The global overuse of antibiotics has accelerated the spread of antibiotic resistance genes (ARGs) in water and food systems. While quantitative PCR is widely used to monitor ARG degradation, its limitations in quantifying fragmented and unculturable ARGs hinder accurate comparisons. To address this, we implemented hydrogel digital loop-mediated isothermal amplification (LAMP)-a novel visual quantification platform enabling single-molecule detection by spatially confining LAMP reactions in hydrogel microchambers. This method was applied to evaluate the degradation kinetics of two critical ARGs (blaNDM from carbapenem-resistant Escherichia coli and mecA from methicillin-resistant Staphylococcus aureus) under food-relevant treatments: heat, ultrasound, UV254, TiO2 photocatalysis, and light exposure (red/blue/white). Results revealed that mecA exhibited higher thermal sensitivity than blaNDM, with degradation rates escalating at elevated temperatures. Ultrasound degraded most of both ARGs within 20 min, while UV254 achieved near-complete inactivation in 2 min. TiO2 photocatalysis showed moderate efficacy, with mecA being more susceptible. Red light induced degradation via thermal radiation, whereas blue/white light had negligible effects within 60 min. Our hydrogel digital LAMP system provides rapid, visual, and precise ARG quantification, offering significant potential for on-site monitoring in food safety control. [GRAPHICS]

Bacteria play important roles in tumor formation, growth and metastasis through downregulating immune response and initiating drug resistance. Herein, size-tunable nanogels (NGs) have been developed to address the existing size paradox in tumor accumulation, intratumoral penetration and intracellular release of therapeutics for the treatment of Fusobacterium nucleatum (F. nucleatum)-infected colorectal cancer. Zinc-imidazolate frameworks with doxorubicin (DOX) loading and folate grafting (f-ZIFD) were mixed with metronidazole (MET) and encapsulated in NGs through thiol-ene click crosslinking of sulfhydryl hyaluronan, sulfhydryl alginate and 4-arm poly(ethylene glycol) acrylate. Hyaluronidase-initiated matrix degradation causes NG swelling to release sufficient MET and maintains a large size for an extended time period, and the gradually discharged f-ZIFD nanoparticles (NPs) from NGs exhibit acid-responsive intracellular release of DOX after folate-mediated internalization into tumor cells. The encapsulation into NGs significantly enhances the bioavailability and increases half-lives of MET and DOX by around 20 times. In the F. nucleatum-infected tumor model, the extended retention of swollen NGs and the efficient tumor infiltration and cellular uptake of the discharged f-ZIFD NPs cause 6 times higher DOX levels in tumors than that of free DOX administration. F. nucleatum promotes tumor cell proliferation and tumor growth, and the cascaded releases of MET and f-ZIFD NPs eliminate F. nucleatum to effectively inhibit tumor growth with a significant extension of animal survival. Thus, the hyaluronidase-mediated NG expansion and dual-responsive cascaded drug release have overcome challenges in the release regimen and size paradox of drug delivery carriers to combat bacteria-infected cancer.

In the harsh gastrointestinal tract, helical bacteria with hierarchical chiral architectures possess strong abilities. Taking inspirations from nature, we developed a multichiral mesoporous silica nanoscrew (L/D-MCNS) as an efficient oral drug delivery platform by modifying the structural chiral silica nanoscrew (CNS) with L/D-alanine (L/D-Ala) enantiomers via the sequential application of a chiral template and postmodification strategies. We demonstrated that L-MCNS showed differential biological behaviors and superior advantages in oral adsorption compared to those of CNS, D-MCNS, and DL-MCNS. During the delivery, helical L/D-MCNS presenting distinctive topological structures, including small section area, large rough external surface, and a screw-like body, displayed multiple superiorities in mucus diffusion and mucosal adhesion. Meanwhile, the grafted chiral enantiomers enabled positive or negative chiral recognition with the biosystems. Once racemic flurbiprofen (FP) was encapsulated into the nanopores of L/D-MCNS (FP@L/D-MCNS), L/D-MCNS providing highly cross-linked and mesoscopic chiral nanochannels was beneficial for controlling the drug loading/release kinetics with chiral microenvironment sensitivity. Particularly, we noticed enantioselective absorption of FP in vivo, which could be attributed to the differential biological behaviors of L/D-MCNS. By simple design and regulation of the multilevel chirality of nanocarriers, L/D-MCNS can be employed for efficient oral drug delivery from the perspectives of material science, pharmacy, and bionics.

Precious metal-supported catalytic materials have been widely used in the solvent-free aerobic oxidation of alcohols. However, their high cost limits their industrial application because high catalytic performance is usually obtained at high precious metal loading and O-2 pressure. In this work, an ultralow Pd-loading 0.03 wt % Pd/1D-TiO2 catalyst with smaller Pd particles (1.68 nm) is prepared and exhibits a turnover frequency (TOF) value as high as 68191.2 h(-1) for the benzyl alcohol oxidation, which is much higher than that of previously reported TiO2-supported catalysts under similar reaction conditions. The results also show that the catalytic activity is strongly dependent on the Pd particle size and light radiation, and a smaller Pd particle size can obviously improve the catalytic reaction rate. Compared to thermocatalytic oxidation, photothermocatalytic oxidation can also obviously improve the catalytic activity and change the main product from >90% benzaldehyde to >60% benzoic acid. A smaller Pd size is beneficial for the benzoic acid formation and decreases the difference in activity between thermocatalytic and photothermocatalytic oxidation. This work provides a valuable method in the green photothermal catalytic synthesis of high value-added chemicals over the ultralow loading precious metal-supported catalysts.

Enhancing the stiffness, energy absorption, and load-carrying capacity of conventional re-entrant honeycombs (CRH) is crucial for adapting complex loading conditions. To address this issue, a folded stiffener design is introduced into the CRH structure, known as improved re-entrant honeycomb (IRH). By utilizing the variational asymptotic method, the intricate 3D model is deconstructed into microscopic unit-cell constitutive modeling and macroscopic analysis via 2D equivalent plate model or 3D equivalent Cauchy model. The equivalent models aid in incorporating macroscopic responses into the recovery relationships, thereby facilitating the identification of local field distributions. Quasi-static uniaxial compression tests and numerical simulations on 3D printed multi-cellular structures reveal that the auxetic effect of IRH structures arises from constrained rotation of both folded stiffeners and inclined struts. Parameter analysis highlights the significant impact of slenderness ratio on in-plane and out-of-plane modulus, while adjusting the re-entrant angle affects Poisson’s ratio markedly. Compared to the CRH, the inclusion of folded stiffeners in the IRH results in a fourfold increase in the elastic modulus and reduces the negative Poisson’s ratio to 15%. Furthermore, the IRH demonstrates a more uniform stress distribution and improved energy absorption efficiency. Incorporating folded stiffeners to reduce porosity and enhance structural performance provides valuable insights for re-entrant auxetic honeycomb design.

Wound healing and infection remain significant challenges due to the ineffectiveness against multidrug-resistant (MDR) bacteria and the complex oxidative wound microenvironments. To address these issues, thymoquinone-reinforced injectable and thermosensitive TQ@PEG-PAF-Cur hydrogels with dual functions of microenvironment reshaping and photodynamic therapy are developed. The hydrogel comprises natural compound thymoquinone (TQ) and poly (ethylene glycol)-block-poly (alanine-co-phenyl alanine) copolymers (PEG-PAF) conjugated with natural photosensitizer curcumin (Cur). The incorporation of TQ and Cur reduces the sol-to-gel transition temperature of TQ@PEG-PAF-Cur to 30°C, compared to PEG-PAF hydrogel (37°C), due to the formation of strong hydrogen bonding, matching the wound microenvironment temperature. Under blue light excitation, TQ@PEG-PAF-Cur generates significant amounts of reactive oxygen species such as H2O2, 1O2, and ·OH, exhibiting rapid and efficient bactericidal capacities against methicillin-resistant Staphylococcus aureus and broad spectrum β-lactamases Escherichia coli via photodynamic therapy (PDT). Additionally, Cur effectively inhibits the expressions of proinflammatory cytokines in skin tissue-forming cells. As a result, the composite hydrogel can rapidly transform into a gel to cover the wound, reshape the wound microenvironment, and accelerate wound healing in vivo. This collaborative antibacterial strategy provides valuable insights to guide the development of multifunctional materials for efficient wound healing.

This research details the synthesis of biogenic silver nanoparticles utilizing Origanum majorana as a supporting material. The leaves of the plant served as a natural reducing agent and a proficient stabilizer for the generated silver nanoparticles (Ag NPs). The formulated NPs were analyzed through the analysis of their physicochemical properties utilizing UV-Vis, FT-IR, and FE-SEM techniques. The investigation of the antioxidant characteristics of the Ag NPs was followed using the DPPH assay. The results revealed that the silver nanoparticles exhibited considerable antioxidant activity, as evidenced by the IC50 value. Recent findings suggest that the effectiveness of nanoparticles in treating human breast cancer can be linked to their antioxidant characteristics. An evaluation was followed on the ability of biologically synthesized Ag NPs to combat breast cancer in various cell lines. The silver nanoparticles exhibited significant anti-breast cancer properties, successfully eradicating the MCF10 cancer cell line in a manner that was influenced by both time and concentration, as assessed through the MTT assay. Ag NPs facilitate cell apoptosis, a process linked to elevated levels of pro-apoptotic markers, including Bax and cleaved caspase-8, while concurrently reducing the concentration of the anti-apoptotic marker Bcl-2. Furthermore, silver nanoparticles exhibit a decrease in colony formation in comparison to the respective control group. The examination of molecular pathways in cells exposed to Ag NPs revealed a significant elevation in p53 expression, coupled with a decrease in both total and phosphorylated levels of Signal Transducer and Activator of Transcription 3 (STAT3) in the studied cell lines. This suggests that p53 and STAT3 play essential roles in the biological responses triggered by the extract in human breast cancer cells.

SCOPE:Bile acids (BAs) are closely associated with obesity and non-alcoholic fatty liver disease (NAFLD). How BAs change within the enterohepatic circulation during the onset and progression of NAFLD as biomarkers deserves to be explored.METHODS AND RESULTS:Four-week-old young mice were fed with high-fat diet plus 4% v/w fructose drinking water (HFF) or normal diet with tap water (ND) for 4 and 12 weeks. In comparison, eight-week-old adult mice were fed with HFF or ND for 12 weeks. BAs were measured in six different anatomical sites to evaluate the systematic changes of BAs within the enterohepatic circulation. The dysregulated BA metabolism had occurred in HFF after 4-week intervention, represented by increased primary BAs and decreased hyocholic acid (HCA) species. After 12 weeks, the impact was more significant with increased secondary BA synthesis and excretion, particularly for lithocholic acid (LCA) species. More interestingly, the BA changes were more significant in younger mice in response to 12-week diet intervention.CONCLUSIONS:The enterohepatic circulation of BAs changed with the development of NAFLD, and the younger mice were more susceptible to the unhealthy diet. HCA and LCA species may be potential biomarkers for predicting the development of NAFLD.

Ferroptosis is a newly identified type of programmed cell death that has been shown to contribute to the progression of septic cardiomyopathy. Although the role of miR-130b-3p as an oncogene that accelerates cancer progression by suppressing ferroptosis has been demonstrated, its role in the regulation of ferroptosis and cardiac injury in Lipopolysaccharide (LPS)-induced cardiomyopathy has not been fully clarified. In this study, we demonstrated that miR-130b-3p remarkably improved cardiac function and ameliorated morphological damage to heart tissue in LPS-induced mice. miR-130b-3p also improved cell viability and mitochondrial function and reduced the production of lipid ROS and ferroptosis in LPS-treated H9c2 cells. In addition, miR-130b-3p significantly upregulated GPX4 expression and suppressed ACSL4 activity in LPS-induced mouse heart tissue and H9c2 cells. Mechanistically, we used database analysis to locate miR-130b-3p and confirmed its inhibitory effects on the ferroptosis-related gene ACSL4 and autophagy-related gene PRKAA1 using a dual-luciferase reporter assay. In addition, we found that miR-130b-3p inhibited the activation of autophagy by downregulating the expression of the AMPK/mTOR signaling pathway. Meanwhile, our results show that RAPA (an autophagy activator) reverses the protective effect of miR-130b-3p mimic against LPS-induced ferroptosis, while CQ (an autophagy inhibitor) plays a facilitative role, suggesting that miR-130b-3p plays an important role in the development of ferroptosis by regulating autophagy in vitro. The findings reveal a novel function of miR-130b-3p in attenuating ferroptosis in cardiomyocytes, providing a new therapeutic target for ameliorating septic cardiomyopathy injury.

Kidney organoids can be generated from induced pluripotent stem cells (iPSCs) through various approaches. These organoids hold great promise for disease modeling, drug screening, and potential therapeutic applications. This article presents a step-by-step procedure to create kidney organoids from iPSCs, starting from the posterior primitive streak (PS) to the intermediate mesoderm (IM). The approach relies on the APEL 2 medium, which is a defined, animal component-free medium. It is supplemented with a high concentration of WNT agonist (CHIR99021) for a duration of 4 days, followed by fibroblast growth factor 9 (FGF9)/heparin and a low concentration of CHIR99021 for an additional 3 days. During this process, emphasis is given to selecting the optimal cell density and CHIR99021 concentration at the start of iPSCs, as these factors are critical for successful kidney organoid generation. An important aspect of this protocol is the suspension culture in a low adherent plate, allowing the IM to gradually develop into nephron structures, encompassing glomerular, proximal tubular, and distal tubular structures, all presented in a visually comprehensible format. Overall, this detailed protocol offers an efficient and specific technique to produce kidney organoids from diverse iPSCs, ensuring successful and consistent results.

Peptide-based supramolecular photodynamic therapy (PDT) is an emerging modality for cancer treatments. This strategy exploits the biological activities and self-assembling potential of peptides to enhance the delivery selectivity of photosensitizers to the lesion site, ultimately achieving the improved treatment efficacy upon the light irradiation. In this review, we survey the recent advancements in the rational design and preparation of peptide-based PDT systems for targeted delivery and anti-tumor treatments. We carefully analyze the different synthetic approaches for the preparation of PDT systems in terms of the associative interactions between peptides and photosensitizers, i.e., non-covalent interactions and covalent ones. Both mono-therapeutic approaches and PDT-based combination therapies are examined in this review. Future outlooks and upcoming challenges in peptide-based PDT systems with an eye toward translational medicine are also discussed at the end.

Guangsangon E (GSE) is a natural product separated from Morus alba L. It has been reported to treat lung cancer through autophagy. However, whether GSE is effective in repressing triple-negative breast cancer (TN BC) cells is yet to be elucidated. In the present study, GSE inhibited cell growth of MDA-MB-231, MDA-MB-453, and MDA-MB-468 cells. Moreover, GSE induced mitochondrial dysfunction, including membrane potential loss, mitochondria fission, and reactive oxygen species accumulation, and finally led to mitophagy-related non-apoptotic cell death. In the xenograft tumor nude mice, GSE treatment significantly reduced the size and weight of MDA-MB-231 tumors. The tumor inhibition rates of GSE treatment were 49.68% (low-dose) and 48.73% (high-dose). In summary, GSE is a potential anticancer drug available for treating TNBC with apoptosis resistance.

Most multiplex nucleic acids detection methods require numerous reagents and high-priced instruments. The emerging clustered regularly interspaced short palindromic repeats (CRISPR)/Cas has been regarded as a promising point-of-care (POC) strategy for nucleic acids detection. However, how to achieve CRISPR/Cas multiplex biosensing remains a challenge. Here, an affordable means termed CRISPR-RDB (CRISPR-based reverse dot blot) for multiplex target detection in parallel, which possesses the advantages of high sensitivity and specificity, cost-effectiveness, instrument-free, ease to use, and visualization is reported. CRISPR-RDB integrates the trans-cleavage activity of CRISPR-Cas12a with a commercial RDB technique. It utilizes different Cas12a-crRNA complexes to separately identify multiple targets in one sample and converts targeted information into colorimetric signals on a piece of accessible nylon membrane that attaches corresponding specific-oligonucleotide probes. It has demonstrated that the versatility of CRISPR-RDB by constructing a four-channel system to simultaneously detect influenza A, influenza B, respiratory syncytial virus, and SARS-CoV-2. With a simple modification of crRNAs, the CRISPR-RDB can be modified to detect human papillomavirus, saving two-thirds of the time compared to a commercial PCR-RDB kit. Further, a user-friendly microchip system for convenient use, as well as a smartphone app for signal interpretation, is engineered. CRISPR-RDB represents a desirable option for multiplexed biosensing and on-site diagnosis.

The application of nanocarriers as drug delivery system for chemotherapeutic drugs has become a research hotspot in cancer treatment. Chemotherapy with high tumor-targeting accuracy and drug release specificity is the key to improve the efficacy of tumor chemotherapy and reduce the side effects caused by repeated doses drugs.Here, we synthesized a redox-sensitive nano-micelle formed by hyaluronic acid (HA) conjugated with d-α-tocopherol succinate (TOS) using a disulfide bond as the linker (HA-SS-TOS, HSST), which could actively accumulate to the tumor sites and metastasis cancer cells with high expression of CD44. The micelles could dissociate under the high GSH level in cancer cells, triggering a release of paclitaxel (PTX). Surprisingly, the precise chemotherapy instead induced a suppressive tendency of immune system, manifested by a significant increase in TGF-β, which weakened the therapeutic effect of micelles. Moreover, the high levels of TGF-β might be related to the increased drug-resistance of cancer cells. Research has shown that PD-1 pathway blockade can result in reduction in TGF-β expression, thus, a PLGA microsphere encapsulating PD-1 antagonist peptides A12 (A12@PLGA) was further prepared to activate the host immune response. Our data indicated that PTX-loaded HSST could accurately “find” the tumors as well as metastasis cancer cells, and efficiently kill most of them. The joining of a durable PD-1 blockage significantly boosted the efficacy of PTX@HSST on multiple tumor models, including lung metastatic tumors and even multidrug-resistant tumors. Thus, our work presented an optimal chemo-immunotherapy combined system, which shows profound significance for future cancer therapy in clinic.

The broad application of strobilurin fungicide led to pathogen resistance, and toxic effects have been reported for several species. Benzene kresoxim-methyl (BKM) is a novel strobilurin fungicide mainly used to control the cucumber powdery mildew. However, information about the fate of BKM in agrofood systems and related human exposure is limited. In this study, greenhouse experiments were conducted to investigate the distribution, translocation, and residual of the 10% suspension concentrate (SC) commercial BKM formulations on mature cucumber plants using C-14 tracer technology. After foliage and fruit application, 25.84% of the applied C-14-labeled BKM can be absorbed into mature cucumber plants at 21 days after treatment. The absorbed BKM transferred through-out the plant acropetally and basipetally, although over 81.13% of absorbed BKM remained in the labeled leaves. In the edible parts, 14.35% of the absorbed BKM remained in the pericaip of labeled fruits, only 0.027 mg kg(-1) accumulated in the sarcocarp. The concentration of BKM in newborn fruits was 0.005 mg kg(-1), indicating low dietary exposure. These findings develop a better understanding of the fate of BKM in the cucumber plants, provide guidance in the rational use of BKM and can be incorporated into food and environmental assessments of BKM. (C) 2020 Elsevier B.V. All rights reserved.

As a refractory fibrosis disease, intrauterine adhesions (IUAs) is defined as fibrosis of the physiological endometrium. Although hysteroscopic adhesiolysis is widely recommended as an effective treatment, prognosis and recurrence remain poor in severe cases. Recently, stem cell therapy has been promoted as a promising treatment for IUAs. The ability of human amniotic epithelial cells (hAECs), emerging as a new candidate for stem cell therapy, to treat IUAs has not been demonstrated. To study the potential effects of hAECs on IUAs, we created an IUA rat model using mechanical injury and injected cultured primary hAECs into the rats’ uteri. Next, we observed the morphological structure of endometrial thickness and glands using hematoxylin and eosin staining, and we detected extracellular-matrix collagen deposition using Masson staining. In addition, we performed immunohistochemical staining and reverse-transcription polymerase chain reaction (RT-PCR) to investigate potential fibrosis molecules and angiogenesis factors 7 d after hAECs transplantation. Finally, we detected estrogen receptor (ER) and growth factors via RT-PCR to verify the molecular mechanism underlying cell therapy. In the IUA rat models, endometrial thickness and endometrial glands proliferated and collagen deposition decreased significantly after hAEC transplantation. We found that during the recovery of injured endometrium, the crucial fibrosis marker transforming growth factor-β (TGF-β) was regulated and angiogenesis occurred in the endometrial tissue with the up-regulation of vascular endothelial growth factor. Furthermore, hAECs were shown to promote ER expression in the endometrium and regulate the inflammatory reaction in the uterine microenvironment. In conclusion, these results demonstrated that hAEC transplantation could inhibit the progression of fibrosis and promote proliferation and angiogenesis in IUA rat models. The current study suggests hAECs as a novel stem cell candidate in the treatment of severe IUA.

In this work, single-crystal BiFeO3/PbTiO3 (BFO/PTO) nanoplates have been successfully synthesized via a hydrothermal method. It is interesting to find that BFO films selectively grew on the negative polar surface of PTO nanoplates with a saturation thickness of about 18-20 nm. Below such saturation size, the thickness of BFO films can be controlled in the range of 4-18 nm. An atomically sharp interface was identified to be formed between BFO and PTO with an epitaxial relationship of {012}(BFO)//{001}(PTO). In particular, a room-temperature ferromagnetism in the nanoplates has been observed, where the saturation magnetization significantly decreased from to 0.029 to 0.003 emu g(-1) upon the film thickness decreasing to 4 nm. Electron energy-loss spectroscopy (EELS) analysis on the interface suggests that the positive polar surface of BFO could be screened by both of the negative polarization and addition electrons in PTO. A mechanism of "screening balance broken and rebuilt" on the negative interface was proposed to explain the existence of selective growth, the saturation thickness and magnetism of the nanoplates. The findings suggest that a polar surface could be a powerful platform to realize the controllable epitaxial growth of perovskite oxide heterostructures with the intriguing interface functionality.

Understanding the bioavailability and phytotoxicity of Carbendazim (MBC) bound residues (BR) in soils incubated with different Superabsorbent polymer (SAP) amendment on succeeding crops is essential to assess their environmental fate and risks. In our research, we studied the morphological characteristics and 14C-accumulation of Chinese cabbage and released BR in three typical cultivated soils. The plant dry weight was in order of superabsorbent-hydrogels formulations (HMBC) > MBC > MBC and SAP (MBC-SAP) at 35 d in basic soil 3 (S3), with 675.40 ± 29.07 mg/plant.d.w, 575.93 ± 25.35 mg/plant.d.w and 427.86 ± 18.79 mg/plant.d.w. The whole plant accumulated 2-fold more BR when grew in neutral soil 2 (S2) treated with SAP than MBC at 7 d. The root accumulated a greater proportion of 14C-MBC residue than shoot, with order of MBC-SAP > MBC > HMBC at 21d. The results indicate MBC-BR could be released and accumulated in plant. HMBC promoted the Chinese cabbage growth with lowest 14C accumulation, while MBC-SAP inhibited plant growth with the highest 14C uptake. The released BR rate was 61.43 ± 3.75% of initial BR in MBC-SAP, with 2-fold higher than MBC and HMBC. It is assumed HMBC could be a potential environmentally friendly measure for rational use of pesticides in future.

Oxide heterointerface is a platform to create unprecedented two-dimensional electron gas, superconductivity and ferromagnetism, arising from a polar discontinuity at the interface. In particular, the ability to tune these intriguing effects paves a way to elucidate their fundamental physics and to develop novel electronic/magnetic devices. In this work, we report for the first time that a ferroelectric polarization screening at SrTiO3/PbTiO3 interface is able to drive an electronic construction of Ti atom, giving rise to room-temperature ferromagnetism. Surprisingly, such ferromagnetism can be switched to antiferromagnetism by applying a magnetic field, which is reversible. A coupling of itinerant electrons with local moments at interfacial Ti 3d orbital was proposed to explain the magnetism. The localization of the itinerant electrons under a magnetic field is responsible for the suppression of magnetism. These findings provide new insights into interfacial magnetism and their control by magnetic field relevant interfacial electrons promising for device applications.