Ting-Xuan Lin

Graphene oxide (GO), a two dimensional material with high aspect ratio and polar functional groups, can physically adsorb single-strand DNA through ππ stacking, hydrogen bonding and hydrophobic interactions, making it became an attractive nanocarrier for nucleic acids. In this work, we demonstrate a strategy to target exosites I and II of thrombin simultaneously by using programmed hybrid-aptamers for enhanced anticoagulation efficiency and stability. The targeting ligand is denoted as Supra-TBA15/29 (supramolecular TBA15/29), containing TBA15 (a 15-base nucleotide, binding to the exosite I of thrombin), TBA29 (29-base nucleotide, binding to the exosite II of thrombin) and is designed to allow consecutive hybridization of TBA15 and TBA29 to form a network of TBAs (i.e., supra-TBA15/29). The programmed hybrid-aptamers (Supra-TBA15/29) were self-assembled on GO to further boost the anticoagulation activity. The Supra-TBA15/29GO composite was formed mainly through multivalent interaction between poly(adenine) from Supra-TBA15/29 and GO. We controlled the assembly of Supra-TBA15/29 on GO by regulating the preparation temperature and the concentration ratio of Supra-TBA15/29 to GO to optimize the distance between TBA15 and TBA29 units, aptamer density, and aptamer orientation on the GO surfaces for improving the inhibition of thrombin activity toward the formation of fibrin from fibrinogen. The dose-dependent thrombin clotting time (TCT) delay caused by Supra-TBA15/29–GO was >10 times longer than that of commercially available drugs including heparin, argatroban, hirudin, and warfarin. The in vitro cytotoxicity and hemolysis analyses revealed the high biocompatibility of Supra-TBA15/29–GO. In addition, the thromboelastography of whole-blood coagulation and rat-tail bleeding assays indicate the anticoagulation ability of Supra-TBA15/29–GO is superior to the most widely used anticoagulant (heparin). Our highly biocompatible Supra-TBA15/29–GO with strong multivalent binding affinity toward thrombin (dissociation constant (Kd) = 1.9  1011 M) shows great potential as an effective direct thrombin inhibitor for treatment of hemostatic disorders.

Copper/zinc superoxide dismutase (SOD1) can clear cisplatin‐ (CP‐) induced excessive reactive oxygen species (ROS), but exogenous SOD1 cannot enter cells because of its low biomembrane permeability. Cell‐penetrating peptides (CPPs) can rapidly cross plasma membranes. This study is aimed at identifying an efficient and stable CPP‐SOD1 and investigating its effects on CP‐induced nephrotoxicity. We recombined SOD1 with 14 different CPPs and purified them using an NTA‐Ni 2+ column. In in vitro experiments, CPPs‐SOD1 cell membrane penetration ability and JNK/p38 MAPK signaling pathway were evaluated using Western blotting. ROS production, mitochondrial membrane potential (MMP), and cell apoptosis were determined using flow cytometry and immunofluorescence staining in VERO and HK‐2 cells. For in vivo experiments, mice were administered PSF‐SOD1 for 2 h before cotreatment with a single CP injection for an additional 4 days. Blood and kidney samples were collected for renal function assessment (creatinine, urea nitrogen, histopathology, TUNEL assay, and JNK/p38 MAPK signaling pathway). Compared with TAT‐SOD1, we found that PSF‐SOD1 is more efficient at crossing the cell membrane and is stable after transduction into cells. Pretreatment with PSF‐SOD1 inhibited CP‐induced apoptosis, ROS generation, and JNK/p38 MAPK activation and restored CP‐induced MMP loss in VERO and HK‐2 kidney cells. Treatment of mice with PSF‐SOD1 inhibited CP‐induced serum creatinine, blood urea nitrogen elevation, and JNK/p38 MAPK activation. H&E staining and TUNEL assay indicated that kidney tissue damage was alleviated following PSF‐SOD1 pretreatment. Overall, PSF‐SOD1 ameliorated CP‐induced renal damage by partially reducing oxidative stress and cell apoptosis by regulating JNK/p38 MAPK signaling pathway and might be a better cytoprotective agent than TAT‐SOD1.

Background: XueBiJing injection (XBJ) is renowned for its multi-target pharmacological effects, including immunomodulatory, antithrombotic, and antioxidant activities, offering potential therapeutic benefits for patients with severe infections such as sepsis and Coronavirus disease 2019 (COVID-19). Despite its clinical effectiveness, the molecular targets and mechanisms of XBJ remain unclear, warranting further investigation. Purpose: This study aimed to identify the key bioactive compounds in XBJ and elucidate their molecular targets and mechanisms. Methods: The zebrafish model was first used to evaluate the anti-inflammatory and antioxidant effects of XBJ, and the differentially expressed genes (DEGs) were identified by RNA sequencing and network analysis. Network pharmacology was used to analyze the relationship between bioactive compounds and molecular targets, and molecular docking and kinetic simulation were used to explore the target binding ability of key compounds. Cellular Thermal Shift Assay-Western Blot (CETSA-WB) and Surface Plasmon Resonance (SPR) further verified the interaction between compounds and targets; finally, the key pathways were confirmed by gene silencing experiments. Results: The zebrafish model results reveal that XBJ significantly reduced neutrophil and macrophage counts in a dose-dependent manner, emphasizing its potent anti-inflammatory effects. A transcriptomic analysis highlighted the differential expression of key genes in the KEAP1/NRF2 pathway, including HMOX1, SLC7A11, NQO1, and TXNRD1. A network analysis further pinpointed KEAP1 as a central molecular target, with tanshinone IIA, baicalein, and luteolin identified as key active compounds modulating this pathway. Among these, tanshinone IIA and baicalein exhibited strong binding interactions with KEAP1, which were confirmed through molecular docking and kinetic simulations. Further validation showed that baicalein directly targets KEAP1, as demonstrated by CETSA-WB and SPR analysis. Additionally, the gene silencing experiments of KEAP1 and NRF2 reinforced their crucial roles in activating the KEAP1/NRF2 pathway. Conclusion: These findings collectively establish baicalein as a critical bioactive compound in XBJ, driving its antioxidant and anti-inflammatory effects via KEAP1/NRF2 pathway activation through direct binding to KEAP1, providing new insights into the mechanism of action of XBJ.

As an agri-food by-product, the rice bran of pigmented rice, encompassing varieties such as red, black, and purple rice, has garnered increasing attention due to its richness in terms of bioactive compounds. Being mainly composed of the pericarp, aleuron, seed coat, and germ, the brown outer layer of the rice kernel offers potential health benefits and has applications in skincare. Human skin serves as the primary barrier against external threats, including pathogens, pollutants, and ultraviolet (UV) radiation. Notably, UV radiation accelerates the aging process and contributes to various skin issues. Recent trends suggest a heightened interest in incorporating pigmented rice into skincare regimens, motivated by its potential to mitigate oxidative stress, inflammation, and pigmentation, which are pivotal factors in skin aging and photodamage. With increasing consumer demand for natural and sustainable ingredients, pigmented rice has emerged as a promising candidate within the skincare and personal care sectors, effectively bridging the gap between nutrition and dermatological health. This review examines the applications of pigmented rice in skincare, with a particular focus on its bioactive components and potential mechanisms of action that contribute to skin health. The unique chemical composition of pigmented rice, which includes compounds such as anthocyanins, flavonoids, phenolic acids, and vitamin E, underlies its antioxidant, anti-inflammatory, and skin-protective properties. Despite the increasing recognition of its benefits, a comprehensive understanding of the underlying mechanisms remains limited, underscoring the necessity for further research to exploit the potential of pigmented rice in skincare applications fully.

Bloodstream infections (BSIs) pose a significant diagnostic challenge, largely due to the limitations of traditional methods such as blood cultures. These methods often yield low positive rates, have lengthy processing times that delay treatment, and are limited in detecting only a narrow range of pathogens. Such delays and inaccuracies can critically impede timely clinical interventions, potentially compromising patient outcomes. Next-generation sequencing (NGS) is a powerful tool for rapid, precise pathogen identification. While metagenomic NGS (mNGS) offers broad pathogen coverage, it is often costly and complex. Targeted NGS (tNGS), however, focuses on key regions of clinically relevant pathogens, reducing costs and simplifying workflows while maintaining high sensitivity, making it more practical for routine diagnostics. In this study, we introduce a novel approach combining a human cell-specific filtration membrane with a multiplex tNGS panel to overcome these challenges. The filtration membrane, designed with surface charge properties to be electrostatically attractive to leukocytes for the selective capture of specific cells, demonstrated high efficiency in removing host cells and nucleic acids, achieving over a 98% reduction in host DNA and thereby minimizing background interference in pathogen detection. Additionally, we developed an effective multiplex tNGS panel targeting over 330 clinically relevant pathogens and verified its consistency with mNGS and blood culture results, demonstrating a significant improvement in detection sensitivity. By integrating these two methods, we achieved a synergistic enhancement in diagnostic capability, boosting pathogen reads by 6- to 8-fold, which enabled reliable identification even in cases of low-abundance pathogens. This approach provides faster, more accurate, and more sensitive detection of BSIs, enabling earlier identification of infections. This facilitates timely and targeted treatment, ultimately improving patient outcomes in critical care settings. Given the unique properties of the filtration membrane and the strengths of the tNGS panel, this approach shows promising applications in prenatal and genetic health support, as well as in advancing early cancer screening strategies.