Dual regulation of osteosarcoma hypoxia microenvironment by a bioinspired oxygen nanogenerator for precise single-laser synergistic photodynamic/photothermal/induced antitumor immunity therapy.

The hypoxic tumor microenvironment (TME) of osteosarcoma (OS) is the Achilles' heel of oxygen-dependent photodynamic therapy (PDT), and tremendous challenges are confronted to reverse the hypoxia. Herein, we proposed a "reducing expenditure of O2 and broadening sources" dual-strategy and constructed ultrasmall IrO2@BSA-ATO nanogenerators (NGs) for decreasing the O2-consumption and elevating the O2-supply simultaneously. As O2 NGs, the intrinsic catalase (CAT) activity could precisely decompose the overexpressed H2O2 to produce O2 in situ, enabling exogenous O2 infusion. Moreover, the cell respiration inhibitor atovaquone (ATO) would be at the tumor sites, effectively inhibiting cell respiration and elevating oxygen content for endogenous O2 conservation. As a result, IrO2@BSA-ATO NGs systematically increase tumor oxygenation in dual ways and significantly enhance the antitumor efficacy of PDT. Moreover, the extraordinary photothermal conversion efficiency allows the implementation of precise photothermal therapy (PTT) under photoacoustic guidance. Upon a single laser irradiation, this synergistic PDT, PTT, and the following immunosuppression regulation performance of IrO2@BSA-ATO NGs achieved a superior tumor cooperative eradicating capability both in vitro and in vivo. Taken together, this study proposes an innovative dual-strategy to address the serious hypoxia problem, and this microenvironment-regulable IrO2@BSA-ATO NGs as a multifunctional theranostics platform shows great potential for OS therapy.

Bi2S3/Ti3C2-TPP nano-heterostructures induced by near-infrared for photodynamic therapy combined with photothermal therapy on hypoxic tumors.

BACKGROUND: Photodynamic therapy (PDT) efficacy of bismuth sulfide (Bi2S3) semiconductor has been severely restricted by its electron-hole pairs (e--h+) separation inefficiency and oxygen (O2) deficiency in tumors, which greatly hinders reactive oxygen species (ROS) generation and further clinical application of Bi2S3 nanoparticles (NPs) in biomedicine. RESULTS: Herein, novel Bi2S3/titanium carbide (Ti3C2) two-dimensional nano-heterostructures (NHs) are designed to realize multimode PDT of synchronous O2 self-supply and ROS generation combined with highly efficient photothermal tumor elimination for hypoxic tumor therapy. Bi2S3/Ti3C2 NHs were synthesized via the in situ synthesis method starting from Ti3C2 nanosheets (NSs), a classical type of MXene nanostructure. Compared to simple Bi2S3 NPs, Bi2S3/Ti3C2 NHs significantly extend the absorption to the near-infrared (NIR) region and enhance the photocatalytic activity owing to the improved photogenerated carrier separation, where the hole on the valence band (VB) of Bi2S3 can react with water to supply O2 for the electron on the Ti3C2 NSs to generate ·O2- and ·OH through electron transfer. Furthermore, they also achieve 1O2 generation through energy transfer due to O2 self-supply. After the modification of triphenylphosphium bromide (TPP) on Bi2S3/Ti3C2 NHs, systematic in vitro and in vivo evaluations were conducted, revealing that the synergistic-therapeutic outcome of this nanoplatform enables complete eradication of the U251 tumors without recurrence by NIR laser irradiation, and it can be used for computed tomography (CT) imaging because of the strong X-ray attenuation ability. CONCLUSION: This work expands the phototherapeutic effect of Bi2S3-based nanoplatforms, providing a new strategy for hypoxic tumor theranostics.

Synthesis and anti-SARS-CoV-2 evaluation of lipid prodrugs of ß-D-N4-hydroxycytidine (NHC) and a 3'-fluoro-substituted analogue of NHC.

ß-D-N4-hydroxycytidine (NHC, EIDD-1931) is a nucleoside analogue that exhibits broad spectrum antiviral activity against a variety of RNA viruses. Herein, we report the synthesis of a series of lipid prodrugs of NHC and a novel 3'-fluoro modified NHC analogue, and evaluation of their antiviral activity against five variants of SARS-CoV-2. All lipid prodrugs showed potent antiviral activity against the tested SARS-CoV-2 variants with EC50 values in the range of 0.31-3.51 µM, which were comparable to those of NHC or higher than those of remdesivir and molnupiravir. An increase in the cytostatic activity of the lipid prodrugs was found, but prodrug 2d proved equally selective as molnupinavir. The 3'-F analogue of NHC (6) only displayed minor antiviral activity against the SARS-CoV-2 Omicron variant (EC50 = 29.91 µM), while no activity was found for other variants at the highest concentration tested. The promising antiviral data of the lipid prodrugs of NHC suggest that they deserve further investigation as new anti-SARS-CoV-2 drugs.

An Immune-Related Long Noncoding RNA Pair as a New Biomarker to Predict the Prognosis of Patients in Breast Cancer.

Background: Immune-related long non-coding RNAs (irlncRNAs) might remodel the tumor immune microenvironment by changing the inherent properties of tumor cells and the expression of immune genes, which have been used to predict the efficacy of immunotherapy and the prognosis of various tumors. However, the value of irlncRNAs in breast cancer (BRCA) remains unclear. Materials and Methods: Initially, transcriptome data and immune-related gene sets were downloaded from The Cancer Genome Atlas (TCGA) database. The irlncRNAs were extracted from the Immunology Database and Analysis Portal (ImmPort) database. Differently expressed irlncRNAs (DEirlncRNAs) were further identified by utilizing the limma R package. Then, univariate and multivariate Cox regression analyses were conducted to select the DEirlncRNAs associated with the prognosis of BRCA patients. In addition, the univariate and least absolute shrinkage and selection operator (LASSO) Cox regression analyses were performed to determine the DEirlncRNA pairs with the independent prediction capability of prognosis in BRCA patients. Finally, the chosen DEirlncRNA pair would be evaluated in terms of survival time, clinicopathological characteristics, tumor-infiltrating immune cells, immune checkpoints (ICs), signaling pathways, and potential small-molecule drugs. Results: A total of 21 DEirlncRNA pairs were extracted, and among them, lncRNA MIR4435-2HG and lncRNA U62317.1 were chosen to establish a risk signature that served as an independent prognostic biomarker in BRCA patients. Patients in the high-risk group had a worse prognosis than those in the low-risk group, and they also had an abundance of infiltration of CD4+ T and CD8+ T cells to enhance the immune response to tumor cells. Furthermore, the risk signature showed a strong correlation with ICs, signaling pathways, and potential small-molecule drugs. Conclusion: Our research revealed that the risk signature independent of specific DEirlncRNA pair expression was closely associated with the prognosis and tumor immune microenvironment in BRCA patients and had the potential to function as an independent prognostic biomarker and a predictor of immunotherapy for BRCA patients, which would provide new insights for BRCA accurate treatment.

Synthesis and biological evaluation of BU-4664L derivatives as potential anticancer agents.

BU-4664L is a naturally occurring N-farnesylated dibenzodiazepinone with important biological activities. Herein, we report the synthesis and antitumor evaluation of two series of BU-4664L derivatives bearing different substituent patterns on the dibenzodiazepinone core and with diverse side chains. All of the derivatives displayed micromolar activity against the human prostate cancer PC-3 cells, while lower or no activity against the human lung H460 cells. The most active derivatives were 10a and 16c which exerted antiproliferative activity against PC-3 cells with GI50 values of 5.66 and 5.94 µM, respectively, and thus represent promising lead compounds for further development.

A biodegradable, mechanically tunable micro-arc oxidation AZ91D-based composite implant with calcium phosphate/chitosan coating promotes long-term bone tissue regeneration.

BACKGROUND: To reduce the biodegradable rate and develop the long-term osteogenic ability of magnesium (Mg) alloy, we prepared a new biodegradable micro arc oxidation AZ91D-based composite implant with calcium phosphate/chitosan coating (CaP-CS/MAO/AZ91D) and investigated its mechanical property and long-term bone tissue regeneration ability. MAIN METHODS AND MAJOR RESULTS: The results showed that the binding force and bioactivity of CaP-CS/MAO/AZ91D was better when the ratio of water to ethanol was 4:6 and MAO constant current was 0.1 A cm-2 . Compressive strengths of 4:6 sample were more than 1300 N when the soaking time was increased to 21 days. CaP-CS/MAO/AZ91D extracts promoted differentiation and proliferation of rat mesenchymal stem cells (RMSC), which achieved higher proliferation rates over 16 days of culture and exhibited early alkaline phosphatase activity and late bone sialoprotein markers. CONCLUSIONS AND IMPLICATIONS: CaP-CS/MAO/AZ91D was established to promote RMSC osteogenic differentiation within a proper range for at least 90 days through Wnt/ß-catenin pathway activation, which would allow sufficient time for bone healing. Collectively, our findings suggest that the CaP-CS/MAO/AZ91D coating could not only reduce the corrosion rate and lead to better long-term biocompatibility but also promote osteogenic mineralization.

CNTs-CaP/chitosan-coated AZ91D magnesium alloy extract promoted rat dorsal root ganglia neuron growth via activating ERK signalling pathway.

Increasing attention has been paid on the application of biodegradable materials such as magnesium and its alloys in neuron repair. AZ91D magnesium alloy coated with carbon nanotubes (CNTs) and/or calcium phosphate (CaP)/chitosan (CS) was fabricated in this study. To evaluate the bioactivity of these AZ91D-based composites, the extracts were prepared by immersing samples in modified simulated body fluid (m-SBF) for 0, 2, 8, 16, 24, 34, 44, 60, or 90 days. Immunofluorescence staining for neuronal class III ß-tubulin (TUJ1) revealed that both CNTs-CaP/CS-AZ91D and CaP/CS-AZ91D extracts promoted axon outgrowth of dorsal root ganglia (DRG) neurons, accompanied with increased expression of phosphorylated focal adhesion kinase (p-FAK) and growth associated protein-43 (GAP-43). Besides, the extracts increased the expression and the release of neurotrophic factors including nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). ERK signalling was activated in DRG neurons after treating with either CNTs-CaP/CS-AZ91D or CaP/CS-AZ91D extracts, and its inhibition with U0126 counteracted the beneficial effects of these extracts on DRG neuron. Overall, the extracts from these AZ91D-based composites might promote DRG neuron growth via activating ERK signalling pathway. Notably, CNTs-CaP/CS-AZ91D extracts showed a better promoting effect on neuron growth than CaP/CS-AZ91D. Assessment of ion elements showed that the addition of CNTs coating enhanced magnesium corrosion resistance and reduced the deposition of calcium and phosphorus on the surface of CaP/CS-AZ91D alloy. These findings demonstrate that CNTs-CaP/CS-AZ91D likely provide a more suitable environment for neuron growth, which suggests a potential implantable biomaterial for the treatment of nerve injury. SIGNIFICANCE: AZ91D magnesium alloy coated with carbon nanotubes (CNTs) and/or calcium phosphate (CaP)/chitosan (CS) was fabricated and their immersion extracts were prepared using modified simulated body fluid in this study. Both extracts from CNTs-CaP/CS and CaP/CS-coated AZ91D magnesium alloy promotes rat dorsal root ganglia (DRG) neuron growth via activating ERK signalling pathway. Notably, the addition of CNTs improves the performance of CaP/CS-AZ91D. For the first time, our research demonstrates that CNTs-CaP/CS-AZ91D likely provide a suitable environment for neuron growth, suggesting these AZ91D-based composites as potential implantable biomaterials for the treatment of nerve injury.

Evaluation of long-term biocompatibility and osteogenic differentiation of graphene nanosheet doped calcium phosphate-chitosan AZ91D composites.

To improve the biodegradable performance and osteoinductivity of magnesium (Mg) alloy, a novel graphene nanosheet (GNS) doped micro-arc oxidized-AZ91D (MAO-AZ91D) based calcium phosphate (CaP)-chitosan (CS) (GNS-CaP-CS/AZ91D) composite was fabricated. Their long-term in vitro biocompatibility, osteoinductivity and the related signaling pathway were evaluated. The electrochemical test, Mg concentration and SEM results showed that the corrosion rate of GNS-CaP-CS/AZ91D became much slower, whose corrosion degree with immersion time being 90 d was similar to that of AZ91D being 16 d. Good biocompatibility and non-cytotoxicity were observed during the whole immersion process of GNS-CaP-CS/AZ91D. Extracts from GNS-CaP-CS/AZ91D especially in the early stage (16 days and before) could significantly enhance alkaline phosphatase (ALP) activity, calcium mineral deposition and osteoblast-related genes expression of human bone marrow mesenchymal stem cells (hBMSCs). GNS released from GNS-CaP-CS/AZ91D composites might serve a critical role to induce osteogenic differentiation through ERK/MAPK pathway in the long-term immersion process, while, PI3K/Akt signaling only significantly in the early stage. Moreover, Mg element could promote osteogenic differentiation through ERK/MAPK pathway and the maximum concentration was about the range between 200 and 500 ppm.

Targeting property and toxicity of a novel ultrasound contrast agent microbubble carrying the targeting and drug-loaded complex FA-CNTs-PTX on MCF7 cells.

The application of ultrasound contrast agents not only is confined to the enhancement of ultrasound imaging but also has started to be used as a drug system for diagnosis and treatment. In this paper, Span60 and PEG1500 were used as membrane materials, and a new targeting and drug-loading multifunctional ultrasound contrast agent microbubble enveloping the FA-CNTs-PTX complex was successfully prepared by acoustic cavitation. With the breast cancer cell line MCF7 as the research target, the effects of the microbubble with FA-CNTs-PTX on the proliferation and toxicity of MCF7 cells were studied using a CCK-8 and AO/EB double-staining method. The influences of the microbubbles with FA-CNTs-PTX on the cellular morphology and apoptosis period of the MCF7 cells were detected using an inverted fluorescence microscope. The apoptosis of MCF7 cells induced by the microbubbles with FA-CNTs-PTX was investigated with flow cytometry and an annexin and PI double staining fluorescence quantitative analysis. The results indicated that the ultrasound contrast agent microbubble with FA-CNTs-PTX remarkably inhibited the proliferation of MCF7 cells, which was mainly controlled by the drug loading rate and the nanometer size of the microbubbles. Moreover, the proliferative inhibition rate of the microbubbles with FA-CNTs-PTX was related to the cell apoptosis period of MCF7 cells. Its inhibition degree on the proliferation of MCF7 cells was higher than that of the hepatoma HepG2 cells. The apoptosis rate of MCF7 cells induced by the microbubbles with FA-CNTs-PTX was higher than that of normal human umbilical vein endothelial cells (HUVECs), and the microbubbles with FA-CNTs-PTX could target the MCF7 cells.

Effect of the addition CNTs on performance of CaP/chitosan/coating deposited on magnesium alloy by electrophoretic deposition.

CaP/chitosan/carbon nanotubes (CNTs) coating on AZ91D magnesium alloy was prepared via electrophoretic deposition (EPD) followed by conversion in a phosphate buffer solution (PBS). The bonding between the layer and the substrate was studied by an automatic scratch instrument. The phase compositions and microstructures of the composite coatings were determined by using X-ray diffraction (XRD), Fourier-transformed infrared spectroscopy (FTIR), Raman spectroscopy and scanning electron microscope (SEM). The element concentration and gentamicin concentration were respectively determined by inductively coupled plasma optical emission spectrometer (ICP-OES) test and ultraviolet spectrophotometer (UV). The cell counting kit (CCK) assay was used to evaluate the cytotoxicity of samples to SaOS-2 cells. The results showed that a few CNTs with their original tubular morphology could be found in the CaP/chitosan coating and they were beneficial for the crystal growth of phosphate and improvement of the coating bonding when the addition amount of CNTs in 500 ml of electrophoretic solution was from 0.05 g to 0.125 g. The loading amount of gentamicin increased and the releasing speed of gentamicin decreased after CNTs was added into the CaP/chitosan coating for immersion loading and EPD loading. The cell viability of Mg based CaP/chitosan/CNTs was higher than that of Mg based CaP/chitosan from 16 days to 90 days.

Microdochium tabacinum, Confirmed as a Pathogen of Alfalfa in Gansu Province, China.

A crown and root rot complex was detected in the alfalfa (Medicago sativa 'Longdong') fields of Huanxian County. The symptoms of the diseased plants were characterized, and 11 fungal species were obtained from the roots. These fungi included isolates that resembled the genus Microdochium. An isolate of this type, designated MP313, was proven to infect alfalfa, fulfilling Koch's postulates. Isolate MP313 was examined by microscopy and the morphological characteristics indicated that it was similar to members of the genus Microdochium. Sequence analyses of the 28S large subunit as well as the internal transcribed spacer (ITS) region of the ribosomal DNA (rDNA) of MP313 revealed 98 to 100% similarity to the corresponding regions of M. tabacinum. A polymerase chain reaction assay based on the ITS region of the rDNA was developed to amplify a 304-bp fragment from DNA concentrations as low as 20 fg/µl, which was sensitive enough to detect isolate MP313 in diseased root samples. Taken together, these results confirmed that M. tabacinum was one of a complex of fungi associated with crown and root rot in the alfalfa samples collected in Gansu Province. This is the first report of M. tabacinum being a pathogen of alfalfa in China.

A construction of novel iron-foam-based calcium phosphate/chitosan coating biodegradable scaffold material.

Slow corrosion rate and poor bioactivity restrict iron-based implants in biomedical application. In this study, we design a new iron-foam-based calcium phosphate/chitosan coating biodegradable composites offering a priority mechanical and bioactive property for bone tissue engineering through electrophoretic deposition (EPD) followed by a conversion process into a phosphate buffer solution (PBS). Tensile test results showed that the mechanical property of iron foam could be regulated through altering the construction of polyurethane foam. The priority coatings were deposited from 40% nano hydroxyapatite (nHA)/ethanol suspension mixed with 60% nHA/chitosan-acetic acid aqueous solution. In vitro immersion test showed that oxidation-iron foam as the matrix decreased the amount of iron implanted and had not influence on the bioactivity of this implant, obviously. So, this method could also be a promising method for the preparation of a new calcium phosphate/chitosan coating on foam construction.

Degradable behavior and bioactivity of micro-arc oxidized AZ91D Mg alloy with calcium phosphate/chitosan composite coating in m-SBF.

The degradation behavior of a MAO-AZ91D with a calcium phosphate/chitosan composite coating in the modified simulated human body fluid (m-SBF) was investigated by immersion experiments and electrochemical impedance spectroscopy. The compositions of the composite coating soaked in the m-SBF for different time intervals were studied by X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, thermo-gravimetric analysis and differential thermal analysis. The microstructures of the composite coating at different soaking stages were observed by using a scanning electron microscope. Results show that the as-prepared composite coating could slow down the corrosion rate of the AZ91D alloy and it demonstrated good bioactivity in the m-SBF. The coating's morphology changed from a flake-like one into a spherical-shaped one with the increase of immersion time in the m-SBF.

[Detection of tofu hardness by near infrared spectroscopy].

The possibility of determination of tofu hardness by near infrared spectroscopy (NIR, 833-2500 nm) was studied. The influences of water content and protein content on the spectra were discussed, in order to detect the hardness. The models of Tofu hardness were calibrated by partial least squares regression (PLS) after eliminating outline, spectra pretreatment, and wavelength optimization (r = 0.994, RMSEC = 0.391%, RMSEP = 0.416%, RSD = 2.27%). All of the results were better than those by sense assessing method.