Cannabidiol alleviates carbon tetrachloride-induced liver fibrosis in mice by regulating NF-κB and PPAR-α pathways.

Liver fibrosis has become a serious public health problem that can develop into liver cirrhosis and hepatocellular carcinoma and even lead to death. Cannabidiol (CBD), which is an abundant nonpsychoactive component in the cannabis plant, exerts cytoprotective effects in many diseases and under pathological conditions. In our previous studies, CBD significantly attenuated liver injury induced by chronic and binge alcohol in a mouse model and oxidative bursts in human neutrophils. However, the effects of CBD on liver fibrosis and the underlying mechanisms still need to be further explored. A mouse liver fibrosis model was induced by carbon tetrachloride (CCl4) for 10 weeks and used to explore the protective properties of CBD and related molecular mechanisms. After the injection protocol, serum samples and livers were used for molecular biology, biochemical and pathological analyses. The results showed that CBD could effectively improve liver function and reduce liver damage and liver fibrosis progression in mice; the expression levels of transaminase and fibrotic markers were reduced, and histopathological characteristics were improved. Moreover, CBD inhibited the levels of inflammatory cytokines and reduced the protein expression levels of p-NF-κB, NF-κB, p-IκBα, p-p38 MAPK, and COX-2 but increased the expression level of PPAR-α. We found that CBD-mediated protection involves inhibiting NF-κB and activating PPAR-α. In conclusion, these results suggest that the hepatoprotective effects of CBD may be due to suppressing the inflammatory response in CCl4-induced mice and that the NF-κB and PPAR-α signaling pathways might be involved in this process.

Resilience and its influencing factors after emergency percutaneous coronary intervention in young and middle-aged patients with first acute myocardial infarction.

Mental health after acute myocardial infarction (AMI) influences the prognosis of patients. Resilience may contribute to improving a patient's mental health. However, no study has investigated resilience and its associated factors in young and middle-aged patients undergoing emergency percutaneous coronary intervention (PCI) after the first AMI. This study aimed to identify critical associated factors influencing resilience in these patients. This cross-sectional study recruited 161 young and middle-aged patients with first-episode AMI using a purposive sampling method. These patients were assessed 48 h after emergency PCI using the General Information Questionnaire, the Connor-Davidson Resilience Scale-10, the Perceived Social Support Scale, the General Self-Efficacy Scale, and the Post-traumatic Stress Disorder Scale Civilian Version. Stepwise and logistic regression were conducted to analyze the factors influencing resilience. Receiver operating characteristics (ROC) were used to compare the area under the curves (AUC) for each indicator. The resilience of the 161 participants was 29.50 ± 4.158. Monthly household income, self-efficacy, social support, and post-traumatic stress disorder explained 51.4% of the variance in resilience. Self-efficacy (OR 0.716, CI 0.589-0.870, P < 0.01) and social support (OR 0.772, CI 0.635-0.938, P < 0.01) were protective factors for psychological resilience, while post-traumatic stress disorder (OR 1.278, CI 1.077-1.515, P < 0.01) was a risk factor. ROC curve revealed that self-efficacy, social support, and PTSD had an AUC of 0.822, 0.855, and 0.889, respectively. Self-efficacy and social support improve, and PTSD degrades psychological resilience in young and middle-aged AMI patients undergoing emergency PCI.

Polarized emission of Cs3Cu2I5 nanowires embedded in nanopores of an anodic aluminum oxide template.

Due to the intrinsic polarized emission property, polarized emissive materials with anisotropic nanostructures are expected to be potential substitutes for polarizers. Herein, by the template-assisted strategy, well-aligned lead-free metal halide Cs3Cu2I5 nanowire (NW) arrays are fabricated by evaporating the precursor ink in the anodic aluminum oxide (AAO) for polarized emission. The Cs3Cu2I5/AAO composite film emits highly polarized light with a degree of polarization (DOP) of 0.50. Furthermore, by changing the molar ratio of CsI/CuI, the stability of Cs3Cu2I5 precursor inks is improved. Finally, an ultraviolet (UV) light-emitting diode (LED) is adopted to pump the composite film to achieve a blue LED device. The reported Cs3Cu2I5/AAO composite film with highly polarized light emissions will have great potential for polarized emission applications such as liquid crystal display backlights, waveguides, and lasers.

Electrochemiluminescence Analysis of Multiple Glycans on Single Living Cell with a Closed Bipolar Electrode Array Chip.

The profiling of multiple glycans on a single cell is important for elucidating glycosylation mechanisms and accurately identifying disease states. Herein, we developed a closed bipolar electrode (BPE) array chip for live single-cell trapping and in situ galactose and sialic acid detection with the electrochemiluminescence (ECL) method. Methylene blue-DNA (MB-DNA) as well as biotin-DNA (Bio-DNA) codecorated AuNPs were prepared as nanoprobes, which were selectively labeled on the cell surface through chemoselective labeling techniques. The individual cell was captured and labeled in the microtrap of the cathodic chamber, under an appropriate potential, MB molecules on the cellular membrane underwent oxidation, triggering the reduction of [Ru(bpy)3]2+/TPA and consequently generating ECL signals in the anodic chamber. The abundance of MB groups on the single cell enabled selective monitoring of both sialic acid and galactosyl groups with high sensitivity using ECL. The sialic acid and galactosyl content per HepG2 cell were detected to be 0.66 and 0.82 fmol, respectively. Through comprehensive evaluation of these two types of glycans on a single cell, tumor cells, and normal cells could be effectively discriminated and the accuracy of single-cell heterogeneous analysis was improved. Additionally, dynamic monitoring of variations in galactosyl groups on the surface of the single cell was also achieved. This work introduced a straightforward and convenient approach for heterogeneity analysis among single cells.

Method for multi-task learning fusion network traffic classification to address small sample labels.

In the context of the proliferated evolution of network service types and the expeditious augmentation of network resource deployment, the requisition for copious labeled datasets to facilitate superior performance in traffic classification methods, particularly those hinging on deep learning, is imperative. Nonetheless, the procurement and annotation of such extensive datasets necessitate considerable temporal and human resource investments. In response to this predicament, this work introduces a methodology, termed MTEFU, leveraging a deep learning model-based multi-task learning algorithm, strategically designed to mitigate the reliance on substantial labeled training samples. Multiple classification tasks, encompassing duration, bandwidth size, and business traffic category, are incorporated, with a shared parameter strategy implemented amongst tasks to assure the transference of information across disparate tasks. Employing CNN, SAE, GRU, and LSTM as multi-task learning classification models, training validation and experimental testing were conducted on the QUIC dataset. A comparative analysis with single-task and ensemble learning methods reveals that, in the context of predicting network traffic types, the accuracy derived from the multi-task learning strategy, even with a mere 150 labeled samples, can emulate the 94.67% accuracy achieved through single-task learning with a fully labeled dataset of 6139 samples.

A Blind Source Separation Method Based on Bounded Component Analysis Optimized by the Improved Beetle Antennae Search.

Currently, the widely used blind source separation algorithm is typically associated with issues such as a sluggish rate of convergence and unstable accuracy, and it is mostly suitable for the separation of independent source signals. Nevertheless, source signals are not always independent of one another in practical applications. This paper suggests a blind source separation algorithm based on the bounded component analysis of the enhanced Beetle Antennae Search algorithm (BAS). Firstly, the restrictive assumptions of the bounded component analysis method are more relaxed and do not require the signal sources to be independent of each other, broadening the applicability of this blind source separation algorithm. Second, the objective function of bounded component analysis is optimized using the improved Beetle Antennae Search optimization algorithm. A step decay factor is introduced to ensure that the beetle does not miss the optimal point when approaching the target, improving the optimization accuracy. At the same time, since only one beetle is required, the optimization speed is also improved. Finally, simulation experiments show that the algorithm can effectively separate independent and dependent source signals and can be applied to blind source separation of images. Compared to traditional blind source separation algorithms, it has stronger universality and has faster convergence speed and higher accuracy compared to the original independent component analysis algorithm.

Synergistic ROS generation and directional overloading of endogenous calcium induce mitochondrial dysfunction in living cells.

Taking advantage of endogenous Ca2+ to upregulate intramitochondrial Ca2+ level has become a powerful mean for mitochondrial dysfunction-mediated tumor therapy. However, the Ca2+ entered into mitochondria is limited ascribing to the uncontrollability and non-selectivity of endogenous Ca2+ transport. It remains a great challenge to make the maximum use of endogenous Ca2+ to ensure sufficient Ca2+ overloading in mitochondria. Herein, we smartly fabricate an intracellular Ca2+ directional transport channel to selectively transport endogenous Ca2+ from endoplasmic reticulum (ER) to mitochondria based on cascade release nanoplatform ABT-199@liposomes/doxorubicin@FeIII-tannic acid (ABT@Lip/DOX@Fe-TA). In tumor acidic microenvironment, Fe3+ ions are firstly released and reduced by tannic acid (TA) to Fe2+ for ROS generation. Subsequently, under the NIR light irradiation, the released ABT-199 molecules combine with ROS contribute to the formation of IP3R-Grp75-VDAC1 channel between ER and mitochondria, thus Ca2+ ions are directionally delivered and intramitochondrial Ca2+ level is significantly upregulated. The synergetic ROS generation and mitochondrial Ca2+ overloading effectively intensifies mitochondrial dysfunction, thereby achieving efficient tumor inhibition. This work presents a new insight and promising avenue for endogenous Ca2+-involved tumor therapies.

Predictive model for severe thrombocytopenia after transfemoral transcatheter aortic valve replacement.

The aim of this study was to develop a predictive model for severe thrombocytopenia after transfemoral transcatheter aortic valve replacement (TAVR). A total of 155 patients treated with TAVR at our center were retrospectively enrolled in this study. The incidence of severe thrombocytopenia after TAVR was 25.16%, and most patients suffered from severe thrombocytopenia on 4 days after procedure. Multivariate regression analysis showed that weight <60 kg, New York Heart Association Functional Classification (NYHAFC IV), major vascular complications, and lower first post-procedural platelet count were independent risk factors for severe thrombocytopenia after TAVR. The c-statistic for the area under the curve was 0.758, the sensitivity was 0.744, the specificity was 0.784, and the negative predictive value of the model was 91.38%. The overall predictive value was 76.77%. The predictive model developed from this cohort data could effectively identify patients at high risk of severe thrombocytopenia after TAVR, and might be applicable to patients with aortic regurgitation (AR) and severe thrombocytopenia with different definitions.

Efficiently targeted therapy of glioblastoma xenograft via multifunctional biomimetic nanodrugs.

BACKGROUND: Glioblastoma multiforme (GBM) is a fatal malignant primary brain tumor in adults. The therapeutic efficacy of chemotherapeutic drugs is limited due to the blood-brain barrier (BBB), poor drug targeting, and short biological half-lives. Multifunctional biomimetic nanodrugs have great potential to overcome these limitations of chemotherapeutic drugs. METHODS: We synthesized and characterized a biomimetic nanodrug CMS/PEG-DOX-M. The CMS/PEG-DOX-M effectively and rapidly released DOX in U87 MG cells. Cell proliferation and apoptosis assays were examined by the MTT and TUNEL assays. The penetration of nanodrugs through the BBB and anti-tumor efficacy were investigated in the orthotopic glioblastoma xenograft models. RESULTS: We showed that CMS/PEG-DOX-M inhibited cell proliferation of U87 MG cells and effectively induced cell apoptosis of U87 MG cells. Intracranial antitumor experiments showed that free DOX hardly penetrated the BBB, but CMS/PEG-DOX-M effectively reached the orthotopic intracranial tumor through the BBB and significantly inhibited tumor growth. Immunofluorescence staining of orthotopic tumor tissue sections confirmed that nanodrugs promoted apoptosis of tumor cells. This study developed a multimodal nanodrug treatment system with the enhanced abilities of tumor-targeting, BBB penetration, and cancer-specific accumulation of chemotherapeutic drugs by combining chemotherapy and photothermal therapy. It can be used as a flexible and effective GBM treatment system and it may also be used for the treatment of other central nervous systems (CNS) tumors and extracranial tumors.

Monitoring the electrochemical reactions on a gold nanoelectrode array via fluorescence-enabled electrochemical imaging.

High-spatial resolution fluorescence imaging on a high-density gold nanoelectrode array was achieved by anchoring fluorescent molecules to limit diffusion. Meanwhile, an ionic liquid as the electrolyte with a refractive index matching with the optical system could effectively improve the sensitivity of fluorescence imaging. Moreover, spatially resolved imaging of the electrochemical reactions on a single nanoelectrode modified with a single Pt nanoparticle was achieved.

2D Copper(II) Metalated Metal-Organic Framework Nanocomplexes for Dual-enhanced Photodynamic Therapy and Amplified Antitumor Immunity.

The immunosuppressive tumor microenvironment (TME) poses tremendous challenges for efficient immunotherapy. Smart nanomedicine is designed to modulate immunosuppressive TMEs based on the combination of dual-enhanced photodynamic therapy (PDT) triggered immunogenic cell death (ICD) and relieved hypoxic microenvironment. Copper(II) metalated metal-organic framework nanosheets (Cu-TCPP(Al)) are the foundation of the nanomedicine, and platinum nanoparticles (Pt NPs) and folate are subsequently introduced onto the Cu-TCPP(Al) surface (Cu-TCPP(Al)-Pt-FA). Upon targeted cellular uptake, intracellular GSH concentration is decreased because of the specific adsorption between GSH and CuII; meanwhile, Pt NPs possess catalase-like activity, which can continuously depose intracellular H2O2 to O2 to alleviate the hypoxic TME. The two factors synergistically improve the ROS concentration for dual-enhanced PDT. The highly toxic ROS can correspondingly cause amplified oxidative stress and then trigger the ICD. The ICD process stimulates antigen-presenting cells and activates the systemic antitumor immune response. Furthermore, the relieved hypoxic TME increases the infiltration of cytotoxic T lymphocytes (CTLs) at the tumor site, which can promote the transformation of the immunosuppressive M2 macrophage to immunoactive M1 phenotype. The easily prepared yet versatile nanomedicine possesses an excellent antitumor effect with the cooperation of dual-enhanced PDT and immunotherapy.

TfR Aptamer Enhanced Blood-Brain Barrier Penetration of Biomimetic Nanocomplexes for Intracellular Transglutaminase 2 Imaging and Silencing in Glioma.

Engineering a versatile nanocomplex integrating effective penetration of the blood-brain barrier (BBB), accurate diagnosis, and boosting therapy has always been an intractable challenge in glioblastoma multiforme (GBM). Herein, biomimetic nanocomplexes (TMPsM) for single intracellular transglutaminase 2 (TG2)-triggered self-assembly imaging and RNAi therapy for GBM are subtly developed. To prove the concept, transferrin receptor (TfR) aptamer-modified brain metastatic tumor cell membrane is prepared as the shell for dual BBB targeting capability and prolonged blood retention time. Upon targeting entering into GBM, hollow MnO2 is decomposed to release KKGKGQQ-tetraphenylethene (Pep-TPE) and siRNA. Owing to TG2 dependence, the non-emissive Pep-TPE would be self-aggregated to induce the emission turn-on in GBM that contain overexpressed TG2. The resulting aggregation-induced emission fluorescence imaging with a high signal-to-noise ratio can achieve the precise localization of the tumor and dynamic detection of TG2 activity, thereby allowing the GBM accurate diagnosis. Notably, the TG2 can be silenced by the released siRNA to cause cell apoptosis and increase chemotherapeutic sensitivity, ultimately realizing excellent antitumor efficacy. In vitro and in vivo results demonstrate that the as-prepared TMPsM indeed possess superior BBB penetration, precise diagnosis, and effective therapy of GBM. The proposed strategy may pioneer a new path for the theranostics of brain tumors.

In Situ Imaging of Endogenous Hydrogen Peroxide Efflux from Living Cells via Bipolar Gold Nanoelectrode Array and Electrochemiluminescence Technology.

The integration of a closed bipolar electrode (c-BPE) array and electrochemiluminescence (ECL) detection received a boost in applications in the detection of cell adhesion and disease-related biomarkers. This work proposed a gold nanorod array based c-BPE-ECL system to realize an in situ image of endogenous hydrogen peroxide (H2O2) efflux from living cells and parallel analysis of endogenous H2O2 released from multiple cells by converting electrochemical signals into optical signals. The gold nanorod array with high density was prepared by a repeating chronopotentiometry procedure with anodic aluminum oxide (AAO) membrane as a template. The c-BPE array was fabricated by assembling poly(dimethylsiloxane) (PDMS) chips on both sides of the gold nanorod array. When an appropriate driving potential is applied, H2O2 generated from living cells at the sensing pole was reduced on the gold nanorod, triggering the oxidation of the ECL reagent at the reporting pole, which allowed the detection of H2O2 released from living cells. Under phorbol myristate acetate (PMA) stimulation, H2O2 released from living HeLa, HepG2, MCF-7, and LO2 cells was determined to be 47, 32.4, 25.7, and 6.3 µM, respectively. This indicated that the amount of H2O2 released from PMA-stimulated cancer cells was significantly higher than that from the stimulated normal cells. This work presented a new approach for in situ imaging of H2O2 released from living cells and could also be used to detect other electrochemically active or non-electrochemically active molecules through simple cell surface modification, which may have potential applications in cell apoptosis study and disease diagnosis.

Gold Nanowires Array-Based Closed Bipolar Nanoelectrode System for Electrochemiluminescence Detection of α-Fetoprotein on Cell Surface.

Inspired by the promising applications of a closed bipolar electrodes (c-BPEs) system in electrochemiluminescence (ECL) detection of cell adhesion and disease-related biomarkers, here, a gold nanowires array-based c-BPEs system was constructed for cell surface protein detection. Regular and uniform gold nanowires array were prepared by intermittent potentiostatic deposition. Then, two poly(dimethylsiloxane) (PDMS) chips with a hole diameter of 2 mm as a reservoir were placed at both sides of Au nanowires array to construct c-BPEs system. Thionine-functionalized silicon dioxide nanoparticles conjugated to antibody (Ab2-Th@SiO2) were used as the electrochemical probe, while [Ru(bpy)3]2+-wrapped SiO2 nanoparticles (Ru(II)@SiO2) were employed as the ECL signal readout. Taking α-fetoprotein (AFP) as model, the gold nanowires array-based c-BPEs system allowed sensitive detection of AFP at a linear range from 0.002 to 50.0 ng/mL and at least 6 living cells ascribing to the synergetic amplification effect at both sensing and reporting chambers. Besides, the amount of AFP expressed by HepG2 cells was calculated to be 6.71 pg/cell. The presented strategy with high sensitivity provided a promising and universal platform for the detection of other cancer cells and disease-related biomarkers (such as proteins, glycan, miRNA).

Prediction Model of Hot Metal Silicon Content Based on Improved GA-BPNN.

The inconsistency of the detection period of blast furnace data and the large time delay of key parameters make the prediction of the hot metal silicon content face huge challenges. Aiming at the problem that the hot metal silicon content is not consistent with the detection period of time series of multiple control parameters, the cubic spline interpolation fitting model was used to realize the data integration of multiple detection periods. The large time delay of the blast furnace iron making process was analyzed. Moreover, Spearman analysis was combined with the weighted moving average method to optimize the data set of silicon content prediction. Aiming at the problem of low prediction accuracy of the ordinary neural network model, genetic algorithm was used to optimize parameters on the BP neural network model to improve the convergence speed of the model to achieve global optimization. Combined with the autocorrelation analysis of the hot metal silicon content, a modified model for the prediction of hot metal silicon content based on error analysis was proposed to further improve the accuracy of the prediction. The model comprehensively considers problems such as data detection inconsistency, large time delay, and inaccuracy of prediction results. Its average absolute error is 0.05009, which can be used in actual production.

The protective effect of cannabinoid type II receptor agonist AM1241 on ConA-induced liver injury in mice via mitogen-activated protein kinase signalling pathway.

INTRODUCTION: The endocannabinoid system plays an important role in regulating the immune responses in inflammation. At present, there are no good clinical drugs for many immune liver diseases. METHODS: We explored the protective effect of the cannabinoid type II (CB2) receptor agonist AM1241 on the liver of mice with acute liver injury caused by concanavalin from the perspective of inflammation and immunity. Pathological evaluation in hepatic tissue was examined by haematoxylin and eosin (HE) staining and the levels of biochemical parameters in the serum were measured by automatic biochemical analysis. The content of inflammatory factors was measured by enzyme-linked immunosorbent assay and real-time quantitative reverse transcription polymerase chain reaction (real-time PCR). The liver apoptosis-related proteins were observed by immunohistochemistry. The expression of liver injury-related proteins was analysed by Western blot. Immune cells were isolated from the liver of mice and studied in vitro. RESULTS: Reduced levels of alanine transaminase and aspartate transaminase were observed in ConA-induced liver injury mice treated with AM1241, together with attenuated liver damage evidenced by H&E staining. Moreover, AM1241 inhibited the protein and gene expression levels of TNF-α, IL-6 and IFN-γ in the livers of mice. The phosphorylation levels of p38, JNK, ERK1/2, P65 and cAMP response element-binding protein (CREB) in the mouse were significantly reduced in AM1241 pretreatment, while the level of p-JNK increased. In addition, the P/T-P65 and P/T-CREB of the AM1241 pretreatment group were significantly reduced. The results of immunohistochemistry measurement are consistent with those of Western blotting. The CB2-mediated effect is through macrophage-like Kupffer cells. CONCLUSION: Our study suggests that the ConA-induced liver injury model in mice is protected by CB2 agonist AM1241 by modulation of CB2 receptor-rich immune cells, for example, Kupffer cells. Reduced inflammatory responses regulate apoptosis/cell death in the liver particularly hepatocytes and other parenchymal cells.

Knockout of the Cannabinoid Receptor 2 Gene Promotes Inflammation and Hepatic Stellate Cell Activation by Promoting A20/Nuclear Factor-κB (NF-κB) Expression in Mice with Carbon Tetrachloride-Induced Liver Fibrosis.

BACKGROUND This study aimed to investigate the effect of deleting the cannabinoid receptor 2 (CB2) gene on the development of hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice via regulating inflammation. MATERIAL AND METHODS The DNA was extracted from the tails of mice to identify whether the cannabinoid receptor 2 gene was successfully knocked out. A liver fibrosis model was established by an intraperitoneal injection of CCl4 into mice. Hepatic damage and hepatic fibrosis were evaluated by detecting serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), and staining paraffin sections of liver tissue with hematoxylin-eosin (HE). The secretion and distribution of collagen in liver tissue were observed by Masson staining. Western blot analysis was performed to detect the expression of a-smooth muscle actin (alpha-SMA), transforming growth factor-ß1 (TGF-ß1), tumor necrosis factor alpha-induced protein 3 (A20), phosphorylated nuclear factor-kB p65 (p-NF-kappaB p65), tumor necrosis factor alpha (TNF-alpha), and interleukin-6 (IL-6) in liver tissue. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of IL-6 and TNF-alpha mRNA in liver tissue. RESULTS Compared with the control mice, the mice with CB2 knockout that were exposed to CCl4 exhibited increased liver damage, liver fibrosis, and upregulated alpha-SMA, TGF-ß1, A20, and p-NF-kappaB p65 protein levels. IL-6 and TNF-alpha protein levels and mRNA levels were upregulated. CONCLUSIONS The deletion of the CB2 gene promoted the activation of hepatic stellate cells in mice with liver fibrosis and aggravated liver fibrosis by up-regulating the protein expression of A20 and p-NF-kappaB p65 and inducing inflammatory response, potentially providing new insight into the treatment of liver fibrosis.

Etiological characteristics of "tail blister disease" of Australian redclaw crayfish (Cherax quadricarinatus).

In November 2019, an acute disease outbreak in Australian redclaw crayfish (Cherax quadricarinatus) occurred in a farm in Hubei, China, with a cumulative mortality rate of over 80%. One of the characteristic symptoms of the disease was blisters on the tail. This symptom is also common in diseased Procambarus clarkii every year in this country, but the causative agent has not been determined. This study analyzed the etiological characteristics of this disease. Bacterial isolation and identification combined with high-throughput sequencing analysis were conducted to obtain the microbiota characteristics in the hemolymph, hepatopancreas, and intestines. Results showed that this outbreak was caused by infection from Aeromonas hydrophila and Aeromonas veronii. The underlying cause was stress imposed on crayfish during transferring from outdoor pond to indoor pond because of temperature drops. Aeromonas infection caused remarkable changes in the structure of the microbial composition in the hemolymph, hepatopancreas, and intestines of the crayfish. The abundance of Aeromonas in the hemolymph of the sick crayfish was as high as 99.33%. In particular, KEGG metabolic pathway analysis showed that some antibiotic synthesis, enterobactin biosynthesis, and myo-inositol degradation pathways were abundant in healthy crayfish hemolymphs, which may be the mechanism of maintaining crayfish health. Conversely, inhibition of these pathways led to the disorder of microbiota structure, finally leading to the occurrence of diseases. To the knowledge of the authors, this study was the first to use high-throughput amplicon sequencing targeting the 16S rRNA gene to find the causative bacteria in aquatic animals. This protocol can provide more comprehensive and reliable evidence for pathogen identification, even if the pathogenic bacteria are anaerobes or other hard-to-culture bacteria.

Review: Multiplexed profiling of biomarkers in extracellular vesicles for cancer diagnosis and therapy monitoring.

Extracellular vesicles (EVs) are nanoscale vesicles secreted by normal and pathological cells. The types and levels of surface proteins and internal nucleic acids in EVs are closely related to their original cells, tumor occurrence, and development. Thus, the sensitive and accurate detection of EV biomarkers is a reliable approach for noninvasive disease diagnosis and treatment response monitoring. However, the purification and molecular profiling of these EVs are technically challenging. Much effort has been dedicated to developing new methods for the detection of multiple EV biomarkers. In this review, we summarize the recent progress in EV protein and nucleic acid biomarker analysis. Additionally, we systematically discuss the advantages of multiplexed EV biomarker detection for accurate cancer diagnosis, therapy monitoring, and cancer screening. This article aims to present an overview of all kinds of analytical technologies for assessing EVs and their applications in clinical settings.

Biomimetic nanoreactor for targeted cancer starvation therapy and cascade amplificated chemotherapy.

Consuming glucose by glucose oxidase (GOx) has attracted great interest in cancer starvation therapy, but the therapeutic effect is severely limited by the tumor hypoxia environment. Herein, to overcome such limitation, cancer cell membranes disguised biomimetic nanoreactors were elaborately established for synergetic cancer starvation therapy and cascade amplificated hypoxia activated chemotherapy. Via a metallothionein-like self-assembly and infiltration approach, GOx and hypoxia activated prodrug banoxantrone (AQ4N) were efficiently loaded into metal-organic framework ZIF-8 nanocarriers to yield nanoreactor AQ4N/GOx@ZIF-8. Subsequently, the biomimetic nanoreactor (AQ4N/GOx@ZIF-8@CM) was obtained by camouflaging the nanoreactor with cancer cell membrane, which endowed the biomimetic nanoreactor homotypic targeting, immune escape and prolonged blood circulation features. Once targeted accumulating into tumor sites, the acid environment triggered the decomposition of ZIF-8, then encapsulated GOx and AQ4N were released. GOx would rapidly exhaust endogenous glucose and O2 to shut off the energy supply of tumor cells for starvation treatment. Furthermore, the aggravated tumor intracellular hypoxia environment would activate the cytotoxicity of AQ4N for chemotherapy. In vitro and in vivo results demonstrated that the designed biomimetic nanoreactor exhibited negligible systemic toxicity, besides, the combination of starvation therapy and cascade amplified hypoxia activated chemotherapy significantly inhibited the tumor growth and improved the therapeutic efficacy.