and toxicity of MTX, suggesting a potential avenue for personalized treatment strategies. Future research should aim to further validate and refine this correlation. Additionally, attention should also be directed toward other metabolites of MTX, which may hold clinical significance.

Early diaphragm dysfunction assessed by ultrasonography after cardiac surgery: a retrospective cohort study.

Objective: Approximately 10%-70% of patients may develop diaphragmatic dysfunction after cardiac surgery, which may lead to delayed weaning from mechanical ventilation, increased ICU stays, postoperative hospitalization stays, and respiratory complications. However, its impact on prognosis and risk factors remain controversy. Therefore, we conducted a retrospective cohort study in which we evaluated diaphragmatic dysfunction in patients who underwent cardiac surgery via bedside diaphragm ultrasound to investigate its prognosis and possible risk factors. Methods: Data from the electronic medical records system included case records and ultrasound images of the diaphragm for 177 consecutive patients admitted to the ICU following cardiac thoracotomy surgeries performed between June and September 2020. Diaphragmatic dysfunction was defined as a diaphragmatic excursion of less than 9 mm in women and less than 10 mm in men at rest, with an average thickening fraction of less than 20%. SPSS 25.0 software was used to analyse the relationships between patients' general information, intraoperative and postoperative factors and diaphragmatic dysfunction, as well as the impact on patients' hospitalization days, mechanical ventilation time and respiratory system complications. Results: The incidence of early postoperative diaphragmatic dysfunction after cardiac surgery was 40.7%. Patients with diaphragmatic insufficiency were more likely to sequentially use noninvasive ventilation within 24 h after weaning off mechanical ventilation (3.8% vs. 12.5%, P = 0.029) and to require more oxygen support (23.8% vs. 40.3%, P = 0.019). Although there was no significant difference, the diaphragmatic dysfunction group tended to have longer ICU stays and postoperative hospital stays than did the normal diaphragmatic function group (P = 0.119, P = 0.073). Univariate and multivariate logistic regression analyses both revealed that chest tube drainage placed during surgery accompanied by bloody drainage fluid was an independent risk factor for diaphragmatic dysfunction (univariate analysis: 95% CI: 1.126-4.137, P = 0.021; multivariate analysis: 95% CI: 1.036-3.897, P = 0.039). Conclusion: Eearly diaphragmatic dysfunction after cardiac surgery increased the proportion of patients who underwent sequential noninvasive ventilation after weaning from mechanical ventilation and who required more oxygen. Chest tube drainage placed during surgery accompanied by bloody drainage fluid was an independent risk factor for diaphragmatic dysfunction, providing evidence-based guidance for respiratory rehabilitation after cardiac surgery.

Effects of water immersion on immune, intestinal flora and metabolome of Chinese mitten crab (Eriocheir sinensis) after air exposure.

Air exposure stress can induce stress response of Eriocheir sinensis and affect its normal life activities. The goal of this study was to investigate the effects of water immersion on the recovery of hepatopancreas immune-related enzyme activity, intestinal microbial diversity and metabolic level of Chinese mitten crabs after exposure to air. The results show that immersion can effectively alleviate the adverse effects of air exposure on the antioxidant capacity and immune capacity of Chinese mitten crabs, and the longer the time of immersion, the more obvious the recovery effect. Among them, the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase significantly increased after exposure to air (P < 0.05), reached a peak at 3 h, began to decline after immersion, and returned to a level close to the initial value at 24 h (P < 0.05). In addition, after exposure to air, the glucose and total cholesterol in haemolymph of Eriocheir sinensis were significantly different from the initial values (P < 0.05), gradually recovered to the initial level after re-immersion. However, changes in intestinal flora and hepatopancreas metabolism caused by air exposure did not fully recover after water exposure, and its negative effects did not completely disappear. The sequencing results showed that the species composition and diversity of intestinal microorganisms of Chinese mitten crab changed after air exposure and immersion treatment. The relative abundance of Actinomycetes increased significantly, while that of Proteobacteria and Firmicutes decreased significantly. Metabolomics analysis showed that air exposure and immersion destroyed the metabolic balance of amino acids and carnitine, reduced the level of carnitine metabolism, hindered the absorption of nutrients, and led to the accumulation of harmful substances.

Impact of Surface Enhanced Raman Spectroscopy in Catalysis.

Catalysis stands as an indispensable cornerstone of modern society, underpinning the production of over 80% of manufactured goods and driving over 90% of industrial chemical processes. As the demand for more efficient and sustainable processes grows, better catalysts are needed. Understanding the working principles of catalysts is key, and over the last 50 years, surface-enhanced Raman Spectroscopy (SERS) has become essential. Discovered in 1974, SERS has evolved into a mature and powerful analytical tool, transforming the way in which we detect molecules across disciplines. In catalysis, SERS has enabled insights into dynamic surface phenomena, facilitating the monitoring of the catalyst structure, adsorbate interactions, and reaction kinetics at very high spatial and temporal resolutions. This review explores the achievements as well as the future potential of SERS in the field of catalysis and energy conversion, thereby highlighting its role in advancing these critical areas of research.

TCCA/RSeSeR-Mediated Selenoalkoxy of Allenamides via a Radical Process: Synthesis of Selanyl-allylic N,O-Aminals.

An efficient TCCA (trichloroisocyanuric acid)/RSeSeR-mediated selenoalkoxy of allenamides for the construction of selanyl-allylic N,OA-aminal derivatives was developed. The reaction exhibits good functional group tolerance and high efficiency, affording the products in good to excellent yields. Mechanistic investigations indicated that a selanyl-allylic radical intermediate was first formed via the RSe radical added to the central carbon of allenamides, which subsequently furnished highly stable selanyl-allylic carbocation intermediate by abstraction of an electron by the chlorine radical. Moreover, this is the first report of using selenium reagent (RSeCl) to activate allenamides via a radical process.

Experimental species removal reveals species contributions to positive pollinator-mediated reproductive interactions.

Pollinator-mediated reproductive interactions among co-flowering plant species provide a canonical example of how biotic factors may contribute to species coexistence, yet we lack understanding of the exact mechanisms. Flowering-dominant and unusually attractive "magnet species" with disproportionate contributions to pollination may play key roles in such reproductive interactions, but their relative roles within the same community have rarely been assessed. We experimentally removed either a flowering-dominant or a highly attractive magnet species and compared effects on visitation frequency, pollinator richness, and seed set of co-flowering plants. Removal of either the flowering-dominant species or the magnet species reduced community-level pollinator visitation. Removal of the magnet species had the most consistent effect, including reduced pollinator visitation and richness, and reduced seed set of most co-flowering plants. These results suggest that the magnet species, which interacts with a wider range of pollinator species than does the dominant species, promotes the visitation and reproductive fitness of most other species. Removal of the flowering-dominant species affected only certain species, perhaps because these plants had floral traits similar to the dominant species. Our results highlight the role of attractive magnet species within a community in structuring reproductive interactions and identify potential mechanisms involved in coexistence facilitated by reproductive interactions.

Oilseed flax cultivation: optimizing phosphorus use for enhanced growth and soil health.

Introduction: Oilseed flax (Linum usitatissimum L.) yields are phosphate (P) fertilizer-limited, especially in the temperate semiarid dryland regions of North China. However, there are limited studies on the effects of P-fertilizer inputs on plant growth and soil microorganisms in flax planting systems. Methods: To address this gap, a field experiment was conducted with four treatments: no P addition and application of 40, 80, and 120 kg P ha-¹, respectively. The aim was to investigate the influence of various P fertilizer inputs on yield, plant dry matter, P use efficiency, as well as the population of soil arbuscular mycorrhizal fungi (AMF) and bacteria in dryland oilseed flax. Results: Our results show that the P addition increased the dry matter, and the yield of oilseed increased by ~200% at 120 kg P ha-1 addition with inhibition on the growth of AMF hyphae. The moderate P supply (80 kg ha-1) was adequate for promoting P translocation, P use efficiency, and P recovery efficiency. Soil pH, available P, and available K significantly (p< 0.05) promoted the abundance of the dominant taxa (Acidobacteria_GP6, Sphingobacteria and Bacteroidetes). In addition, it is imperative to comprehend the mechanism of interaction between phosphorus-fertilizer inputs and microbiota in oilseed flax soil. Discussion: This necessitates further research to quantify and optimize the moderate phosphorus supply, regulate soil microbes to ensure high phosphorus utilization, and ultimately establish a sustainable system for oilseed flax cultivation in the local area.

Effects of low temperature air exposure and immersion on antioxidant, immune, intestinal flora and metabolome of Chinese mitten crab (Eriocheir sinensis).

The aim of this study was to investigate the effects of immersion on immune enzyme activity, haemolymph index, intestinal microbiome and metabolome of E. sinensis after low temperature air exposure. The results showed that low temperature air exposure induced stress response, which led to hepatopancreas injury and increased membrane permeability, but this situation was reversible and alleviated after immersion. In addition, after exposure to low temperature air, haemolymph metabolism-related substances such as glucose and total cholesterol were significantly different from the initial value (P < 0.05), and gradually returned to the initial level after immersion. The changes of intestinal flora and hepatopancreas metabolism caused by low temperature air exposure did not fully recover after immersion, and its negative effects did not completely disappear. The sequencing results showed that the species composition and diversity of intestinal microorganisms of Chinese mitten crabs were changed after low temperature air exposure and immersion treatment. The relative abundance of Bacteroidetes and Proteobacteria were increased, while the relative abundance of Firmicutes was decreased (P < 0.05). Metabolomics analysis showed that lysine levels increased significantly, taurocholic acid levels decreased significantly, and amino acid metabolism and lipid metabolism balance were disturbed in hepatopancreas of E. sinensis after exposure to low temperature air and immersion (P < 0.05). This study will provide new insights into the recovery mechanism of water immersion on Chinese mitten crabs after exposure to air.

Nursing of forearm hematoma after transradial coronary intervention.

Background: Complications such as forearm hematoma after coronary intervention through the radial artery are a common complication. Material and methods: By observing, describing, and analyzing the pictures taken during clinical diagnosis and consultation, we summarize the prevention, treatment, and nursing of forearm hematoma after percutaneous coronary intervention, to provide reference for the nursing of patients with forearm hematoma. Results: We have innovatively summarized the risk classification of forearm hematoma and the three key time points for preventing hematoma. Conclusion: Complications such as forearm hematoma after coronary intervention through the radial artery are a common complication. We have innovatively summarized the risk classification of forearm hematoma and the three key time points for preventing hematoma, providing reference for the prevention and management of forearm hematoma in clinical practice. For patients undergoing transradial coronary intervention, the three key time points for preventing hematoma and symptomatic management based the risk classification of forearm hematoma are crucial.

Impact of competition on reimbursement decisions for cancer drugs in China: an observational study.

Background: Annual Chinese National negotiations for including innovative drugs in the National Reimbursement Drug List (NRDL) reveal an increasing number of new drugs with overlapping action mechanisms of action and similar indications. Yet, it is unclear if competition affects reimbursement decisions. Thus, we explored the impact of competition on reimbursement decisions for cancer drugs in China. Methods: We identified the cancer drugs involved in NRDL negotiations from 2017 to 2022 and focused on the initial reimbursement decision for eligible newly negotiated drugs. Drugs were classified as within-class competitors based on their equivalent biological mechanisms of action and approved indications, including identified and potential competitors. Other variables included drug type, clinical benefit and safety, monthly drug cost, and disease incidence rate. We employed traditional univariate and multivariate Firth's penalized logistic regression to assess the association between reimbursement decisions and variables at the indication and drug levels. Findings: Between 2017 and 2022, 102 cancer drugs corresponding to 141 indications were studied, and 66 drugs (64.7%) covering 95 indications (67.4%) were added to the NRDL. The proportion of reimbursements for indications with identified competition was significantly higher than that for indications without identified competition (84.6% vs 52.6%, p < 0.0001). However, the difference in reimbursement proportions between groups with and without potential competition was not statistically significant (66.7% vs 68.3%, p = 0.84). Firth's penalized logistic regression showed that identified competition was positively correlated with successful NRDL inclusion, whereas potential competition had no significant effect on negotiation outcomes. Improved overall survival or progression-free survival were positively associated with NRDL inclusion, whereas disease incidence negatively impacted reimbursement decisions. Interpretation: Improved clinical benefit and identified competition were positively correlated with NRDL inclusion. In China's value-based negotiation model, clinical benefits served as a crucial foundation of price negotiation for cancer drugs, and market competition helped these drugs enter the NRDL at more reasonable prices. This has important implications for reimbursement decisions and accessibility and affordability improvement for innovative drugs worldwide. Funding: National Natural Science Foundation of China (No. 72104151).

Cost-effectiveness analysis of 11 pharmacotherapies for recurrent glioblastoma in the USA and China.

Background: Several studies have systematically assessed the efficacy and safety of progressive or recurrent glioblastoma multiforme (GBM). However, the discernible limitations of efficacy and the elevated costs of interventions instigate an investigation into the cost-effectiveness of these treatments. Objectives: This study aimed to evaluate cost-effectivenesses of 11 pharmacotherapeutic interventions for recurrent GBM from the perspective of healthcare payers in the United States (US) and China. Design: A model-based pharmacoeconomic evaluation. Methods: A partitioned survival model was employed to evaluate the cost-effectiveness of 11 distinct drug-based treatments. The clinical efficacy and safety data were obtained from a network meta-analysis, while the medical expenditure and health utility were primarily derived from published literature. One-way sensitivity analyses, scenario analyses, and probabilistic sensitivity analyses (PSA) were performed to scrutinize the impact of potential uncertainties to ensure the robustness of the model. The primary endpoint was the incremental cost-effectiveness ratio. Results: Among the therapeutic interventions evaluated, lomustine emerged as the cheapest option, with costs amounting to $78,998 in the United States and $30,231 in China, respectively. Regorafenib displayed the highest quality-adjusted life years at 0.475 in the United States and 0.465 in China. The one-way sensitivity analyses underscored that drug price was a key factor influencing cost-effectiveness. Both scenario and PSA consistently demonstrated that, considering the willingness-to-pay thresholds, lomustine was a cost-effective treatment with probability of more than 94%. Conclusion: In comparison to the alternative antitumor agents, lomustine was likely to be a cost-effective option for relapsed GBM patients from the perspective of healthcare payers in both the United States and China.

Rationality and implication of catheter-based adrenal ablation for bilateral primary aldosteronism.

Mineralocorticoid receptor antagonists (MRAs) for bilateral primary aldosteronism (PA) are the mainstay option recommended by guidelines, but poor compliance occurs due to numerous side effects. We aimed to examine whether catheter-based adrenal ablation could be an alternative treatment for bilateral PA.644 PA patients were included from a total of 6054 hypertensive patients. Adrenal CT scan and adrenal venous sampling (AVS) were both performed for PA subtype classification. Clinical and biochemical outcomes were assessed at 6 months after treatment according to the Primary Aldosteronism Surgical Outcome (PASO) criteria.93 patients with PA were recruited to be treated by adrenal ablation, including 25 bilateral PA and 68 unilateral PA according to AVS results. Office SBP and DBP significantly decreased from baseline levels, serum potassium levels increased and ARR significantly decreased (p < 0.01) in both the bilateral and unilateral groups. In the bilateral group, complete, partial and absent clinical success was achieved in 6 (24.0%), 11 (44.0%) and 8(32.0%) patients, respectively. In the unilateral group, complete, partial and absent clinical success was achieved in 12 (17.6%), 37 (54.4%), and 19 (27.9%) patients, respectively. The numbers of patients achieving complete, partial, and absent biochemical success were 15 (60.0%), 6 (24.0%), and 4 (16.0%), respectively, in the bilateral group versus 37 (54.4%), 9 (13.2%), and 22 (32.3%), respectively, in the unilateral group. In conclusion, we provide evidence for the beneficial outcomes of unilateral adrenal ablation for patients with bilateral PA. Our findings provide insight into an alternative option for patients with bilateral excess aldosterone.

High-precision high-stability acousto-optic pulse picking driver with a direct digital synthesis technique.

A method for maintaining a fixed phase relationship between the driving signal of acousto-optic modulator (AOM) and the original mode-locked seed laser is proposed and realized, which stabilizes the amplitude of diffracted signal output from the AOM for subsequent amplification. A field-programmable gate array (FPGA), combined with external summing amplifiers, is used to directly synthesize high-timing-precision driving signals that are synchronized with the seed laser pulses, and the accuracy of signal timing control reaches 160 ps. Using this driver, the standard deviation of the diffracted signal output from the AOM is significantly decreased from 0.52% to 0.18%. This pulse-picking solution also includes a control system that can flexibly control the frequency, gating width, etc., of the driving signal, which makes it more convenient for subsequent laser amplification and makes it suitable for a variety of mode-locked lasers.

Fixed parameters in the population pharmacokinetic modeling of valproic acid might not be suitable: external validation in Chinese adults with epilepsy or after neurosurgery.

PURPOSE: This study aims to assess the predictive performance of published valproic acid (VPA) population pharmacokinetic (PPK) models using an external data set in Chinese adults with epilepsy or after neurosurgery. METHODS: A total of 384 concentrations from 290 Chinese adults with epilepsy or after neurosurgery were used for external validation. Data on published VPA PPK models were extracted from the literature. Prediction-based diagnostics (such as F20 and F30), simulation-based diagnostics, and Bayesian forecasting were used to evaluate the predictability of models. RESULTS: The results of prediction-based diagnostics of all models were unsatisfactory. Models B, F, and H showed the best prediction performance in simulation-based diagnostics and Bayesian forecasting, demonstrating superior precision and accuracy. Bayesian forecasting demonstrated significant improvements in the model predictability. CONCLUSION: The published PPK models showed extensive variation in predictive performance for extrapolation among Chinese adults with epilepsy or after neurosurgery patients. Fixed parameters of Vd and Ka in the PPK modeling of VPA might be the reason for the unsatisfied predictive performance. Bayesian forecasting significantly improved model predictability and may help to individualize VPA dosing.

Causal impact of statins on susceptibility to osteoarthritis: insights from a two-sample Mendelian randomization analysis.

BACKGROUND: Osteoarthritis is a widely prevalent cause of pain and disability among older adults. It is an incurable condition, and most treatments are aimed at alleviating symptoms. AIM: This study aimed to investigate the impact of statins on osteoarthritis by using a two-sample Mendelian randomization approach, using genetic variants associated with statin use as instrumental variables. METHOD: Information on single nucleotide polymorphisms associated with statin medication was obtained from the FinnGen study, and data on osteoarthritis were sourced from the UK Biobank. The inverse variance weighted method was used as the primary analytical approach for the Mendelian randomization analysis. Sensitivity analyses were conducted to evaluate horizontal pleiotropy and heterogeneity. To examine the genetic relationship between statins and osteoarthritis, linkage disequilibrium score regression-based estimates were used. RESULTS: Mendelian randomization analysis indicated a positive effect of statin use on the treatment of osteoarthritis (odds ratio 0.951, 95% confidence interval 0.914-0.99, p < 0.05). This conclusion was supported by various Mendelian randomization methods. Sensitivity analyses revealed no significant directional pleiotropy or influential single nucleotide polymorphisms that could compromise the overall causal inference. Linkage disequilibrium score regression-based estimates suggested a modest genetic correlation between statin use and osteoarthritis (Rg = 0.098, Se = 0.034, p < 0.05), thus reinforcing the robustness of the Mendelian randomization analysis. CONCLUSION: Statins reduce the risk of osteoarthritis, aligning with the results of observational studies. Further research is essential to validate these results and explore the underlying mechanisms in detail.

Reappraisal and refined diagnosis of ultrasonography and histological findings for hydatidiform moles: a multicentre retrospective study of 821 patients.

AIMS: Specific identification of a hydatidiform mole (HM) and subclassification of a complete hydatidiform mole (CHM) or partial hydatidiform mole (PHM) are critical. This study aimed to reappraise the diagnostic performance of ultrasonography and histology with a refined diagnosis. METHODS: This was a retrospective, multicentre cohort study of 821 patients with histologically suspected HM specimens. Refined diagnostic algorithms with p57 immunohistochemistry and short tandem repeat (STR) genotyping were performed and used as the true standard for assessing the diagnostic performance of the original ultrasonography and morphology methods. The diagnostic performance was calculated using accuracy, agreement rate, sensitivity and the positive predictive value (PPV) compared with refined diagnostic results. RESULTS: Of the 821 histologically suspected HM cases included, 788 (95.98%) were successfully reclassified into 448 CHMs, 213 PHMs and 127 non-molar (NM) abortuses. Ultrasonography showed an overall accuracy of 44.38%, with a sensitivity of 44.33% for CHM and 37.5% for PHM. The overall classification accuracy of the original morphological diagnosis was 65.97%. After exclusion of the initially untyped HMs, the overall agreement rate was 59.11% (κ=0.364, p<0.0001) between the original and refined diagnoses, with a sensitivity of 40.09% and PPV of 96.05% for diagnosing CHMs and a sensitivity of 84.98% and a PPV of 45.59% for diagnosing PHMs. The interinstitutional variability of morphology in diagnosing HMs was significant among the 15 centres (range, 8.33%-100.00%, p<0.0001). CONCLUSION: The current diagnosis of HM based solely on ultrasound or morphology remains problematic, and ancillary techniques, particularly p57 immunohistochemistry and DNA genotyping, should be integrated into routine practice as much as possible.

Effect of Filling Material Properties on 1-3 Piezoelectric Composite Performance.

The 1-3 piezoelectric composite is the key component of the acoustic transducer, which is widely used in detection, due to the high energy conversion efficiency, cheap raw material, and low aging. To reveal the influence of epoxy mixture, used to connect the piezoelectric column, on the composite performance, a 1-3 piezoelectric composite model was built. The effects of mixture properties on the impedance curves, vibration mode, and deformation displacement of the composite were determined. Six 1-3 piezoelectric composites with different filling mixture properties, by changing the glass microspheres' mass ratio in the epoxy, were prepared and measured to validate the model. The results showed that with the increase in the proportion of the glass microsphere in the epoxy mixture, the vibration coupling of the piezoelectric composites was gradually eliminated. The acoustic impedance was reduced by 12%. The electromechanical coupling coefficient and effective electromechanical coupling coefficient were increased by 5.4% and 8.3%, respectively. The density and Young's modulus decrease in filling mixture can significantly improve piezoelectric composite performance.

Development of two novel on-site detection visualization methods for murine hepatitis virus based on the multienzyme isothermal rapid amplification.

Murine hepatitis virus (MHV) infection is one of the most prevalent types of mice infection in laboratory. MHV could cause death in mice and even interfere with the results in animal experiments. Herein, we developed two isothermal approaches based on the Multienzyme Isothermal Rapid Amplification (MIRA), for rapid detection of MHV in conserved M gene. We designed and screened several pairs of primers and probes and the isothermal fluorescence detector was applied for the exonuclease Ⅲ reverse transcription MIRA (exo-RT-MIRA) assay. To further simplify the workflow, the portable fluorescence visualization instrument, also as a palm-sized handheld system, was used for the naked-eye exo-RT-MIRA assay. The amplification temperature and time were optimized. The assay could be processed well at 42 °C 20 min for the exo-RT-MIRA and the naked-eye exo-RT-MIRA assay. The limit of detection (LoD) of the exo-RT-MIRA assay was 43.4 copies/µL. The LoD of the naked-eye exo-RT-MIRA assay was 68.2 copies/µL. No nonspecific amplifications were observed in the two assays. A total of 107 specimens were examined by qPCR and two assays developed. The experimental results statistical analysis demonstrated that the exo-RT-MIRA assay with the qPCR yielded sufficient agreement with a kappa value of 1.000 (p < 0.0001). The results also exhibited a good agreement (kappa value, 0.961) (p < 0.0001) between the naked-eye exo-RT-MIRA assay and the qPCR assay. In our study, the exo-RT-MIRA assay and the naked-eye exo-RT-MIRA assay presented the possibility of new methods in MHV point-of-testing diagnosis.

Integrated analyses of single-cell transcriptome and Mendelian randomization reveal the protective role of FCRL3 in multiple sclerosis.

Background: Multiple sclerosis (MS) represents a multifaceted autoimmune ailment, prompting the development and widespread utilization of numerous therapeutic interventions. However, extant medications for MS have proven inadequate in mitigating relapses and halting disease progression. Innovative drug targets for preventing multiple sclerosis are still required. The objective of this study is to discover novel therapeutic targets for MS by integrating single-cell transcriptomics and Mendelian randomization analysis. Methods: The study integrated MS genome-wide association study (GWAS) data, single-cell transcriptomics (scRNA-seq), expression quantitative trait loci (eQTL), and protein quantitative trait loci (pQTL) data for analysis and utilized two-sample Mendelian randomization study to comprehend the causal relationship between proteins and MS. Sequential analyses involving colocalization and Phenome-wide association studies (PheWAS) were conducted to validate the causal role of candidate genes. Results: Following stringent quality control preprocessing of scRNA-seq data, 1,123 expression changes across seven peripheral cell types were identified. Among the seven most prevalent cell types, 97 genes exhibiting at least one eQTL were discerned. Examination of MR associations between 28 proteins with available index pQTL signals and the risk of MS outcomes was conducted. Co-localization analyses and PheWAS indicated that FCRL3 may exert influence on MS. Conclusion: The integration of scRNA-seq and MR analysis facilitated the identification of potential therapeutic targets for MS. Notably, FCRL3, implicated in immune function, emerged as a significant drug target in the deCODE databases. This research underscores the importance of FCRL3 in MS therapy and advocates for further investigation and clinical trials targeting FCRL3.