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A histidine (His)-tag is composed of six His residues and typically exerts little influence on the structure and solubility of expressed recombinant fusion proteins. Purification methods for recombinant proteins containing His-tags are relatively well-established, thus His-tags are widely used in protein recombination technology. We established a one-step enzyme-linked immunosorbent assay (ELISA) for His-tagged recombinant proteins. We analyzed variable heavy and light chains of the anti-His-tag monoclonal antibody 4C9 and used BLAST analyses to determine variable zones in light (VL) and heavy chains (VH). VH, VL, and alkaline phosphatase (ALP) regions were connected via a linker sequence and ligated into the pGEX-4T-1 expression vector. Different recombinant proteins with His tags were used to evaluate and detect ALP-scFv activity. Antigen and anti-His-scFv-ALP concentrations for direct ELISA were optimized using the checkerboard method. ZIKV-NS1, CHIKV-E2, SCRV-N, and other His-tag fusion proteins demonstrated specific reactions with anti-His-scFv-ALP, which were accurate and reproducible when the antigen concentration was 50 µg mL−1 and the antibody concentration was 6.25 µg mL−1. For competitive ELISA, we observed a good linear relationship when coating concentrations of recombinant human anti-Müllerian hormone (hAMH) were between 0.78 and 12.5 µg mL−1. Our direct ELISA method is simple, rapid, and accurate. The scFv antibody can be purified using a prokaryotic expression system, which provides uniform product quality and reduces variations between batches.

This study explores the prognostic value of immune escape-related genes in acute myeloid leukemia (AML) patients. Using TARGET_AML and GSE37642 datasets, we identified CEP55, DNAJC13, and EMC2 as significant prognostic indicators, with high transcript abundance correlating with poor outcomes. Consensus clustering divided patients into two groups, with Cluster 1 showing worse prognosis. A prognostic signature based on these genes stratified patients into high- and low-risk groups, with the high-risk group experiencing worse outcomes. The risk score was an independent prognostic factor. Functional analysis revealed that high-risk genes could promote cell cycle progression. The selected genes were strongly associated with immune cells, particularly mast cells and CD8+ T cells. This study enriches the prognostic evaluation system for AML and suggests a new therapeutic direction. [ABSTRACT FROM AUTHOR]

The purpose of this research was to examine the relationship between the hemoglobin-to-red blood cell distribution width ratio (HRR) and cardiovascular disease (CVD)-related mortality in people who have diabetes.Data derived from the National Health and Nutrition Examination Survey (NHANES), between the years 1999 to 2018, were meticulously analyzed. Mortality data, encompassing events until December 31, 2019, were systematically collected. A comprehensive group comprising of 8,732 participants were subjected to scrutiny, and subsequently, classified into four distinct groups predicated upon quartiles of baseline HRR levels: Q1 (n = 2,183), Q2 (n = 2,181), Q3 (n = 2,185), and Q4 (n = 2,183). The correlation between HRR and CVD-related mortality was examined through the use of survival curves and Cox proportional hazard regression models, the latter incorporating weights as advised by NHANES.Among the 8,732 participants in the study cohort, CVD-related mortality was identified in 710 cases. The Kaplan-Meier analysis demonstrated a significant association, indicating that a decreased HRR was correlated with a reduction in survival in cases with CVD. Both univariate and multivariable Cox proportional hazard regression analyses consistently indicated that patients exhibiting a lower HRR exhibited a markedly elevated risk of CVD-related mortality in comparison to those with higher HRR. Notably, the correlation between HRR and decreasing CVD-related mortality was discerned to be non-linear.In patients with diabetes, a decreased HRR was associated with an increased risk of CVD-related mortality. [ABSTRACT FROM AUTHOR]

Aim: To explore the trajectories of consciousness recovery and prognosis-associated predictors in children with prolonged disorder of consciousness (pDoC)., Method: This single-centre, retrospective, observational cohort involved 134 (87 males, 47 females) children diagnosed with pDoC and hospitalized at the Department of Rehabilitation at the Children's Hospital of Chongqing Medical University in China. The median onset age was 30 (interquartile range [IQR] 18-54) months, with onset ages ranging from 3 to 164 months. Least absolute shrinkage and selection operator (LASSO) regression and logistic regression analyses were performed to identify the independent predictors of consciousness recovery at 1 year after brain injury. Discrimination and calibration were assessed using 1000 bootstrap resamples. The potential predictors of resultant living independence were also explored., Results: The predictors for consciousness recovery at 1-year postinjury were: traumatic brain injury (odds ratio [OR]: 3.26, 95% confidence interval [95% CI]: 1.21-9.46), electroencephalogram (EEG) grade IV or below based on Young's classification (OR: 3.41, 95% CI: 1.38-8.70), and no bilateral impairments in the basal ganglia (OR: 3.75, 95% CI: 1.50-9.91) or posterior cingulate (OR: 5.61, 95% CI: 2.20-15.54). A nomogram was constructed with the area under the curve of 0.845 (95% CI: 0.780-0.911). Additionally, EEG grade IV or below, and the absence of bilateral impairments in the frontal lobes and occipital lobes were associated with favorable functional outcomes., Interpretation: These findings underscore the importance of comprehensive early-stage assessments in evaluating consciousness and function, assisting clinicians and families in making clinical decisions., (© 2025 Mac Keith Press.)

The growing threat of electromagnetic pollution has become a pressing safety concern. Metal-organic framework (MOF) derivatives are considered ideal candidates for mitigating electromagnetic radiation. However, due to the limitations imposed by complex post-processing and disruption of pristine crystal structures, the mechanisms of electromagnetic wave absorption remain unclear, let alone achieving atomic-level regulation in MOF derivatives. Moreover, research on MOF-based electromagnetic wave absorbers (EMWA) has predominantly focused on 2D and 3D structures, leaving 1D MOFs largely unexplored. To address these challenges, a bottom-up polarization gene weaving strategy is proposed to integrate polarizable conjugated groups, thieno(3,2-b)thiophene (TBTT), into two types of conductive MOFs by fine-tuning self-assembly conditions. As expected, both MOFs exhibited strong natural polarization effects. Among them, the 1D linear coordination mode of CuTBTT-1D demonstrated enhanced charge carrier mobility and geometric effects compared to the 2D structure, CuTBTT-2D. The synthesized 1D molecular polarization wire, with a thickness of 2.2 mm, achieved ultra-high reflection loss (-77 dB) and super-wide absorption bandwidth (6.52 GHz). Its performance surpasses that of all known MOF-based EMWAs. This study provides a valuable strategy for the rational design of next-generation 1D MOF EMWA with atomic precision., (© 2024 Wiley‐VCH GmbH.)

The XBB.1.5 subvariant has garnered significant attention due to its exceptional immune evasion and transmissibility. Significantly, the evolutionary trajectory of SARS-CoV-2 has shown continual progression, with a recent global shift observed from XBB to BA.2.86, exemplified by the emergence of the predominant JN.1 subvariant. This phenomenon highlights the need for vaccines that can provide broad-spectrum antigenic coverage. In this study, we utilized a NS1-deleted (dNS1) influenza viral vector to engineer an updated live-attenuated vectored vaccine called dNS1-XBB-RBD. This vaccine encodes the receptor-binding domain (RBD) protein of the XBB.1.5 strain. Our findings demonstrate that the dNS1-XBB-RBD vaccine elicits a similar systemic and mucosal immune response compared to its prototypic form, dNS1-RBD. In hamsters, the dNS1-XBB-RBD vaccine provided robust protection against the SARS-CoV-2 immune-evasive strains XBB.1.9.2.1 and Beta. Remarkably, nasal vaccination with dNS1-RBD, which encodes the ancestor RBD gene, also effectively protected hamsters against both the XBB.1.9.2.1 and Beta strains. These results provide valuable insights about nasal influenza-vectored vaccine and present a promising strategy for the development of a broad-spectrum vaccine against COVID-19 in the future., (© The Author(s) 2024. Published by Oxford University Press on behalf of National Academy of Sciences.)

Background: The correlation of clinical characteristics of cerebral palsy (CP) and the magnetic resonance imaging classification system (MRICS) for (CP) is inconsistent. Specifically, the variance in rehabilitation potential across MRICS remains underexplored., Aims: To investigate the clinical characteristics and potential for rehabilitation in children with CP based on MRICS., Materials and Methods: Children with CP admitted to the Department of Rehabilitation, Children's Hospital of Chongqing Medical University between 2017 and 2021 were included in the study. Qualified cases underwent a follow-up period of at least one year. The clinical characteristics of CP among different MRICS were analyzed, then the rehabilitation potential was explored by a retrospective cohort study., Results: Among the 384 initially enrolled children, the male-to-female ratio was 2.3:1, and the median age of diagnosis was 6.5 months (interquartile range: 4-12). The most prevalent MRICS categorization was predominant white matter injury (40.6%), followed by miscellaneous (29.2%) and predominant gray matter injury (15.6%). For the predominant white matter injury and miscellaneous categories, spastic diplegia emerged as the leading subtype of CP, with incidences of 59.6% and 36.6%, respectively, while mixed CP (36.7%) was the most common type in children with predominant gray matter. Notably, 76.4% of children with predominant white matter injury were classified as levels I-III on the gross motor function classification system (GMFCS), indicating significantly less severity than other groups ( χ 2   = 12.438, p  = 0.013). No significant difference across MRICS categories was observed for the manual ability classification system (MACS) ( H  = 8.176, p =  0.085). Rehabilitation potential regarding fine motor function and adaptability based on Gesell assessment was dependent on MRICS over the follow-up period. Children with normal MRI scans exhibited superior rehabilitation outcomes. Commencing rehabilitation at an earlier stage produced consistent and beneficial results in terms of fine motor function and adaptability across all MRICS categories. Moreover, participants below 2 years of age demonstrated enhanced rehabilitation potential regarding fine motor outcomes and adaptability within the MRICS framework., Conclusion: MRICS displayed a significant association with clinical characteristics and rehabilitation efficacy in children with CP., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2024 Yang, Chen, Chen, Zheng, Chen, Li, Jia and Li.)

Objective To prepare human anti-Mullerian hormone (AMH) immunomagnetic beads and HRP-labeled antibodies and establish a rapid double-antibody sandwich ELISA based on nanometer magnetic beads. Methods The expression vector of human AMH protein was constructed, and the recombinant AMH protein was expressed and purified. BALB/c mice were immunized with the recombinant protein to prepare the polyclonal antibody. Spleen cells were fused with myeloma Sp2/0 cells by PEG. Hybridoma cell lines which could stably secret monoclonal antibodies against AMH were screened out by ELISA. Monoclonal antibodies were produced from the ascites fluid of mice injected intraperitoneally with hybridoma cells and evaluated by Western blotting. Polyclonal antibodies purified from protein A were coupled to nano-magnetic' beads and used as capture antibodies, while HRP-labeled monoclonal antibody was prepared by sodium periodate method and used as probe antibody. A double antibody sandwich ELISA based on the nano-magnetic beads was established and optimized. Results A monoclonal antibody with good specificity for AMH was obtained,' and its subtype was IgG2b. The titers of purified polyclonal antibodies and monoclonal antibodies were up to 1:51 200. The capture antibody coupled with magnetic beads and the probe antibody labeled with HRP kept their good activity. The established method could detect AMH antigen within 1 hour and the detection limit was 50 ng/mL. Conclusion The prepared AMH immunomagnetic beads can be used for the fast and visualized detection of recombinant AMH.