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.

[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.

Galectin-9 as a new biomarker of acute-on-chronic liver failure.

Galectin-9 (Gal-9) expression in patients with acute-on-chronic liver failure and its correlation with prognosis remain unclear. This study investigated the relationship between liver failure prognosis and Gal-9 expression analysis in patients with acute-on-chronic liver failure. Patients with acute-on-chronic liver failure attributable to hepatitis B and those with chronic hepatitis B were included in this single-center prospective cohort study. The Gal-9 levels in the acute-on-chronic liver failure group were significantly higher than those in the chronic hepatitis B group, and there was an upregulation of Gal-9 and T-cell immunoglobulin domain and mucin domain-3 expressions in peripheral blood T cells. Gal-9 was localized in the regenerative areas of liver tissues in patients with acute-on-chronic liver failure, co-localizing with Kupffer cells. Kaplan-Meier survival curves showed that patients with Gal-9 levels < 9.6 ng/ml had a worse prognosis, with the area under the receiver operating characteristic curve (AUC-ROC) being similar to that of the Model for End-Stage Liver Disease score. The combined ROC curve of the two had better predictive performance, with an AUC of 0.945. High Gal-9 levels in liver regenerative areas can serve as a prognostic marker, indicating a better prognosis for patients with hepatitis B virus-acute-on-chronic liver failure.

In vitro and in vivo synergistic inhibition of Malassezia furfur targeting cell membranes by Rosa rugosa Thunb. and Coptidis Rhizoma extracts.

Background: Malassezia furfur (M. furfur) is a prevalent dermatophyte that significantly impairs patients' quality of life. This study aimed to evaluate the synergistic antifungal effects of combined extracts from Rosa rugosa Thunb. (MG) and Coptidis Rhizoma (HL) against M. furfur, both in vitro and in vivo. Methods: High-performance liquid chromatography (HPLC) was used to identify the major active compounds present in MG and HL. The antifungal activity of the combined Meilian extract (ML) was assessed using the checkerboard method and time-kill curves. Microstructural alterations in the fungi were observed using scanning electron microscopy (SEM) and transmission electron microscopy (TEM). The impact of the extracts on the fungal cell membrane was investigated through propidium iodide staining, protein concentration assays, and ergosterol quantification. Transcriptomic analysis was conducted to elucidate the molecular mechanisms underlying the effects of the extracts. Furthermore, the synergistic antifungal effects of ML were evaluated in a mouse model of seborrheic dermatitis induced by M. furfur. Results: The study demonstrated that the combined application of MG and HL significantly affected the integrity of the M. furfur cell membrane and potentially modulated its formation processes. In the M. furfur-induced seborrheic dermatitis model, ML exhibited synergistic antifungal effects and effectively alleviated skin inflammation. These findings provide an important theoretical basis for understanding the antifungal mechanisms of ML and its potential application in dermatological therapy.

Serum dsDNA is environmentally contingent.

Psoriasis is a recurrent autoimmune disease characterized by seasonal and latitudinal variations. Double-stranded DNA (dsDNA) is a crucial component of nucleic acids and nucleosomes that provoke innate immune responses. Given the potential influence of climate on immunity and the development of autoimmune diseases, a comprehensive quantitative analysis of dsDNA levels in the population is warranted. In this case-control study conducted from 2016 to 2020, 10,110 psoriasis patients and matched controls from 12 regions in China were included. This study examined variations in serum dsDNA levels based on season and latitude. The results revealed significant associations between geographical location, climatic conditions, and season with serum dsDNA concentration. Individuals residing in Northern China exhibited significantly higher serum dsDNA levels compared to those in the South (1.00 vs. 0.96 ng/ml), and those in medium latitude regions had higher levels than their counterparts in areas with extreme latitudes (0.98 vs. 0.96 ng/ml). Furthermore, individuals in regions with low to medium ultraviolet exposure demonstrated higher serum dsDNA concentrations than those in areas with high ultraviolet levels (1.03 vs. 0.93 ng/ml), and individuals in winter showed higher levels than those in summer (1.03 vs. 0.92 ng/ml). Factors such as sex, UV index, humidity, and sunshine duration were inversely related to serum dsDNA levels, while age and daylight hours showed a positive association. These findings suggest that meteorological and climatic factors play a role in influencing serum dsDNA levels.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

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.

Myeloid-derived suppressor cells in the tumor microenvironment reduce uncoupling protein 1 expression to boost immunosuppressive activity.

Uncoupling protein 1 (UCP1) is located at the inner membrane of mitochondria and mediates nonshivering thermogenesis. Its abnormal expression is associated with metabolic diseases, cancer, and acute kidney injury. Myeloid-derived suppressor cells (MDSCs) with immunosuppressive activity accumulate in the tumor microenvironment (TME). Here, decreased UCP1 expression in MDSCs was observed in the peripheral blood of patients with colorectal cancer and transplanted mouse tumors. Aggravated tumor progression was observed in UCP1-knockout mice and conditional knockout mice (UCP1fl/fl-S100A8cre). The number of G-MDSCs and M-MDSCs increased in the transplanted tumor tissues from UCP1-deficient mice compared with those from wild-type mice. The tumor-promoting effect disappeared when the tumor-bearing mice were depleted of MDSCs by the α-DR5 administration. Adoptive transfer of tumor-derived MDSCs sharply promoted the tumor growth in vivo. Furthermore, these tumor-derived MDSCs enhanced the proliferation, reduced death, inhibited IFN-γ production of CD4+ and CD8+T cells, and induced Treg cells ex vivo. In conclusion, MDSCs in the TME alter the metabolic pattern by decreasing UCP1 expression to enhance immunosuppressive activity for tumor escape.