A non-viral DNA delivery system consisting of multifunctional chimeric peptide fused with zinc-finger protein.

Non-viral gene delivery systems have received sustained attention as a promising alternative to viral vectors for disease treatment and prevention in recent years. Numerous methods have been developed to enhance gene uptake and delivery in the cytoplasm; however, due to technical difficulties and delivery efficiency, these systems still face challenges in a range of biological applications, especially in vivo. To alleviate this challenge, we devised a novel system for gene delivery based on a recombinant protein eTAT-ZF9-NLS, which consisted of a multifunctional chimeric peptide and a zinc-finger protein with sequence-specific DNA-binding activity. High transfection efficiency was observed in several mammalian cells after intracellular delivery of plasmid containing ZF9-binding sites mediated by eTAT-ZF9-NLS. Our new approach provides a novel transfection strategy and the transfection efficiency was confirmed both in vitro and in vivo, making it a preferential transfection reagent for possible gene therapy.

Orchestrated Codelivery of Peptide Antigen and Adjuvant to Antigen-Presenting Cells by Using an Engineered Chimeric Peptide Enhances Antitumor T-Cell Immunity.

The intrinsic pharmacokinetic limitations of traditional peptide-based cancer vaccines hamper effective cross-presentation and codelivery of antigens (Ag) and adjuvants, which are crucial for inducing robust antitumor CD8+ T-cell responses. In this study, we report the development of a versatile strategy that simultaneously addresses the different pharmacokinetic challenges of soluble subunit vaccines composed of Ags and cytosine-guanosine oligodeoxynucleotide (CpG) to modulate vaccine efficacy via translating an engineered chimeric peptide, eTAT, as an intramolecular adjuvant. Linking Ags to eTAT enhanced cytosolic delivery of the Ags. This, in turn, led to improved activation and lymph node-trafficking of Ag-presenting cells and Ag cross-presentation, thus promoting Ag-specific T-cell immune responses. Simple mixing of eTAT-linked Ags and CpG significantly enhanced codelivery of Ags and CpG to the Ag-presenting cells, and this substantially augmented the adjuvant effect of CpG, maximized vaccine immunogenicity, and elicited robust and durable CD8+ T-cell responses. Vaccination with this formulation altered the tumor microenvironment and exhibited potent antitumor effects, with generally further enhanced therapeutic efficacy when used in combination with anti-PD1. Altogether, the engineered chimeric peptide-based orchestrated codelivery of Ag and adjuvant may serve as a promising but simple strategy to improve the efficacy of peptide-based cancer vaccines.

Biomolecules-mediated electrochemical signals of Cu2+: Y-DNA nanomachines enable homogeneous rapid one-step assay of lung cancer circulating tumor cells.

This study presents a straightforward efficient technique for extracting circulating tumor cells (CTCs) and a rapid one-step electrochemical method (45 min) for detecting lung cancer A549 cells based on the specific recognition of mucin 1 using aptamers and the modulation of Cu2+ electrochemical signals by biomolecules. The CTCs separation and enrichment process can be completed within 45 min using lymphocyte separation solution (LSS), erythrocyte lysis solution (ELS), and three centrifugations. Besides, the influence of various biomolecules on Cu2+ electrochemical signals is comprehensively discussed, with DNA nanospheres selected as the medium. Three single-stranded DNA sequences were hybridized to form Y-shaped DNA (Y-DNA), creating DNA nanospheres. Upon specific capture of mucin 1 by the aptamer, most DNA nanospheres could form complexes with Cu2+ (DNA nanosphere-Cu2+), significantly reducing the concentration of free Cu2+. Our approach yielded the limit of detection (LOD) of 2 ag/mL for mucin 1 and 1 cell/mL for A549 cells. 39 clinical blood samples were used for further validation, yielding results closely correlated with pathological, computed tomography (CT) scan findings and folate receptor-polymerase chain reaction (FR-PCR) kits. The receiver operating characteristic (ROC) curve displayed an area under the curve (AUC) value of 0.960, demonstrating 100% specificity and 93.1% sensitivity for the assay. Taken together, our findings indicate that this straightforward and efficient pretreatment and rapid, highly sensitive electrochemical assay holds great promise for liquid biopsy-based tumor detection using CTCs.

Endoplasmic Reticulum Peroxynitrite Fluctuations in Hypoxia-Induced Endothelial Injury and Sepsis with a Two-Photon Fluorescence Probe.

Sepsis is a serious systemic inflammatory disease that frequently results in death. Early diagnosis and timely targeted interventions could improve the therapeutic effect. Recent work has revealed that the reactive oxygen species (ROS) in the endoplasmic reticulum (ER) and hypoxia-induced endothelial injury play significant roles in sepsis. However, the relationship between the levels of peroxynitrite (ONOO-) and hypoxia-induced endothelial injury as well as different states of sepsis remain unexplored. Herein, we developed a unique two-photon fluorescent probe (ER-ONOO-) for detecting ONOO- in aqueous solution that has high sensitivity, high selectivity, and ultrafast response time. In addition, ER-ONOO- was successfully used to evaluate the levels of ONOO- at the ER with three kinds of methods in a hypoxia-induced endothelial injury model. Furthermore, ER-ONOO- is capable of monitoring the changes in organ fluorescence through ONOO- variation in different stages of a cecum ligation and puncture (CLP) mouse model. Moreover, we also confirmed that the endoplasmic reticulum stress and oxidative stress participated in the CLP model. Consequently, this research can provide a reliable tool for studying ONOO- fluctuation in sepsis and provide new insights into the pathogenic and therapeutic mechanisms involved.

Association between the rs4784227-CASC16 polymorphism and the risk of breast cancer: A meta-analysis.

OBJECTIVE: Although several studies have identified an association between the rs4784227-cancer susceptibility candidate gene 16 (CASC16) polymorphism and breast cancer, the results remain inconclusive. Therefore, we conducted a meta-analysis to assess the relationship between the rs4784227-CASC16 polymorphism and breast cancer risk. METHODS: Studies were searched in the PubMed, Web of Science, Embase, Google Scholar, and Cochran Library databases until June 10, 2021, to identify all potential literature on rs4784227-CASC16 polymorphism and breast cancer risk association. Fixed-effect or random-effect models were used to calculate odds ratios (ORs) and their corresponding 95% confidence intervals (95% CIs). Subgroup analyses, publication bias, and sensitivity analyses were also conducted. RESULTS: Seventeen eligible studies involving 34,719 subjects (18,445 cases and 16,274 healthy controls) from 7 articles were included in the current meta-analysis. The pooled ORs regarding the association between the rs4784227-CASC16 polymorphism and breast cancer risk were statistically significant [T vs C: OR = 1.244, 95% CI = 1.202-1.287; TT vs CT + CC: OR = 1.407, 95% CI = 1.296-1.528; CC vs CT + TT: OR = 0.777, 95% CI = 0.745-0.811; TT vs CC: OR = 1.544, 95% CI = 1.419-1.681; CT vs CC: OR = 1.244, 95% CI = 1.189-1.301]. On subgroup analysis, the rs4784227-CASC16 T/C gene has a certain correlation with breast cancer susceptibility in Asian and North American populations, but no significant risk in the Australian population. CONCLUSION: Our pooled analysis showed a significant association between the rs4784227- (T) allele and breast cancer susceptibility in Asian and North American populations, and intervention with this mutation might be a new therapeutic strategy for breast cancer. However, large-scale and well-designed studies are needed in different populations to further evaluate the role of the rs4784227-CASC16 polymorphism in breast cancer.

Multi-Classifier Fusion Based on MI-SFFS for Cross-Subject Emotion Recognition.

With the widespread use of emotion recognition, cross-subject emotion recognition based on EEG signals has become a hot topic in affective computing. Electroencephalography (EEG) can be used to detect the brain's electrical activity associated with different emotions. The aim of this research is to improve the accuracy by enhancing the generalization of features. A Multi-Classifier Fusion method based on mutual information with sequential forward floating selection (MI_SFFS) is proposed. The dataset used in this paper is DEAP, which is a multi-modal open dataset containing 32 EEG channels and multiple other physiological signals. First, high-dimensional features are extracted from 15 EEG channels of DEAP after using a 10 s time window for data slicing. Second, MI and SFFS are integrated as a novel feature-selection method. Then, support vector machine (SVM), k-nearest neighbor (KNN) and random forest (RF) are employed to classify positive and negative emotions to obtain the output probabilities of classifiers as weighted features for further classification. To evaluate the model performance, leave-one-out cross-validation is adopted. Finally, cross-subject classification accuracies of 0.7089, 0.7106 and 0.7361 are achieved by the SVM, KNN and RF classifiers, respectively. The results demonstrate the feasibility of the model by splicing different classifiers' output probabilities as a portion of the weighted features.

A Canopy Information Measurement Method for Modern Standardized Apple Orchards Based on UAV Multimodal Information.

To make canopy information measurements in modern standardized apple orchards, a method for canopy information measurements based on unmanned aerial vehicle (UAV) multimodal information is proposed. Using a modern standardized apple orchard as the study object, a visual imaging system on a quadrotor UAV was used to collect canopy images in the apple orchard, and three-dimensional (3D) point-cloud models and vegetation index images of the orchard were generated with Pix4Dmapper software. A row and column detection method based on grayscale projection in orchard index images (RCGP) is proposed. Morphological information measurements of fruit tree canopies based on 3D point-cloud models are established, and a yield prediction model for fruit trees based on the UAV multimodal information is derived. The results are as follows: (1) When the ground sampling distance (GSD) was 2.13-6.69 cm/px, the accuracy of row detection in the orchard using the RCGP method was 100.00%. (2) With RCGP, the average accuracy of column detection based on grayscale images of the normalized green (NG) index was 98.71-100.00%. The hand-measured values of H, SXOY, and V of the fruit tree canopy were compared with those obtained with the UAV. The results showed that the coefficient of determination R2 was the most significant, which was 0.94, 0.94, and 0.91, respectively, and the relative average deviation (RADavg) was minimal, which was 1.72%, 4.33%, and 7.90%, respectively, when the GSD was 2.13 cm/px. Yield prediction was modeled by the back-propagation artificial neural network prediction model using the color and textural characteristic values of fruit tree vegetation indices and the morphological characteristic values of point-cloud models. The R2 value between the predicted yield values and the measured values was 0.83-0.88, and the RAD value was 8.05-9.76%. These results show that the UAV-based canopy information measurement method in apple orchards proposed in this study can be applied to the remote evaluation of canopy 3D morphological information and can yield information about modern standardized orchards, thereby improving the level of orchard informatization. This method is thus valuable for the production management of modern standardized orchards.

Circulating miR-10a as Predictor of Therapy Response in Rheumatoid Arthritis Patients Treated with Methotrexate.

BACKGROUND: Anti-inflammatory drugs and DMARDs, especially methotrexate, have dramatically improved the outlook of RA patient. However, in some patients the treatment effect is still not satisfactory and needs to change therapeutic regimen in time. So, timely prediction of patient treatment effect is particularly important. METHODS: miRNAs are a large family of highly conserved noncoding genes and some of them become potential biomarker of disease diagnosis. Circulating miR-10a was detected by qRT-PCR. Receiver operating characteristic (ROC)curve analyses were performed for circulating miR-10a in RA diagnosis and predictor of therapy effectiveness in RA patients treated with methotrexate. RESULTS AND CONCLUSION: In the present study, we found that the circulating miR-10a was significantly decreased in RA patients compared to osteoarthritic patients and healthy people. Importantly, circulating miR-10a was up-regulated in RA patients treated with MTX. Moreover, we identified circulating miR-10a that may serve as a biomarker of RA diagnosis and predictor of therapy effectiveness in rheumatoid arthritis patients treated with methotrexate.

Characterization of Staphylococcus aureus Isolated from Non-Native Patients with Skin and Soft Tissue Infections in Shanghai.

BACKGROUND: Staphylococcus aureus is one predominant cause of skin and soft-tissue infections (SSTIs), but little information exists regarding the characterization of S. aureus from non-native patients with SSTIs in China. METHODS: In this study, we enrolled 52 non-native patients with S. aureus SSTIs, and 65 native control patients with S. aureus SSTIs in Shanghai. 52 and 65 S. aureus isolates were collected from both groups, respectively. S. aureus isolates were characterized by antimicrobial susceptibility testing, toxin gene detection, and molecular typing with sequence type, spa type, agr group and SCCmec type. RESULTS: Methicillin-resistant S. aureus (MRSA) was detected in 8 non-native patients and 14 native patients with SSTIs. Overall, antimicrobial susceptibilities of S. aureus isolated from non-native patients were found higher than those from native patients. CC59 (ST338 and ST59) was found in a total of 14 isolates (4 from non-native patients; 10 from native patients), 9 of which were carrying lukS/F-PV (3 from non-native patients; 6 from native patients). ST7 was found in 12 isolates and all 12 isolates were found in native patients. The livestock-associated clone ST398 was found in 11 isolates (6 from non-native patients; 5 from native patients), and 5 ST398 lukS/F-PV-positive methicillin-susceptible S. aureus (MSSA) were all discovered among non-native patients. The molecular epidemiology of S. aureus isolated from non-native patients was quite different from those from native patients. lukS/F-PV was more frequent in isolates originating from non-native patients with SSTIs compared to native patients (31 vs. 7, P <0.0001). CONCLUSIONS: CC59 was the most common clonal complex among patients with SSTIs in Shanghai. The other most common sequence types were ST7 and Livestock ST398. The molecular epidemiology of S. aureus isolated from non-native patients was quite different from those from native patients. S. aureus isolated from non-native patients was more likely to carry lukS/F-PV.

Molecular epidemiologic analysis of Staphylococcus aureus isolated from four burn centers.

Staphylococcus aureus has been a major cause of hospital-acquired infections. Methicillin-resistant Staphylococcus aureus (MRSA) has emerged since 1980s as an epidemiologic problem in hospitals. This old pathogen brings a new challenge to all physicians and bacteriologists. Hence, effective measures of MRSA control are in critical need. S. aureus or MRSA is one of the leading causes of infection among burn centers, resulting in a number of poor outcomes and even death. The present study performed a molecular epidemiologic analysis of S. aureus isolated from four burn centers in the southeast of China. A total of 85 isolates were collected, and molecular characters were determined for further investigation. In this study, the prevalent clone of MRSA among four burn centers was found to be SCCmec III (spa-type t030, agr I), which is resistant to 4 kinds of antimicrobials including erythromycin, clindamycin, kanamycin and mupirocin. Discrepancy between mecA detection and conventional tests used for MRSA identification was observed unintentionally. Our data demonstrated that the overall prevalence rate of MRSA was 55.3%, and drugs such as sulfamethoxazole/trimethoprim, linezolid and fusidic acid are efficient antibiotic options for treating S. aureus or MRSA infections among four burn centers studied in present investigation.