Tailor-Made Cell-Based Biomimetic Nanoprobes for Fluorescence Imaging Guided Colorectal Cancer Chemo-immunotherapy.

Colorectal cancer has become one of the malignant tumors with a high rate of morbidity and mortality. Therefore, how to effectively treat colorectal cancer is crucial. Although nanodelivery system has been applied to the therapy of colorectal cancer, the majority of existing nanodelivery systems still have drawbacks such as low biocompatibility and poor targeting ability. In this work, tailor-made cell-based biomimetic nanoplatform was prepared to enhance the targeting and therapeutic effect for colorectal cancer chemo-immunotherapy. First, hollow long persistence luminescence nanomaterials were synthesized with superior background-free bioimaging effect and high drug-loading content. After loaded with cisplatin, the nanoplatform was camouflaged with tailor-made erythrocyte and programmed cell death receptor 1 (PD-1) expressed hybrid cell membrane. In vivo animal imaging experiment showed that the nanoplatform camouflaged with hybrid cell membrane not only had excellent immune escapability but also had excellent tumor active targeting ability. In vivo anticancer experiments showed that combined chemotherapy and immunotherapy of the nanoplatform could significantly inhibit tumor growth in tumor-bearing mice. In summary, the tailor-made cell-based membrane camouflage produced excellent immune escapability and cancer active targeting ability, providing a modality for biomimetic nanodelivery systems.

Newly Constructed NiCo2O4 Derived from ZIF-67 with Dual Mimic Enzyme Properties for Colorimetric Detection of Biomolecules and Metal Ions.

Integration of novel bio-/nanostructures as effective sensing platforms is still of great significance for robust and rapid analysis. Herein, a novel metal-organic framework-derived NiCo2O4 was synthesized via a feasible templating method. Significantly, redox couples of both Ni3+/Ni2+ and Co3+/Co2+ provided richer oxidation-reduction reactions, thereby leading to an enhanced catalytic activity. Furthermore, NiCo2O4 as an enzyme mimic with peroxidase-like activity and oxidase-like activity could oxidize colorless thylbenzidine (TMB) to blue oxTMB in the absence of H2O2. Thus, a sensitive chromogenic sensing platform for detecting Fe2+, thiourea, cysteine (Cys), and epigallocatechin-3-gallate (EGCG) was proposed. The colorimetric detection methods exhibited great features of low limit of detection (LOD) and broad linear range. Owing to the complexation reaction, the chromogenic sensing system of TMB + NiCo2O4 + Cys achieved effective detection of Cu2+ and Mn2+ with the LODs of 0.0022 and 0.0181 mM, respectively. Developed detection methods with wide linear ranges of 0.008-0.1 mM for Cu2+ and 0.08-1 mM for Mn2+ had excellent practical potential. Similarly, the reaction system of TMB + NiCo2O4 + EGCG could achieve the colorimetric detection of Cu2+ and Fe3+. The great chromogenic sensing performance for detecting Cu2+ and Fe3+ with a broad linear range and a low LOD could be also realized.

Promotion Effect of EGCG on the Raised Expression of IL-23 through the Signaling of STAT3-BATF2-c-JUN/ATF2.

Tea polyphenol of epigallocatechin-3-gallate (EGCG) has been verified to possess multiple biological activities. Interleukin-23 (IL-23) is a heterodimeric cytokine consisting of two subunits of IL-23p19 and IL-12p40, with the functionality in regulating the production of cytokines under physiological or pathological conditions. By serendipity, the raised expression of IL-23 was observed after treating cells with EGCG, whereas the detailed mechanism remains poorly understood. This study was proposed to investigate the signaling related to EGCG-induced IL-23. The raised expression of IL-23 was confirmed primarily by intraperitoneally injecting with different concentrations of EGCG (0, 20, 50, 80 mg/kg) into BALB/c mice, and the raised expression was confirmed by quantitative real-time polymerase chain reaction (qRT-PCR) and western blot. Results from enzyme-linked immunosorbent assay (ELISA) revealed the increase of IL-23 in serum from 116.09 to 153.90 pg/mL after treating with EGCG. The same results were also observed in RAW264.7 and peritoneal macrophages after treating with EGCG (0, 1, 5, 10, 25 µM) with the increased tendency of IL-23 in cultural medium (7.98 to 25.38 pg/mL for RAW264.7; 3.64 to 260.93 pg/mL for peritoneal macrophages). After preliminary exploration of the signaling related to the increased IL-23, the classical signaling pathways and key transcription factors, such as nuclear factor kappa-B (NF-κB), mitogen-activated protein kinase (MAPK) signaling pathways, and interferon regulatory factor 5 (IRF5), were demonstrated with no relevant contribution. A further study revealed the involvement of the key transcription factor of BATF2, which could antagonistically modulate the transcription and translation of IL-23. The signaling of STAT3-BATF2-c-JUN/ATF2-IL-23 has been further verified in RAW264.7 macrophages using the STAT3 inhibitor of AG490 and the activator of Colivelin TFA. The results indicated that EGCG inhibits the phosphorylation of STAT3 to facilitate the decreased level of BATF2, which contributed to the increased level of IL-23 by the enhancing heterodimerization of c-JUN and ATF2.

Construction of pH-Dependent Nanozymes with Oxygen Vacancies as the High-Efficient Reactive Oxygen Species Scavenger for Oral-Administrated Anti-Inflammatory Therapy.

It is of great significance to eliminate excessive reactive oxygen species (ROS) for treating inflammatory bowel disease (IBD). Herein, for the first time, a novel nanozyme NiCo2 O4 @PVP is constructed via a step-by-step strategy. Noticeably, the existence of oxygen vacancy in the NiCo2 O4 @PVP is helpful for capturing oxygenated compounds, while both redox couples of Co3+ /Co2+ and Ni3+ /Ni2+ will offer richer catalytic sites. As expected, the obtained NiCo2 O4 @PVP exhibits pH-dependent multiple mimic enzymatic activities. Benefiting from the introduction of polyvinylpyrrolidone (PVP), the NiCo2 O4 @PVP possesses good physiological stability and excellent biosafety in stomach and intestines' environment. Meanwhile, the NiCo2 O4 @PVP also presents strong scavenging activities to ROS in vitro, including • O2- , H2 O2 , as well as • OH. Furthermore, a dextran sodium sulfate (DSS)-induced colitis model is established for evaluating the anti-inflammatory activity of NiCo2 O4 @PVP in vivo. Based on the size-mediated and charge-mediated mechanisms, the nanozyme can pass through the digestive tract and target the inflamed site for oral-administrated anti-inflammatory therapy. More interestingly, compared with the model group, the expression levels of inflammatory factors (e.g., Interleukin- 6 (IL-6), Interleukin- 1ß (IL-1ß), tumor necrosis factor-α (TNF-α), and inducible nitric oxide synthase (iNOS)) in colon of mice show a significant decrease after nanozyme intervention, thereby inhibiting the development of IBD. In short, current work provides an alternative therapy for patients suffering from IBD.

Selection of specific nanobodies to develop an immuno-assay detecting Staphylococcus aureus in milk.

The interaction between conventional immunoglobulins (Igs) and the Ig-binding surface proteins of Staphylococcus aureus (S. aureus) have obstructed the development of immuno-assays to detect these bacteria. The current study aimed to select nanobodies (Nbs) recognizing specifically S. aureus and to establish an immuno-assay to uncover S. aureus contaminations in foods. An alpaca was immunized with an inactivated S. aureus strain followed by the construction of a Nb library from which four target-specific Nbs were retrieved. Subsequently, a sandwich ELISA employing the Nb147 and biotinylated-Nb147 pair to capture and to detect S. aureus, respectively, was established to possess a detection limit of 1.4 × 105 colony forming units (CFU)/mL. The dedicated immuno-assay has been verified by detecting 10 CFU/mL of S. aureus in milk samples after an 8 h-enrichment step. This study provides the basis of an easy, reproducible and effective immuno-assay to screen for S. aureus contaminations in foods.

Selection of Specific Nanobodies against Lupine Allergen Lup an 1 for Immunoassay Development.

The declaration of lupine supplements is mandatory to avoid lupine allergy for sensitive individuals. However, reliable detection methods against lupine allergen remain critical to prevent the unintended consumption of allergen contaminated food. In this study, we have immunized an alpaca with lupine protein extracts and retrieved nanobodies (Nbs). Nevertheless, the target antigen has been recognized as Lup an 1, which has been classified as ß-conglutin, and confirmed to connect with lupine allergy. After selection of the best Nb-pair, a sandwich enzyme-linked immunosorbent assay (ELISA) has been developed providing a linear range of 0.036-4.4 µg/mL with detection limit of 1.15 ng/mL. This immunoassay was confirmed by detecting the samples with spiked allergen, and a recovery from 86.25% to 108.45% with coefficient of variation (CV) less than 4.0% has been determined. Generally, this study demonstrated the selection of Nbs against allergen with crude protein content to develop the immunoassay for lupine surveillance in foods.

Unbiased Immunization Strategy Yielding Specific Nanobodies against Macadamia Allergen of Vicilin-like Protein for Immunoassay Development.

Macadamia nut contains important food allergens that potentially cause allergic reactions with severe adverse effects in infants and adults. Reliable and accurate detection of macadamia is critical to avoid allergic reactions. However, knowledge on macadamia allergen is scarce and a reliable detection method has not been reported, yet. In this study, an unbiased immunization and selection strategy was employed to select nanobodies (Nbs) recognizing specifically macadamia allergen, as well as to establish a detection method to unveil a macadamia protein contamination. An alpaca was immunized with a crude protein extract of macadamia followed by construction of a Nb library from its lymphocytes. The panning and screening of this immune Nb repertoire resulted in the selection of six target-specific Nbs. Nb-mediated immuno-capturing combined with mass spectrometry allowed us to identify the target as the macadamia vicilin-like antimicrobial peptides 2-3 (MiAMP2), a novel food allergenic protein abbreviated as Mac i 1. Later on, an immunoassay of a heterologous sandwich ELISA method based on the selected Nb-pairs was established, providing a linear response in the range of 0.442-2,800 µg/mL and with a limit of detection of 27.1 ng/mL. The dedicated immunoassay has been verified by detecting the antigen spiked in food samples. Our study provided evidence for the successful application of the unprejudiced strategy to retrieve Nbs against a priori undefined macadamia allergen. These target-specific Nbs were used to design a highly reliable and effective immunoassay.

A Novel Mediation Strategy of DSS-Induced Colitis in Mice Based on an Iron-Enriched Probiotic and In Vivo Bioluminescence Tracing.

Iron deficiency (ID) caused by blood loss and/or reduced iron absorption is a serious problem influencing health in inflammatory bowel disease (IBD). However, traditional iron supplements may fail to meet no side effect demands for ID of IBD; thus, a new iron supplementation is highly desired to be developed. Herein, for the first time, probiotic Lactobacillus alimentarius NKU556 with an iron-enriching ability was screened from Chinese traditional fermented food then employed to intervene DSS-induced colitis with bioluminescence tracing in mice. As expected, oral administration with NKU556-Fe can remarkably enhance the expression of tight junction proteins and effectively reduce the pro-inflammatory cytokines as well as the oxidative stress on DSS-induced colitis in mice. Meanwhile, in comparison with the FeSO4 group, the intake of NKU556-Fe could suppress the expression of hepcidin derived from the liver and reduce the degradation of FPN1, thereby leading to the increase in the iron absorption of colitis in mice. According to the bioluminescence result, it was believed that the beneficial effects of oral administration with NKU556/NKU556-Fe on DSS-induced colitis in mice were hardly related to its metabolites but associated with its own function. These results concluded that the oral administration of NKU556-Fe could relieve colitis inflammation and increase iron absorption. In summary, current work not only proposed a novel mediation strategy for IBD but also offered some inspirations for future treatment of extraintestinal complications.

Assessment of migration regularity of phthalates from food packaging materials.

Phthalate acid esters (PAEs) are one of the essential plastic additives which may lead to plenty of harmful effects, including reproductive toxicity, teratogenicity, and carcinogenicity. Increasing attention has been paid to the migration of plasticizer. In this article, the disposable plastic lunch boxes were taken as the research object. The result showed that dibutyl phthalate (DBP) and diisobutyl phthalate (DIBP) have been mainly found, whose content was 1.5 mg/kg and 2.4 mg/kg, respectively. The LOD was 2 ng/g, and LOQ was 6.7 ng/g. We further investigated the migration of PAEs into the simulated liquid at different temperature conditions. Then, the linear fitting performing by first-order kinetic migration model revealed that the lower the polarity of the simulated liquid, the larger the rate constant K 1 and initial release rate V 0. The higher the temperature, the bigger the K 1 and V 0.