Novel platform for engineering stable and effective vaccines against botulinum neurotoxins A, B and E.

Botulinum neurotoxin (BoNT), produced by Clostridium botulinum, is the most toxic protein known, capable of causing severe paralysis and posing a significant bioterrorism threat due to its extreme lethality even in minute quantities. Despite this, there are currently no FDA-approved vaccines for widespread public use. To address this urgent need, we have developed an innovative vaccine platform by fusing the neuronal binding domain of BoNT/E (Hc/E) with core-streptavidin (CS), resulting in a stable CS-Hc/E vaccine. Mice vaccinated with CS-Hc/E exhibited superior antibody titers compared to those receiving Hc/E alone. To develop a trivalent vaccine against BoNT/A, BoNT/B, and BoNT/E- key contributors to the vast majority of human botulism-we conjugated CS-Hc/E with a biotinylated atoxic chimeric protein incorporating neutralizing epitopes from BoNT/A and BoNT/B. This chimeric protein includes the binding domain of BoNT/A, along with the protease-inactive light chain and translocation domains of BoNT/B. The interaction between CS and biotin formed a stable tetrameric antigen, EBA. Vaccination with EBA in mice elicited robust antibody responses and provided complete protection against lethal doses of BoNT/A, BoNT/B, and BoNT/E. Our findings highlight EBA's potential as a stable and effective broad-spectrum vaccine against BoNT. Moreover, our technology offers a versatile platform for developing multivalent, stable vaccines targeting various biological threats by substituting the BoNT domain(s) with neutralizing epitopes from other life-threatening pathogens, thereby enhancing public health preparedness and biodefense strategies.

Sleep pattern, genetic risk, and the risk of incident rheumatoid arthritis: A cohort study.

OBJECTIVE: To investigate the associations of sleep behaviors with the risk of rheumatoid arthritis, and whether the associations differ among individuals with low, intermediate, or high genetic risk. METHODS: We included participants who were free of rheumatoid arthritis at baseline based the UK Biobank. We evaluated the associations of five sleep behaviors with the risk of rheumatoid arthritis using Cox proportional hazard regression models. We then generated a sleep risk score which combined five sleep behaviors and assessed its association with the risk of rheumatoid arthritis. We finally generated a genetic risk score and examined the joint effects of sleep patterns and genetic susceptibility on the risk of rheumatoid arthritis. RESULTS: Of the 375,133 participants at baseline, 4913 incident rheumatoid arthritis cases were identified over a median follow-up of 11.73years. We found that insomnia and daytime sleepiness were associated with a 33% and a 38% increased risk of rheumatoid arthritis. A U-shaped association was observed between sleep duration and the risk of rheumatoid arthritis, with a 29% higher risk for those with short sleep and a 30% higher risk for those with long sleep. Participants with unfavorable sleep patterns had a 63% increased risk of rheumatoid arthritis compared with those with favorable sleep patterns. Participants with unfavorable sleep patterns and high genetic risk showed the highest risk of rheumatoid arthritis although no statistically significant multiplicative or additive interaction was found. CONCLUSIONS: Our study suggested that insomnia, daytime sleepiness, and short or long sleep duration, as well as sleep risk score were associated with an increased risk of rheumatoid arthritis.

[Outcome-based education enhances the innovative capabilities of biological science undergraduates in the context of new engineering].

In the context of new engineering, it is of great importance to enhance the innovative capabilities of biological science undergraduates receiving higher education. We conducted a questionnaire survey at the School of Life and Environmental Science, Hangzhou Normal University to examine the current situation of undergraduates' innovative capabilities. We explored the teaching reform in biological majors based on outcome-based education (OBE) from teaching innovation, tutorial system establishment, and dual integration of research-education and production-education. Furthermore, an empirical study adopting the Williams's Creativity Assessment Packet highlighted that the OBE-oriented teaching reform effectively fostered students' creativity and laid a foundation for enhancing their innovative capabilities. This study provides valuable insights for cultivating innovative talents majoring in biological sciences in universities.

Resistance-based directed evolution of nanobodies for higher affinity in prokaryotes.

A prokaryotic resistance-based directed evolution system leveraging protein-fragment complementation assay (PCA) was devised, and its proficiency in detecting protein-protein interactions and discriminating varying degrees of binding affinity was demonstrated by two well-characterized protein pairs. Furthermore, we constructed a random mutant library based on the GBPR36K/E45K mutant, characterized by almost no affinity towards EGFP. This library was subjected to PCA-based prokaryotic directed evolution, resulting in the isolation of back-mutated variants. In summary, we have established an expedited, cost-effective, and structural information-independent PCA-based prokaryotic directed evolution platform for nanobody affinity maturation, featuring tunable screening stringency via modulation of antibiotic concentrations.

Thiourea enhanced oxidase-like activity of CeO2/CuxO nanozyme for fluorescence/colorimetric detection of thiourea and glutathione.

A novel fluorescence/colorimetric dual-mode sensor, based on enhancement of the oxidase-like activity of CeO2/CuxO nanozyme towards the oxidation of o-phenylenediamine (OPD) induced by thiourea (TU), has been proposed for TU detection. The catalytic activity enhancement on CeO2/CuxO can be attributed to the strong electron-donation ability of TU, which promoted hydroxyl radical generation and amplified OPD oxidization with enhanced dual-signal readout. By integrating a portable paper-chip and smartphone system, this CeO2/CuxO-OPD system achieved on-site visual colorimetric analysis of TU. The dual-mode sensor demonstrated high sensitivity and specificity in recognizing TU, with a detection limit (LOD) of 1.90 µM and a linear range (LR) 2.5-80 µM in fluorescent mode; as well as an LOD of 6.69 µM and an LR 10-250 µM in colorimetric mode. Furthermore, the CeO2/CuxO-TU-OPD system has been designed for dual-mode glutathione (GSH) detection with enhanced sensitivity, achieving an LOD of 0.19 µM and an LR 0.5-10 µM in fluorescent mode; as well as an LOD of 1.24 µM and an LR 1.25-25 µM in colorimetric mode. Additionally, GSH discrimination (fluorescent mode) was successfully achieved in different biological samples, showing good consistency with the standard method. The recoveries ranged from 96.8 % to 116.7 % in serum samples and from 97.3 % to 107.7 % in cell lysates, with RSDs less than 2 %. This work not only introduced a novel approach to enhance oxidase-like activity of nanozymes but also provided an efficient field-suitable tool for enhanced dual-mode response towards TU and GSH.

Label-free detection of the cytotoxicity effect of cisplatin in human leukemic cells using Raman spectroscopy in conjunction with multivariate analysis.

The single-cell Raman spectra of human leukemic Jurkat cells can be obtained by confocal microscopy Raman spectroscopy, including cell groups treated with different doses of cisplatin (0, 3.5, 7, 10.5 and 14 µmol L-1) for 24 hours and those treated with 10.5 µmol L-1 cisplatin for different times (0, 6, 12, 24 and 36 hours). The structure and amount of protein, nucleic acid and other major molecules from different cell groups show special changes in the percentage of biochemical constituents. Compared with the control group, the two protein Raman bands (1449 and 1659 cm-1) and two DNA bands (1303 and 1338 cm-1) in the treatment groups decrease in intensity with the increase of the drug dose and treatment time of cisplatin. Partial least squares combined with support vector machines was used to develop diagnostic algorithms for distinguishing between control and treatment groups. The support vector machines for classification between the control group (0 µmol L-1) and cell groups treated with 10.5 and 14 µmol L-1 cisplatin for 24 hours have achieved good diagnostic results with a high sensitivity of 100%, specificity of 100% and accuracy of 100%, respectively, indicating that 10.5 µmol L-1 can be used as an appropriate therapeutic dose. Using the same method, the diagnostic sensitivity, specificity and accuracy between the control group (0 hours) and cell groups treated with 10.5 µmol L-1 cisplatin for 24 and 36 hours are all 100%, showing that 24 hours can be used as an appropriate therapeutic time. These results showed that Raman spectroscopy in conjunction with multivariate statistical analysis could be a useful tool for evaluating the cytotoxicity induced by cisplatin in human leukemic cells.

Double-Needle Bidirectional Barbed Wire Continuous Layered Suture Technique for Laparoscopic Stage I Common Bile Duct Surgery.

Background: Laparoscopic common bile duct exploration (LCBDE) proves a safe and effective treatment for choledochal stones. After LCBDE, preferred choledochal closure is favored for short- and long-term outcomes compared with t-tube drainage. However, there are no relevant studies on the technique of layered closure of the common bile duct with double-needle bidirectional barbed suture at home and abroad. Materials and Methods: A retrospective study of 37 patients who underwent laparoscopic choledochotomy from January 2021 to October 2023 in our hospital was performed. A continuous layered one-stage suture using two-needle bidirectional barb wire. The primary outcomes were stone clearance, operative time, blood loss, and complications. Secondary outcomes were complications, length of hospitalization, and time to drain removal. Results: During the study period, laparoscopic surgery was successful in all cases, and the initial stones were removed without complications. Conclusion: The treatment of choledocholithiasis with continuous layered one-stage suture with double-needle bidirectional barbed wire after LCBDE is a new convenient and effective treatment in selected patients.

Pharmacological Mechanism of Chinese Medicine in Systemic Lupus Erythematosus: A Narrative Review.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disorder affecting multiple systems, characterized by the development of harmful autoantibodies and immune complexes that lead to damage in organs and tissues. Chinese medicine (CM) plays a role in mitigating complications, enhancing treatment effectiveness, and reducing toxicity of concurrent medications, and ensuring a safe pregnancy. However, CM mainly solves the disease comprehensively through multi-target and multi-channel regulation process, therefore, its treatment mechanism is often complicated, involving many molecular links. This review introduces the research progress of pathogenesis of SLE from the aspects of genetics, epigenetics, innate immunity and acquired immunity, and then discusses the molecular mechanism and target of single Chinese herbal medicine and prescription that are commonly used and effective in clinic to treat SLE.

A hippocampal circuit mechanism to balance memory reactivation during sleep.

Memory consolidation involves the synchronous reactivation of hippocampal cells active during recent experience in sleep sharp-wave ripples (SWRs). How this increase in firing rates and synchrony after learning is counterbalanced to preserve network stability is not understood. We discovered a network event generated by an intrahippocampal circuit formed by a subset of CA2 pyramidal cells to cholecystokinin-expressing (CCK+) basket cells, which fire a barrage of action potentials ("BARR") during non-rapid eye movement sleep. CA1 neurons and assemblies that increased their activity during learning were reactivated during SWRs but inhibited during BARRs. The initial increase in reactivation during SWRs returned to baseline through sleep. This trend was abolished by silencing CCK+ basket cells during BARRs, resulting in higher synchrony of CA1 assemblies and impaired memory consolidation.

Mobile Network Coverage Prediction Using Multi-Modal Model Based on Deep Neural Networks and Semantic Segmentation.

A coverage prediction model helps network operators find coverage gaps, plan base station locations, evaluate quality of service, and build radio maps for spectrum sharing, interference management, localization, etc. Existing coverage prediction models rely on the height and transmission power of the base station, or the assistance of a path loss model. All of these increase the complexity of large-scale coverage predictions. In this paper, we propose a multi-modal model, DNN-SS, which combines a DNN (deep neural network) and SS (semantic segmentation) to perform coverage prediction for mobile networks. Firstly, DNN-SS filters the samples with a geospatial-temporal moving average filter algorithm, and then uses a DNN to extract numerical features. Secondly, a pre-trained model is used to perform semantic segmentation of satellite images of the measurement area. Thirdly, a DNN is used to extract features from the results after semantic segmentation to form environmental features. Finally, the prediction model is trained on the dataset consisting of numerical features and environmental features. The experimental results on campus show that for random location prediction, the model achieves a RMSE (Root Mean Square Error) of 1.97 dB and a MAE (Mean Absolute Error) of 1.41 dB, which is an improvement of 10.86% and 10.2%, respectively, compared with existing models. For the prediction of a test area, the RMSE and MAE of the model are 4.32 dB and 3.45 dB, respectively, and the RMSE is only 0.22 dB lower than that of existing models. However, the DNN-SS model does not need the height, transmission power, and antenna gain of the base station, or a path loss model, which makes it more suitable for large-scale coverage prediction.

Silicon Waveguide-Integrated Platinum Telluride Midinfrared Photodetector with High Responsivity and High Speed.

The detection of mid-infrared light, covering a variety of molecular vibrational spectra, is critical for both civil and military purposes. Recent studies have highlighted the potential of two-dimensional topological semimetals for mid-infrared detection due to their advantages, including van der Waals (vdW) stacking and gapless electronic structures. Among them, mid-infrared photodetectors based on type-II Dirac semimetals have been less studied. In this paper, we present a silicon waveguide integrated type-II Dirac semimetal platinum telluride (PtTe2) mid-infrared photodetector, and further improve detection performance by using PtTe2-graphene heterostructure. For the fabricated silicon waveguide-integrated PtTe2 photodetector, with an external bias voltage of -10 mV and an input optical power of 86 nW, the measured responsivity is 2.7 A/W at 2004 nm and a 3 dB bandwidth of 0.6 MHz is realized. For the fabricated silicon waveguide-integrated PtTe2-graphene photodetector, as the external bias voltage and input optical power are 0.5 V and 0.13 µW, a responsivity of 5.5 A/W at 2004 nm and a 3 dB bandwidth of 35 MHz are obtained. An external quantum efficiency of 119% can be achieved at an input optical power of 0.376 µW.

Association of serum 25-hydroxyvitamin D concentrations with all-cause and cause-specific mortality among individuals with gout and hyperuricemia.

BACKGROUND: We aimed to probe the association of serum 25-hydroxyvitamin D [25(OH)D] concentrations with all-cause and cause-specific mortality among patients with gout and hyperuricemia (HUA). METHODS: The study included 1169 gout patients and 7029 HUA patients from the National Health and Nutrition Examination Survey (NHANES) 2007-2018 and 2001-2018, respectively. The association between serum 25(OH)D and mortality was evaluated by Cox proportional hazard and restricted cubic spline models. RESULTS: Among participants with gout and HUA, the weighted mean concentrations of serum 25(OH)D were 71.49 ± 30.09 nmol/L and 64.81 ± 26.92 nmol/L, respectively. Vitamin D deficiency occurred in 29.68% of gout patients and 37.83% of HUA patients. During 6783 person-years of follow-up among gout patients, 248 all-cause deaths occurred, among which 76 died from cardiovascular disease (CVD) and 49 died from cancer. 1375 HUA patients were recorded for all-cause mortality during 59,859 person-years of follow-up, including 427 CVD deaths and 232 cancer deaths. After multifactorial adjustment, per one-unit increment in natural log-transformed 25(OH)D was associated with lower risk of 55% all-cause mortality and 61% CVD mortality among gout patients, and a 45% reduced risk of cancer mortality among HUA patients. Restricted cubic splines showed a U-shaped relationship with all-cause and CVD mortality among HUA patients, with inflection points of 72.7 nmol/L and 38.0 nmol/L, respectively. The results were robust in subgroup and sensitivity analyses. CONCLUSIONS: Serum 25(OH)D was negatively linearly correlated with mortality among gout patients, whereas U-shaped correlated with mortality in HUA patients. These results indicate that adequate vitamin D status could prevent premature death.

Genome-wide association study of carotenoids in maize kernel.

In this study, the contents of four carotenoids in 244 maize inbred lines were detected and about three million single nucleotide polymorphisms (SNPs) for genome-wide association study to preliminarily analyze the genetic mechanism of maize kernel carotenoids. We identified 826 quantitative trait loci (QTLs) were significantly associated with carotenoids contents, and two key candidate genes Zm00001d029526 (CYP18) and Zm00001d023336 (wrky91) were obtained. In addition, we found a germplasm IL78 with higher carotenoids. The results of this study can provide a theoretical basis for screening genes that guide kernel carotenoids selection breeding.

Regulation and Therapeutic Application of Long non-Coding RNA in Tumor Angiogenesis.

Tumor growth and metastasis rely on angiogenesis. In recent years, long non-coding RNAs have been shown to play an important role in regulating tumor angiogenesis. Here, we review the multidimensional modes and relevant molecular mechanisms of long non-coding RNAs in regulating tumor angiogenesis. In addition, we summarize new strategies for tumor anti-angiogenesis therapies by targeting long non-coding RNAs. The aim of this study is to provide new diagnostic targets and treatment strategies for anti-angiogenic tumor therapy.

Tailoring cell sheets for biomedical applications.

Cell sheet technology has emerged as a novel scaffold-free approach for cell-based therapies in regenerative medicine. Techniques for harvesting cell sheets are essential to preserve the integrity of living cell sheets. This review provides an overview of fundamental technologies to fabricate cell sheets and recent advances in cell sheet-based tissue engineering. In addition to the commonly used temperature-responsive systems, we introduce alternative approaches, such as ROS-induced, magnetic-controlled, and light-induced cell sheet technologies. Moreover, we discuss the modification of the cell sheet to improve its function, including stacking, genetic modification, and vascularization. With the significant advances in cell sheet technology, cell sheets have been widely applied in various tissues and organs, including but not limited to the lung, cornea, cartilage, periodontium, heart, and liver. This review further describes both the preclinical and clinical applications of cell sheets. We believe that the progress in cell sheet technology would further propel its biomedical applications.

Low-loss and low-cross talk polarization-insensitive multimode silicon waveguide crossing.

A polarization-insensitive multimode silicon waveguide crossing is investigated and experimentally characterized in this Letter. By employing the particle swarm optimization (PSO) algorithm and finite difference time domain (FDTD) method, the lengths and widths of the waveguides in the proposed device are optimized for attaining wide bandwidth, small insertion loss (IL), low cross talk (CT), and compact size. Measurement results reveal that the footprint of the presented device is 11.92 µm × 11.92 µm. From 1520 to 1600 nm, the measured insertion loss and cross talk are smaller than 0.67 dB and -28.6 dB in the case of the TE0 mode, lower than 0.65 dB and -28.7 dB in the case of the TE1 mode, less than 0.48 dB and -36.3 dB in the case of the TM0 mode, and lower than 0.62 dB and -28 dB in the case of the TM1 mode.

Low Expression of SCN4B Predicts Poor Prognosis in Non-Small Cell Lung Cancer.

BACKGROUND: Sodium voltage-gated channel beta subunit 4 (SCN4B) plays a suppressive role in various tumors. However, the role of SCN4B in non-small cell lung cancer (NSCLC) is not yet clear. This study aims to investigate the expression of SCN4B in NSCLC patients and its correlation with prognosis. METHODS: Firstly, the expression of SCN4B in non-small cell lung cancer (NSCLC) was analyzed using The Cancer Genome Atlas (TCGA) database. Then, differential expression genes (DEGs) were identified using R software. Next, DEG enrichment pathways were analyzed using the R package clusterProfiler. Protein-protein interaction networks were revealed through STRING analysis. A heatmap showed significant differential expression of SCN4B. Further analysis included examining SCN4B expression in a pan-cancer context and its correlation with 24 types of immune cells in NSCLC. Subsequently, quantitative real-time polymerase chain reaction (qRT-PCR), Western Blot, immunohistochemistry, and clinical data were used to validate SCN4B expression and prognostic value in NSCLC patients. RESULTS: SCN4B mRNA expression in non-small cell lung cancer tissues was significantly lower than in adjacent normal tissues (p < 0.001). Clinical correlation analysis confirmed its association with clinical pathology. Gene set enrichment analysis (GSEA) and tumor immune-related analyses indicated that SCN4B is involved in NSCLC-related Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways and participates in immune infiltration. qRT-PCR, Western Blot, and immunohistochemistry also confirmed that SCN4B is downregulated in NSCLC patients and is associated with poor prognosis. CONCLUSION: SCN4B is downregulated in tumor tissues of NSCLC patients and is associated with a poor prognosis.

Functional network reorganization after endovascular thrombectomy in patients with anterior circulation stroke.

BACKGROUND: Endovascular thrombectomy has been confirmed to be an effective therapy for acute ischemic stroke (AIS). However, how functional brain networks reorganize after restoration of blood supply in AIS patients, and whether the degree of reperfusion associates with functional network changes remains unclear. METHODS: Resting-state fMRI data were collected from 43 AIS patients with anterior circulation occlusion after thrombectomy and 37 healthy controls (HCs). Both static and dynamic functional connectivity (FC) within four advanced functional networks including dorsal attention network (DAN), ventral attention network (VAN), executive control network (ECN) and default mode network (DMN), were calculated and compared between post-thrombectomy patients and HCs, and between two subgroups of post-thrombectomy patients with different reperfusion conditions. RESULTS: As compared to HCs, patients showed significant differences in static FC of four functional networks, and in dynamic FC of DAN, ECN and DMN. Furthermore, patients with better reperfusion conditions exhibited increased static FC with precuneus, and altered dynamic FC within precuneus. Moreover, these alterations were associated with clinical assessments of stroke severity and functional recovery in post-thrombectomy patients. CONCLUSIONS: Collectively, these findings may provide the potential imaging markers for assessment of thrombectomy efficacy and help establish the specific rehabilitation treatments for post-thrombectomy patients.

MicroRNA-19b exacerbates systemic sclerosis through promoting Th9 cells.

Systemic sclerosis (SSc) is a chronic autoimmune disease characterized by fibrosis of the skin and multiple vital organs, but the immunological pathogenesis of SSc remains unclear. We show here that miR-19b promotes Th9 cells that exacerbate SSc. Specifically, miR-19b and interleukin (IL)-9 increase in CD4+ T cells in experimental SSc in mice induced with bleomycin. Inhibiting miR-19b reduces Th9 cells and ameliorates the disease. Mechanistically, transforming growth factor beta (TGF-ß) plus IL-4 activates pSmad3-Ser213 and TRAF6-K63 ubiquitination by suppressing NLRC3. Activated TRAF6 sequentially promotes TGF-ß-activated kinase 1 (TAK1) and nuclear factor κB (NF-κB) p65 phosphorylation, leading to the upregulation of miR-19b. Notably, miR-19b activated Il9 gene expression by directly suppressing atypical E2F family member E2f8. In patients with SSc, higher levels of IL9 and MIR-19B correlate with worse disease progression. Our findings reveal miR-19b as a key factor in Th9 cell-mediated SSc pathogenesis and should have clinical implications for patients with SSc.

Plasma-assisted synthesis of ultra-fine NiCo2O4/NiCo-layered double hydroxides nanoparticles-decorated electrospun carbon nanofibers with enhanced electrochemical performance.

Nickel cobaltate/NiCo-layered double hydroxides (NiCo2O4/NiCo-LDH) as energy storage materials offer considerable potential for various applications. However, many of current methods for synthesizing NiCo2O4/NiCo-LDH suffer from long synthesis times, complex preparation process, and high temperatures and high pressures. In this study, we present a green, simple, and efficient approach known as assisted liquid-phase plasma electrolysis, which realizes the rapid fabrication of ultra-fine NiCo2O4/NiCo-LDH nanoparticle-decorated electrospun carbon nanofibers (NiCo2O4/NiCo-LDH/CNFs) composites. Ultra-fine NiCo2O4/NiCo-LDH nanoparticles (<70 nm) are uniformly deposited on the CNF surface. The CNFs are intertwined to form a highly conductive three-dimensional mesh structure, which synergizes the NiCo2O4/NiCo-LDH nanoparticles with a high specific capacitance in favor of ion/electron transport efficiency. In addition, the cooperative effect between the two phases of NiCo2O4 and NiCo-LDH further improves the electrochemical properties. The NiCo2O4/NiCo-LDH/CNFs composites exhibit a high specific capacitance of 1534.7 F/g at 1 A/g and a capacitance retention of 93.9 % after 5000 cycles. An assembled asymmetric supercapacitor using activated carbon and NiCo2O4/NiCo-LDH/CNFs composites achieves an energy density of 33.8 Wh/kg at a power density of 400 W/kg and a capacitance retention of 93.0 % after 5000 cycles. Notably, two series-connected NiCo2O4/NiCo-LDH/CNFs ASC supercapacitors can light up an LED bulb, which maintains a certain brightness even after 50 min. Hence, this work provides a new and efficient route for synthesizing carbon-based NiCo2O4/NiCo-LDH composites for use as advanced energy storage materials.