Affiliate stigma and caregiver burden in parents of children with epilepsy.

Objective: This study aimed to investigate the current status of affiliated stigma and caregiver burden among parents of children with epilepsy, analyze their correlation, and identify factors influencing affiliated stigma. Methods: A cross-sectional survey was conducted among 194 parents of children with epilepsy who met the inclusion and exclusion criteria in Shenzhen City, Guangdong Province, China. Data were collected through questionnaires, including a demographic information sheet, an affiliated stigma scale, and a caregiver burden scale. Results: The results revealed that parents of children with epilepsy experienced a moderate level of affiliated stigma, with an average score of 54.92 ± 10.44. Similarly, caregiver burden scores fell within the moderate range, with an average score of 44.14 ± 16.02. Factors influencing affiliated stigma scores included the frequency of epileptic seizures in children, the types of anti-epileptic medications taken by children, and the place of residence. The total caregiver burden score and scores in various dimensions (emotional, cognitive, and behavioral) of caregivers for epilepsy patients were positively correlated with the affiliated stigma score. Affiliated stigma was found to independently explain 21.3 % of the variation in caregiver burden. Conclusion: In the future, healthcare professionals should develop targeted interventions for children with epilepsy and their parents to reduce affiliated stigma, decrease caregiver burden, and enhance the caregiving capabilities of parents of children with epilepsy. These measures are essential to improve the overall well-being of both parents and children affected by epilepsy.

A DNA/Upconversion Nanoparticle Complex Enables Controlled Co-Delivery of CRISPR-Cas9 and Photodynamic Agents for Synergistic Cancer Therapy.

Photodynamic therapy (PDT) depends on the light-irradiated exciting of photosensitizer (PS) to generate reactive oxygen species (ROS), which faces challenges and limitations in hypoxia and antioxidant response of cancer cells, and limited tissue-penetration of light. Herein, a multifunctional DNA/upconversion nanoparticles (UCNPs) complex is developed which enables controlled co-delivery of CRISPR-Cas9, hemin, and protoporphyrin (PP) for synergistic PDT. An ultralong single-stranded DNA (ssDNA) is prepared via rolling circle amplification (RCA), which contains recognition sequences of single guide RNA (sgRNA) for loading Cas9 ribonucleoprotein (RNP), G-quadruplex sequences for loading hemin and PP, and linker sequences for combining UCNP. Cas9 RNP cleaves the antioxidant regulator nuclear factor E2-related factor 2 (Nrf2), improving the sensitivity of cancer cells to ROS, and enhancing the synergistic PDT effect. The G-quadruplex/hemin DNAzyme mimicks horseradish peroxidase (HRP) to catalyze the endogenous H2O2 to O2, overcoming hypoxia condition in tumors. The introduced UCNP converts NIR irradiation with deep tissue penetration to light with shorter wavelength, exciting PP to transform the abundant O2 to 1O2. The integration of gene editing and PDT allows substantial accumulation of 1O2 in cancer cells for enhanced cell apoptosis, and this synergistic PDT has shown remarkable therapeutic efficacy in a breast cancer mouse model.

Optimal delivery strategies for nanoparticle-mediated mRNA delivery.

Messenger RNA (mRNA) has emerged as a new and efficient agent for the treatment of various diseases. The success of lipid nanoparticle-mRNA against the novel coronavirus (SARS-CoV-2) pneumonia epidemic has proved the clinical potential of nanoparticle-mRNA formulations. However, the deficiency in the effective biological distribution, high transfection efficiency and good biosafety are still the major challenges in clinical translation of nanomedicine for mRNA delivery. To date, a variety of promising nanoparticles have been constructed and then gradually optimized to facilitate the effective biodistribution of carriers and efficient mRNA delivery. In this review, we describe the design of nanoparticles with an emphasis on lipid nanoparticles, and discuss the manipulation strategies for nanoparticle-biology (nano-bio) interactions for mRNA delivery to overcome the biological barriers and improve the delivery efficiency, because the specific nano-bio interaction of nanoparticles usually remoulds the biomedical and physiological properties of the nanoparticles especially the biodistribution, mechanism of cellular internalization and immune response. Finally, we give a perspective for the future applications of this promising technology. We believe that the regulation of nano-bio interactions would be a significant breakthrough to improve the mRNA delivery efficiency and cross biological barriers. This review may provide a new direction for the design of nanoparticle-mediated mRNA delivery systems.

Influence of continuity of care on self-management ability and quality of life in outpatient maintenance hemodialysis patients.

INTRODUCTION: The objective of this study was to evaluate the effect of continuity of care on self-management ability and quality of life (QOL) in patients undergoing maintenance hemodialysis (MHD). METHODS: One hundred patients were randomly assigned to the observation group and the control group. In the observation group, patients received a 12-month continuity of care. In the control group, patients were given with routine nursing. Evaluate the patients' self-management ability and QOL between two groups 1 week before discharge and 6 and 12 months outpatient MHD. RESULTS: Observation group had higher Hemodialysis Self-Management Instrument (HD-SMI) scores and Kidney Disease Quality of Life-Short Form (KDQOL-SF™) scores than control group at 6 and 12 months outpatient MHD. But patients in observation group had a much lower systolic blood pressure than those in control group at 12 months outpatient MHD. CONCLUSIONS: Our study suggested that continuity of care in the form of online education, telephone visit, and outpatient visit could improve self-management ability and QOL of patients undergoing MHD.

Construction of Organelle-Like Architecture by Dynamic DNA Assembly in Living Cells.

The design of controllable dynamic systems is vital for the construction of organelle-like architectures in living cells, but has proven difficult due to the lack of control over defined topological transformation of self-assembled structures. Herein, we report a DNA based dynamic assembly system that achieves lysosomal acidic microenvironment specifically inducing topological transformation from nanoparticles to organelle-like hydrogel architecture in living cells. Designer DNA nanoparticles are constructed from double-stranded DNA with cytosine-rich stick ends (C-monomer) and are internalized into cells through lysosomal pathway. The lysosomal acidic microenvironment can activate the assembly of DNA monomers, inducing transformation from nanoparticles to micro-sized organelle-like hydrogel which could further escape into cytoplasm. We show how the hydrogel regulates cellular behaviors: cytoskeleton is deformed, cell tentacles are significantly shortened, and cell migration is promoted.

The Protection Role of Magnesium Ions on Coupled Transcription and Translation in Lyophilized Cell-Free System.

Cell-free protein synthesis (CFPS) is a promising platform for protein engineering and synthetic biology. The storage of a CFPS system usually involves lyophilization, during which preventing the conformational damage of involved enzymes is critical to the activity. Herein, we report the protection role of magnesium ions on coupled transcription and translation in a lyophilized cell-free system. Mg2+ prevents the inactivation of the CFPS system from direct colyophilization of enzymes and substrates (nucleotides, and amino acids), and furthermore activates the CFPS system. We propose two-metal-ion regulation of Mg2+: Mg2+ (I) acts as an allosteric role for enzymes to prevent the conformational damage of enzymes from direct binding with substrates during lyophilization which locks up inactive enzyme-substrate complex; Mg2+ (II) consequently binds to enzymes to activate the CFPS system. Our work provides important implications for maximizing protein yields by using a cell-free system in protein engineering and understanding the functions of Mg2+ in biological systems.

Chiral Carbon Dots Mimicking Topoisomerase†I To Mediate the Topological Rearrangement of Supercoiled DNA Enantioselectively.

Nanomaterials with enzyme-mimetic activities are possible alternatives to natural enzymes. Mimicking enzymatic enantioselectivity remains a great challenge. Herein, we report that cysteine-derived chiral carbon dots (CDs) can mimic topoisomerase I to mediate topological rearrangement of supercoiled DNA enantioselectively. d-CDs can more effectively catalyze the topological transition of plasmid DNA from supercoiled to nicked open-circular configuration than l-CDs. Experiments suggest the underlying mechanism: d-CDs intercalatively bind with DNA double helix more strongly than l-CDs; the intercalative CDs can catalyze the production of hydroxyl radicals to cleave phosphate backbone in one strand of the double helix, leading to topological rearrangement of supercoiled DNA. Molecular dynamics (MD) simulation show that the stronger affinity for hydrogen-bond formation and hydrophobic interaction between d-cysteine and DNA than that of l-cysteine is the origin of enantioselectivity.

Gene Circuit Compartment on Nanointerface Facilitatating Cascade Gene Expression.

Cellular genes that are functionally related to each other are usually confined in specialized subcellular compartments for efficient biochemical reactions. Construction of spatially controlled biosynthetic systems will facilitate the study of biological design principles. Herein, we fabricated a gene circuit compartment by coanchoring two function-related genes on surface of gold nanoparticles and investigated the compartment effect on cascade gene expression in a cell-free system. The gene circuit consisted of a T7 RNA polymerase (T7 RNAP) expression cassette as regulatory gene and a fluorescent protein expression cassette as regulated reporter gene. Both the expression cassettes were attached on a Y-shaped DNA nanostructure whose other two branches were mercapto-modified in order to steadily anchor the gene expression cassettes on the surface of gold nanoparticles. Experimental results demonstrated that both the yield and initial expression rate of the fluorescent reporter protein in the gene circuit compartment system were enhanced compared with those in free gene circuit system. Mechanism investigation revealed that the gene circuit compartment on nanoparticle made the regulatory gene and regulated reporter gene spatially proximal at nanoscale, thus effectively improving the transfer efficiency of the regulatory proteins (T7 RNAP) from regulatory genes to the regulated reporter genes in the compartments, and consequently, the biochemical reaction efficiency was significantly increased. This work not only provided a simplified model for rational molecular programming of genes circuit compartments on nanointerface but also presented implications for the cellular structure-function relationship.

Saccharides Create a Crowding Environment for Gene Expression in Cell-Free Systems.

Cellular physical microenvironment such as crowding shows great influence on enzymatic reactions. Herein, we report a new finding that saccharides with low molecular weight create an effective crowding microenvironment for gene expression in cell-free protein synthesis, which provides valuable implications for living systems. Four saccharides including sorbose, galactose, sucrose, and cellobiose are screened out as effective crowders. At a low concentration range of saccharides, both the mRNA and protein amounts present an upward trend with the concentration increment of saccharides; when the concentrations exceed a critical value, the mRNA and protein amounts decrease. A mechanism is proposed that at low concentrations of saccharides, the effective concentrations of reactants increase due to the coexistence of crowders and reactants in a finite volume; when the concentrations exceed a critical value, the molecular diffusion of reactants is dominantly restricted due to the increased viscosity. Our finding opens a new view that saccharides with low molecular weight could be crowders and provides a new insight that substances with low molecular weight in cells would produce a crowding effect on biochemical reactions in living systems.

Branched DNA Architectures Produced by PCR-Based Assembly as Gene Compartments for Cell-Free Gene-Expression Reactions.

The physical distance between genes plays important roles in controlling gene expression reactions in vivo. Herein, we report the design and synthesis of a branched gene architecture in which three transcription units are integrated into one framework through assembly based on the polymerase chain reaction (PCR), together with the exploitation of these constructs as "gene compartments" for cell-free gene expression reactions, probing the impact of this physical environment on gene transcription and translation. We find that the branched gene system enhances gene expression yields, in particular at low concentrations of DNA and RNA polymerase (RNAP); furthermore, in a crowded microenvironment that mimics the intracellular microenvironment, gene expression from branched genes maintains a relatively high level. We propose that the branched gene assembly forms a membrane-free gene compartment that resembles the nucleoid of prokaryotes and enables RNAP to shuttle more efficiently between neighboring transcription units, thus enhancing gene expression efficiency. Our branched DNA architecture provides a valuable platform for studying the influence of "cellular" physical environments on biochemical reactions in simplified cell-free systems.

Encapsulating Microorganisms inside Electrospun Microfibers as a Living Material Enables Room-Temperature Storage of Microorganisms.

Room-temperature storage and transportation of microorganisms maximize the power of microorganisms in healthcare, energy, and environment. Recently, paper-based biotechnologies have been developed to enable room-temperature storage of a variety of nonliving biosystems such as diagnostic devices and cell-free systems. Herein, room-temperature storage of living microorganisms is realized by an electrospun nonwoven paper containing convex region, which is composed of coiled microfibers with dense distribution of microorganisms. Microorganisms are encapsulated into the microfibers and remain intact after electrospinning. Poly(ethylene oxide) is used as polymer matrix, and glycerol and dextran are used as additives. When the contents of glycerol and dextran are optimized as 5 and 0.4%, the room-temperature time is prolonged to 2 days, more than 8 folds as compared with the control group. Upon demand, the microorganisms can be activated by adding water and used for culturing microorganisms directly. Furthermore, mechanisms which account for microbial activity and storage are studied. Our microfiber-based strategy is universal for the room-temperature storage of prokaryotic and eukaryotic microorganisms in the solid formulation. Besides, our microorganism/polymer complex structures represent novel living materials via a bottom-up strategy, which are of great potential for new biomedical applications.

Discovery of Indolinone-Based Multikinase Inhibitors as Potential Therapeutics for Idiopathic Pulmonary Fibrosis.

Idiopathic pulmonary fibrosis (IPF) is a serious and deadly disease for which treatment options are limited. The recent approval of antifibrosis agent nintedanib represents one of the first therapeutic approaches for the treatment of IPF. Here, we report novel indolinone-based multikinase inhibitors that target angiogenesis and fibrosis pathways and may serve as potential therapeutics for IPF. KBP-7018 is a novel, tyrosine kinase-selective inhibitor with potent effects on three fibrotic kinases (c-KIT, PDGFR, and RET). The pharmacokinetics (PK) properties of KBP-7018 were favorable in mice, rats, and dogs. In a bleomycin (BLM)-induced mouse pulmonary fibrosis model, 10, 30, and 100 mg/kg daily doses (q.d.) of KBP-7018 improved the 28-day survival rate in a dose-dependent manner. The improved efficacy of KBP-7018 compared to nintedanib provided a certain level of chemical validation for the involvement of PDGFR, c-KIT, and RET in IPF. Thus, KBP-7018 represents a novel multikinase inhibitor with differentiated activity, highly enhanced selectivity, and acceptable PK profiles that will enter phase I clinical trials.