Supramolecular assembly of polycation/mRNA nanoparticles and in vivo monocyte programming.

Size-dependent phagocytosis is a well-characterized phenomenon in monocytes and macrophages. However, this size effect for preferential gene delivery to these important cell targets has not been fully exploited because commonly adopted stabilization methods for electrostatically complexed nucleic acid nanoparticles, such as PEGylation and charge repulsion, typically arrest the vehicle size below 200 nm. Here, we bridge the technical gap in scalable synthesis of larger submicron gene delivery vehicles by electrostatic self-assembly of charged nanoparticles, facilitated by a polymer structurally designed to modulate internanoparticle Coulombic and van der Waals forces. Specifically, our strategy permits controlled assembly of small poly(ß-amino ester)/messenger ribonucleic acid (mRNA) nanoparticles into particles with a size that is kinetically tunable between 200 and 1,000 nm with high colloidal stability in physiological media. We found that assembled particles with an average size of 400 nm safely and most efficiently transfect monocytes following intravenous administration and mediate their differentiation into macrophages in the periphery. When a CpG adjuvant is co-loaded into the particles with an antigen mRNA, the monocytes differentiate into inflammatory dendritic cells and prime adaptive anticancer immunity in the tumor-draining lymph node. This platform technology offers a unique ligand-independent, particle-size-mediated strategy for preferential mRNA delivery and enables therapeutic paradigms via monocyte programming.

Associations between insufficient sleep, skipping breakfast and depressive symptoms in children and adolescents: A school-based cross-sectional study in China.

OBJECTIVE: Insufficient sleep and skipping breakfast are increasingly prevalent among children and adolescents. Both behaviors are associated with the onset of depression. This study aims to examine the independent and joint associations of these two behaviors with depressive symptoms, and investigate whether these associations varied by age or sex. METHODS: The Center for Epidemiological Studies Depression Scale (CES-D) was used to evaluate the depressive symptoms. This cross-sectional study including 11,887 students aged 11-19 years using a stratified cluster, multistage sampling method in Ningbo, China. Multiple logistic regressions were conducted to evaluate the independent and joint association between insufficient sleep, skipping breakfast and depressive symptoms. Sensitivity analyses and stratified analyses by age and sex were performed using the same modelling strategies. RESULTS: The overall prevalence of depressive symptoms was 15.27%. Skipping breakfast (Odds Ratio (OR) = 2.557, 95% Confidence Interval (CI) = 2.236-2.925) and insufficient sleep (OR = 1.547, 95%CI = 1.390-1.723) was independently associated with depressive symptoms. Compared to students with "sufficient sleep and breakfast every day", the OR was 4.385 (95%CI = 3.649-5.271) for those with "insufficient sleep and skipping breakfast". Meanwhile, the joint association was moderated by age group, with a more apparent association observed in the 11-15-year-old group compared to the 16-19-year-old group. CONCLUSIONS: These findings indicated that insufficient sleep and skipping breakfast were independently and jointly associated with depressive symptoms. Insufficient sleep and skipping breakfast could be considered as two of the predictors of depression.

Ring-Opening Polymerization of Cyclohexene Oxide and Cycloaddition with CO2 Catalyzed by Amine Triphenolate Iron(III) Complexes.

A series of novel amine triphenolate iron complexes were synthesized and characterized using UV, IR, elemental analysis, and high-resolution mass spectrometry. These complexes were applied to the ring-opening polymerization (ROP) of cyclohexene oxide (CHO), demonstrating excellent activity (TOF > 11050 h-1) in the absence of a co-catalyst. In addition, complex C1 maintained the dimer in the presence of the reaction substrate CHO, catalyzing the ring-opening polymerization of CHO to PCHO through bimetallic synergy. Furthermore, a two-component system consisting of iron complexes and TBAB displayed the ability to catalyze the reaction of CHO with CO2, resulting in the formation of cis-cyclic carbonate with high selectivity. Complex C4 exhibited the highest catalytic activity, achieving 80% conversion of CHO at a CHO/C4/TBAB molar ratio of 2000/1/8 and a CO2 pressure of 3 MPa for 16 h at 100 °C, while maintaining >99% selectivity of cis-cyclic carbonates, which demonstrated good conversion and selectivity.

[Advances in vestibular function and rehabilitation of large vestibular aqueduct syndrome].

Large vestibular aqueduct syndrome（LVAS） is a common recessive hereditary hearing loss disease, and some patients may also experience vestibular dysfunction. With the wide application of cochlear implant（CI） and the development of vestibular medicine, the pathophysiological mechanism of LVAS and the influence mechanism of CI on vestibular function are gradually elucidated. Consequently, the evaluation and rehabilitation of vestibular dysfunction function have also become research hotspots. This article reviews studies on vestibular function and related rehabilitation in patients with large vestibular aqueduct syndrome.

Impact of Long COVID on Health-Related Quality of Life Among Patients After Acute COVID-19 Infection: A Cross-Sectional Study.

Concerns have been raised globally regarding the long-term effects of the novel coronavirus disease 2019 (COVID-19). This study aimed to investigate the impact of long COVID on the health of patients recovering from acute COVID-19 in China. We conducted a cross-sectional questionnaire survey from 1 February to 9 March 2023. Propensity score matching (PSM) was used to understand the differences in health utility values between individuals with and without long COVID. Factors associated with health-related quality of life (HRQoL) were determined using a multiple linear regression model. A chi-square test was used to compare differences between the 2 groups for each dimension of the EuroQoL-5 Dimension-5 Level (EQ-5D-5L) scale. In total, 307 participants were included in the analysis, of which 40.39% exhibited at least 1 persistent symptom. The common symptoms of long COVID were fatigue/weakness, coughing, memory decline, poor concentration, and phlegm in the throat. Most patients with long COVID reported mild effects from their symptoms. After propensity score matching, the long-COVID group had lower health utility scores than the non-long-COVID group (0.94 vs 0.97). In the multivariable linear regression analysis, persistent symptoms and low annual household income were associated with lower health utility values (P < .05). Anxiety/depression and pain/discomfort were the major problems experienced by the participants with long COVID. Long-COVID symptoms following acute COVID-19 infection have a serious impact on health-related quality of life. Therefore, it is necessary to implement interventions to improve patient health after the recovery from acute COVID-19.

Effects of oat ß-glucan on the retrogradation behavior of rice starch and its potential mechanism.

In this study, to minimize the quality deterioration caused by the retrogradation of starch-based food, the effect and mechanism of oat ß-glucan (OG) on the retrogradation of rice starch was investigated. OG effectively decreased storage modulus (G'), syneresis, and retrogradation enthalpy, indicating the inhibition of short-term and long-term retrogradation of rice starch. The competition for water molecules between the OG and rice starch resulted in partial swelling of the starch granules, consequently reducing particle size, lowering amylose leaching, and decreasing the proportion of short-amylose chains. The microstructure characterization showed that the OG-treated rice starch group (ST-OG) exhibited a smoother and denser surface. Particularly, no notable alterations were observed in the structure of the ST-OG sample during storage, owing to the improved water-holding capacity of starch gel and reduced proportion of free water caused by OG. Furthermore, the ordered structure results confirmed the occurrence of hydrogen bonding between OG and rice starch, which hindered the rearrangement of starch molecules. Therefore, OG is an effective natural additive for controlling the retrogradation of starch-based foods.

Automated and miniaturized screening of antibiotic combinations via robotic-printed combinatorial droplet platform.

Antimicrobial resistance (AMR) has become a global health crisis in need of novel solutions. To this end, antibiotic combination therapies, which combine multiple antibiotics for treatment, have attracted significant attention as a potential approach for combating AMR. To facilitate advances in antibiotic combination therapies, most notably in investigating antibiotic interactions and identifying synergistic antibiotic combinations however, there remains a need for automated high-throughput platforms that can create and examine antibiotic combinations on-demand, at scale, and with minimal reagent consumption. To address these challenges, we have developed a Robotic-Printed Combinatorial Droplet (RoboDrop) platform by integrating a programmable droplet microfluidic device that generates antibiotic combinations in nanoliter droplets in automation, a robotic arm that arranges the droplets in an array, and a camera that images the array of thousands of droplets in parallel. We further implement a resazurin-based bacterial viability assay to accelerate our antibiotic combination testing. As a demonstration, we use RoboDrop to corroborate two pairs of antibiotics with known interactions and subsequently identify a new synergistic combination of cefsulodin, penicillin, and oxacillin against a model E. coli strain. We therefore envision RoboDrop becoming a useful tool to efficiently identify new synergistic antibiotic combinations toward combating AMR.

Effects of traditional Chinese exercises in fibromyalgia syndrome: A meta-analysis of randomized controlled trials.

OBJECTIVES: To explore the efficacy and safety of five traditional Chinese exercises (TCEs) in patients with fibromyalgia syndrome (FMS). METHODS: The PubMed, Embase, Scopus, ProQuest, Web of Science, Cochrane, CNKI, WanFang, and VIP databases were comprehensively searched for randomized controlled trials (RCTs) related to TCEs published from inception until February 2023. Standardized mean differences (SMD) and 95% confidence intervals (CI) were used to determine the combined effects of the intervention, and the Cochrane risk-of-bias assessment tool and Review 5.2 software were used to assess methodological quality. The data were extracted and analyzed by the Stata15.0 random effects model. RESULTS: Nineteen RCTs including 1315 participants were included in the analysis. The studies were found to be heterogeneous (I2 =86.2, P = 0.000), and thus a random effects model was used to combine the data. The results showed that traditional Chinese exercises had potentially beneficial effects on reducing pain (SMD =-0.66,95% CI [-1.08, -0.25], P = 0.002), improving sleep (SMD = -0.35,95% CI [-0.68,0. 01], P = 0.041) and relieving depression (SMD= -0.24,95% CI [-0.47, -0.02], P = 0.034) in FMS patients. However, no significant effects were found on improved quality of life (SMD =-0.20,95% CI [-0.48,0.09], P = 0.176). CONCLUSIONS: TCEs can improve pain, sleep quality and depression in patients with FMS and are safe. However, they do not improve the quality of life significantly. Further large-scale, high-quality, and multi-center RCTs are required to verify the efficacy of TCEs.

Identification of multiomics map and key biomarkers in uveal melanoma with chromosome 3 loss.

Purpose: Chromosome 3 loss is an independent risk factor for uveal melanoma (UM), but its exact molecular mechanisms remain unclear. This study was designed to investigate the relationship between chromosome 3 loss and molecular alterations at multiple levels to construct a prognostic model. Methods: Forty-four UM cases with chromosome 3 loss (chr3 del group) and 36 UM cases without copy number variation on chromosome 3 (chr3 wt group) were collected from the Cancer Genome Atlas (TCGA). The TCGA dataset was subjected to a univariate Cox regression analysis to identify different expressed genes, and a subsequent random forest algorithm analysis revealed significant changes in different expressed genes, which were used to develop key biomarkers for UM. Following that, the immune cell infiltration analysis and drug sensitivity analyses were carried out. The UM cell line was then utilized to investigate the potential functions of the key biomarker via cell apoptosis, proliferation, cycle assays, WB, and RT-qPCR. Results: By analyzing the 80 cases data in TCGA, the authors unveiled molecular changes relevant to loss of chromosome 3 in UM as well as their poor survival. In addition, machine learning analysis identified three hub genes (GRIN2A, ACAN, and MMP9) as potential therapeutic targets. The differentially enriched pathways between the two groups were mainly about immune-system activity, and hub genes expression was also highly correlated with immune infiltration levels. Conclusion: Chromosome 3 loss has considerable clinical significance for UM, and GRIN2A may be useful in diagnosing, treating, and prognosticating the condition.

Liter-scale manufacturing of shelf-stable plasmid DNA/PEI transfection particles for viral vector production.

The transfection efficiency and stability of the delivery vehicles of plasmid DNA (pDNA) are critical metrics to ensure high-quality and high-yield production of viral vectors. We previously identified that the optimal size of pDNA/poly(ethylenimine) (PEI) transfection particles is 400-500 nm and developed a bottom-up assembly method to construct stable 400-nm pDNA/PEI particles and benchmarked their transfection efficiency in producing lentiviral vectors (LVVs). Here, we report scale-up production protocols for such transfection particles. Using a two-inlet confined impinging jet (CIJ) mixer with a dual syringe pump set-up, we produced a 1-L batch at a flow rate of 100 mL/min, and further scaled up this process with a larger CIJ mixer and a dual peristaltic pump array, allowing for continuous production at a flow rate of 1 L/min without a lot size limit. We demonstrated the scalability of this process with a 5-L lot and validated the quality of these 400-nm transfection particles against the target product profile, including physical properties, shelf and on-bench stability, transfection efficiency, and LVV production yield in both 15-mL bench culture and 2-L bioreactor runs. These results confirm the potential of this particle assembly process as a scalable manufacturing platform for viral vector production.

Single-Particle Spectroscopic Chromatography Reveals Heterogeneous RNA Loading and Size Correlations in Lipid Nanoparticles.

Lipid nanoparticles (LNP) have emerged as pivotal delivery vehicles for RNA therapeutics. Previous research and development usually assumed that LNPs are homogeneous in population, loading density, and composition. Such perspectives are difficult to examine due to the lack of suitable tools to characterize these physicochemical properties at the single-nanoparticle level. Here, we report an integrated spectroscopy-chromatography approach as a generalizable strategy to dissect the complexities of multicomponent LNP assembly. Our platform couples cylindrical illumination confocal spectroscopy (CICS) with single-nanoparticle free solution hydrodynamic separation (SN-FSHS) to simultaneously profile population identity, hydrodynamic size, RNA loading levels, and distributions of helper lipid and PEGylated lipid of LNPs at the single-particle level and in a high-throughput manner. Using a benchmark siRNA LNP formulation, we demonstrate the capability of this platform by distinguishing seven distinct LNP populations, quantitatively characterizing size distribution and RNA loading level in wide ranges, and more importantly, resolving composition-size correlations. This SN-FSHS-CICS analysis provides critical insights into a substantial degree of heterogeneity in the packing density of RNA in LNPs and size-dependent loading-size correlations, explained by kinetics-driven assembly mechanisms of RNA LNPs.

Association between e-cigarette use and susceptibility to tobacco product use: findings from the 2019 China National Youth Tobacco Survey.

Background: There is an ongoing debate about whether e-cigarettes act as a gateway to tobacco smoking or contribute to smoking cessation, and relevant studies are limited among Chinese adolescents. This cross-sectional study therefore aimed to explore the relationship between e-cigarette use and susceptibility to tobacco product use among Chinese high school students. Methods: The study population comprised 107,633 never smokers and 19,377 former smokers, generated from the 2019 China National Youth Tobacco Survey. The primary independent variables of interest were ever e-cigarette use, current e-cigarette use, and the frequency of current e-cigarette use. The main outcome was the susceptibility to tobacco product use. Multilevel logistic regression was used to estimate the association between the primary independent variables of interest and the outcome variable. Moreover, two additional multilevel logistic regression models were fitted using two alternative definitions of the outcome as the sensitivity analyses. Results: Among never smokers, students who ever used e-cigarettes were more likely to be susceptible to tobacco product use compared to students who never used e-cigarettes (AOR = 2.83, 95%CI = 2.59-3.08). Students who currently used e-cigarettes were more likely to be susceptible to tobacco product use than those who did not currently use e-cigarettes (AOR = 3.89, 95%CI = 3.21-4.72). Among former smokers, with the same settings of modeling, the AORs were 1.76 (95%CI = 1.62-1.91) and 3.16 (95%CI = 2.52-3.97), respectively. Similar results were obtained from the two sensitivity analyses. Conclusion: Among Chinese high school students, both never smokers and former smokers, e-cigarette use, especially current e-cigarette use, was positively associated with susceptibility to tobacco product use. It is recommended to strengthen the monitoring of e-cigarettes and to provide targeted health education to adolescents.

Skipping breakfast and physical fitness among school-aged adolescents

OBJECTIVES: This study explored the relationship between skipping breakfast and physical fitness in a group of school-aged adolescents in China. METHODS: This cross-sectional study from the Chinese National Surveillance on Students' Constitution and Health (CNSSCH) survey in Ningbo, China, used a standardized questionnaire to assess the frequency of breakfast consumption. Physical fitness was measured through standing long jump, 50-m sprint, 1,000 (or 800)-m run, and vital capacity tests. Multiple linear regression analysis was used to investigate the relationship between the frequency of breakfast consumption and physical fitness. RESULTS: Our study included a total of 1,849 school-aged adolescents (aged 15.53±1.80 years). Among boys, non-breakfast-skippers had good scores for 50-m sprints, 1,000-m run, and vital capacity tests when compared with breakfast skippers (all p<0.05). Among girls, non-breakfast-skippers had a good scores for the standing long jump test compared with breakfast skippers (p=0.003). The multiple linear regression model showed that not skipping breakfast was positively associated with vital capacity (β=-173.78, p=0.004) and inversely associated with 50-m sprint (β=-0.12, p=0.018) and 1,000-m run times (β=-8.08, p=0.001) in boys. CONCLUSION: The results of this cross-sectional study revealed that skipping breakfast might be associated with lower physical fitness in Chinese adolescents aged 13-18 years, especially boys. Breakfast consumption should be promoted among Chinese school-aged boys.