Radiofrequency affects the decrystallization efficiency and physicochemical properties of rape honey via crystal structure modification and inactivating enzyme.

Crystallization degrades the physicochemical properties of honey and reduces consumer acceptance. To address this issue, radiofrequency was developed to investigate the decrystallization efficiency and quality impact mechanism of rape honey. The results showed that radiofrequency significantly decreased the number and size of crystals, leading to shortening the decrystallization time to less than 10 min. The response surface optimization methodology further indicated that the highest decrystallization rate (98.72 ± 0.34 %) and lower 5-Hydroxymethylfurfural (2.45 ± 0.12 mg/kg) contents were obtained. Furthermore, radiofrequency changed the honey from a pseudoplastic into a Newtonian fluid efficiently due to the volumetric heating feature. It is worth noting that the inactivation of glucose oxidase reduced the antibacterial capacity, while the increase in total phenolic and flavonoid contents improved the antioxidant capacity of rape honey. In summary, current findings indicated that radiofrequency is a potential alternative decrystallization technology for water baths.

Assessment of TENS-Evoked Tactile Sensations for Transradial Amputees via EEG Investigation.

Most of current prostheses can offer motor function restoration for limb amputees but usually lack natural and intuitive sensory feedback. Many studies have demonstrated that Transcutaneous Electrical Nerve Stimulation (TENS) is promising in non-invasive sensation evoking for amputees. However, the objective evaluation and mechanism analysis on sensation feedback are still limited. This work utilized multi-channel TENS with diverse stimulus patterns to evoke sensations on four non-disabled subjects and two transradial amputees. Meanwhile, electroencephalogram (EEG) was collected to objectively assess the evoked sensations, where event-related potentials (ERPs), brain electrical activity maps (BEAMs), and functional connectivity (FC) were computed. The results show that various sensations could be successfully evoked for both amputees and non-disabled subjects by customizing stimulus parameters. The ERP confirmed the sensation and revealed the sensory-processing-related components like N100 and P200; the BEAMs confirmed the corresponding regions of somatosensory cortex were activated by stimulation; the FC indicated an increase of interactions between the regions of sensorimotor cortex. This study may shed light on how the brain responds to external stimulation as sensory feedback and serve as a pilot for further bidirectional closed-loop prosthetic control.

Propolis ethanol extract functionalized chitosan/Tenebrio molitor larvae protein film for sustainable active food packaging.

The application of novel insect proteins as future food resources in the food field has attracted more and more attention. In this study, a biodegradable antibacterial food packaging material with beneficial mechanical properties was developed using Tenebrio molitor larvae protein (TMP), chitosan (CS) and propolis ethanol extract (PEE) as raw materials. PEE was uniformly dispersed in the film matrix and the composite films showed excellent homogeneity and compatibility. There are strong intermolecular hydrogen bond interactions between CS, TMP, and PEE in the films, which exhibit the structure characteristics of amorphous materials. Compared with CS/TMP film, the addition of 3 % PEE significantly enhanced the elongation at break (34.23 %), water vapor barrier property (22.94 %), thermal stability (45.84 %), surface hydrophobicity (20.25 %), and biodegradability of the composite film. The composite film has strong antioxidant and antimicrobial properties, which were enhanced with the increase of PEE content. These biodegradable films offer an eco-friendly end-of-life option when buried in soil. Composite films can effectively delay the spoilage of strawberries and extend the shelf life of strawberries. Biodegradable active packaging film developed with insect protein and chitosan can be used as a substitute for petroleum-based packaging materials, and has broad application prospects in the field of fruits preservation.

Combined Impact of Heart Rate Sensor Placements with Respiratory Rate and Minute Ventilation on Oxygen Uptake Prediction.

Oxygen uptake (V˙O2) is an essential metric for evaluating cardiopulmonary health and athletic performance, which can barely be directly measured. Heart rate (HR) is a prominent physiological indicator correlated with V˙O2 and is often used for indirect V˙O2 prediction. This study investigates the impact of HR placement on V˙O2 prediction accuracy by analyzing HR data combined with the respiratory rate (RESP) and minute ventilation (V˙E) from three anatomical locations: the chest; arm; and wrist. Twenty-eight healthy adults participated in incremental and constant workload cycling tests at various intensities. Data on V˙O2, RESP, V˙E, and HR were collected and used to develop a neural network model for V˙O2 prediction. The influence of HR position on prediction accuracy was assessed via Bland-Altman plots, and model performance was evaluated by mean absolute error (MAE), coefficient of determination (R2), and mean absolute percentage error (MAPE). Our findings indicate that HR combined with RESP and V˙E (V˙O2HR+RESP+V˙E) produces the most accurate V˙O2 predictions (MAE: 165 mL/min, R2: 0.87, MAPE: 15.91%). Notably, as exercise intensity increases, the accuracy of V˙O2 prediction decreases, particularly within high-intensity exercise. The substitution of HR with different anatomical sites significantly impacts V˙O2 prediction accuracy, with wrist placement showing a more profound effect compared to arm placement. In conclusion, this study underscores the importance of considering HR placement in V˙O2 prediction models, with RESP and V˙E serving as effective compensatory factors. These findings contribute to refining indirect V˙O2 estimation methods, enhancing their predictive capabilities across different exercise intensities and anatomical placements.

Preparation of chitosan/Tenebrio molitor larvae protein/curcumin active packaging film and its application in blueberry preservation.

With growing concerns about postharvest spoilage of fruits, higher requirements have been placed on high-performance and sustainable active packaging materials. In this study, we prepared curcumin-based functional composite films using chitosan (CS) and Tenebrio molitor larvae protein (TMP) as the substrates. The effects of curcumin concentration on the structural and physicochemical properties of the composite films were determined. Curcumin was equally distributed in the polymer film through physical interactions. Furthermore, the curcumin composite film with 0.3 % addition exhibited a 27.39 % increase in elongation at break (EBA), a 37.04 % increase in the water vapor barrier, and strong UV-blocking properties and antioxidant activity compared with the control film (CS/TMP). The degradation experiment of the composite film on natural soil revealed that the composite film exhibited good biodegradability and environmental protection. Furthermore, the applicability of functional composite films for preserving blueberries was investigated. Compared with the control film and polyethylene (PE) films, the prepared composite films packaging treatment reduced the decay rate and weight loss rate of blueberries during storage, delayed softening and aging, and maintained the quality of blueberries. Using sustainable protein resources (TMP) and natural polysaccharides as packaging materials provides an economically, feasible and sustainable way to achieve the functional preservation of biomass materials.

The correlation analysis between incisal guidance angle and occlusal plane angles and temporomandibular joint morphology.

OBJECTIVES: The correlations between the incisal guidance angle (IGA) and occlusal plane angles and temporomandibular joint (TMJ) morphology were investigated in adults with skeletal Class II division II malocclusion. MATERIALS AND METHODS: CBCT images of 37 patients were analyzed. It included 19 cases of skeletal Class II division II malocclusion with low angle (study group) and 18 cases of skeletal Class I average angle (control group). The Invivo Dental 5 software was employed to acquire the data of the incisal guidance angle (IGA), occlusal plane angle (FH-OP), anterior occlusal plane angle (FH-AOP) and the TMJ measurement items. RESULTS: The results of IGA, FH-AOP angle and FH-OP angle showed the study group > the control group (P < 0.05). There were statistically difference in the condylar mediolateral diameters, articular eminence inclination and height, and posterior joint spaces between two groups. No differences were revealed in the condylar anteroposterior diameters, the condylar inclination angle, condylar head width and height, condylar length, glenoid fossa depth and width between two groups. In the study group, IGA showed a moderate correlation with FH-AOP, a weak correlation with FH-OP and condylar mediolateral diameters. Meanwhile, there was a correlation between FH-AOP, FH-OP, and TMJ indicators. CONCLUSIONS: The IGA was not only related to FH-AOP and FH-OP, but also to the condylar mediolateral diameters. In addition, there was a correlation between the occlusal plane angles and TMJ morphology in skeletal Class II division II low angle malocclusion. CLINICAL RELEVANCE: For patients with skeletal Class II division II low angle malocclusion, adjusting the IGA and the occlusal plane angles could improve the esthetic appearance of the anterior teeth, occlusal function, and TMJ morphology.

Effects of ginsenoside Rh2 on cisplatin-induced nephrotoxicity in renal tubular epithelial cells by inhibiting endoplasmic reticulum stress.

Nephrotoxicity remains a major adverse reaction of the anticancer drug cisplatin (CDDP) chemotherapy, which is an important risk factor for chronic renal disease. Ginsenoside Rh2 from Panax ginseng has been shown to protect against CDDP-induced nephrotoxicity in vivo, but its pharmacological effect on renal tubular epithelial cells is not clearly understood. This study examined the molecular mechanisms underlying the nephroprotective effects of Rh2 on CDDP-induced HK-2 cells and acute kidney injury (AKI) mice. As a result of Rh2 treatment, CDDP-induced HK-2 cells showed increased cell viability and reduced lactate dehydrogenase release. Moreover, Rh2 ameliorated CDDP-induced mitochondrial membrane potential, increased antioxidant enzyme activities, and reduced pro-inflammatory cytokine expression to reduce damage. Rh2 inhibited apoptosis and enhanced the antioxidant capacity of HK-2 cells by reducing proteins associated with endoplasmic reticulum (ER) stress, as well as by attenuating tunicamycin-induced ER stress. In addition, treatment of CDDP-induced AKI mice with Rh2 substantially reduced blood urea nitrogen and serum creatinine levels, attenuated histological damage of kidney. Further, Rh2 also improved kidney function by inhibiting ER stress to support in vitro findings. These results consistently demonstrated that Rh2 protects renal tubular epithelial cells from CDDP-induced nephrotoxicity and apoptosis by restoring ER homeostasis, which might suggest a therapeutic potential and providing new insights into AKI alternative therapies.

FCER1G as a novel immune-associated blood biomarker in cardiogenic stroke.

Background: Cardioembolic stroke (CE) exhibits the highest recurrence rate and mortality rate among all subtypes of cerebral ischemic stroke (CIS), yet its pathogenesis remains uncertain. The immune system plays a pivotal role in the progression of CE. Growing evidence indicates that several immune-associated blood biomarkers may inform the causes of stroke. The study aimed to identify new immune-associated blood biomarkers in patients with CE and create an online predictive tool in distinguishing CE from noncardioembolic stroke (non-CE) in CIS. Methods: Gene expression profiles that were publicly available were obtained from the Gene Expression Omnibus (GEO). The identification of differentially expressed genes (DEGs) was conducted using the Limma package. The hub module and hub genes were identified through the application of weighted gene coexpression network analysis (WGCNA). In order to identify potential diagnostic biomarkers for CE, both the random forest (RF) model and least absolute shrinkage and selection operator (LASSO) regression analysis were employed. Concurrently, the CIBERSORT algorithm was employed to evaluate the infiltration of immune cells in CE samples and examine the correlation between the biomarkers and the infiltrating immune cells. The diagnostic gene expression in blood samples was confirmed using qRT-PCR in a self-constructed dataset. Univariate and multiple logistic regression analyses were used to identify the risk factors for CE. Subsequently, the mathematical model of the nomogram was employed via Java's "Spring Boot" framework to develop the corresponding online tool, which was then deployed on a cloud server utilizing "nginx". Results: Eleven differentially expressed genes (DEGs) that were upregulated and seven DEGs that were downregulated were identified. Through bioinformatics analysis and clinical sample verification, it was discovered that Fc Fragment of IgE Receptor Ig (FCER1G) could serve as a novel potential blood biomarker for CE. FCER1G, along with other risk factors associated with CE, were utilized to develop a nomogram. The training and validation sets, which consisted of 65 CIS patients, yielded areas under the curve (AUCs) of 0.9722 and 0.9689, respectively. These results indicate a high level of precision in risk delineation by the nomogram. Furthermore, the associated online predictive platform has the potential to serve as a more efficacious and appropriate predictive instrument (https://www.origingenetic.com/CardiogenicStroke-FCER1G) for distinguishing between CE and non-CE. Conclusion: Blood biomarker FCER1G has the potential to identify patients who are at a higher risk of cardioembolism and direct the search for occult AF.The utilization of this online tool is anticipated to yield significant implications in terms of distinguishing between CE and non-CE, as well as enhancing the optimization of treatment decision support.

Microneedle Array Electrodes Fabricated With 3D Printing Technology for High-Quality Electrophysiological Acquisition.

Electrophysiological recordings are vital in assessing biological functions, diagnosing diseases, and facilitating biofeedback and rehabilitation. The applications of conventional wet (gel) electrodes often come with some limitations. Microneedle array electrodes (MAEs) present a possible solution for high-quality electrophysiological acquisition, while the prior technologies for MAE fabrication have been either complex, expensive, or incapable of producing microneedles with uniform dimensions. This work employed a projection stereolithography (P µ SL) 3D printing technology to fabricate MAEs with micrometer-level precision. The MAEs were compared with gel and flat electrodes on electrode-skin interface impedance (EII) and performances of EMG and ECG acquisition. The experimental results indicate that the P µ SL 3D printing technology contributed to an easy-to-perform and low-cost fabrication approach for MAEs. The developed MAEs exhibited promising EII and enabled a stable EMG and ECG acquisition in different conditions without inducing skin allergies, inflammation, or injuries. This research lies in the development of a type of customizable MAE with considerable biomedical application potentials for ultra-minimally invasive or non-invasive electrophysiological acquisition.

MLLT6/ATF2 Axis Restrains Breast Cancer Progression by Driving DDIT3/4 Expression.

Epigenetic deregulation is strongly associated with tumor progression. The identification of natural tumor suppressors to overcome cancer metastasis is urgent for cancer therapy. We investigate whether myeloid/lymphoid or mixed-lineage leukemia translocated (MLLT) family members contribute to breast cancer progression and found that high MLLT6 expression predicted a better prognosis and that gradually decreased MLLT6 expression was accompanied by breast cancer malignancy. MLLT6 was downregulated by hypoxia-induced enrichment of DNMT1 at the MLLT6 promoter. The results of in vitro functional experiments indicated that MLLT6 depletion promoted colony formation and cell migration, probably by hampering apoptosis. RNA profiling revealed that the apoptotic pathway was downregulated following stable knockdown of MLLT6. DNA damage-inducible transcript 3/4 (DDIT3/4) were among the top 10 downregulated genes and may have expression patterns similar to that of MLLT6. Restoring DDIT3/4 expression in cells with MLLT6 depletion blocked colony formation and cell migration and attenuated the successful colonization of breast cancer cells in vivo. We also determined that the transcription factor activating transcription factor 2 is a binding partner of MLLT6 and participates in the MLLT6/ATF2 axis, which was reinforced by inhibition of AKT signaling, in turn inducing DDIT3/4 expression by establishing an active chromatin structure at the DDIT3/4 gene promoters. As MLLT6 promotes breast cancer cell apoptosis by inducing DDIT3/4 expression during metastasis, it could be a novel tumor suppressor. Implications: Control of MLLT6 expression via inhibition of PI3K/AKT kinase activity is a potential therapeutic approach for the management of metastatic breast cancer.

Exploring the effect of high-pressure processing conditions on the deaggregation of natural major royal jelly proteins (MRJPs) fibrillar aggregates.

High pressure processing is a safe and green novel non-thermal processing technique for modulating food protein aggregation behavior. However, the systematic relationship between high pressure processing conditions and protein deaggregation has not been sufficiently investigated. Major royal jelly proteins, which are naturally highly fibrillar aggregates, and it was found that the pressure level and exposure time could significantly promote protein deaggregation. The 100-200 MPa treatment favoured the deaggregation of proteins with a significant decrease in the sulfhydryl group content. Contrarily, at higher pressure levels (>400 MPa), the exposure time promoted the formation of disordered agglomerates. Notably, the inter-conversion of α-helix and ß-strands in major royal jelly proteins after high pressure processing eliminates the solvent-free cavities inside the aggregates, which exerts a 'collapsing' effect on the fibrillar aggregates. Furthermore, the first machine learning model of the high pressure processing conditions and the protein deaggregation behaviour was developed, which provided digital guidance for protein aggregation regulation.

Dual-Frequency Impedance Matching Network Design Using Genetic Algorithm for Power Ultrasound Transducer.

Dual-frequency ultrasounds have demonstrated significant potential in augmenting thermal ablation efficiency for tumor treatment. Ensuring proper impedance matching between the dual-frequency transducer and the power amplifier system is imperative for equipment safety. This paper introduces a novel dual-frequency impedance matching network utilizing L-shaped topology and employing a genetic algorithm to compute component values. Implementation involved an adjustable capacitor and inductor network to achieve dual-frequency matching. Subsequently, the acoustic parameters of the dual-frequency HIFU transducer were evaluated before and after matching, and the effects of ultrasound thermal ablation with and without matching were compared. The proposed dual-frequency impedance matching system effectively reduced the standing wave ratio at the two resonance points while enhancing transmission efficiency. Thermal ablation experiments with matching circuits showed improved temperature rise efficiencies at both frequencies, resulting in an expanded ablation zone. The dual-frequency impedance matching method significantly enhances the transmission efficiency of the dual-frequency ultrasound system at two operational frequencies, thereby ensuring equipment safety. It holds promising prospects for application in dual-frequency ultrasound treatment.

A Polymer Nanocomposite with Strong Full-Spectrum Solar Absorption and Infrared Emission for All-Day Thermal Energy Management and Conversion.

Realizing efficient energy utilization from the heat source of the sun and the cold source of outer space is of great significance for addressing the global energy and environmental crisis. Materials with ideal full-spectrum solar absorption and infrared emission are highly desirable for adapting to the continuous weather dynamic throughout the day, nonetheless, their development remains challenging. Here, a polymer nanocomposite with full-spectrum strong solar (280-2500 nm) absorption ranging from 88.8% to 94.8% with an average value of 93.2% and full-spectrum high infrared (8-13 µm) emission ranging from 81.3% to 90.0% with an average value of 84.2%, is reported by melt-processing polypropylene and uniformly dispersed low-loading MXene nanosheets (1.9 vol%). The nanocomposite can achieve daytime photothermal enhancement of ≈50 °C and nighttime radiative cooling of 8 °C. The temperature difference throughout the day ensures all-day uninterrupted thermoelectric generation, yielding a power density output of 1.5 W m-2 (daytime) and 7.9 mW m-2 (nighttime) in real outdoor environment without any additional energy consumption. This work provides an impressive polymer nanocomposite with ideal full-spectrum solar absorption and infrared emission for all-day uninterrupted thermal energy management and conversion.

Multi-Frequency Ultrasound Imaging Fusion Method Based on Wavelet Transform for Guided Screw Insertion.

Intraosseous ultrasound imaging can serve as a guiding tool for the placement of pedicle screws during spinal fusion surgery; thus far, there has been limited scholarly exploration of methods for intraosseous multifrequency ultrasound imaging, which can achieve simultaneous high resolution and deep penetration. The proposed method introduced a dynamic fusion strategy grounded in wavelet transformation for multifrequency image decomposition. This strategy accomplished the effective amalgamation of high-frequency ultrasound images and low-frequency ultrasound images, enabling the obtaining of fused images with enhanced details and better overall image quality. A novel near-field effect elimination method was also proposed to improve the quality of ultrasound imaging in the near-field region. Experimental evaluations were conducted on isolated bovine circle bone and sheep spine with pedicle screw tracks. The fusion images are capable of effectively detecting areas within the pedicle screw track that have either ruptured or are in close proximity to rupture, even measuring the size of breaches. Evaluation criteria, including information entropy (IE), spatial frequency (SF), average gradient (AG), mutual information (MI), structural similarity index (SSIM), and edge information-based image fusion quality metric (QAB/F), were employed to assess the fusion performance; moreover, the influence of mother wavelet function selection and decomposition levels on computational complexity and fusion image quality was thoroughly discussed. The proposed method exhibited promising potential for intraosseous imaging navigation, which can aid in accurate diagnosis, treatment planning, and monitoring in fields such as orthopedics, surgery, and interventional procedures.

A novel unsupervised dynamic feature domain adaptation strategy for cross-individual myoelectric gesture recognition.

Objective.Surface electromyography pattern recognition (sEMG-PR) is considered as a promising control method for human-machine interaction systems. However, the performance of a trained classifier would greatly degrade for novel users since sEMG signals are user-dependent and largely affected by a number of individual factors such as the quantity of subcutaneous fat and the skin impedance.Approach.To solve this issue, we proposed a novel unsupervised cross-individual motion recognition method that aligned sEMG features from different individuals by self-adaptive dimensional dynamic distribution adaptation (SD-DDA) in this study. In the method, both the distances of marginal and conditional distributions between source and target features were minimized through automatically selecting the optimal feature domain dimension by using a small amount of unlabeled target data.Main results.The effectiveness of the proposed method was tested on four different feature sets, and results showed that the average classification accuracy was improved by above 10% on our collected dataset with the best accuracy reached 90.4%. Compared to six kinds of classic transfer learning methods, the proposed method showed an outstanding performance with improvements of 3.2%-13.8%. Additionally, the proposed method achieved an approximate 9% improvement on a publicly available dataset.Significance.These results suggested that the proposed SD-DDA method is feasible for cross-individual motion intention recognition, which would provide help for the application of sEMG-PR based system.

MiR-499 enhances Porphyromonas gingivalis LPS-induced inflammatory response in macrophages by targeting NRIP1 via JAK/STAT pathway.

MicroRNA functions as an important part of the activity and development of immune cells. miR-499 has been demonstrated to play a significant role in the activity and development of immune cells. The precise mechanism by which miR-499 regulates the inflammatory response, however, remains unclear. This study was aimed to examine the role of microRNA miR-499 in the regulation of the inflammatory response in macrophages. RAW 264.7 macrophages were used as a cell model. The levels of miR-499 were measured in Porphyromonas gingivalis LPS-stimulated macrophages using qRT-PCR, and the levels of inflammatory cytokines (IL-6, IL-1ß, and TNF-α) were determined using both qRT-PCR and ELISA. StarBase was used to predict the binding sites between NRIP1 and miR-499, and the mRNA expression of NRIP1 was measured using qRT-PCR. The regulation of inflammatory factors controlled by miR-499 was also evaluated by using miR-499 inhibitor and sh-NRIP1. The activation of the JAK/STAT pathway was determined using western blotting to measure the levels of phosphorylated JAK2 and STAT1. Porphyromonas gingivalis LPS caused a high expression of miR-499, which promoted the inflammatory response in macrophages. miR-499 targeted the NRIP1 3' UTR and regulated the mRNA expression of inflammatory cytokines, including IL-6, IL-1ß, and TNF-α. The positive correlation between miR-499 and the expression of inflammatory factors and the negative correlation between NRIP1 and miR-499 suggests that the regulation of inflammatory factors controlled by miR-499 was associated with NRIP1. The phosphorylated proteins of the JAK/STAT pathway (p-JAK2 and p-STAT1) were activated by miR-499 through its regulation of NRIP1. These findings suggest that miR-499 regulates the P. gingivalis LPS-induced inflammatory response in macrophages and activates the JAK/STAT pathway through the regulation of NRIP1.

A Novel Transformer-Based Approach for Simultaneous Recognition of Hand Movements and Force Levels in Amputees Using Flexible Ultrasound Transducers.

Accurate hand motion intention recognition is essential for the intuitive control of intelligent prosthetic hands and other human-machine interaction systems. Sonomyography, which can detect the changes in muscle morphology and structure precisely, is a promising signal source for fine hand movement recognition. However, sonomyography measured by traditional rigid ultrasound probes may suffer from poor acoustic coupling because the rigid probe surfaces cannot accommodate the curvilinear shape of the human body, particularly in the case of small and irregular residual limbs in amputees. In this study, we used a self-designed lightweight, flexible, and wearable ultrasound transducer to acquire muscle ultrasound images, and proposed a sonomyography transformer (SMGT) model for simultaneous recognition of hand movements and force levels. The performance of SMGT was systematically compared to two commonly used image processing methods, HOG and Gray Gradient, as well as a deep CNN model, in simultaneously recognizing ten classes of hand/finger movements and three force levels. Additionally, ten subjects including seven non-disabled subjects and three trans-radial amputees who are the end users of prosthetic hands were recruited to evaluate the effectiveness of SMGT. Results showed that our proposed method achieved average classification accuracies of 98.4% ± 0.6% and 96.2% ± 3.0% in non-disabled subjects and amputee subjects, respectively, which are much higher than those of other methods. This study provided a valuable approach for ultrasound-based hand motion recognition that may promote the applications of intelligent prosthetic hands.

Metatranscriptome of human lung microbial communities in a cohort of mechanically ventilated COVID-19 Omicron patients.

The Omicron variant of the severe acute respiratory syndrome coronavirus 2 (SARS­CoV­2) infected a substantial proportion of Chinese population, and understanding the factors underlying the severity of the disease and fatality is valuable for future prevention and clinical treatment. We recruited 64 patients with invasive ventilation for COVID-19 and performed metatranscriptomic sequencing to profile host transcriptomic profiles, plus viral, bacterial, and fungal content, as well as virulence factors and examined their relationships to 28-day mortality were examined. In addition, the bronchoalveolar lavage fluid (BALF) samples from invasive ventilated hospital/community-acquired pneumonia patients (HAP/CAP) sampled in 2019 were included for comparison. Genomic analysis revealed that all Omicron strains belong to BA.5 and BF.7 sub-lineages, with no difference in 28-day mortality between them. Compared to HAP/CAP cohort, invasive ventilated COVID-19 patients have distinct host transcriptomic and microbial signatures in the lower respiratory tract; and in the COVID-19 non-survivors, we found significantly lower gene expressions in pathways related viral processes and positive regulation of protein localization to plasma membrane, higher abundance of opportunistic pathogens including bacterial Alloprevotella, Caulobacter, Escherichia-Shigella, Ralstonia and fungal Aspergillus sydowii and Penicillium rubens. Correlational analysis further revealed significant associations between host immune responses and microbial compositions, besides synergy within viral, bacterial, and fungal pathogens. Our study presents the relationships of lower respiratory tract microbiome and transcriptome in invasive ventilated COVID-19 patients, providing the basis for future clinical treatment and reduction of fatality.

Immunogenicity and Antigenicity of the Ectodomain of Rabies Virus Glycoprotein Stably Expressed in HEK293T Cells.

Rabies continues to be a huge threat to public health. The rabies virus envelope glycoprotein (RABV G) is a major rabies virus antigen and contains neutralizing epitopes, which are primary candidates for subunit vaccines and diagnostic antigens. However, the production and purification of rRABV G while retaining its antigenic and immunogenic remains to be a challenge. Here, we aimed to establish a platform for rRABV G production and purification, and determine the immunogenicity and antigenicity of rRABV G. The cDNA fragment encoding the soluble form of RABV G was synthesized and cloned into a lentiviral expressing vector. Recombinant lentiviral vector LV-CMV-RABV G-eGFP was packaged, titered, and then transduced into HEK 293T cells. The cell culture supernatant was purified using nickel affinity chromatography and subsequently confirmed through Western Blot analysis and indirect enzyme-linked immunosorbent assay (ELISA). The ELISA utilized human sera obtained from individuals who had been vaccinated with the human commercial Purified Vero Cells Rabies Vaccine (PVRV). Notably, we observed a neutralizing antibody response in immunized pigs rather than in mice. This discrepancy could potentially be attributed to factors such as the instability of the rRABV G protein, variations in host responses, and variances in the adjuvant used. Taking all these findings into account, the rRABV G protein generated in this study exhibits promise as a potential vaccine candidate for the prevention of rabies.

Tim-3 facilitates immune escape in benzene-induced acute myeloid leukemia mouse model by promoting macrophage M2 polarization.

Benzene poisoning can cause acute myeloid leukemia (AML) through a variety of passways. Tim-3 has gained prominence as a potential candidate in mediating immunosuppression in tumor microenvironments. The macrophage polarization is also related to immune escape. Herein, we reported that Tim-3 and macrophage M2 polarization play a vital role in benzene-induced AML. First, the benzene-induced AML C3H/He mouse model was constructed by subcutaneously injecting 250 mg/kg of benzene. After six months, macrophage phenotype, cytokines, and Tim-3 expression levels were investigated. Flow cytometry assay revealed that the T-cell inhibitory receptor Tim-3 was significantly upregulated in both bone marrow and spleen of the benzene-induced AML mouse model. Elisa's results displayed a decreased serum level of IL-12 while increased TGF-ß1. Mechanistically, changes in cytokine secretion promote the growth of M2-type macrophages in the bone marrow and spleen, as determined by immunofluorescence assay. The increased levels of PI3K, AKT, and mTOR in the benzene-exposure group further proved the crucial role of Tim-3 in regulating the functional status of macrophages in the AML microenvironment. These results demonstrate that Tim-3 and macrophage polarization may play a vital role during the immune escape of the benzene-induced AML. This study provides a new potential intervention site for immune checkpoint-based AML therapeutic strategy.