Glutathione hybrid poly (beta-amino ester)-plasmid nanoparticles for enhancing gene delivery and biosafety.

INTRODUCTION: CRISPR/Cas9 gene editing technology has significantly advanced gene therapy, with gene vectors being one of the key factors for its success. Poly (beta-amino ester) (PBAE), a distinguished non-viral cationic gene vector, is known to elevate intracellular reactive oxygen species (ROS) levels, which may cause cytotoxicity and, consequently, impact gene transfection efficacy (T.E.). OBJECTIVES: To develop a simple but efficient strategy to improve the gene delivery ability and biosafety of PBAE both in vivo and in vitro. METHODS: We used glutathione (GSH), a clinically utilized drug with capability to modulating intracellular ROS level, to prepare a hybrid system with PBAE-plasmid nanoparticles (NPs). This system was characterized by flow cytometry, RNA-seq, Polymerase Chain Reaction (PCR) and Sanger sequencing in vitro, and its safety and efficacy in vivo was evaluated by imaging, PCR, Sanger sequencing and histology analysis. RESULTS: The particle size of GSH-PBAE-plasmid NPs were 168.31 nm with a ζ-potential of 15.21 mV. An enhancement in T.E. and gene editing efficiency, ranging from 10 % to 100 %, was observed compared to GSH-free PBAE-plasmid NPs in various cell lines. In vitro results proved that GSH-PBAE-plasmid NPs reduced intracellular ROS levels by 25 %-40 %, decreased the total number of upregulated/downregulated genes from 4,952 to 789, and significantly avoided the disturbance in gene expression related to cellular oxidative stress-response and cell growth regulation signaling pathway compared to PBAE-plasmid NPs. They also demonstrated lower impact on the cell cycle, slighter hemolysis, and higher cell viability after gene transfection. Furthermore, GSH hybrid PBAE-plasmid NPs exhibited superior safety and improved tumor suppression ability in an Epstein-Barr virus (EBV)-infected murine tumor model, via targeting cleavage the EBV related oncogene by delivering CRISPR/Cas9 gene editing system and down-regulating the expression levels. This simple but effective strategy is expected to promote clinical applications of non-viral vector gene delivery.

Depth-dependent responses of soil bacterial communities to salinity in an arid region.

Soil salinization adversely affects soil fertility and plant growth in arid region worldwide. However, as the drivers of nutrient cycling, the response of microbial communities to soil salinization is poorly understood. This study characterized bacterial communities in different soil layers along a natural salinity gradient in the Karayulgun River Basin, located northwest of the Taklimakan desert in China, using the 16S rRNA Miseq-sequencing technique. The results revealed a significant filtering effect of salinity on the bacterial community in the topsoil. Only the α-diversity (Shannon index) in the topsoil (0-10 cm) significantly decreased with increasing salinity levels, and community dissimilarity in the topsoil was enhanced with increasing salinity, while there was no significant relationship in the subsoil. BugBase predictions revealed that aerobic, facultatively anaerobic, gram-positive, and stress-tolerant bacterial phenotypes in the topsoil was negatively related to salinity. The average degree and number of modules of the bacterial co-occurrence network in the topsoil were lower under higher salinity levels, which contrasted with the trends in the subsoil, suggesting an unstable bacterial network in the topsoil caused by higher salinity. The average path length among bacterial species increased in both soil layers under high salinity conditions. Plant diversity and available nitrogen were the main drivers affecting community composition in the topsoil, while available potassium largely shaped community composition in the subsoil. This study provides solid evidence that bacterial communities adapt to salinity through the adjustment of microbial composition based on soil depth. This information will contribute to the sustainable management of drylands and improved predictions and responses to changes in ecosystems caused by climate change.

Dialdehyde starch cross-linked aminated gelatin sponges with excellent hemostatic performance and biocompatibility.

Developing a hemostatic material suitable for rapid hemostasis remains a challenge. This study presents a novel aminated gelatin sponge cross-linked with dialdehyde starch, exhibiting excellent biocompatibility and hemostatic ability. This aminated gelatin sponge features hydrophilic surface and rich porous structure with a porosity of up to 80 %. The results show that the aminated gelatin sponges exhibit superior liquid absorption capacity and can absorb up to 30-50 times their own mass of simulated body fluid within 5 min. Compared with the commercial gelatin hemostatic sponge and non-aminated gelatin hemostatic sponge, the aminated gelatin hemostatic sponge can accelerate the hemostatic process through electrostatic interactions, demonstrating superior hemostatic performance in both in vitro and in vivo hemostasis tests. The aminated gelatin sponge can effectively control the hemostatic time within 80 s in the in vivo rat femoral artery injury model, significantly outperforming both commercial and non-aminated gelatin sponges. In addition, the aminated gelatin sponge also exhibits good biocompatibility and certain antibacterial properties. The proposed aminated gelatin sponge has very good application prospects for the management of massive hemorrhage.

Unsupervised underwater shipwreck detection in side-scan sonar images based on domain-adaptive techniques.

Underwater object detection based on side-scan sonar (SSS) suffers from a lack of finely annotated data. This study aims to avoid the laborious task of annotation by achieving unsupervised underwater object detection through domain-adaptive object detection (DAOD). In DAOD, there exists a conflict between feature transferability and discriminability, suppressing the detection performance. To address this challenge, a domain collaborative bridging detector (DCBD) including intra-domain consistency constraint (IDCC) and domain collaborative bridging (DCB), is proposed. On one hand, previous static domain labels in adversarial-based methods hinder the domain discriminator from discerning subtle intra-domain discrepancies, thus decreasing feature transferability. IDCC addresses this by introducing contrastive learning to refine intra-domain similarity. On the other hand, DAOD encourages the feature extractor to extract domain-invariant features, overlooking potential discriminative signals embedded within domain attributes. DCB addresses this by complementing domain-invariant features with domain-relevant information, thereby bolstering feature discriminability. The feasibility of DCBD is validated using unlabeled underwater shipwrecks as a case study. Experiments show that our method achieves accuracy comparable to fully supervised methods in unsupervised SSS detection (92.16% AP50 and 98.50% recall), and achieves 52.6% AP50 on the famous benchmark dataset Foggy Cityscapes, exceeding the original state-of-the-art by 4.5%.

Combination therapy with oncolytic virus and T cells or mRNA vaccine amplifies antitumor effects.

Antitumor therapies based on adoptively transferred T cells or oncolytic viruses have made significant progress in recent years, but the limited efficiency of their infiltration into solid tumors makes it difficult to achieve desired antitumor effects when used alone. In this study, an oncolytic virus (rVSV-LCMVG) that is not prone to induce virus-neutralizing antibodies was designed and combined with adoptively transferred T cells. By transforming the immunosuppressive tumor microenvironment into an immunosensitive one, in B16 tumor-bearing mice, combination therapy showed superior antitumor effects than monotherapy. This occurred whether the OV was administered intratumorally or intravenously. Combination therapy significantly increased cytokine and chemokine levels within tumors and recruited CD8+ T cells to the TME to trigger antitumor immune responses. Pretreatment with adoptively transferred T cells and subsequent oncolytic virotherapy sensitizes refractory tumors by boosting T-cell recruitment, down-regulating the expression of PD-1, and restoring effector T-cell function. To offer a combination therapy with greater translational value, mRNA vaccines were introduced to induce tumor-specific T cells instead of adoptively transferred T cells. The combination of OVs and mRNA vaccine also displays a significant reduction in tumor burden and prolonged survival. This study proposed a rational combination therapy of OVs with adoptive T-cell transfer or mRNA vaccines encoding tumor-associated antigens, in terms of synergistic efficacy and mechanism.